Sinh Khoi Nam Men

SX sinh khi VSV giu protein cho gia sc
GVHD: PGS.TS L Vn Vit Mn
Chng 1
M U
I. GII THIU Thut ng protein n bo mi hnh hnh trong gii khoa hc t nhng nm 50 ca th k trc. Thc t loi ngi bit s dng loi protein ny v cc cht c trong t bo VSV t rt lu. Protein n bo (SPC-Single cell protein) l thut ng ch mt loi cht dinh dng c trong t bo v ch sn xut t vi sinh vt (VSV), c s dng lm thc n cho ngi v ng vt. Thut ng ny khng ch n gin l protein t t bo ca c th n bo, v rt nhiu VSV khng phi l c th n bo m ngi ta vn khai thc chng. Do , thut ng ny nn hiu l ngun dinh dng cha nhiu protein t VSV khc nhau, c n bo ln a bo (t vi khun, nm men, nm si v to). c s dng trc ht l ngun protein trong dinh dng ng vt, ch yu l trong chn nui. C s khoa hc ca phng php sinh tng hp protein nh VSV l li dng kh nng sinh trng nhanh v s phong ph v protein cng nh cc acid amin hp phn ca n trong t bo VSV lm ngun cung cp protein cho gia sc v thc n cho ngi. II. C IM CA SN XUT SINH KHI VI SINH VT Chi ph lao ng thp hn nhiu so vi sn xut nng nghip. C th sn xut nhng a im bt k trn tri t, khng chu nh hng ca kh hu, thi tit, qu trnh cng ngh d c kh ho v t ng ho. Nng sut cao. S dng cc ngun nguyn liu r tin v hiu sut chuyn ho cao. Hm lng protein trong t bo rt cao. Cht lng protein cao. Kh nng tiu ho ca protein tt An ton v c t. Nhng vn k thut.
III. C IM CA SN XUT PROTEIN N BO u im VSV l c th c tc sinh trng rt mnh, kh nng tng trng nhanh. Ch trong mt thi gian rt ngn ta c th thu nhn c mt khi lng sinh khi rt ln; thi gian ny c tnh bng gi, cn TV hay V th thi gian ny c tnh bng thng hay hng chc nm. http://www.ebook.edu.vn Trang 1
Hm lng protein rt cao, cao hn rt nhiu so vi protein c trong TV hay V (hm lng protein VSV khong 20-80% trng lng kh). Vi khun 60-70% tnh theo cht kh, c loi ti 87% Nm men 40-60% Nm mc v x khun <30%. Cht lng protein ca vi khun l cao nht, v cc thnh phn acid amin cn i hn nm men. Nhng v kch thc t bo vi khun nh v cc iu kin nui cy phc tp hn, nn vic sn xut sinh khi VSV lm ngun protein trong cng nghip vi sinh ch yu l t nm men. Tc sinh tng hp protein trong t bo VSV cng rt cao, t 100-10.000 ln so vi b. V d v thi gian tng i khi lng ca vi sinh vt thi k pht trin cc i c so snh vi mt s sinh vt nh sau: Bng 1. Tc sinh tng hp protein trong t bo VSV Sinh vt Vi khun, Nm men Nm v To Chlorella C v cc thc vt khc G mi G con Ln con Cc loi gia sc n c Thi gian tng i khi lng 0.2-2 gi 2-6 gi 144-288 gi 288-576 gi 500 gi 576-864 gi 720-1.500 gi
Protein ca VSV c cht lng tng ng protein V v hn hn protein TV( V protein cha y v rt cn i cc acid amin, TV thng thiu loi acid amin ny hay acid amin khc). Thnh phn cu to v gi tr dinh dng ca protein VSV c th iu khin bng cch thay i thnh phn mi trng v iu kin nui cy hoc bng cch tc ng lm thay i c cu di truyn ca chng, ging. VSV c kh nng hp th, phn gii nhiu loi nguyn liu r tin, d kim, thm ch c cht thi, nc thi ca mt qu trnh sn xut no . Hon ton c th sn xut theo quy m cng nghip (sn xut hng lot, c th kim sot v cht lng sn phm ng nht). Nui cy VSV khng ph thuc vo kh hu cng nh thi tit trong nm, qu trnh nui cy c tin hnh trong cc ni ln d dng n nh cc iu kin k thut nh thnh phn mi trng, nhit , pH, bng cc h thng hiu chnh t ng.
http://www.ebook.edu.vn
Trang 2
Nui cy VSV ch cn mt din tch khng ng k xy dng x nghip (trng trt v chn nui thng chim din tch rng ln). Sinh khi VSV l mt khi thng nht, do c th thu hoch mt cch n gin v d dng (khc vi cc loi cy trng trong sn xut nng nghip) C th phn lp v la chn VSV c ch v thch hp cho cc qu trnh cng ngh, cho tng loi nguyn liu tng i nhanh v khng kh khn lm. Thnh phn cu to v gi tr dinh dng ca protein VSV c th iu chnh c bng cch thay i thnh phn mi trng, iu kin nui cy, hoc bng cch tc ng lm thay i c cu di truyn ca chng, ging Trong protein ca VSV c y cc acid amin thnh phn v t bit l cc acid amin khng thay th c gi tr dinh dng cao. Mt u vit cn nhc ti l trong sn xut protein t VSV li s dng nguyn liu VSV l loi ph liu, ph phm ca mt s ngnh cng nghip khc. Ngun nguyