B GIO DC V O TO I HC NNG TRNG I HC BCH KHOA

BI GING MN HC CNG NGH SN XUT PROTEIN, AXIT AMIN V AXIT HU C

BIN SON: TRNG TH MINH HNH B MN CNG NGH THC PHM SINH HC

NNG, NM 2006

MC LC PHN I: CNG NGH SN XUT PROTEIN T VI SINH VT

M u
- Gii thiu chung v ng hng sn xut protein - Nhu cu protein v kh nng sn xut protein trn th gii

Chng 1: Khi nim chung v vi sinh vt

1.1. Cc vi sinh vt tng hp protein v a xit amin - To - Nm men v vi khun - Nm mc v x khun 1.2. Qu trnh dinh dng t bo vi sinh vt 1.3. C ch sinh tng hp protein 1.4.Cc yu t t6ng hp protein

Chng 2: S dy chuyn cng ngh thu nhn cc sn phm protein

2.1. Nguyn liu v phng php x l 2.2. Nui cy vi sinh vt 2.3. Tch protein, c c v sy

Chng 3: Sn xut protein t cc ngun hydrat cacbon

3.1. Nui cy vi sinh vt trn dch thy phn cc nguyn liu thc vt 3.2. Nui cy vi sinh vt trn dch thy phn than bn 3.3. Nui cy vi sinh vt trn dch thy phn g 3.4. Nui cy vi sinh vt trn nguyn liu polysacarit cha thy phn 3.5. Nui cy vi sinh vt trn b ru t nguyn liu ht v r ng - c tnh nguyn liu - X l nguyn liu - S dy chuyn cng ngh

Chng 4: Cng ngh sn xut protein t ngun cacbua du m, kh t

4.1. Nui cy vi sinh vt trn nguyn liu cacbua hydro lng 4.2. Nui cy vi sinh vt trn kh cacbua hidro

Chng 5: Sn xut thc n protein t vi sinh vt

5.1. Protein t nm men 5.2. Protein t to v vi khun 5.3. Protein t nm si PHN II: CNG NGH SN XUT CC AXIT AMIN

Chng 1: Khi qut chung v axit amin

1.1. c tnh ca cc axit amin, vai tr v ng dng 1.2. C ch iu chnh sinh tng hp cc axit amin 1.3. Cc phng php sn xut cc axit amin

Chng 2: Sn xut lizin

2.1. Tng hp lizin t t bo vi sinh vt 2.2.Nguyn liu v phng php x l 2.3. Qu trnh sinh tng hp lizin 2.4. Tch v sy lizin 2.5. S cng ngh sn xut lizin

Chng 3: Sn xut axit glutamic

3.1. Mt s phng php sn xut axit glutamic 3.2. Tng hp axit glutamic t vi sinh vt 3.3. Cc yu t nh hng n qu trnh sinh tng hp axit glutamic t r ng

Chng 4 : Sn xut valin v triptophan

4.1. Ngun nguyn liu 4.2. Ngun vi sinh vt tng hp 4.3. S dy chuyn cng ngh PHN III: CNG NGH SN XUT CC AXIT HU C

M u Chng 1: Axit xitric

1.1. Mt s khi nim chung 1.2. C s l thuyt ca qu trnh ln men axit xitric 1.3. Ging vi sinh vt v phng php nui cy 1.4. Chun b mi trng nui cy 1.5. Ln men 1.5.1. Phng php ln men b mt 1.5.2. Phng php ln men b su 1.6. Cc yu t nh hng n qu trnh 1.7. X l dch ln men bng phng php ha hc v thu nhn sn phm L: Trung ha Phn gii xitrat caxi - Lc - Kt tinh - Sy

Chng 2: Axit lactic

2.1.Khi nim chung 2.2.Vi sinh vt v nguyn liu 2.3. C s l thuyt ca qu trnh ln men lactic 2.4. S cng ngh sn xut axit lactic 2.4.1. Ln men lactic 2.4.2.X l dch ln men - lc 2.4.3. Phn gii lactac canxi 2.4.4. C c

Chng 3: Axit axetic

3.1. M u - Khi nim chung 3.2. Nguyn liu v vi sinh vt 3

3.3. C s l thuyt ca qu trnh ln men axetic 3.4. Cc phng php ln men axetic 3.5. Chng ct axit axetic 3.6. S cng ngh sn xut axit axetic TI LIU THAM KHO 1. Nguyn c Lng, Cng ngh vi sinh tp 2, Nh xut bn i hc Quc gia thnh ph H Ch Minh, 2002

2. Lng c Phm, H Xng, Vi sinh tng hp, Nh xut bn khoa hc v k thut, H Ni,1978
3. TS Nguyn Hu Phc, Gio trnh cng ngh vi sinh, Thnh ph H Ch Minh, 2001 4. PGS. TS Trn Minh Tm, Cng ngh vi sinh ng dng, Nh xut bn nng nghip, Thnh ph H Ch Minh, 2000 5. Robert Noyes , Protein food supplement, Noyes Development corporation, Park Ridge, New Jerbey, USA (1969) 6. Richard I Matelles and Steven, Single - Cell Protein, R. Tanneebaum Editors, Cambrige, Massachusettes and London, England (1978)

PHN 1 CNG NGH SN XUT PROTEIN

M U
1. Vai tr ca protein i vi con ngi: - C th ngi v ng vt thng xuyn i hi cung cp cc cht dinh dng c trong thc n c th tin hnh trao i cht, trc ht nhm duy tr s sng, tng cng sinh trng v pht trin. - Thc n, ngoi nc cn gm nhng nhm cht: protein, cht bo, gluxit, vitamin, mui khong, cc cht gia v, trong phn qu him nht l protein. - Protein l ngun nit duy nht cho ngi v ng vt. Trong qu trnh tiu ho ca ngi v ng vt, protein phn gii thnh khong 20 axit amin thnh phn, trong c 8 axit amin khng thay th (hoc 9 i vi tr em, 10 i vi ln v 11 i vi gia cm) cn phi c sn trong thc n. Nu khng nhn c cc axit amin ny c th s b bnh hoc cht. - Thiu protein s dn n nhiu bnh tt ht sc him ngho:

+ Bnh thiu protein ln u tin c pht hin Chu Phi, c tn gi quc t l Kwashiokor, hin ny l bnh ph bin nhiu vng trn th gii. Tr em mc bnh ny chm ln, ci cc, km pht trin v tr tu. Bnh ny c th iu tr bng cch thm vo khu phn bnh nhn mt lng thch ng cc loi protein c phm cht tt nh cazein. Tuy nhin nhiu ti liu cho thy s km pht trin v tr tu v bnh ny khng phc hi c v nh hng n ton b cuc i ca bnh nhn. + V mt sinh l, thiu protein dn n gim th trng. Hng ngy c th ngi trng thnh c ti 100 t t bo cht v cn thay th. Thiu protein th trc ht protein ca gan, mu v cht nhy nim mc, rut c huy ng b p. V nh vy s dn n suy gan, s lng khng th trong mu gim i, sc khng ca c th i vi bnh b yu. + V nhu cu protein ca ngi, nhiu nh nghin cu cho bit dao ng trong khong 80 120g/ngy. 2. nh ngha v sinh khi: Sinh khi l ton b t bo vi sinh vt (biomas) thu nhn c trong qu trnh ln men. N c s dng nh mt ngun dinh dng protein cho ngi v ng vt, i khi ng ngha vi protein n bo (single cell protein SCP). 3. Protein n bo v a bo: Cm t protein n bo c dng ch ngun protein mi tm ra t nhng c th n bo (t vi sinh vt), phn bit n vi protein t ng vt v thc vt (protein a bo v protein truyn thng). 3.1. Protein a bo: l ngun dinh dng quan trng nui sng loi ngi t trc ti nay. y l ngun cung cp protein quan trng nht. Tuy nhin, do tc pht trin dn s qu nhanh nn ngun protein ny khng cn cung cp cho nhu cu ngy cng tng ca con ngi. Hin nay trn th gii c khong 2/3 dn s ang ng trc thc trng thiu v i protein, cn 1/3 dn s li c cp s lng protein d tha so vi nhu cu. Nguyn nhn: - S phn phi khng ng u ngun protein a bo gia cc quc gia v gia cc vng dn c trong mt quc gia. - Trnh k thut v pht trin ngun protein a bo khng ng u. - S khc nhau v iu kin a l: nhng vng sa mc t nhin hoc vng c iu kin kh hu khng thun li cho trng trt v chn nui. - Do chnh con ngi gy ra nh tnh trng nhim mi trng, nhim ngun nc, rng tha, i trc, sng con, s khai thc thiu khoa hc lm cc ngun thy hi sn ngy cng cn kit v .v.. Cc gii php tng nhanh ngun protein a bo: - Ci bin h thng di truyn ca cy trng v vt nui: thc phm c ch bin t ngun ng vt v thc vt bin i gen gi l thc phm bin i gen. Chng trnh GMO (chng trnh c th bin i gen) gp nhiu kin phn i ch trch v cho rng thc phm bin i gen c th to ra nhng bnh tt cho ngi v

ng vt. Tuy nhin cho n nay nhiu nc nh M, Trung Quc v mt s nc vn pht trin mnh cc loi u, c chua, bp bin i gen. - Pht trin k thut di truyn nhng vn khng ngng nghin cu nng cao hn na k thut truyn thng trong trng trt v chn nui. 3.2. Protein n bo: Protein n bo l thut ng ch mt loi cht dinh dng c trong t bo v ch c sn xut t vi sinh vt. Thut ng ny khng ch n gin l protein t t bo ca c th n bo, v rt nhiu vi sinh vt khng phi l c th n bo m vn khai thc chng. Do , thut ng ny nn hiu l ngun dinh dng cha nhiu protein t vi sinh vt (t vi khun, nm men, nm si v to). Protein n bo l hng nghin cu mnh m hin nay gii quyt vn thiu ht protein. 3.2.1. Lch s pht trin: Thut ng protein n bo c t nhng nm 50 ca th k 20 nhng thc t loi ngi bit s dng loi protein ny v cc cht c trong t bo vi sinh vt t rt lu: lm bnh m, sa chua, phomat, bia bng hot ng sng ca vi sinh vt d khng hiu vi sinh vt l g. Mi n th k 17, ngi ta mi bit n vi sinh vt l mt sinh vt th ba sau ng vt v thc vt. Trc th k 20, vic s dng vi sinh vt trong cc qu trnh ch bin thc phm hon ton mang tnh truyn thng v iu kin t nhin. Vic nghin cu v sn xut protein n bo cn xa l vi loi ngi, nht l vi qui m cng nghip. u th k th I, nh my sn xut sinh khi nm men c coi l nh my u tin sn xut protein n bo ti c vi phng php nui Candida utilis cn gi l nm men Torula. Sau , mi quan tm ca c gim i nhng n nm 1930, c m phc hi v m rng sn xut, nng sut nm men l 15.000 Tn/nm, trn c s nui trn dch kim sunfit, dch thi ca cng nghip xenluloza, lm thc phm phc v trong qun i v dn thng, ch yu l nu canh v lm xc xch. Sau nm 1950, phong tro sn xut SCP lan rng khp Chu u, M. Tuy nhin tt c vn qui m va v nh, ch yu cho chn nui v c th chit tch tinh sch protein lm thc n nhn to hoc b sung vo cc ngun ch bin TP. Vo lc din ra hi ngh ln th I v SCP ti Vin K thut Massachusett (MIT) nm 1967, a s cc d n ch mi nm trong thc nghim, ch s hng British Petroleum (BP) l c bo co v nhng kt qu ca qu trnh ln men SCP qui m cng nghip (CNG NGHIP). Nhng n hi ngh ln th II hp vo nm 1973 th nhiu hng ca nhiu nc khc nhau bt u sn xut SCP qui m CNG NGHIP. Cng bt u t nm 1973, CNG NGHIP sn xut SCP c nhng bc pht trin nhy vt do vic s dng hidrocabon ca du m, kh t lm ngun cabon v nng lng rt c hiu qu. Vy nguyn nhn no dn n vic nhiu nc phi sn xut SCP? Sn xut SCP l ngun protein c cht lng cao thay th cc loi bt dinh dng lm t cc ht cha du nh u tng hoc bt c dnh cho ng vt s gii quyt c 2 vn : + Tng ngun u tng c, v c ng cc cho dinh dng ngi. + Cc nc Chu u, Nga, Nht v mt s vng khc khng trng c u tng, do SCP s gip cho nc khng ph thuc vo vic nhp khu protein.
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+ Trong t bo vi sinh vt, ngoi hm lng protein tng i ln cn c cht bo, vitamin v cc cht khong, nng sut ca vi sainh vt vt xa nng sut cy trng v vt nui trong cng nghip nhiu ln. 3.2.2. c im ca sn xut Protein n bo: - Chi ph lao ng t hn nhiu so vi sn xut nng nghip. - C th sn xut bt k a im no trn tri t, khng chu nh hng ca kh hu thi tit, cc qu trnh cng nghip , d c kh ho v t ng ho. - Nng sut cao: vi sinh vt c tc sinh sn mnh, kh nng tng trng nhanh. Ch trong mt thi gian ngn c th thu nhn c mt khi lng sinh khi rt ln; thi gian ny c tnh bng gi, cn ng vt v thc vt, tnh bng thng hoc hng chc nm. - S dng cc ngun nguyn liu r tin v hiu sut chuyn ho cao. Cc nguyn liu thng l ph phm, ph phm ca cc ngnh khc nh r ng, dch kim sufit, parafin du m v..v.. , thm ch c nc thi ca mt qu trnh sn xut no . Hiu sut chuyn ho cao: hidrat cacbon c chuyn ho ti 50%, cacbuahidro ti 100% thnh cht kh ca t bo. - Hm lng protein trong t bo rt cao: vi khun l 60 -70%, nm men l 40-50% cht kh v..v Hm lng ny cn ph thuc vo loi v chu nhiu nh hng ca iu kin nui cy. Cn ch rng hm lng protein y ch bao hm protein ch khng gm c thnh phn nit phi protein khi xc nh theo phng php nit tng s ca Kjeldal, nh axit nucleic, cc peptit ca thnh phn t bo. - Cht lng protein cao: Nhiu axit amin c trong vi sinh vt vi hm lng cao, ging nh trong sn phm ca tht, sa v hn hn protein ca thc vt. Protein vi sinh vt c bit giu lizin, l mt li th ln khi b sung thc n v chn nui, v trong thc n thng thiu axit amin ny. Tri li, hm lng cc axit amin cha lu hunh li thp. - Kh nng tiu ho ca protein: c phn hn ch bi thnh phn phi protein nh axit nucleic, peptit ca thnh t bo, hn na, chnh thnh v v t bo vi sinh vt kh cho cc enzim tiu ho i qua. - An ton v mt c t: Trong sn xut protien n bo khng dng vi sinh vt gy bnh cng nh loi cha thnh phn c hoc nghi ng. V vy n nay hu nh SCP ch dng trong dinh dng ng vt. - Nhng vn k thut: Sinh khi vi sinh vt phi tch v x l. Vn ny ph thuc ch yu vo kch thc t bo. Sinh khi nm men d tch bng li tm hn vi khun. Ngoi ra, vi sinh vt no c kh nng sinh trng mt cao s cho nng sut cao, sinh trng tt nhit cao (c tnh cht a nhit v chu nhit) s gim chi ph v lm ngui trong sn xut, t mn cm vi tp nhim v..v.. s dng cc ngun cacbon r tin, chuyn ho cng nhiu cng tt .. th s c dng trong sn xut. V vy nm men c s dng ch yu trong sn xut protein n bo. Nh vy u im ca sn xut protein n bo l c th phn lp v la chn cc chng vi sinh vt c ch v thch hp cho cc qui trnh cng ngh, cho tng nguyn liu 1 cch tng i nhanh v d dng.

CHNG 1 KHI NIM CHUNG V VI SINH VT

Protein ca vi sinh vt ch yu c tng hp hnh thnh cc enzim. V vy phn ln nm trong t bo, mt s rt t c tch ra ngoi mi trng. Yu cu ca cc chng vi sinh vt dng trong sn xut: - Thi gian nhn i ngn. - C kh nng to thnh 40-70% protein. - Tiu ho ti a cc cht dinh dng ca mi trng. - Khng gy bnh v em vo mi trng c t. - C sc bn cao v chu c iu kin nui cy khng v trng. - D tch khi dch nui cy trong iu kin tuyn ni (flotation) v li tm tch. 1. Cc nhm vi sinh vt tng hp protein: 1.1. To n bo v a bo 1.1.1. Vai tr ca to trong i sng To theo ting Latin l Algue c ngha l c bin, nhng thc ra trong nc ngt cng nh trong t, trong thn v l cy, trong bo hoa du v..v u c to. Trong t nhin c nhiu loi to c hm lng protein cao nhng khng s dng cho ngi v gia sc v c c t. Mt s to l mn n dn gian nhiu a phng nh: Trung v ng , Nam M hay dng to lam, Bolovi v mt s nc Nam M dng loi Nostae commune (Sphaeronostos commune), Trung Quc dng loi Nematonostos Flagelliforme, Chu Phi vt loi to lam a bo Spirulina maxima cc ao h giu mui canxi lm thc n bi b v dng lm mt s thuc cha bnh nh ph chn, au rng v ng tiu ho. T , to Spirulina c nhiu nc trn th gii a vo sn xut cng nghip. Khong nm 1970, nhng nh khoa hc ngi Php pht hin ra to c kh nng pht trin nhanh v c hm lng protein cao nn h nghin cu v xy dng c nhng qui nh cng ngh sn xut to. n nay ch c 3 loi to n bo sn xut qui m ln v c kinh t cao l: + Chlorella + Spirulina + Scenedesmus. trong hai loi Chlorella v Spirulina c sn xut nhiu hn c. 1.1.2. u im ca to n bo: - Gi tr dinh dng ca to cao v phm vi ng dng rng ri: + To n bo c hm lng protein rt cao (chim khong 40-55% cht kh), ring to Spirrlina c cha ti 70%.

+ Protein ca to thuc loi protein hon ho v c cht lng cao. Hm lng axit amin ca nhng protein trong to gn vi qui nh protein tiu chun, c bit l lizin trong protein ca to cao hn hn lizin ca la mch. Tng s axit amin khng thay th trong protein rt cao, c khi ln n 42% (bng 1 v bng 2). + To cha nhiu protein v vitamin (VTM) (nht l VTM B12 v C) nn c sn xut lm thc n cho ngi, gia sc, gia cm v tm c. + Gi tr dinh dng ca to cn th hin cht lng v s lng ca cc VTM c trong . To Chlorella c nhiu VTM A, nhm VTM B, trong t bo ti c rt nhiu VTM C. Ngoi ra c rt nhiu VTM B, K, axit aconitic, axit pantotenic, biotin, lencophorin trong cc loi to. - Cho n nay cha tm thy c t no nguy him tn ti trong sinh khi to. - c im ca t bo cc loi to l c cht dip lc (chlorophyll). Cht ny c vai tr quan trng trong vic c nh nng lng nh sng mt tri ca to. V vy to l loi sinh vt t dng, chng hon ton c kh nng quang hp m cc gii hin vi sinh vt khc khng c. - To c kch thc t bo ln, hon ton c th p ng ti mi yu cu k thut, c bit thun li trong giai on thu nhn. - Khng b virus tn cng, sng trong nhng iu kin n gin. - To c kh nng lm sch cc ngun nc bn, gi v sinh mi trng. To lam c th tham gia qu trnh c nh nit ca khng kh v nh nhng tnh cht c bit ca mnh, to lam li cun s ch ca cc nh khoa hc trong lnh vc di truyn, t bo, ho sinh, l sinh. 1.1.3. So snh 2 loi to Chlorella v Spirulina - To Spirulina cha VTM B12, caroten nhiu hn hn to Chlorella, cha nhiu xantophin l cht rt cn thit cho gia cm ( g CNG NGHIP cho trng g c lng ti, tht g vng v ngon), Spirulina cn cha nhiu loi cht khng sinh chng vi khun v cc loi nm, nn c th bo qun rt lu m khng b mc. - Hm lng protein trong to Spirulina cao hn nhiu so vi to Chlorella. Protein ca t bo Spirulina l 60-70%, Chlorella l 40-50%. - Kch thc ca to Spirulina ln hn kch thc ca to Chlorella. Mt khc, to Spirulina trong qu trnh pht trin c xu hng ni ln b mt trong khi to Chlorella c kch thc nh li c xu hng lng chm khi khng khuy trn. Thu hoch to Spirulina bng nhng phong php n gin, trong khi vi to Chlorella th phc tp ging nh thu hoch sinh khi nm men hoc sinh khi vi khun. - Thnh t bo to Spirulina mng, thnh t bo ca Chlorella dy hn. Do h s tiu ho khi ta dng to Spirulina cao hn to Chlorella. To Spirulina pht trin trong mi trng kim cn Chlorella pht trin trong mi trng axit yu. - Khi dng CO2 nh ngun cacbon, m ngun cacbon ny trong iu kin kim t d chuyn ho sang dng d hp th theo phn ng sau: HCO3 + OH CO32- + CO2 + H2O
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CO32- + H2O 2HCO3

Spirulina hp th CO2 theo chiu hng ny tt hn to Chlorella. V vy, hin nay trong sn xut cng nghip, to Spirulina chim mt v tr u th. 1.2. Nm men v vi khun: 1.2.1. Nm men: - Trong cc ngun protein sn xut bng con ng vi sinh vt, nm men c nghin cu sm nht v c p dng rng ri trn th gii. Con ngi s dng nm men hoc cc sn phm hot ng sng ca chng t hng nghn nm nay. - Nm men l tn chung ch nhm nm c cu to n bo, sinh sn bng cch ny chi. Nm men khng c dip lc v khng th s dng nng lng mt tri. V vy chng dinh dng bng cc hydratcbon, cc hydrocacbua, trc ht l ng. - Trong t bo nm men c cha hu ht cc cht cn thit cho s sng (protein, gluxit, lipit, cc enzim, cc VTM, cc axit nucleic, cc cht khong). - Khng mt sn phm thc vt hoc ng vt no c trong thnh phn ca mnh mt lng cc cht c tc dng c hiu nh trong nm men. Tuy nhin thnh phn cc cht c hiu ca nm men khng ph hp hon ton vi nhng nhu cu sinh l ca ng vt. - Nm men c ch nhiu, v khng nhng trong t bo ca chng c nhiu cht dinh dng c gi tr, m chng li c kh nng tng sinh khi v cc c im sinh l ph hp vi iu kin sn xut cng nghip. - V c im lch s: Men gia sc c sn xut u tin c vo khong nm 1880. Lc ngi ta dng men bia (Saccharomyces cerevisiae). Trong th chin th I, men gia sc v men thc phm c sn xut ch yu c l ging Torula utilis. M, t nm 1946 mi t chc sn xut sinh khi nm men. Lc u, ngi ta nui cy nm men trn sacaroza thu hi sinh khi lm thc n cho ngi. Sau v l do kinh t, dn dn ngi ta thay sacaroza bng dch thy phn t tinh bt v xenluza, ph liu cng nghip ng, bia, ru Nm 1968, Lin X l nc u tin xy dng nh my sn xut nm men t paraphin du m, sau Anh, Php , Nht vv.. tin hnh rt nhanh trong lnh vc s dng ngun nguyn liu di do v r tin ny vo mc ch thu protein ca nm men v a sn lng nm men trn th gii ngy cng tng. - V gi tr dinh dng: + Nm men rt giu protein v VTM, c bit l cc VTM nhm B. + Sinh khi nm men cha khong 75-80% nc, 20-25% cht kh trong : cacbon 45-50%, nit 7-10% (tng ng vi 40-60% protein, hydro 5-7%, oxy 2530%, cc nguyn t v c 5-10% (photpho v kali chim ti 95-97%) tng lng tro, s cn li l canxi, magi, nhm, lu hunh, clo, st, silic. Ngoi ra cn c mt lng rt nh cc nguyn t mangan, km, molipden, bo, cacbon ..). + Trong thnh phn qu nht l protein. Hm lng protein tu thuc vo tng loi ging, vo thnh phn mi trng v iu kin nui cy. Dao ng trong khong 40-60%.
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+ V tnh cht protein ca nm men gn ging protein ngun gc ng vt. Protein ca nm men cha khong 20 axit amin khng thay th (bng 5). Thnh phn cc axit amin ca nm men cn i hn so vi la m v cc ht ng cc khc, km cht t so vi sa, bt c, bt xng tht v cc sn phm ng vt ni chung. S thay i thnh phn cc axit amin trong thi gian nui cy c nghin cu cho thy thnh phn ca cc axit amin thay i mt giai on pht trin: giai on tim pht. Sau 3 gi pht trin, tng hm lng cc axit amin trong protein tng ln 17% so vi thi im ban u. Sau tng hp axit amin gim xung v gi mc trn 40%. n cui, t bo gi, cc cht d tr, trc ht l glucogen tiu hao nhiu nn gim trng lng, do t l gia cc axit amin so vi trng lng chung ca cc t bo tng ln gn 50% (tng khng thc cht). - Cc ging nm men dng lm thc phm cho ngi v thc n gia sc l: Endomyces vernalis, Hansenula anomala, Hansenula suaveolens, Saccharomyces cerevisiae, Candida arbores, Candida tropicalis, Mycotorula lipolytica, Mycotorula japonica, Torulopis utilis, Torulopis utilis var, major, Torulopsis utilis var thermophilis, Monilia candia, Oidium lactic. - Cc tiu chun la chn ging nm men sn xut protein t cc ngun hydrocacon: + C kh nng ng ho nhiu ngun cacbon khc nhau, nht l cc loi pentoza (xiloza, arabinoza) v cc axit hu c. + C th pht trin tt trn mi trng c nng cht kh cao. + C kh nng pht trin nhanh, c sc khng cao i vi nng CO2. + Sn lng cao, sinh khi cha nhiu cht dinh dng c gi tr (hm lng protein cao, c nhiu axit amin khng thay th, vitamin ..) + Kch thc t bo tng i ln d tch bng li tm. + Chu ng c nhit tng i cao, t lm bin i pH mi trng. - Trong sn xut nm men thng dng cc chng thuc ba ging Saccharmyces, Candida v Torulopsis. Kh nng chuyn ho ca ba ging ny rt cao v a dng, qui trnh cng ngh tng i n gin. 1.2.2. Vi khun: - Vi khun sn xut protein thng c nui trn cacbua hidro. Thng s dng cc ging Pseudomonas, Flavobacterium, Mycobacterium v Nocardia. - Cc ging vi khun ny c kh nng ng ho cc ankal (C6-C18) , cacbua hydro bo v thm khc. - i vi nguyn liu s dng l metan, s dng cc ging Methylomonas, Methyllococens capsulatus. - Ngoi ra nhiu ni cn s dng vi khun kh n c cc i din ca ging Hydrogenomonas (H. facilia, H. entropha). - c im ca vi khun: + Tc sinh trng nhanh
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+ Dng c nhiu c cht. + pH cn gi 5-7, nu khng c th c nguy c nhim cc vi khun gy bnh. + Thu hi bng li tm: kh + Thnh phn cc axit amin cn i nhng hm lng cc axit amin cha S hi thp. + Khi dng cc vi khun Gram m sn xut SCP cn lu kh nng sn sinh c t ca chng. 1.3. Nm mc v x khun: - Ni chung ngi ta t dng nm mc v x khun sn xut protein. V mt dinh dng, protein ca cc vi sinh vt ny km gi tr hn so vi protein ca vi khun, nm men ... V k thut nui cy, do h si pht trin thnh bi chng cht nn tr ngi n vic sc kh v khuy trn. - Nm mc l nhng c th a bo, giu vitamin nhm B, cha chng 30-60% protein. Hm lng metionin v tryptophan thp, cn c cc axit amin khc tng t nh protein tiu chun ca FAO. Cc ging nm mc c hm lng protein cao l Fusarium, Rhizopus, Penicillium, Aspergillus. Trong nhng nghin cu thu nhn protein t nm mc, ngi ta ch nhiu n cng trnh ca B.Volesky v H.Zajic. Hai ngi ny phn lp c t nc t chng mc thuc Graphium, chng ny c cha ti 52% protein, trong c 16 axit amin, metionin chim 1% so vi protein th, lizin chim n 7,7%, cc axit amin khng thay th khc u c hm lng tng ng vi protein tiu chun, tr izolxin. Chng mc ny c kh nng ng ho etan, metan v c nui trong mi trng cha hn hp hai nguyn liu ny thu sinh khi. - Gi tr dinh dng protein mt s nm mc c th xem bng 14. - Nh ni, nm mc t c dng trong sn xut protein. Hin nay ch c mt s c s sn xut nh United Parer rills Phn Lan, cng sut 10.000tn/nm, nguyn liu chnh l nc sunfit, RHM Foods ( 10.000tn/nm ) v Tate anotty1 (4.000tn/nm) u Anh. - Cho n nay x khun cha c dng trong sn xut protein. Tuy vy, ngi ta vn thng thu h si ca chng v ca nm mc, trong qu trnh sn xut cc cht khng sinh, cc enzim, axit xitric di dng sn phm ph ca nh my, nhm s dng protein, vitamin, enzim c trong vo nhng mc ch khc nhau. Nhc im ca sinh khi x khun v nm mc thu theo phng php ny l chng b h hng, v vy phi ch khu sy ngy sau khi tch sinh khi ra khi dy chuyn cng ngh. Trong cng nghip khng sinh, ngi ta c th thu c sinh khi h si gn 17% cc cht cha nit, trong s cc cht cha nit ng ho khong 14%, gn 10% protein tiu ho, 2% cht bo, 2,5% cht x sinh khi ny c th s dng trong chn nui. 2. Qu trnh dinh dng ca t bo vi sinh vt Trong qu trnh sng, t bo vi sinh vt tin hnh trao i cht khng ngng vi mi trng chung quanh. Cc cht dinh dng qua mng t bo v c chuyn ho to thnh nhng cht ring bit cn thit xy dng t bo. Cc cht dinh dng ny khi i qua mng t bo s tham gia vo hai loi phn ng sinh ho:

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- Bin i d ho: lm xut hin nhng sn phm c cu trc n gin hn, Mt s c thi i, mt s khc lm vt liu hoc lm tin cht cho cc phn ng ng ho. Nhng bin i ny cung cp cho vi sinh vt nng lng chuyn ho dng ATP hoc nhng hp cht giu nng lng khc. - Bin i ng ho: m bo s tng hp ca thnh phn mi c cu trc phc tp hn v phn t lng cao hn. Qu trnh ny gi l ng ho hoc phn ng sinh tng hp. Khi trong mi trng c nhng hp cht - vt liu th vi sinh vt s trc tip s dng. Nhng khng phi bao gi trong mi trng cng c sn nhng hp cht vt liu cn cho qu trnh sinh tng hp. Mun c t bo vi sinh vt bt buc phi t sn xut bng cch t bin i d ho nhng thnh phn c trong mi trng nui cy. Cc cht dinh dng ca vi sinh vt ch yu ly mi trng chung quanh cc mi trng dinh dng nhn to cn cung cp y nng lng, cc vt liu xy dng t bo v m bo hiu sut sinh tng hp cao. Thnh phn ca mi trng gm cc ngun thc n cacbon, nit, cht khong, cc nguyn t vi lng v cc cht kch thch sinh trng. Vic la chn cc ngun dinh dng v nng ca chng trong mi trng ph thuc vo c tnh sinh l ca tng chng, tng loi vi sinh vt v iu kin nui cy chng. 2.1. Dinh dng cacbon: Ngun v s ngun cacbon: Cacbon c trong t bo cht, thnh t bo, trong tt c cc phn t enzim, axit nucleic v cc sn phm trao i cht. S ngun cacbon i vi sinh vt v cng ln. Hu nh khng c hp cht cacbon no (tr kim cng, than ch) m khng c nhm vi sinh vt nht nh s dng. Gi tr dinh dng v kh nng hp th ca cc ngun cacbon ph thuc vo: - Thnh phn v cu to ho hc, c bit l mc oxi ho ca nguyn t cacbon. - c im sinh l ca vi sinh vt: + vi cc hp cht c phn t thp nh mt s ng th vi sinh vt c th ng ho trc tip. + Vi cc hp cht hu c cao phn t (tinh bt, protein ) s c phn hu nh cc enzim to thnh cc hp cht phn t thp m vi sinh vt c th ng ho c. + Vi cc hp cht khng tan trong nc (lipit, xenluloza, parafin ..) th vi sinh vt hp th quanh b mt ca chng v phn gii chng dn dn. Ngun thc n cacbon ch yu ca vi sinh vt: l hydrat cacbon trc ht phi k n glucoza. Trao i hydrat cacbon p ng 3 nhu cu ca t bo: + Sn sinh nng lng + To thnh nhng tin cht + To ra cc qu trnh oxi ho-kh bin i nhng tin cht ny thnh nhng sn phm trung gian hay sn phm cui cng xy dng t bo, ng thi tch t trong mi trng mt hoc vi sn phm sinh tng hp.
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Trong cng nghip ln men ni chung, tr trng hp thu sinh khi vi sinh vt n thun, ngi ta c gng to iu kin cho vi sinh vt c th s dng ngun dinh dng cacbon tng hp cc sn phm cn thit nhiu hn l tng sinh khi v to thnh CO2. Nh vy, c cht dinh dng lm ngun cacbon trong qu trnh trao i cht v trong sn xut ln men l cc loi ng sacaroza, maltoza, lactoza, glucoza, cc ng hexoza khc v cc loi bt ng cc nh bt go, bt ng, bt i mch cha ch yu l tinh bt. ng ho c tinh bt, cc vi sinh vt phi tit vo mi trng cc enzim amilaza nh -amilaza, -amilaza, -glucosidaza. H enzim ny c sinh ra trong t bo ri tit ra ngoi mi trng phn hu c cht cm ng l tinh bt. Qu trnh ng ho tinh bt vi sinh vt c gii thiu trong s sau (theo V.Lilli v G.Banettu, 1953): Cc qu trnh ngoi bo Tinh bt , - amilaza Maltoza T bo vi sinh vt Cc qu trnh ni bo Glucoza Cc enzim CO2, ru v cc sn phm trao i hiu k khc 2.2. Dinh dng nit: Vi sinh vt cng nh tt c cc c th sng khc rt cn nit trong qu trnh sng xy dng t bo. Tt c cc loi protein u cu to t axit amin. Cc axit amin dng t do l nguyn liu tng hp cc phn t protein. Cc axit amin c to thnh do qu trnh trao i cacbon v nit. Vic tng hp cc axit amin tri qua nhng hng lot nhng phn ng phc tp vi s xc tc ca nhiu loi enzim khc nhau, nhng c th qui v hai phn ng c trong t bo vi sinh vt l phn ng amin ho v phn ng chuyn amin. Ngun nit + Nit trong khng kh rt phong ph, song n rt bn vng v mt ho hc, kh b oxi ho hoc kh. Ch c mt s vi sinh vt c nh nit mi c kh nng ng ho nit trong khng kh. + Trong tt c cc mi trng nui cy cn thit phi c cc loi hp cht nit m vi sinh vt c th ng ho c m bo hiu sut ln men cao. Cc ngun nit dng trong cng nghip ln men l cc hp cht nit hu c v v c. Cc enzim CO2, ru v cc axit hu c v cc sn phm trao i hiu kh khc Glucoza - glucosidaza

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* Cc axit amin c mt trong mi trng thng khng c vi sinh vt s dng trc tip m phi tin hnh 2 loi phn ng trao i cht: phn ng kh amin v phn ng kh cacboxy1. * Cc axit amin dng hp cht thng l cc protein ca u tng, kh lc .. v pepton. Mun ng ho c cc hp cht ny, Vi sinh vt phi tit vo mi trng h enzim proteaza thy phn cc axit amin thnh cc axit amin. Rt nhiu loi nm mc, vi khun, x khun c hot tnh proteaza cao: Asperillus, Penicillium, Fusarium, Rhizopus, Actinomyces, Clostridium, Bacillus ..v..v.. Nhng axit amin, purin v pirimidin l nhng thc n thch hp hay c Vi sinh vt s dng. S d ho ca purin v pirimidin l hai hp cht c to thnh trong qu trnh thu phn axit nucleic, nucleotit hoc nucleozit thnh cacbonic, amoniac, axit focmioc, axetic hoc lactic v chng c th tham gia vo cc chui chuyn ho khc nhau. * Ur c dng tronuwg cng tng hp c hai tc dng: Lm ngun N v cht iu chnh pH. Di tc dng ca ereaza, uree phn hu thnh CO2 v NH3. (NH2)2CO + H2O
ereaza

2NH3 + CO2

* Nitrat: Vi sinh vt thng khng trc tip ng ho c nitrat m phi qua cc qu trnh bin i: 4AH2 + HNO3 HNO3 Axit nitric HNO2 Axit nit NH3 + 3H2O (HNO)2 Hyponitrit NH2OH Hydrolamin NH3 AH2 - cht kh c trong mi trng.

Qu trnh ny thc hin nh h enzim nitratreductaza. Mui amon: Tt c cc loi vi sinh vt u ng ho c mui amon. Vic s dng ngun N hu c, ure v cc mui amon u gn lin vi vic tch NH3 ra ri hp th vo t bo. Nh vy, NH3 l trung tm ca cc con ng dinh dng nit ca Vi sinh vt. nh hng ca ngun nit n kh nng sinh tng hp ca vi sinh vt khng nhng ch ph thuc vo cc ngun N m cn ph thuc vo t s C:N trong mi trng. T s ny c nhiu ngha. N to cho vi sinh vt c kh nng trao i cht thch hp, kh nng tch t cao cc sn phm sinh tng hp v to thnh cc h enzim tin hnh cc phn ng ho sinh theo chiu hng c li. 2.3. Dinh dng khong 2.3.1. Cc hp cht photpho S c mt ca cc hp cht photpho v nng ca chng trong mi trng c nh hng rt ln n qu trnh trao i cht trong t bo vi sinh vt. Ngoi ra, photpho trong mi trng cn c tc dng iu chnh hot tnh h enzim ng ho cc loi thc n cacbon. Ngun photpho c mt trong mi trng nui cy vi sinh vt thng l cc loi hp cht photpho hu c c trong bt u, cao ng, b ru, kh du v cc hp cht photpho v c, cc mui photpho mono hoc dibazic ca K hoc Na, amon v super photpho.
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Yu cu v photpho ca vi sinh vt ph thuc vo chng loi, vo t l thnh phn mi trng trc ht l t l C:N v iu kin nui cy. Nng cc ngun photpho qu cao cng lm cho vi sinh vt km pht trin v gim hiu sut sinh tng hp. Nu trong mi trng c cacbonat canxi, khi thanh trng, cc cht photpho v c kt hp vi ion Ca2+ v to thnh kt ta. Vi sinh vt thng s dng nhanh nht cc photpho v c ho tan, cn cc hp cht photpho v c khng tan trong mi trng thng s dng t v chm. 2.3.2. Cc cht khong khc Trong t bo vi sinh vt c hng lot cc cht khong khc nh: magi, natri, st, nhm, kali, liti, rubidi, mangan, ch v..v.. Vi sinh vt ly cht khong t mi trng dinh dng, c trng hp phi b sung vo mi trng mt s mui khong hoc c khi chng c sn trong nguyn liu pha mi trng (ng, bt, cao ng, r ng, cacbonnat canxi) v trong nc. Nhng hp cht khong trong mi trng c nhiu ngha sinh l khc nhau: - Lm thay i trng thi ho keo ca cc t bo cht. - Lm thay i tc cc phn ng enzim trong t bo cht. V d nh mui n (NaCL) trong mi trng ln men cc cht khng sinh, ngoi tc dng cung cp ngun ion Cl-, cn c tc dng lm thay i sc thm thu ca t bo, to iu kin tit cht khng sinh t cc si mc, x khun vo mi trng d dng. Mt s kim loi (km, st, mangan, magi ..) l cc cht hot ho enzim. Mt s kim loi nh Zn, Cu, Mn, Mo, B, K, Mg, Ca cng c nh hng ln n hot tnh sinh tng hp ca vi sinh vt. Nm cht u cn vi mt lng rt t nn gi l nguyn t vi lng v thng c sn trong cc nguyn liu pha trong mi trng. C khi cn phi pha thm vo trong mi trng ny dng mui. 3. C ch sinh tng hp protein 3.1. Vai tr iu khin s tng hp protein ca ADN. Protein c phn t rt ln, trong ho hc ngi ta gi l i phn t. i phn t protein c cu to t nhng phn t n gin hn l cc axit amin nh kt k tip nhau. S lng cc axit amin trong phn t protein c n hng trm hoc hng nghn n v, nhng tt c cng ch thuc trong s 20 axit amin khc nhau. Mt loi protein c th khng c y c 20 loi axit amin (thng l khong trn 10), do thnh phn cc protein ca cc vi sinh vt khc nhau th khc nhau. Gi tr dinh dng ca cc loi protein cng hon ton ph thuc vo thnh phn v s lng ca cc axit amin trong vic hnh thnh cc chng loi protein khc nhau. Do t 20 axit amin, c th sng c th hnh thnh v s cc loi protein khc nhau. Trong t bo sng thng xuyn c 2 loi axit nucleic: Ribonucleic (ARN) v dexoxyribonucleic (ADN). Chng khc nhau v thnh phn, cu to ha hc v v tr ca chng trong t bo. ADN ch c hoc ch yu trong nhn cn ARN thng c thy trong t bo cht nhiu hn trong nhn.

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Theo cc thuyt v sinh tng hp protein, cc axit nucleic quyt nh cu trc ho hc v xc nh cc v tr cc axit amin trong chui protein tng hp trong vai tr ca ADN rt quan trng. N quyt nh thnh phn v cu to cc kiu ARN c bit gi l ARN thng tin (ARNt), do quyt nh thnh phn v cu to phn t protein. Nhng ARN i vo riboxm thc hin chc nng lm khun mu. Cc axit amin c xp t vo phn t protein theo trt t ph hp vi cu trc ca ARNt . Qu trnh iu khin sinh tng hp protein ca axit nucleic c th trnh by s hnh 1.1 sau:

Hnh 1.1. S tng hp protein v vai tr nh hng ca axit dexoxyribonucleic (ADN) Theo s ny s tng hp protein xy ra riboxm. y c 2 dng ho li vi nhau: l dng ARNt t nhn ti v dng cc axit amin c hot ho v nh ARN vn ti (ARNv), chuyn n. 3.2. C ch sinh tng hp protein S tng hp protein c th trnh by theo s nh sau: Theo s ny, t nhn t bo, ARNt chui qua mng nhn m i vo t bo cht v nh vo riboxm. Cc axit amin tn ti t do trong t bo cht c gn vo mt loi ARN c bit gi l ARN vn ti (ARNv), ri ARNv chuyn cc axit amin vo mt riboxm. Ti y, ARNt ng vai tr cc khun mu. Trn cc khun mu ny, cc axit amin nh vo mt cch c la chn ti cc riboxm ny v qu trnh tng hp protein c hon thnh. Nh vy, c ch sinh tng hp protein c th tm tt nh sau: - Ni tng hp protein trong t bo vi sinh vt (v c t bo ng thc vt) l cc riboxm.

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- S tng hp protein cn c s tham gia ca cc enzim hot ho v cc ARN vn ti. - riboxm xy ra qu trnh tng hp protein do ADN iu khin, nh vy phi tn ti mt mc xch trung gian gia nhn v t bo cht. l cc ARNt. - Trong t bo vi khun, ARN thng tin rt nhanh chng b ph hu, n ch hon thnh chc nng ca mnh trong mt thi gian rt ngn, ngha l xc nh protein tng hp nn, sau cc phn t ARNt khc li i vo ribxm.

Hnh 1.2. S tng hp protein trong t bo ( theo Lobasov) 4. Cc yu t tng hp protein 4.1. Riboxm Trong t bo cht ca cc vi sinh vt, c 1 loi ht b nh nht trong cc thnh phn cu to nn t bo cht, loi ht ny gi l ribxm, c quan trung tm tng hp nn mi loi protein. Ribxm ca vi khun cha khong 40-60% ARN v 60-40% protein. Ngoi ra, ribxm cn cha mt t lipit, mt s enzim nh ribonucleaza, lexinaminopeptidaza, B-galactozidaza v cht khong (im c bit ca ribxm l giu magi v t canxi hn cc thnh phn khc ca t bo). Ribxm l trung tm tng hp protein ca t bo, nhng khng phi mi ribxm u c kh nng tham gia vo qu trnh ny. S ribxm tham gia tng hp protein thng khng qu 5-10% tng s protein c trong t bo. Nhng ribxm hot

18

ng ny dng nhng tp hp gm 1 s ribxm gi l poliribxm hay l polixm. Cu trc ny khng vng chc v cc ribxm lin kt vi nahu ch bng mt si ARNt. 4.2. ARN thng tin u tin trong nhn t bo xy ra hin tng sao chp nhng on ca phn t ADN v nh s sao chp ny m mt loi ARN c bit c hnh thnh. S sao chp thc hin theo nguyn tc b sung nhau, nhng c mt vi ngoi l: Chui kp gm 2 si ADN tch ri nhau, mt trong 2 si c dng lm khun tng hp nn si ARN. Theo nguyn tc b sung nhau, tng ng vi xitozin (vit tt l X) trong ADN l guanin (G) trong ARN. Tng ng vi timin (T) trong ADN l adenin (A) trong ARN, nhng tng ng vi adenin trong ADN th khng phi l timin na m l uraxin (U) trong ARN. im ngoi l ny khng quan trng lm v v mt ho hc th uraxin v timin cng tng t nhau. Khc vi ADN c cu to chui kp (gm hai si) phn t AND c cu to chui n (ch c mt si). V ADN c sao chp li theo trt t ca cc nucleotit (cc gc kim) trong khun ADN, nn ngi ta ni rng ADN truyn thng tin cho ARN, cn ARN th gi ly thng tin di truyn ca ADN thay mt ADN iu khin s tng hp protein. V th loi ARN c bit ny gi l ARN mi gii hay ARN thng tin. 4.3. ARN vn chuyn ARNv l mt loi axit ribonucleic c bit, c phn t lng thp (khong 25.000 n 30.000). Trong lc , ARNt c phn t lng cao hn trn 10 ln (250.000 n 500.000). Mi mt loi axit amin trong s 20 axit amin thng thng c t nht mt kiu ARNv c th cho mnh, c khi c vi kiu ARNv. Mi mt ARNv, ny c cu trc phn t c bit ring, ch cho php nh kt c vi mt axit amin thch hp v mang n n ARNt t n vo mt ch trn ARNt dnh sn cho axit amin (ch khng cho axit amin khc). Sau khi giao c axit amin ny cho ARNt, ARN v tip tc lm nhim v vn chuyn ln khc v c th vn chuyn axit amin mt ln lin tip nh vy. Trn khun mu (tc l ARNt) c lp y axit amin cn thit th mt chui cc axit amin c hnh thnh. chnh l chui polipeptit. Mt protein c th gm mt chui polipeptit, nhng thng th protein gm mt s chui polipeptit khc nhau. (S d ARNv c kh nng chuyn cc phn t ca mt loi axit amin nht nh n nhng ni nht nh trn ARNt l do trong ARNt c nhng n v m ring bit tc l c nhng b ba ca cc gc kim (nucleotit). Phn t ARNt l mt chui di k tip ca nhng b ba nh th. Mi ARNv cng c mt n v b ba ca cc gc kim c bit. Cc n v b ba nhng gc kim k tip nhau trong phn t ARNt tng ng theo nguyn tc b sung (tc l A trong ARN ny tng ng vi U trong ARN kia v ngc li; G trong ARN ny tng ng vi X trong ARN kia v ngc li) vi n v b ba ca nhng gc kim ca cc ARNv, nh th m ARNv cng vi axit amin nh kt vo n c th tm ch thch hp trn ARNt. 4.4. S hot ho axit amin

19

Trc khi tham gia vo tng hp protein, axit amin phi c hot ho, ngha l c lin kt vi mt ARNv tng ng. Ch sau axit amin ny mi c vn chuyn n ribxm. Qu trnh hot ho axit amin din ra qua 2 bc nh vo xc tc ca cng mt loi enzim axit amin ARNv sintetaza c trng i vi mi axit amin: - Trc ht axit amin phn ng vi ATP thnh phc hp cao nng axit amin AMT. - Tip n axit amin phc hp c chuyn n ARNv tng ng.

20

CHNG 2 S DY CHUYN CNG NGH THU NHN CC SN PHM PROTEIN

1. Sn xut sinh khi nm men t ngun nguyn liu thng thng 1.1. Nguyn liu v x l nguyn liu Cc dng nguyn liu cha hydrat cacbon thng l cc ph phm v ph phm sau: - Cc sn phm cha sacaroza ca cng nghip ch bin ng (r ng ma, r ng c ci, b ma, cn r ng, nc ra th ..) - Nc thi ca nh my sa cn cha nhiu lactoza - Dch kim sunfit c cha nhiu pentoza, hexoza, dch thu phn g. - Cc nguyn liu cha tinh bt v xenluluza khc. im chung nht d nhn thy cc dng nguyn liu trn l ngoi ng, chng cn cha nhiu axit hu c, N.P,S v cc cht khc. S phc tp ny ny sinh hin tng sinh trng kp lm cn tr s dng chng trong nui cy lin tc mt giai on. 1.1.1. R ng V l thuyt: T 1g C6H12O6 c th thu c 0,5 g sinh khi nm men kh (theo nghin cu ca A.J.Forage): C6H12O6 (1g) + O2 (0,4g) + NH3 (0.05g) Hoc theo nghin cu C.L Cooorey C6H12O6 (2kg) + O2 (0,7g) + N,P,K, Mg, S(0,1kg) Sinh khi nm men kh (1kg) + CO2 (1,1g) + H2O (0,7g) CO2 (0,67g) + H2O (0,27g) Q(1,25kcal) Sinh khi nm men kh 0,5g

Cc nguyn liu cha sacaroza (r ng..) l dng nguyn liu l tng nht n sn xut protein n bo, v cc nguyn liu ny cha nhiu yu t kch thch sinh trng, kh,biotin v sn phm protein thu c hu nh sch, khng c. R ng c dng lm cc c cht cho nhiu qu trnh ln men v: - Gi thnh r hn cc nguyn liu cha ng khc. - Ngoi ng sacaroza, r ng cn cha mt s cht v c, hu c v vitamin c gi tr. Thnh phn ca r ng ma v r ng c ci c s khc nhau c bng 2.1.

21

Bng 2.1. Thnh phn ca r ng c ci v r ng ma cha 75% cht kh Thnh phn ng tng s % Cht hu c khng phi ng % Protein (Nx6,25) % K % Ca % Mg % P % Biotin mg/kg Axit pantothenic mg/kg Inozitol mg/kg Tiamin mg/kg R ng c ci 48 - 52 12 - 17 6 - 10 2,0 - 7,0 0,1 - 0,5 0,09 0,02 - 0,07 0,02 - 0,15 50 - 110 5000 - 8000 khong 1,3 R ng ma 48 - 56 9 - 12 2-4 1,5 - 5,0 0,4 - 0,8 0,06 0,6 - 2,0 1,0 - 3,0 15 - 55 2500 - 6000 1,8

S khc bit c bn gia 2 loi nguyn liu ny l: - R ng ma ni chung c pH thp hn (5,5 6,5) do s c mt ca cc axit bo v pH thp dng trong qu trnh lm trong. - R ng ma c mu ti hn ng c ci nn khi dng khng trn vi r ng c ci th nm men thu c s c mu ti hn. - R ng c ci cha nhiu ng sacaroza hn r ng ma v trong r ng c ci hu nh khng c mt loi ng chuyn ho no (c khi ch c khong 1%) trong khi r ng ma c th cha ti 15-25% hidrat cacbon ca n di dng ng chuyn ho. - Ni chung, r ng c ci cha nit hu c nm ln cao hn r ng ma, nhng mt na l betain, mt thnh phn khng c Saccharomyces ng ho, trong khi betain khng c mt trong r ng ma. - S khc bit v hm lng vitamin trong r ng ma v ng c ci cng l tiu chun quan trng: + Cc cht sinh trng c mt trong r ng ma vi hm lng ln: r ng ma cha khong 2,5 g biotin/g gp 20 ln hn r ng c ci. + Trong khi r ng ma ngho cc cht khong v axit amin: r ng c ci cha axit pantothenic gp 2-4 ln so vi r ng ma. Nh vy, r ng dng nui cy nm men khng nhng l ngun ng m cn cung cp cc hp cht hu c khc, cc mui khong cn thit v cc nhn t sinh trng. Tuy nhin, ngoi cc thnh phn c ch cho s sinh trng ca nm men, r ng cng c th cha cc hp cht c hi c th lm h hng qu trnh ln men: hm lng canxi cao ni ln cht lng thp ca r ng v c th gy nn nhng kh khn trong vic sn xut nm men. R ng cng c th d dng nhim cc vi sinh vt v gy nn nhng vn khng c li trong ln men. X l r ng:
22

R ng cn c x l cht t trc khi nui cy. Thng thng n c axit ho bng axit sunfuric ti pH = 4 v un nng ti 120-1500C trong 1 pht kt ta mt s cht v c v cht l lng. Cn phi loi b mt phn cc cht sinh trng, ng thi b sung cc mui khong cn thit (nh ur 0,15%, KH2PO4 0,35%, Mg, Ca) v c th phi thm hn hp cc axit amin dng protein thy phn (dch nm men t phn, dch thi trong sn xut nc chm, dch b ru giai on nhn ging). Khi chun b phi trn, r ng c ci v r ng ma phi c x l tch bit trong cc khu pha long, iu chnh pH, un nng, lm trong, kh trng ri mi c phi trn. Thng pha long n nng ng khong 5-6%. Sau khi chun b xong mi trng dinh dng, tin hnh thanh trng nhit 1200C. 1.1.2. Cc nguyn liu khc: - Dch kim sufit: Nc thi cc nh my giy xenluloza theo phng php sunfit gi l dch kim sunfit (SWL-Sunfit Waste Liquors) cng l ngun nguyn liu tt sn xut nm men. Thnh phn hydrocacbon ca n ch yu l ng pentoza, mt loi ng ch c nm men mi chuyn ho tt. Ngoi ra cn c linhin, phi xenluloza, mt s axit hu c Khi s dng dch kim sunfit cn phi c lm nng v thng kh trc khi nui nm men loi bo cc yu t kim hm (SO2 v furfurol). B sung cht dinh dng vo dch thi trn (nh NH4+ v PO4--), iu chnh pH v khong 5 s c mi trng nui cy nm men kh tt v lng sinh khi nm men sinh ra sau qu trnh ln men c cht lng ng k vi cc thnh phn nh sau: protein (46% cht kh), lipit (7-8%), photpho (1,8%), axit nucleic (10%) Ngi ta tnh rng khong 5 tn bt xenluloza sn xut giy s thi ra mt lng dch kim sunfit cha ti 180 kg ng. Dch ny hp ph nhiu O2 nn khi nui cy nm men c th gim mc cung cp oxi ti 60% so vi bnh thng. - Cc ngun xenluloza thc vt (g, rm, r b ma, li ng..) c ch nhiu trong sn xut nm men. Trc ht cn phi thu phn xenluloza bng axit hoc bng enzim. Nu dng g th thng phi thu phn bng axit sunfuric. - Nc thi ca nh my ch bin sa, cn gi l nh thanh (lactoserum): trong qu trnh ln men lactic ch bin phomat, sau khi kt ta cazein ra khi sa, phn cn li gi l nh thanh c cha lactoza, protein, axit lactic, axit bo, mt s vitamin v mui khong. Ngi ta chn chng nm men thch hp c th thu phn c lin kt -galactozidaza v thu c sinh khi nm men dng kh c thnh phn protein th khong 32%, lipit 4-5%, lacto khong 23%. Chng nm men C.utilis v C.pseudotropical rt thch hp trong mi trng trn y. - Bt ng cc: l ngun sn xut sinh khi nm men rt tt. Bt hoc tinh bt dng vo mc ch ny trc tin phi tin hnh thu phn bng axit hoc bng enzim ca mm m hoc enzim ca vi sinh vt bin cc polysacarit thnh cc dng ng m nm men c th ng ho c. Trong trng hp dng nm men Saccharomysces cerevisiae th c th kt hp chng ct thu ly cn t dch thi sau khi tch sinh khi. Nh vy trong dy chuyn cng ngh cn phi trang b thm b phn chng ct. Dch ly tm c a vo h li tm tch (separator) v dch thi sau khi c tch ra c chuyn n khu chng ct.
23

1.2. Chng nm men: Tu theo tng loi nguyn liu khc nhau, chng ta c th s dng nhng chng nm men ph hp to sinh khi c hiu qu nht. i vi nguyn liu l r ng, dung dch ng, nm men thng dng l Saccharomysces cerevisiae, Candidas tropicalis, Candidas utilis. i vi nguyn liu tinh bt hay nc thi tinh bt, dng chng nm men tng ng l Endomycopis fibuligera hoc phi hp gia Endomycopis vi Candidas tropicalis. Nu nguyn lin l b ru, chng nm men l Candidas utilis. Nu s dng lactoserum (nh thanh sa) th chng nm men c chng l Torula cremoris, T. lactosa. Nguyn liu l kim sunfit, chng nm men s dng l Cryptococus diffluens, Candidas tropicalis, Candidas utilis. Tuy nhin trong trng hp khng c nhng chng nm men ph hp, chng ta c th thay th mt trong cc chng trn y. 1.3. Mt s qui trnh cng ngh tiu biu 1.3.1. Sn xut sinh khi nm men t r ng R ng X l Pha long Thanh trng Cc mui v c Mi trng dinh dng

Nm men Nhn ging

Nui thu sinh khi Li tm Sinh khi Sy kh

24 Thnh phm

X l

Thi b

1.3.2. Sn xut sinh khi vi sinh vt t nguyn lu cha tinh bt hoc xenluloza: Nguyn liu Thu phn To mi trng dinh dng Nui thu sinh khi Li tm Sinh khi Sy kh Thnh phm 2. Sn xut sinh khi vi khun Nguyn liu v vi sinh vt Ngoi nm men, ngi ta cn s dng rng ri vi khun sn xut protein t nguyn liu xenluloza. Protein vi khun c hm lng axit amin cn i hn nm men, t l protein trong t bo vi khun li rt cao, trung bnh l 60-70%, c loi ti 87%. Nhiu nghin cu thnh cng trong vic nui vi khun protein t cy c, rm r nh: Nm 1969, Srinivaan v Han phn lp c hai loi vi khun c kh nng cng sinh l Cellulomonas v Alcaligens. Trong mi trng xenluloza, nu ch ring mt mnh Alcaligens th hu nh vi khun khng pht trin c. Nu ch mt mnh Alcaligens th vi khun pht trin rt km. Nhng nu mt nui cy cng mt lc c hai vi khun ny th sinh khi tng vt ln ( hinh 2.1) - Cc nh bc hc M trng i hc Luisiana phn lp t b ma mt loi vi khun phn hu mnh xenluloza ca ngun nguyn liu ny. Cng trnh nghin cu ny ang c ng dng c kt qu M v Cuba: C 113 136 kg b ma c th sn xut c 18 -23kg protein. Thnh cng ny c mt ngha thc tin v n cho php s dng b ma, li ng, rm r sn xut protein mt cch trc tip m khng phi qua khu thu phn bng H2SO4. Ch bin enzim xenluloza hoc amilaza Nm si Nhn ging Dch li tm Li tm X l Thi b

25

- Hai nh bc hc ngi Austraylia l Roper v Moss a ra mt phng php sn xut protein vi khun t c, rm, b ma, v u, mn ca, dm bo .. vi hiu sut rt cao, c th t n 35% so vi lng rm c s dng. c bit protein do Roper v Moss thu c t rm r c cht lng tng ng vi lng trng g. Gio s Macmilan, nh lnh o phong tro chng i Australia gi cong trnh ca hai nh pht minh ny l Mt ting n k diu trong cuc chin u vi nn i protein ca th gii.

Hnh 2.1. S pht trin ca Cellulomonas v Alcaligenes trong mi trng xenluloza

26

3. Sn xut protein vi sinh vt t du m v kh t 3.1. c im lch s: - Nm 1925, Tauson pht hin kh nng phn gii cacbua hydro ca vi khun. - Nm 1940, nhiu nh khoa hc trn th gii nghin cu sau v vic s dng vi sinh vt trong thm d v khai thc du kh. - Nm 1961, Fush nghin cu thng k c 26 ging trong c 75 loi vi sinh vt c kh nng phn hu mch vng. - Nm 1962, cng trnh u tin v kh nng s dng du m kh t nui cy vi sinh vt thu nhn sinh khi giu protein cho gia sc c cng b ti Hi ngh du m quc t ln th 6. Sau nhiu nh khoa hc phn lp c 498 chng nm men c kh nng phn gii cacbua hidro. V t c nhiu nh my sn xut c sinh khi nm men m sn phm cha ti 60 70% protein. 3.2.Nguyn liu 3.2.1. Du m Ch nhng phn du m nht nh mi c vi sinh vt ng ho nh: - Cc alkan (paraphin) vi chiu di chui C10 - C20 - Cc alkin, anken, hydrocacbon thm. - Cc parafin chui ngn cn li trong phn du m c nhit nng chy thp. - S dng n-parafin tinh khit c tch t m da trn cc nguyn tc sng phn t lm c cht c u im l ngun C b tiu th hon ton v khng li nhng cacbua hidro c. C ch ca s hp th ankal cho n nay cng cha c lm sng t y . So vi cc t bo sinh trng trn glucoza th nm men nui trn cacbua hidro c mng t bo dy hn v c np nhn.. Tuy nhin cc t bo ny khng gp kh khn g trong vic hp th nhng c cht khng tan trong nc c b sung vo mi trng vi nng 2 - 4%. 3.2.2. Kh thin nhin - Me tan: Metan l thnh phn chnh ca kh thin nhin. Tuy nhin metan khng ch l nguyn liu trong lng t m cn c to thnh qua con ng vi sinh vt nh s ln men metan v c sinh ra trong cc b cha bn mc nt trong cc thit b lm sch. Nguyn tc sn xut protein t kh thin nhin l nui vi khun trn dch mui amon v mui khong c thng xuyn thi kh metan v khng kh. u nhc im ca vic s dng metan: u im: - Kh thin nhin r hn du m nhiu ln. - Phn kh khng c vi sinh vt ng ho c loi b mt cch d dng. V vy sn phm rt tinh khit v khng tn km dung mi cho vic ra t bo nh khi s dng du m lm c cht. Nhc im: - Vi sinh vt ng ho kh thin nhin u l cc vi sinh vt hiu kh. Do mi trng dinh dng phi thng xuyn thi hn hp kh metan v oxi hoc l khng kh rt d gy n. Nu nng hn hp kh cao rt d bt la v n, cn nng kh thp th vi sinh vt khng h hp. C hai trng hp khng dinh dng v ngt th, vi sinh vt u pht trin km v hiu sut nui cy thp.

27

- thc hin c qu trnh sinh tng hp protein th oxy v metan phi c chuyn t tng kh sang tng lng bt kh mang nhin liu v cht oxy ho n cc t bo vi sinh vt ang sinh trng mt cch nhanh chng v thc hin qu trnh ng ho. Tuy nhin, ho tan ca metan v oxy trong nc thp. C th khc phc bng cch l tng p sut d trong thit b nhng vic ch to thit b chu p lc cao s phc tp v khng kinh t. Hoc a mt dung mi hu c no vo mi trng dinh dng tng ho tan ca metan, nhng s lm cho vi sinh vt thch dung mi hn metan v nh vy vic dng kh thin nhin mt ht ngha. - Metanol: khc phc nhng nhc im ca vic s dng metan, c th s dng metanol thu c t metan nh s oxy ho ho hc. l nh nhng u im sau ca metanol: + Metanol d tan trong nc nn c th dng nng cao hn (2-3%). + Nhu cu oxy ca s ng ho metanol l thp hn. + C th dng nm men ng ho metanol. M nm men c kch thc t bo ln hn vi khun nn nng lng cn thit cho qu trnh li tm tch sinh khi t hn so vi vi khun s dng ng ho metan. Tnh kinh t cao hn. Tuy nhin dng metanol c nhc im sau: + Metanol t hn nhiu so vi metan hoc kh thin nhin. + Thu hoch t bo t metanol thp hn t metan. - Etan, propan, butan: Vic s dng cc alkal dng kh chui ngn cha trong du m nh etan, propan, butan din ra khng qua vi khun ng ho metan m ch trong hn hp qun th cha cc c th c kh nng ni trn (Mycobacterium, Nocardia, Pseudomonas). 3.3. Cc chng vi sinh vt 3.3.1. Vi sinh vt phn gii cacbua hidro: - Vi khun: Achrobacter, Alkaligenes, Bacillus, Bacterium, Corynebacterium, Micrococcus, Flavobacterium, Pseudomonas, Micromonospora, Mycobacterium, Mycococcus, Nocardia. - Xa khun: Streptomyces, Actinomyces. - Nm men: Candida, Cytomyces, Debaryomyces. Endomyces, Hansemula. Monolia, Scopuloriopsis. - Nm si: Acremonium, Aspergillus, Penicillium. 3.3.2. Vi sinh vt phn gii kh thin nhin: Ch yu l cc vi khun: Mycobacterium, Pseudomonas, Methanomonas, Bacillus, Corynebacterium, Brevibacterium, Flavobacterium, Bacterium. 3.4. C ch chuyn ho Qu trnh ng ho cacbon t du m v kh t c th ra dng tng qut nh sau: (1) Hydro cacbua Ru bc 1 hoc bc 2 andehyt Cht bo (2) i vi n-alkal, c th l:

28

n-alkan

Ru bc 1 andehyt

hoc

Ru bc 2 metylxeton

Sp Di ra hoc ngn bt 1,2n nguyn t Axit bo

(3) i vi cc hp cht khng no (th d nh 1-olefin), ngi ta cho rng qu trnh oxy ho nh vi sinh vt c th i theo con ng sau:

1-olefin aldehyt 1,2 - epoxyt +H2o Oxy ho 1,2 - diol Axit bo ngn bt 1C -CO2 Axit bo

(4) C ch chuyn kh metan: Cc vi sinh vt phn gii kh metan thnh CO2 v H+ hot ng. Vi sinh vt s dng H+ kh tip CO2 to thnh cc hp cht hu c theo nhng phng trnh tm tt sau: + 2 H2O CO2 + 8(H) CH4 (CH2O) + H2O 4(H) + CO2 2H2O 4(H) + O2 + O2 (CH2O + H2O CH4 Cc axit bo to thnh s c li cun vo cc qu trnh ng ho tip theo, tham gia vo cc qu trnh trao i cht t bo vi sinh vt trong chu trnh Kreb. Mt phn cc axit amin c to thnh s kt hp vi NH3 cho ra cc aminoaxit. Nh cc phn ng chuyn amin m mt s loi axit amin c to thnh ngy cng phong ph v cui cng, di s iu khin ca ADN trong t bo vi sinh vt, cc axit amin ny s c t hp li vi nhau thnh cc phn t protein. 3.5. S qui trnh cng ngh sn xut sinh khi nm men S cng ngh sn xut sinh khi nm men t cc sn phm du m cng tng t nh t cc ngun hydrocacbon, tc l gm cc giai on sau: - Chun b mi trng dinh dng.
29

- Nhn ging v ln men. - Tch v ra sinh khi nm men, - Sy kh. Qui trnh cng ngh sn xut sinh khi nm men c th t du m th v parafin tinh khit cng tng t nhau. Tuy nhin dng du m th th i hi qui trnh cng ngh phc tp hn, mc d gi thnh tng i r hn. Dng parafin th khu tch nm men c th b bt khu ty ra bng dung mi hu c v thc t parafin c nm men s dng hon ton

30

CHNG 3 CNG NGH SN XUT PROTEIN T CC NGUN HYDAT CARBON 1.Cng ngh sn xut protein trn nguyn liu polysacarit cha thu phn 1.1. S cng ngh sn xut nm men t cc nguyn liu thc vt thu phn bng H2SO4. Nguyn liu Fucfurol v cc cht bay hi khc, linhin Thu phn Trung ho Lc v lm sch Lm ngui (30 320C) Ln men Tch sinh khi Nui men ging Dch thi Chng ct, tinh ch Cn ethanol Cc loi dch th cha ng c tp trung vo b ln trc khi phn phi vo cc b ln men. Sau c trung ho bng sa vi v lm trong. cc b lm trong thng c cc cch khuy v ng thng kh, nh cc cht c ch dng bay hi nh fucfurol, SO2 s c loi b. Sau khi c trung ho v lm trong, dch lng cn nng s c lm ngui n nhit 30 32o C, ri pha long n mt nng ng thch hp cho nm men v ty theo yu cu m b sung cc mui v c. 1.2. X l nguyn liu v chun b mi trng i vi cc nguyn liu ban u dng sn xut protein n bo t nm men cn phi c x l s b. Sau tin hnh pha ch mi trng. Tu tng loi nguyn liu v chng vi sinh vt nui cy, chng ta s c cc thnh phn mi trng thch Nc + H2SO4 (nng 0,5 0,6%) Hi nhit (179 1900C) Sa vi B lng (CaSO4)

KCl, superfotfat, (NH4)2SO4 Du ph bt Cht iu chnh pH

31

hp. Ni chung, ngoi ngun c cht c bn l ngun cacbon ra, chng ta cn a vo mi trng ngun nit, photpho, kali, magi, cc nguyn t khong khc na. Ngun nit thng l cc mui sunfat, ngun photpho l supephotphat, KKCl, Mg MgSO4. C th dng amoniac gi pH xc nh. Trong qu trnh ln men cn cn ngun cht sinh trng nh cao ng, cao nm men, hoc cc dch thu phn khc vv.. Cc thnh phn mi trng c ho tan, lc b cn, iu chnh pH n 4,85,2 bng axit sunfuric hoc axit clohydric (i vi mi trng r ng th pH l 4,2 4,5). Nui cy nm men trong sn xut SCP chia lm hai giai on: - Giai on nhn ging c lng ging (s lng t bo). Giai on chun b vt liu nui cy cn phi v trng. Mi trng nhn ging v khi tin hnh nhn ging cn phi v trng. - Giai on ln men: Giai on nui ln qui m cng nghip hay iu kin pilot c th thc hin trong thng kn hoc thng h, iu kin khng cn v trng. Trng hp khng cn v trng th khng cn thanh trng p sut d ca hi nc, m ch cn un nng hoc ozon ho, lc kh khun, clo ho, x l qua vi focmalin v..v.. 1.3. Nui cy nhn ging Nui cy nhn ging u tin c thc hin phng th nghim: ging ng nghim c cy chuyn vo bnh tam gic c mi trng v trng, sau cc bnh c ging c nui cy trn my lc vi nhit bnh thng t 25 300C n tui sinh l thch hp s cy vo mi trng nhn ging ca phn xng : nhn ging cp 2 trong cc bnh thp kn c sc kh n khi t c 3,5 5g sinh khi trong 1l dch nui. Qu trnh kt thc sau 12 15 gi. C th nhn ging cp 3 cc ni c th tch ti 4 5 m3 . T l tip ging chuyn cp l 1:10. Trong qu trnh nhn ging dng nc amoniac gi pH v thi kh lin tc. T ni 4 5 m3 s c chuyn sang thng 12 15 m3 v ti vi chc m3 hoc to hn. Nui ln men cng nghip : l nui m rng trong phn xng khng cn phi v trng. Nhiu nh my t cc ni ln men kn hoc h, thng th tch cc ni ln men l vi chc mt khi, c th ti 500m3. Tin hnh nui men theo phng php bn lin tc cho hiu qu kinh t cao: khi t lng sinh khi c trong dch nui cy ly dn ra v cho thm mi trng mi vo ni ln men c hm lng ng khong 1-2%. 1.4. Cc iu kin k thut: sn xut sinh khi nm men giu protein cc dng nguyn liu trn cn m bo cc iu kin k thut c bn sau: Nng ng trong dch nui cy phi m bo t 2 -4 %. Mui ur 3g/l. Suphephotphat 4g/l. Khng kh v trng Thi gian nui t 18 36 gi.

32

Nhit nui cy 28 30oC. pH mi trng 4,5 5,5 .

Qu trnh sn xut CSP l qu trnh hiu kh. V vy bt buc phi thng kh mi trng. Vic cung cp khng kh c mt s tc dng sau: - Cung cp O2 cho vi sinh vt tng hp vt cht t bo. - Tch CO2 ra khi dung dch nui cy. - Xo trn mi trng, to iu kin thun li cho qu trnh trao i cht tt hn. Khng kh cung cp cho qu trnh sinh tng hp protein phi c lm sch trc khi cho vo thit b ln men. Mt yu t cn ch na l nng ng trong qu trnh nui cy. Khng nn nng ng qu cao trong mi trng v s c ch s tng trng t bo s to ra nhiu sn phm ph khng cn thit. Do nng ng cn khng ch < 4 % l thch hp. 1.5.Thu hi sinh khi: Bt v sinh khi trn ra ngoi trong qu trnh ln men c tch trc tin theo phng php to thnh bt cng vi sinh khi tro ra ngoi ri a i li tm tch. Bt v sinh khi trn ra ngoi c thu gom li i x l bng phng php tuyn ni (flotation) ri a i li tm qua cc my li tm tch (Seprator), c c chn khng. Sinh khi nm men thu c dng st c 75-80 % nc, 20-25% cht kh trong c cacbon 40-50%, nit 7-10% tng ng vi 40-60% protein, hydro 5-7%, oxy 25-30%, cc nguyn t v c 5-10% ( photpho v kali chim 95-97% tng lng tro, s cn li l canxi, magi, nhm, lu hunh, clo, st. Ngoi ra cn c mt lng nh nguyn t Mn, Zn, Mo, Bo, Coban...). Sinh khi c a vo sy my sy 2 trc hoc sy phun. Trong t bo nm men k c vi khun, c nhiu vitamin nhm B (tr VTM B12): tiamin, riboflavin, axit niconitic, axit folic, c bit rt giu tin VTM D2 (ergosterin). Di nh sng tia t ngoi (tia cc tm) ergosterin s chuyn thnh VTM D2. V vy trc khi ng gi sn phm sinh khi nm men c chiu tia t ngoi VTM ho sn phm. Qu trnh cng ngh sn xut sinh khi nm men c gii thiu cc s sau :

33

Hnh 3.1. Qu trnh chun b mi trng v nui cy iu kin v trng. 1. Bnh cha dch amon sunfat. 2. Supephotphat 3. Nc nng 4. Sa vi 5. KCl 6. Bnh t phn nm men. 7. Bnh pha mi trng 8a,8b,8c. thanh trng 9. Bnh lng 10. Ni ging cp 2 11. Ni ging cp 3 12. Nui m rng

Hnh 3.2. S nui v thu sinh khi nm men.

34

1. Ni ln men 2. Thng tuyn ni 3. Bm 4. Bnh tch kh 5. v 8. Li tm tch I v II 6. Thng cha men 7. Bm nc 16, 17. Ct xiclon u.

9. Thng cha men c 10. Bnh iu chnh nhit lin tc 11. Thng tp trung men trc khi c c 12. Thit b c c chn khng 13. Thit b to chn khng 14. Thng cha men trc khi sy 15. Sy phun 18. Thng tn tr

1.6. Cng ngh sn xut protein t sn khng qua qu trnh thu phn ban Nghin cu ca Azoulay gip cho hng Adour Entreprise ( Php) phn lp c mt chng nm men Candida tropicalis c th ln men trc tip sn m khng cn qu trnh thu phn ban u theo qui trnh cng ngh nh sau: C sn c ra, thi mng ri nghin nh. Ho tan tinh bt bng cch un nng > 100oC ng thi cng l thanh trng trnh nhim tp khun. Cch x l ny cng lm phn hu cc axit hydroxianic c trong sn (Manihot esculenta) chuyn thnh amon v axit focmic. Ln men: qu trnh ln men c thc hin trong mt ni ln men c sc kh. Dch ln men thu c cha 10 25 kg nm men/m3 . Sau khi li tm, dch trong c thu hi quay tr li ln men m sau, cn sinh khi nm men cha ti 15 % cht kh c a i x l tip thu hi sinh khi. Trong mt s trng hp chn nui gia sc (ln), c th b sung trc tip nm men ti vo thc n m khng cn lm kh (Inchauspe, 1986). 1.7. Sn xut protein t chui: Encuador, nc xut khu chui hng u th gii, v nc Colombia cng l nc ch cht v xut khu chui, lun lun c mt t l ln sn lng chui (> 25 %) khng xut khu c v km cht lng. V vy chui c th l nguyn liu quan trng cho sn xut SCP. Khong 15000 tn chui c th chuyn ho thnh 100000 tn sinh khi mi nm. 2. Cng ngh sn xut protein trn dch thu phn g. M, nm men gia sc c sn xut t dch kim sunfit ca cc nh my bt giy: - Mt s cng ty nh cng ty Enviroson Ltd dng nc thi bt giy em kh trng ( 121oC/1gi) ri lm ngui n 37oC lm c cht cho s pht trin hiu kh ca mt loi vi nm Chactomium cellulolytium. Ngoi ra trong mi trng cn b sung cc cht dinh dng khc cha nit, photpho v kali. Vi nm tn ti nh nhng vt rn dng huyn ph, bm vo si xelluloza trong c cht v tit ra enzim xenluloza lm chuyn ho xenluloza thnh glucoza. Sau khi ng ho c xenluloza, vi nm to sinh khi v thi ra CO2. i vi dch kim sunfit ny, cc chng nm men sn xut cn c lm quen vi nng axit sunfur cao ngay trong cc b tp trung. C mi tn cacbon ca c cht th c th to ra 500kg sinh khi. Sn phm cui cng cha
35

40 % protein, 60% lipit, xenluloza v hydrat cacbon (vi sn phm c m 5%) (theo Chemical Engineering News, 6-2-1984). Mt s nh my khc s dng cng ngh Pekilo ca cng ty Tampella vi chng nm thuc chi Paccilomyces nui cy trn dch sunfit. Trc khi ln men, hu ht SO2 c loi b bng cch sc bng hi nc qua dung dch sunfit. a vo ni ln men cc cht c cht dinh dng khong nh axit photphoric, KCl, kh NH3 v sc u bng khng kh nn. Sau khi ln men, sinh khi vi nm c tch ra v ra trong cc my p lc n Bx = 35%, sau em sy bng khng kh nng ri p v to ht. 3.Cng ngh sn xut protein trn dch thu phn cc nguyn liu thc vt 3.1. Sn xut protein trn nguyn liu chit ng v nc chit la m. 3.1.1. Nguyn liu: l nc chit ng v nc chit la m Thnh phn nc chit t la m v ng ( theo kt qu nghin cu ca Vin nghin cu tinh bt M. V. Plevaco) c cho bng 3.1. Bng 3.1. Thnh phn nc chit t la m v ng Ch s Nng cht kh (%) ng kh g/1 Ni t g/1 Tro g/1 A. amin theo sc k, trong lyzin a. aspactic v glutamic, alanin Biotin /ml Biotin /100g cht kh 10 0,18 360 7 0,75 1025 Nc chit la m 7,1 2,0 3,14 4,79 Nc chit ng 7,5 4,0 0,62 6,46

Trong nc chit la m, thnh phn ng kh c mantoza v glucoza, chim 30% cht kh ca nc chit. Trong nc chit ng, ng kh l mantoza, glucoza, xyloza trong glucoza chim t l cao nht. Hm lng biotin trong nc chit ng gp 4 ln nc chit la m. 3.1.2. Chng nm men: nui nm men, dng chng Candida Tropicalis c kh nng ln men c mono v disacarit. Chng ny cho hiu sut cao v c kh nng pht trin trn nc chit c nng cao. 3.1.3. Nui cy nm men: Gin on v lin tc c sc khng kh. Nng ti thch ca mi trng dinh dng nc chit la m l 1 2%, cn nc chit ng l 11,5%. Theo phng php lin tc vi qui trnh nh sau: Nc chit v mui khong (amon sunfat v i khi supephotphat ) c a vo thit b nui men ( cho nm men ging ) vi tc tng dn trong 6 gi, trong khi mi trng c thng kh lin tc. n cui gi th 6, lng sinh khi nm
36

men sinh ra khong 25 30 % tnh theo nm men p. n gi th 7, mt phn mi trng t thng nui men c chuyn vo thit b ln men ph: 2 gi u 10 %, 2 gi th 2- 15% v 2 gi cui 20% th tch mi trng chung. Sau 12 gi l bt u sang giai on 3, giai on nui cy lin tc. Trong giai on ny, c mi gi thit b nui men li ly ra 20% dung tch, ri b sung vo mi trng nc chit, nc v mui khong. Amon sunfat cho vo tnh theo hm lng cc cht c trong nm men p: nit 2%, photpho P2O5 1,5 -2%. Tc pht trin nm men trn nc chit la m bng 16-20% ( so vi trng lng nm men trong thit b) trong 1h, cn trn nc chit ng 20-22% trong 1h. Hiu sut thu c nh sau: c 100kg cht kh tuyt i ca nc chit la m thu c lng nm men p ( c m 75%) l khong 194kg, cn t nc chit ng l 240 -260 kg. 3.2. Sn xut sinh khi nm men trn nguyn liu nc chit t b khoai ty 3.3.1. Nguyn liu: Trong cc nh my sn xut tinh bt t khoai ty, nc dch chit l nc p c trch ly t b khoai ty, t cc b lng v t cc thit b li tm. Trong nc dch cha khong 96% dch t bo khoai ty, trong c gn 77,8% cht nit, 88% gluxit ho tan, 87% lipit v 63,3% cht khong (tnh theo khi lng ca cc cht ny c trong khoai ty. Trong 1m3 nc dch cha khong 0,54g kali oxit ( K2O) v 0,09 kg axit photphoric. Cht kh cui nc dch ca cc nh my tinh bt c thnh phn (%) nh sau: Thnh phn cht kh - Gluxit tan - Cht khong - Hp cht N2 - Lipit - Cc cht khc % 0,97 0,67 1,76 0,13 0,67

Bn cnh nc dch, nc sa ca cng nghip sn xut tinh bt l nc thu c khi ra tinh bt cc my cha 0,16% tinh bt kh tuyt i so vi s lng khoai ty em ch bin. Lng nc ra chim khong 170 - 270% so vi khi lng khoai ty. Nc ra cha ch yu cc cht v c v hu c ho tan. Thnh phn ho hc ca nc ra rt khc nhau v ph thuc nhiu yu t nh: k thut sn xut, cht lng nguyn liu, iu kin bo qun nguyn liu, kch thc c v...v...Hm lng tinh bt khng vt qu 1g/l. Nc dch v nc ra ny nu khng c tn dng ch bin, hoc x l trc khi thi ra ngoi, s lm nhim bn ngun nc, nu thi ra cc sng, h, ao s lm cht nhiu c. Vi hm lng protein kh ln trong nc dch, nu thi nc ny vo cc cnh ng ti lm sch sinh hc t nhin, cng khng c hiu qu.

37

V vy s dng nc dch thi ny sn xut sinh khi nm men rt c ngha v kinh t v bo v mi trng. Cc nghin cu chng t rng nc dch t bo khoai ty c cha a.aspactic, biotin, D-alanin, l nhng cht rt cn cho nm men sinh trng, pht trin v sinh sn. Nng mi trng thng hp nht l 1,5-4oBx. 3.2.2.Hiu sut tng thu hi. Theo kinh nghim sn xut, c mi tn khoai ty em ch bin c th thu c khng t hn 30kg nm men bnh hoc 7-8 kg nm men gia sc kh. Tnh theo lng protein thu c th n bng 300kg khoai ty. 3.2.3. Qui trnh cng ngh: Sn xut nm men gia sc c th theo s cng nghip sau y ( Hnh 3.3)

Hnh 3.3: S k thut sn xut nm men chn nui. 1. B phn lc 3. Bm pitng 5. Thng cha dch nm men 7. Bm 9. Thng cha cht ph bt 11. Bnh o 13,16. my phn ly 20. My sy hai trc ln 21. Bng chuyn 23. Bn ng gi 25,26. Bng chuyn 29. B ra cc chi tit ca my phn ly. Vic nui nm men theo qui trnh trn c trnh by bng 3.2. 2. Thng trung gian 4. Thit b nui men 6. Thng ho tan cc cht b sung 8. Thng nh lng 10. Bm 12, 15, 18. Bm 14. Thng cha dch c c ln I 17. Thng cha dch c c ln II 22. Phu 24. Cn 27. Qut gi turbin

38

Bng 3.2. Qui trnh nui cy nm men v trng

Gi 1 2 3 4 5 6 7 8 9 10 11 Tng cng sau 11 gi Tng cng sau cc gi tip theo a vo (Mi trng: nc dch) m3 4 4 4 8 8 8 8 8 8 8 8 80 15 a vo (Amon sunfat) kg 5,3 5,3 5,3 10,6 10,6 10,6 10,6 10,6 10,6 10,6 10,6 106 6 avo(Axit octphotphoric) kg 0,45 0,45 0,45 0,90 0,90 0,90 0,90 0,90 0,90 0,90 0,90 9 1,8 Ly ra m3 15

Thuyt minh qui trnh sn xut Nc dch c tch bi tinh bt nh b phn lc 1, chy xung bm pitng 3 qua thng trng gian 2 vo thit b nui nm men 4. Amon sunfat sau khi ho tan trong thng 6, ri cng vi axit octphotphoric c my bm 7 bm vo thng nh lng 8, ri i vo thng nui men 4. S sinh sn ca nm men theo qui trnh lin tc t thng ln men 4 v thng cha sinh khi 5. Cht ph bt t thng cha 9 c bm 10 a v thng nui men qua bnh o 11. Thng nui men lun lun c sc kh nh qut gi turbin 27 Vic nui men theo qui trnh bng 3.2. Sau 11 gi ln men, khi thng 4 cha y mi trng, ngha l 80m3 th bt u tho lin tc nm men xung thng 5 vi lng 15m3/ gi. ng thi a lin tc nc dch vo vi lng bng chng y ( 15m3/ gi) cng vi amon sunfat v axit octphotphoric . Sinh khi ly c t thng 5, nh bm 12, chy lin tc vo my phn ly 13, ri vo thng cha 14, sau tit tc phn ly ln 2 thit b 16 ri cha 17. thng 17,

39

nh my bm 18 vo my sy 20. Men kh c bng chuyn 21 chuyn sang phu 22, vo b phn ng gi trn bn 23 vba c cn trn cn 24. sau qua cc bng vn chuyn 25, 26 i phn ly. 4. Cng ngh sn xut protein trn b ru t r ng Phn ln cc nh my sn xut ru t nguyn liu r ng c mt s lng b thi rt ln. Hin nay lng b thi c c s dng li vi s lng khng ng k nn phn ln phi thi ra ngoi. Nu khng c x l ng mc, b ru phn hu khng hon ton, thng l ngun gy nhim h cha nc. Ngoi ra, do s phn hu cc hp cht hu c cha trong b ru to thnh nhng cht c mi hi thi gy nhim mi trng khng kh trm trng. Ngoi ra cn phi tiu tn mt khong chi ph ln cho vic thi b ( lm sch, m rng, thay th ng ng v..v..).V vy vic nghin cu ng dng cng ngh sn xut cc sn phm thc phm hoc l sinh hc c nguyn liu t b ru ca r ng c mt ngha quan trng v kinh t v bo v m trng. B ru c th c ch bin thnh nhng sn phm sau: - Glyxein v than cc. - Tch t r ng axit glutamic v betain lm thc n gia sc. - Sn xut nm men bnh m v sinh khi nm men cho gia sc trong sn xut sinh khi nm men cho gia sc c quan tm v sn xut nhiu hn c. Qui trnh cng ngh sn xut sinh khi nm men dng cho gia sc tng qut c th trnh by trn s tm tt nh sau: ( S qui trnh cng ngh sn xut) 4.1. Nguyn liu r ng Vi b ru t r ng ma c c thnh phn c cho bi bng 3.3. Bng 3.3. Thnh phn ca b ru t r ng ma Thnh phn Nc Cht kh Protein Tro Cht humin ng kh T l % 54,67 45,33 6,95 10,93 10,4 5,30 Glyerin Axit lactic Cht bo Xenluloza Sap, linhin, gluzzit, phenol, Cc axit hu c 6,15 Thnh phn T l % 2,60 2,70 0,00 0,30

Thnh phn cc vitamin c trong b ru c c c cho bng 3.4

40

B ru

B chn nui

Lc thng quay Nc Dch b R ng Lm ngui (35-370C) O2 Nui men ging

H2SO4, superfotfat (NH4)2SO4 Du ph bt Cht iu chnh pH VTM ho sinh khi nm men Ln men

Tch sinh khi

Sa men c

Sy 2 trc hoc sy phun

Thnh phm

Hnh 3.4. S qui trnh sn xut sinh khi nm men t b ru r ng. Bng 3.4. Thnh phn cc vitamin c trong b ru c c ( mg/g) Thnh phn ca cc vitamin Axit niconitic ( PP) Riboflavin (B2) Piridoxin (B5) Axit pentotenic ( B3) Biotin ( B7) Axit folic Hm lng mg/g 21 8 30 39 1,5 0,3

41

Nh vy b ru t r ng l mi trng c gi tr v y cc cht nui cy nm men to sinh khi. Sinh khi nm men l mt ngun giu protein v cc vitamin l nhng cht quan trng i vi s pht trin ca gia sc, c b sung vo thc n iu chnh, lm cn bng v protein cho thc n gia sc (1 kg ch phm protein c gi tr bng 3,5 kg ht) 4.2. Chng vi sinh vt: Candida Tropicalis, Torulopsis Utilis 4.3. X l nguyn liu v chun b mi trng: Tu theo qui trnh cng nghip ca nh my, nu mun thu nm men thc n gia sc trn b ru khng b sung thm r ng, th s dng chng Candida Tropicalis. B sung thm r ng c th tng hiu sut ca nm men nhng s lm gim h s s dng gluxit ca b ru, do s lm tng gi thnh sn phm. Nu b sung r ng th c t l r ng thanh trng l 1% so vi b ru. B ru trc khi a vo sn xut c bm n thit b lc chn khng thng quay tch nm men cht trong qu trnh chng ct ru trc . Ngoi b ru v r ng, mi trng dinh dng cn c b sung axit photphoric k thut (70%) hm lng 0,5kg/m3 b ru v sunfat amon tinh th (0,5kg/m3). Axit ho dch nui cy bng axit sunfuric hoc HCl n pH mi trng bng 4,5. Lng axit sunfuric dng n 1kg trn 1m3 b ru. Nu pha long vi nc theo t l 1: 1 th cn pha thm 50-70g/m3 magi sunfat. 4.4. Nui cy men ging: Theo qui trnh cng ngh ca Vin nghin cu cng nghip ru Ucraina vi nguyn liu l b ru c tch nm men, vic nhn ging nm men t men ging c tin hnh trong 4 giai on. Trong giai on u ngi ta s dng r ng pha thm 1% super photphat theo khi lng r ng. Nng dch l 2,5% c axit ho thnh axit sunfuric hay axit HCl n pH = 5-5,2 v c thanh trng ni hp p sut 0,5ati trong thi gian 30 pht, sau lm ngui n 30oC (rt 200ml dch vo bnh cu dung tch 700ml ) cy men ging vo dich v bnh trong my lc phng th nghim trong iu kin c sc kh trong 24h. Sau tin hnh nhn ging ln lc vo 3 thit b c th tch l 15,120 v 12001 c trang b my sc kh v my lc khng kh. Khi lng mi trng trong cc thit b l 10,100 v 1000l vi nng dch mi trng l 3,5% thi gian nui cy l 24,18 v 16 gi. T thit b cy men sau cng (thng 1000l) ngi ta a lin tc nm men ging vo thng cha men c dung tch 32m3, cha c 25m3 dung dch sau chuyn vo thit b nui men cng nghip c dung tch tng cng khong 310 m3. 4.5. Nui men cng nghip: Th tch men ging a vo thng ln men c nng 18-20 g/l ( m 75%) bng 10% dung tch c ch ca thng. Qu trnh ln men lin tc c sc kh, thi gian ln men t 5-6 ngy. Sinh khi nm men lng c chiu bng tia t ngoi chuyn ergosterin thnh canxipherol trc khi qua cc thit b ly tm tch v thit b sy. 5. Cng ngh sn xut protein t ngun ph liu xenluloza: T lu ngun xenluloza c ng dng rng ri lm vt liu hu c rn trong nhiu lnh vc. Ngun ph liu xenluloza t nng nghip nh b ma l 36 triu tn.
42

Ring M l 15 triu tn. Thnh cng ca Srinivaane v Han (Louisiana State University) trong vic phn lp c hai loi vi khun c kh nng cng sinh l Cellulomonas v Alcaligens m ra mt hng rt quan trng trong vic s dng cc ngun ph liu xenluloza sn xut protein n bo. Protein ca vi khun li rt cao, trung bnh 60-70% c loi ln n 87%. 5.1. Phn lp vi khun: Hai ng Srinivaane v Han phn lp c VK c hot ng xenluloza cao nh sau: Mi trng phn lp: NaCl 6,0 g/l : K2HPO4 0,5g/l MgSO4 0,1g/l (NH4)2 SO4 1.0g/l KH2PO4 CaCl2 0,5g/l 0,1g/l

0,1 % dch chit men v mt mnh giy lc. Chng 1g ng sacaroza ln men trn vi mi trng . Sau 3 7 ngy nhit 300C trn my trn lc, mt phn giy lc c chuyn thnh mi trng fresh. Qu trnh trn c lp li nhiu ln tng cng s hiu kh v mesophil cha vi khun s dng xenluloza. Giy lc nui cy vi sinh vt c ra ngm trong nc v trng v cy thnh ng trn mi mi trng thch nui cy: Thch cacboxylmetyl xenluloza, thch giy lc. S xut hin khun lc trn mi trng c chuyn sang ng nghim c xenluloza v mui dinh dng. Vi khun cellumonas pht trin tt nhit 25-35oC. 5.2. Qui trnh cng ngh sn xut protein vi khun t b thi xenluloza Mt xng pilot sn xut protein Vi sinh vt t b thi xenluloza (b ma) gm 5 cng on sau: - Cng on gia cng b ma - Cng on ch bin bt b ma - Cng on tit trng - Ln men - Thu hi t bo vi sinh vt v thnh phm Qua nghin cu v sn xut th, ngi ta xy dng nn qui trnh sn xut protein vi khun t xenluloza nh sau (hnh 3.5). u tin nguyn liu xenluloza c qua b phn nghin c bit c 5 cnh nghin c nh. Xenluloza c nghin thnhbt c a qua thit b kim ho bng dung dch NaOH 2-4% o. Sau hn hp rn lng c qua khu li tm tch v qua l oxi ho vi mt cht xc tc oxit ho l clorit coban. Thanh trng 260F 320F qua h thng phun hi Lm ngui: H thng ng ng

43

Ln men; Sau khi lm ngui dch ln men qua van kim tra vo thng ln men. Dch men c th t thng cha hay thng nh tng ho li. iu chnh pH bng NH4OH.

Hnh 3.5. S qu trnh sn xut protein n bo t b thi xenluloza ( theo V.W.Han v cng s 1971) Thnh phn mi trng nh sau: Ngun xenluloza, nc mui v c, mt s cht dinh dng c bit v mt s cht chng bt. Thnh phn C cht: b ma ch bin (trng lng kh) Cht dinh dng: Sunfat amn Mui Photphat MgSO4 CaCl2 NACl Nc chit men Mui khong Polyglycol P -2000 Nc g/1 6,0 3,0 1 0,1 0,1 3,0 0,5 1,0ml 0,1ml 1lit

44

* Thnh phn mui khong CaCl2 FeCl3.6H2O ZnSO4.7H2O CaSO4.7H2O Clorua Coban.6H2O Ethilene dinitriclotetraacetic acid

g/1 0,5 16,7 0,18 0,16 0,18 20,1

45

CHNG 4 CNG NGH SN XUT PRTOTEIN T NGUN HIDRO CACBUA DU M V KH T 1. CNG NGH SN XUT SINH KHI PROTEIN DU M Trong thnh phn cacbua hydro thiu rt nhiu cht dinh dng, trong cc loi mui khong l thiu trm trng nht. V th trong khi nui vi sinh vt trong mi trng ny i hi phi cung cp cc cht dinh dng vi lng v khong cho chng pht trin 1.1. Cc chng vi sinh vt Vn la chn cc chng vi sinh vt c hot lc sinh tng hp cao dng trong sn xut c mt ngha quan trng. Trong cng nghip sn xut protein t du m v kh t, phi chn cc chng p ng c cc yu cu sau: - C kh nng s dng tt ngun nguyn liu hydrocacbua dng trong sn xut. - Sinh trng nhanh chng, cho sn lng cao trong thi gian ngn, khng i hi cc yu t sinh trng b sung trong sn xut ln. - C c im ho hc v nui cy n nh, c hm lng protein cao, cha y cc axit amin cn thit, khng c c t v phi c ng vt ng ho tt. Phn ln cc chng nm men c sn lng cao trn c cht hydrocacbua c phn lp t nhng mu t v bn nhng ni c m du hoc chung quanh cc nh my ch bin du m. Trong hn 500 chng nm men phn lp c, cc nh khoa hc thy cc chng nm men thuc ging Candida cho sn lng cao hn c. Cc chng ny c nui th trong thit b c sc kh trong iu kin phng th nghim trn mi trng nparaphin cho hiu sut khi ti 80-100% (trng lng men kh so vi trng lng parafin c dng). Hm lng protein trong sinh khi khong 50%. Kt qu xc nh trn hai loi nm men Candida cho bng 4.1 nh sau; Bng 4.1. Tn nm men Hiu sut nm men kh (%) Candida Tropicalis Candida Intermedia 94,4 87,1 Hm lng protein (% cht kh) 58,8 51,0

Cc chng nm men thng s dng: - a s cc loi thuc ging Candidas nh: C.Tropicalis, C.Lipolitica, C.pelliculosa. - Torulopsis Famata v..vv. c im ca cc chng vi sinh vt ny ni chung l:

46

- S dng hidrocacbua lm ngun cacbon duy nht trao i cht v nng lng. - Bn vng vi c t ca hidrocacbua vi nng cao. - C kh nng hp th hidrocacbua vo t bo. 1.2. Chun b mi trng dinh dng 1.2.1. Cc cht b sung: Cc hp cht b sung vo mi trng dinh dng: - Axit octophotphoric hoc supephotphat - KCl - MgSO4 - Ngun nit: Nc amoniac c 20-25% NH3 v mt lng nh amon sunfat oxi ha mi trng ban u. NH3 cn dng iu chnh pH trong thi gian nui cy. B sung nguyn t vi lng: Nguyn liu u cc hydrocacbon khng c cc nguyn t vi lng. V vy phi thm vo mi trng dinh dng cc mui sau: - FeCl3.6H2O - MnSO4.H2O - ZnSO4.7H2O - CuSO4.5H2O - KI - Na2MoO4.H2O 1.2.2. Mt s mi trng nui cy vi sinh vt trong du m Sau y gii thiu mt vi mi trng nui cy vi sinh vt trn hidro cacbua lng (theo Nadirop v Popov, 1974) a. Mi trng nui cy nm men Thnh phn n-parafin Supephotphat Amon sunphat Nc amoniac (25%) KCl MgSO4 kg 12,5 2,7 0,45 4,0 0,56 0,28

Nc b sung vo cho 1000 C th thay supephotphat, bng axit octophotphat, amon sunphat bng amon clorua hoc cho ng thi axit sunfuric vi nc amoniac.

47

Hiu sut c th thu c trn 100kg men kh/m3 mi trng b. Mi trng nui cy vi khun Thnh phn n-parafin K2HPO4 KNO3 MgSO4 NaCl FeCl2 kg 10 1 1 0,5 0,1 0,01

Nc b sung vo cho 1000 lt c. Mi trng nui cy nm mc Thnh phn Du diezen NaNO3 K2HPO4 KCl FeSO4 kg 30 3 1 0,5 0,01

Nc b sung vo cho 1000. Cc nguyn t vi lng cn cho Aspergilus niger (mg/l mi trng): Fe 0,2; Zn 0,18; Cu 0,04; Mn 0,22 v Ca 0,02. Vic tuyn chn cc chugr nm men v vi khun c kh nng phn hy du m v parafin ngy nay c ngha rt ln trong vic bo v mi trng khi b nhim du m, sc bit l cc vng t quanh kho cha hoc t ven bin sau cc s c nhim du do cc tu ch b nn. 1.2.3. K thut nui cy: Hin nay nng cao hiu sut s dng c cht v tn dng trit thit b nui cy, nhiu nc ng dng k thut nui cy lin tc trong cng nghip sn xut sinh khi nm men. Qu trnh nui cy nh sau: - Parafin nng (50-600C) c lin tc cho vo thng ln men, nng parafin trong mi trng ban u l 1,5 -2 % th tch. - S tch t sinh khi nm men trong thi gian nui cy c th thc hin trong hai ni ln men: + Ln men chnh: ni th nht; c thi kh mnh. + Ln men ph: c thi kh nhng yu hn.

48

Nu so snh qu trnh ln men sinh khi trn mi trng cha parafin vi mi trng hydratcacbon, chng ta thy c nhng im ging v khc nhau sau y: . Nui cy nm men trn mi trng cha paraphin thng phi thi kh mnh gp 2,6 - 2,8 ln so vi kh nui cy nm men trn mi trng hydrat cacbon. . S sinh trng ca vi sinh vt trn hidrocacbua ph thuc vo pH cng ging nh khi nui trn mi trng sacaroza (pH = 5-6). Tuy nhin, c th tr s pH thp hn trnh tp nhim. . Khi sinh trng trn hydrocacbua, nm men to nhit hn v yu cu v thanh trng khng cht ch nh khi nui trn mi trng sacaroza. 1.4. Ngun c cht: Cht lng ca parafin nh hng ln n sn lng nui cy nm men. Trong n-parafin thng c 93-98% hydro cacbua c to thnh phc cht vi cacbamit, l cc n-ankan c s nguyn t cacbon t 12-24, 2-7% izoparafin naphten v khng qu 0,5% hydro mt hoc hai vng thm. Qua cc s liu cng b trong vic nghin cu la chn cc ngun nguyn liu khc nhau, ta thy: - Sn phm nm men rt ph thuc vo ngun hydrocacbua c trong nguyn liu v phng php lm sch. Nu trong nguyn liu c cha mt s cacbua khc, hm lng ca chng qu mt gii hn nht nh no c th lm c ch sinh trng ca vi sinh vt. - Sn phm oxy ha ca mt s hydrocacbua khc trong nguyn liu c th tc hi n tng sinh khi ca ging nui cy. -c tnh l hc ca c cht ( nng chy, nht, mu sc..) c th lm cho qu trnh sn xut gp kh khn. Th d: parafin khng khuch tn trong mi trng nnuwowoo nhit sinh trng bnh thng ca nm men, d l c s khuy o. khc phc, c th chn cc chng vi sinh vt c nhit sinh trng ti thch cao, tt nht l cc chng vi sinh vt a nhit (trn 400C).

49

1.5. S cng ngh sn xut sinh khi nm men t du m th v t parafin tinh khit: A
Ln men Du th Parafin

B
Ln men

Tch nm men

Du ht parafin

Tch nm men

Ra nc

Ra nc

Lm kh

Lm kh

X l bng dung mi

Bao gi

Thu hi dung mi Nm men thnh phm Ra nc

Lm kh

Bao gi

Nm men thnh phm

2. Cng ngh sn xut sinh khi vi sinh vt t kh t 2.1. u im ca sn xut sinh khi vi sinh vt t kh t: - Kh t thng r hn du m, do gi thnh sinh khi thu c cng r hn. - Sinh khi thu nhn c t kh t thng sch hn rt nhiu so vi sinh khi t du m. 2.2. Nhc im . Cc vi sinh vt ng ho kh thin nhin u l nhng vi sinh vt hiu kh. Chng cn oxy h hp. Khi cho CH4 vo cng O2 s to thnh mt hn hp kh rt d n.

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. ng ho O2 v CH4, chng phi tan trong mi trng v phi tip xc c vi t bo vi sinh vt. Trong khi ho tan ca O2 v CH4 trong iu kin bnh thng rho km. tng ha tan ca metan c th tn p sut d trong thit b, nh vy cn phi ch to thit b chu p lc cao rt phc tp v nh vy s mt tnh kinh t ca phng php. Cch th hai c th l a mt dung mi hu c no vo mi trng dinh dng tng ha tan ca metan, nhng lc s c th lm vi sinh vt thch dung mi hn metan v vic dng kh thin nhin mt ht ngha. 2.3. Cc phng php sn xut sinh khi vi sinh vt bng kh t: Hin nay c 2 phng php sn xut sinh khi vi sinh vt bng kh t - Phng php 1: Ngi ta to ra mi trng dinh dng gm c mui amon v cht khong v a mi trng ny vo cc bnh ln men. Tin hnh nui vi sinh vt c kh nng to sinh khi t kh thin nhin trong bnh ln men ny. Thi kh thin nhin v kh vo bnh dung dch ln men c sn vi sinh vt. Kh khng kh v kh thin nhin vo dung dch ln men s tip xc vi vi sinh vt. Khi vi sinh vt s ng ho kh thin nhin cng vi cc cht dinh dng to thnh sinh khi - Phng php th 2: . Thit k nhng bnh phn ng c cha y cht mang. Cht mang ny cha y vi sinh vt trong . a khng kh v kh thin nhin t di ln. . Khng kh v kh thin nhin i qua cht mang s to c s ng ho ca Vi sinh vt. . Sinh khi oc to thnh nhiu s tch khi cht mang v ri xung pha di . Thu nhn sinh khi t y thit b. Theo phng php ny th hiu sut khng cao nhng c ngha khi x l CH4 trong mi trng khng kh. CH4 c nh hng cao nht n s to thnh hin tng hiu ng nh knh (CH4 lm tng hiu ng nhag knh gp 21 ln so vi CO2. Phng php ny loi tr c CH4 v thu oc sinh khi cng mt lc. y l mt phng php kh l tng trong k thut mi trng.

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CHNG 5 CNG NGH SN XUT PROTEIN T TO N BO 5.1.GII THIU V TO SPIRULINA 5.1.1. c im ca to Spirulina: t lu, to l mn n dn gian ca nhiu a phng trn th gii. Dn min Kanem dng thng mng vt loi to lam a bo Spirulina maxima trong cc ao h giu mui cacbonat lm thc n nhng lc h cha bit c trong to lam cha rt nhiu cc cht c gi tr dinh dng. u nhng nm 70 ca th k XX, Vin nghin cu du m ca Php pht hin ra to c kh nng pht trin nhanh v c hm lng protein rt cao. T nm 1967, Sosa Texcoco( Cng ha Sat, chu Phi) tr thnh c s sn xut cng nghip to Spirulina u tin trn th gii. Trc y ngi ta sn xut nhiu Chlorella nhng dn dn, do nhng im ni bt, to Spirulina chim v tr ch yu. Hin nay rt nhiu nc trn th gii trong c c Vit Nam cng t chc sn xut loi to ny nhng qui m khc nhau. To Spirulina c cu to hnh si a bo, c hnh dng xon l xo, kch thc khong 0.25 0.5 mm, sng t nhin nc kim giu natri bicacbonat.

52

To Spirulina sinh sn bng cch gy ra tng khc, tc sinh trng rt nhanh, c th sng trong mi trng ngho cht dinh dng, iu kin nui cy n gin. c bit iu kin t nhin c cng chiu sng ln v trong mi trng c pH =8.5 9 th tc sinh trng l ln nht. Hiu sut s dng nng lng mt tri cao ti 3 4.5 %. Hiu sut s dng kh CO2 lm ngun cacbon cng rt cao, ti 80 85 % trong khi Chlorella ch t khong 30 %. To Spirulina c kch thc ln, li c xu hng ni ln mt nc v t tp sinh khi nn d dng thu hoch bng cch vt v lc trong khi Chlorella c kch thc nh nn phi thu nhn bng phng php ly tm phc tp. Nng sut tnh trn n v din tch nui trng rt cao do n c gi tr kinh t rt cao. Theo bo Vietnam Net, Long An nui trng to Spirulina bng nh knh, so vi s dng t trng la vi thu nhp 50 triu ng/ha/nm, vic chuyn sang nui to Spirulina s to mc thu nhp khong1.2 t ng/nm, tc tng gp 24 ln so vi trng la. Ngoi nhng u im trn, vic to Spirulina c a vo sn xut vi qui m ln nhiu nc trn th gii cn do gi tr dinh dng to ln ca n. To Spirulina

53

cha hm lng protein rt cao, khong 60 70 % trng lng cht kh c bit l c y cc axit amin khng thay th. Hm lng cc axit amin ca nhng protein ny gn vi qui nh ca protein tiu chun, tng ng vi protein ng vt v cao hn hn protein thc vt.

Bng 5.1.Thnh phn axit amin ca to Spirulina S th t 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 Thnh phn Isoleucin Leucin Lysin Methionin Phenilalanin Theonin Tryptophan Valin Alanin Arginin Axit aspartic Cystin Axit glutamic Glycin Histidin Prolin Serin Tyrosin g/10g 350 540 290 140 280 320 90 400 470 430 610 60 910 320 100 270 320 300 S lng(% tng cht kh) 5.6 8.7 4.7 2.3 4.5 5.2 1.5 6.5 7.6 6.9 9.8 1.0 14.6 5.2 1.6 4.3 5.2 4.8

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Gi tr dinh dng ca to cn th hin cn th hin cht lng v s lng cc vitamin cha trong n nh caroten, vitamin B3, B6, B1, E v c bit rt nhiu vitamin B12. Vin nghin cu ng dng cng ngh chit c caroten dng lm thc n bi dng sc khe, chng suy dinh dng cho tr em. Ngoi ra trong Spirulina cn cha nhiu xantophyl l cht rt cn thit cho gia cm. Nu dng to vi lng 10 % trong khu phn thc n nui c th s c tc dng lm tng cao t l sng ca c bt. Ngy nay ngi ta pht hin v chit tch t to Spirulina cht phycoxianin l mt cht c tc dng cha bnh ung th vng hm, vm hng. Qua nghin cu cho thy rng nu dng phycoxianin kt hp vi gamma Cobalt 60 s hn ch c 70 80 % s pht trin cc t bo ung th. To cn c tc dng bo v c th khi tc hi ca cht phng x v chng suy mn do nhim hi c Chnh v nhng li ch to ln trn ca Spirulina nn hin nay to ang c nui trng rt rng ri nhiu ni trn th gii lm thc n dinh dng cho ngi, l nguyn liu chnh sn xut bt dinh dng cho tr em, s dng trong lnh vc y hc cha bnh ung th, sn xut thuc li sa, thuc chng suy dinh dng ..., s dng lm thc n gia cm, lm m phm... 5.1.2.TNH HNH SN XUT TO SPIRULINA TRN TH GII V VIT NAM 5.1.2.1. Trn th gii: Hin nay trn th gii pht trin rt mnh vic nui cy to Spirulina v Chlorella thu nhn sinh khi cho ngi v ng vt, trong Spirulina c sn xut nhiu hn. Thc t cho thy, 1 ha b mt nui cy to thu nhn c 10- 15 tn to mt nm, cao hn rt nhiu so vi trng la. Mt trong nhng ging c s dng nhiu nht l Spirulina maxima. To lam ny pht trin thnh si, do d thu nhn thm ch bng cc phng php th cng nh co v lc. T thp nin 70, Nht Bn v M to Spirulina c xem l mt loi siu thc phm. n nhng nm 1990, vn tiu th Spirulina tng vt bc ti Trung Quc, n , chu , Bc M lm cho Spirulina ngy cng tr nn ph bin.

55

Mexico: vo nhng nm 70, mt cng ty ca Php pht hin to trong h Texcoco pht trin tt trong mi trng kim c nhiu mui cacbonat. Nh my sn xut cng nghip ln u tin trn th gii c xy dng y.

H Sosa Texcoco -Mexico

Nm 1979, ln u tin Mexico xut khu nhng loi thc n dinh dng t Spirulina sang U.S. Myanmar: Vic sn xut bt u vo nm 1988 h Twin Taung. Nm 1993, 30 tn to c thu hoch v tiu th trong a phng. Vo nm 1999, vic sn xut tng ln 100 tn/ nm. Khong 60 % to c thu hoch bng thuyn bn ngoi ca h v khong 40 % mc dc theo cnh h. trong sut thi k to pht trin mnh vo ma h, khi Spirulina mc thnh nhng thm dy bn trong h, ngi ta dng thuyn thu hoch to vo trong nhng ci thng. Sau khi thu hoch xong c em i lc, ra sch vi nc tinh khit, tch nc v lp qu trnh mt ln na

H Twin Taung- Myanmar

56

Sat: Nhng ci ao c tnh kim xung quanh h Sat chu Phi l mt vng l tng cho to Spirulina pht trin. Ph n bn x thu hoch Spirulina v ch bin thnh mn n gi l dih, sn lng thu hoch khong 30 tn/nm. Thailand: To c trng vo nm 1978 gn Bangkok. Vi iu kin kh hu nhit i, to y pht trin tt v cho nng sut cao 150 tn/nm v lc ny l 170 tn/ nm. Ngoi ra cn mt vi c s sn xut nh Thi Lan. To c bn cho Nht Bn lm thc n dinh dng.

H trng Spirulina ca cng ty Siam- Thi Lan Hawaii, USA: Cyanotech m mt nng tri sn xut Spirulina vo nm 1985 trn b bin Kon thuc o Big- Hawaii, nhng nm gn y, khu vc ny c m rng v sn sinh hn 400 tn Spirulina/nm cng vi to Dunaliella. Trung Quc: Ngy nay Trung Quc c khong 80 loi Spirulina vi nng sut thu hoch khong 500 tn/nm trong o Hainan c sn lng 300 tn/nm. i Loan: Vo thp nin 70, i Loan ch yu trng Chlorella nhng c 5 loi Spirulina c kh nng sn sinh vi trm tn/nm. Ngy nay Spirulina c nui trng rt nhiu v c thu hoch khong 460 tn/nm. n : Vic nghin cu bt u vo cui nhng nm 70, t qui m gia nh h chuyn sang dng canh tc ln hn. Vo nm 1990, n thnh lp nn mt tiu chun quc t v cc loi thc n t Spirulina. Hin nay c 2 c s sn xut ln c tnh sn lng khong vi trm tn/nm. Cuba: C hai c s sn xut khong 40 tn/ nm.

57

Chil: Nm 1991, Solarium bt u sn xut vng Atacama, sn lng khong 3 tn/nm. Israel: Vin nghin cu Desert nghin cu Spirulina c hn 20 nm nhng s sn xut vi qui m ln Israel khng thnh cng. Cc ni khc: Spirulina cn c trng mt s nc nh Bangladesh, Philippiness, Martinique, Peru, Brazil, Spain, Portugal, Australia v mt s nc khc. Vic nui trng Spirulina ang pht trin trn khp th gii 5.1.2.2.Vit Nam: Cng ty c phn nc khong Vnh Ho -Tnh Bnh Thun c c s nui trng to Spirulina Platensis i tr vi qui m ln Vit Nam. Sn lng hin nay t 8 10 tn/nm. D kin tng sn lng ln 15 tn/nm. To Spirulina Platensis nui trng Vnh Ho cha lng m rt cao v nhiu thnh phn sinh ha c gi tr:

Protein: 60 70 % trng lng kh, c y cc axit amin khng thay th. Gluxit: 3 6% Lipit: 2 3 % Cc vitamin: caroten, B1, B2, B3, B6, B12, E Cc nguyn t khong: Na, K, Ca, Mg, Fe Cc sc t: clorophyll, pycobiliproten v carotenoit. Th trng chnh tiu th to l cc cng ty dc , cng ty thc phm cao cp. Mt s hnh nh v vic nui trng to vng sui khong Vnh Ho:

58

Phng th nghim

Khu nhn ging cao tc

H nui trng to

Sn phm

Ngoi ra, nm 2003, m hnh nui to bng nh knh Long An theo qui trnh nui to sch ca Thc s L Vn Lng c sn xut n nh v c hiu qu kinh t. Hin nay m hnh nui trng ny c a vo ng dng vi qui m sn xut 2 3 tn/nm. Gi thnh ca loi to xon ny khong 10 16 USD/kg. n nay, to Spirulina c cng ty thc phm ng Tm dng lm nguyn liu chnh sn xut bt dinh dng cho tr em. To Spirulina c mt s cng ty dc mua bo ch sn xut cc loi thuc li sa, thuc chng suy dinh dng... Ngoi ra, mt s cng ty dc liu nc ngoi t mi quan h v t hng to Spirulina dng kh. Theo Thc s Lng, sp ti s c k hoch m rng qui m sn xut gp 3 ln hin nay tc khong 7500m2 tin ti m rng v xy dng mt trung tm nghin cu sn xut to Spirulina v mt s vi to khc c gi tr nh to Chlorella, to Dualiella,... 5.2.Cng ngh sn xut to Spirulina:
Qui trnh san xut tao noi chung gm co 2 giai oan chnh: - Nui cy tao - Thu nhn sinh khi 5.2.1.Nui cy tao

5.2.1.1. Ging:

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Spirulina l mt chi gm mt s loi c s dng ph bin trong cng ngh nui trng to l Spirulina platensis, Spirulina maxima.Chn Spirulina vi nhng si nh xon cn i thu hoch. Spirulina phi cha t nht 1 % axit - linoleic tnh theo trng lng cht kh. To phi tp trung li vi nhau, ni l lng thnh tng lp. Trong qu trnh nui cy, phi tin hnh khuy trn, nu c th khuy trn lin tc nng ca to c th t n 0,8 g/l. 5.2.2.2. Mi trng c bn nui cy to Spirulina: a. Cc iu kin k thut ca mt qu trnh nui cy to; - Phi c nh sng vi cng chiu cao: v vy, nui to phi c iu kin kh hu thun li, trc ht l mt thi k nh sng mt tri mnh v ko di c y nng lng nh sng.To t b chi phi bi chu k sng/ti nhng nu c chiu sng lin tc th gi tr sinh khi s t c cao nht. Cng nh sng thch hp nht cho Spirulina nm trong khong 25.000 30.000 lux. Nhng si to nh d b ph hy do cng chiu sng mnh v ko di. Bi vy, chng ta cn phi gim bt thi gian chng c chiu sng bi nh sng mt tri. Ma s lm gim bt s bay hi nc nhng vi iu kin khng c y trn ao nui trng to. nh sng v s chiu sng nhn to gip Spirulina pht trin nhanh hn mc d y khng phi l bin php kinh t v rt phin phc v mt cng ngh. Ngi ta dng nhng ci n hunh quang v n halogen nhm va chiu sng va lm nng mi trng nui cy. - pH mi trng phi duy tr= 8,5 -9 (i vi to Spirulina) v trung tnh (i vi to Chlorella) - Phi c cung cp y cc mui vi lng. - Phi c khuy o lin tc, to s tip xc thng xuyn vi nh sng (i vi Spirulina) v phi to ra chu k sng ti thch hp (i vi Chlorella). V vy qu trnh nui cy to i hi nhng thit b c bit. Thng thng ngi ta dng nhng b phng ( b trn), hoc nhng mng phng un khc. Nhng thit b ny c tc dng lt o nhm hn ch s lng ca t bo v a t bo lun tr li b mt chiu sng.

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- Cung cp CO2: ti u khong 4-5% so vi khng kh ( c tc gi cho l 1-3%). Vic cung cp CO2 vi vai tr l ngun cacbon trong qu trnh quang hp rt cn thit. CO2 c th c cung cp bng nhiu cch khc nhau: .Ly trc tip t cc qu trnh ln men khc nh ln men ru etanol, ln men bia v..v hoc ngun kh thi cng nghip. . Sc khng kh c cha CO2 (1-3%) kt hp vi sc CO2 100% ngt qung v..v.. 5.2.1.4. Cc phng php nui to: Hin nay trn th gii c ba hnh thc nui trng Spirulina: thu hoch Spirulina t nhin trong cc h, nui cy trong h hoc trong nh knh c mi che v mi y pht trin h thng nui trong nhng ng trong sut tng s tip xc gia to v nh sng mt tri. Nhng h thng nui cy bn t nhin th cho cht lng tt hn thu hoch to mc t nhin. V qui m c chia lam 3 loai: - Nui qui m thu cng n gian: Nui cac ao t nhin hay cac b ( xy bng xi mng) hay la thung g, nha. Trong trng hp nay thng ngi ta khng suc kh CO2, khng khuy ao -Nui qui m ban cng nghip: Mt m hnh nui qui m ban cng nghip nh m hnh nui trng Chlorella u tin Hoa Ky. Tai y tao uoc nui trong cac ng cht deo trong sut, hnh ch U, dai hn 20m, ng knh 1,2 m. Khi ng nm ngang, cho mi trng bvvao trong ng vi cao khoang 0,625m. Kh CO2 c bm vao mi trng, ng thi mi trng c lun chuyn vn ng tun hoan nh mt may bm khac. Nui tao bng nng lng anh sang mt tri vi nhit mi trng duy tr khoang 25-260C. - Nui qui m cng nghip: nui trng vi tao, vi khun lam qui m cng nghip, co 2 h thng chnh : H thng kn va h thng h. Du la h thng nao i na,vic khuy ao, suc kh tao iu kin cho t bao tip xuc vi anh sang mt tri va kh CO2 la yu cu rt quan trong cac chung ging thc hin qua trnh quang hp. Do o, cac h thng nui trng c thit k gn lin vi h thng khuy ao va suc kh. H kn: h thng nui nay, vi tao, vi khun lam c nui trong cac b ln men chu yu dung anh sang nhn tao ( anh sang en), co cng va h thng suc kh CO2 tuy theo yu cu cng ngh. u im :. Khng phu thuc vao iu kin kh hu , thi tit.
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. iu kin nui cy c kim tra, khng ch mt cach chu ng. . Nng sut cao Nhc im : Gia thanh t nn t c ap dung rng rai. H h: c im :. Qua trnh quang hp cua vi tao va vi khun lam gn lin vi vic s dung anh sang t nhin ( anh sang mt tri). . Chiu cao ct mi trng khoang 15-17 cm, bng 0,7 chiu cao b nui cy. . Khuy ao vi mt ch thch hp tao c tip xuc vi anh sang mt tri va giup cho tao khng b lng xung ay b, am bao cho s phn b u cht dinh dng cho toan b t bao cua h thng nui

ARTISANAL FARM Sn xut th cng

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MEDIUM SIZE FARM Sn xut vi qui m trung bnh

MASS PRODUCTIONFARM San xut qui m cng nghip

5.2.2. Thu nhn sinh khi: Vic thu nhn sinh khi thng qua cac bc sau: - Lam c s b - Loc bng trong lc va chn khng - Pha v t bao - Sy kh - Nghin - ong goi

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Khi hm lng sinh khi t cc i th tin hnh thu hoch to. Vic thu hoch l mt thao tc kh d dng tr khi n tr nn qu gi v dnh li vi nhau th vic thu hoch tr nn rt kh khn. Thi gian thu hoch to tt nht l vo bui sng sm v nhiu l do: Cng vic s d dng hn khi thi tit mt m. Tri nng s d lm kh sn phm. Phn trm protein trong Spirulina cao nht vo bui sng. V c bn, vic thu hoch to c 2 bc: C c s b thu c khong 10 % cht kh v phn cn li cha 50 % mi trng nui cy. Vic loi b phn cn li ca mi trng nui cy trong sinh khi Spirulina ti s c tin hnh trong qu trnh s dng hoc qu trnh sy kh, n s cha khong 20 % vt cht kh v dng nh khng cn mi trng nui cy. Cch lc th c tin hnh n gin bng cch cho c to v mi trng qua mt lp vi lc nh vo trng lc. Lp vi c lm t poliamide hoc poliester vi kch thc mt li c 30 50 m l thch hp nht. Vic h tr thm mt li lc mn s lm tng nhanh qu trnh lc v bo v lp vi lc khng b thng. Nhng cch n gin nht l c th s dng mt ti ln lc. Vic lc c th c tin hnh trc tip ao nui cy to phc hi nc lc. vic thu hoch to s c tin hnh thng qua mt ci sng vi kch thc mt li 200 m gi li bt k nhng cht l no nh su b, u trng, l cy, s vn cc ca cc polysacarit hoc bn. Tht tin li xc nhng mng Spirulina khi chng ni ln trn, c th s dng nhng ci thng mc to. vic thu hoch to khi n ni thnh tng lp s c khuynh hng tng thm phn trm Spirulina, nu n khng ni ln trn th s gy kh khn cho qu trnh thu hoch. Khi hu ht nc c lc, sinh khi s c tp trung li thnh nhng ci cun. Vic tch s thc hin tt hn vi vi lc bng cotton. *B sung mi trng dinh dng: Nhng cht dinh dng b tch ra cng vi sinh khi khi thu hoch cn phi c b sung duy tr mi trng dinh dng.

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Cht dinh dng chnh l cacbon, n c th c ly trc tip t khng kh nh kh CO2 mi khi pH > 10. Tuy nhin trong khng kh ch cha mt lng rt nh kh CO2 nn s hp th n l mt qu trnh rt chm, qu trnh ny ch t cc i khi pH > 10,5. CO2 tinh khit c cung cp t hi t hoc oxi ha cc hp cht hu c nh ng. Lng kh cn thit khi sc chim khong 4 % tng din tch ca h. Vic thm HCO3- l mt cch lm gim bt pH c hiu qu v d thc hin nht nhng n s lm tng mn ca mi trng. Thnh thong phi rt bt mt phn mi trng nui cy v thay th bng mi trng giu HCO3- mi duy tr mt mn nht nh. Hm lng kh, r ng, HCO3- b sung s iu chnh c pH khong 10,4. pH< 10,2 c th gy ra s sn sinh tha khng mong mun nhng khng nguy him. ng c th gy ra mt s bin i ca mi trng dinh dng v vy ch nn s dng mt lng nh hn 0,3 kg/kg v cung cp cng u n cng tt. Ngoi C, Spirulina cn phi c cc cht dinh dng cn thit nh: N, P, K, S, Mg, Ca, Fe v mt s nguyn t vi lng khc. Trong mt s trng hp, cc nguyn t vi lng v canxi c th khng cn cung cp v n c sn trong nc v nhng cht ha hc s dng lm thc n cho Spirulina. Trong mt vi trng hp, nc c cha mt lng ln Ca, Mg, Fe, n s lm c mi trng. Nu s dng phn bn ha hc th chng phi ha tan c phng vic c cc kim loi nng nh Hg, Cd, Pb, Spirulina s d dng hp thu nhng cht v s b kt dnh li. Nitrat l mt ngun cung cp nit tt, n cha ng nhiu cht dinh dng ngoi nit. Ngun nit r nht l ur, ur c to thnh t NH3 v CO2 l mt cht dinh dng tuyt vi cho Spirulina nhng hm lng phi c gi mc thp, khong 60 mg/l. Ur tha cng c th chuyn ha thnh NH3 hoc NO3- trong mi trng. Trong trng hp cn thit, tt c cc cht dinh dng v cc nguyn t vi lng tr Fe c th c cung cp bi nc tiu ly t ngi hoc ng vt c tnh trng sc khe tt, khng dng thuc vi lng khong 15 20 l/kg Spirulina. Fe c th c cung cp bng cch ha tan trong mi trng axit. Phn khc vi ur, n c th c cung cp mt thng mt ln nhng ur th phi c cung cp hng ngy da vo hm lng c xc nh. *Bo qun sn phm:
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Sinh khi ti mi thu hoch s khng gi c lu trong t lnh v khng hn vi gi nhit phng. Thm vo 10 % mui l mt phng php tng thi hn bo qun ln ti vi thng nhng v b ngoi v mi v sn phm b thay i: mu xanh ca phycocyanin b mt i, sn phm s tr nn lng v mi v c phn ging nh bt c Vic lnh ng l mt cch gi Spirulina trong mt thi gian di. N cng lm mt mu xanh ca to nhng khng lm thay i mi v. Sy l mt phng php ph bin bo qun v phn phi Spirulina. Nu Spirulina c sy v ng gi ng k thut th sy c coi l phng php tt gi Spirulina trong vng 5 nm. *. Sy kh: My sy Spirulina dng trong cng nghip l my sy phun, iu ny l ngoi tm vi ca nhng ngi sn xut th cng. Sy kh bng nh nng mt tri l phng php ph bin nht lm kh sn phm i vi nhng ngi sn xut nh. Vic sy trc tip bng nh nng mt tri phi tin hnh nhanh nu khng cholorophyl s b ph hy. D dng ngun nhit no th lp sinh khi to phi mng c th kp kh trc khi n bt u ln men. Khng kh m v kh xuyn qua sinh khi phi vi tc cao vo lc bt u qu trnh sy. Trong qu sy cng nh v sau, sn phm sy phi c bo v chng li s nhim bn t bi, su b v khng c tip xc trc tip vi tay. Nhit sy cn phi nh hn 68oC. S ln men xut hin trong qu trnh sy c th c pht hin bi vic xut hin mi trong v sau qu trnh sy. Tuy nhin, mi s mnh nht khi bt u qu trnh sy.[9] *Vic thu hoch Spirulina Myanmar:
Spirulina thu hoch xong c em i lc, ra bng nc sch v p, qu trnh ny c lp li mt ln na. Bt nho sau khi lc c p thnh tng cy v c sy di nh nng mt tri trn nhng tm nha trong sut.

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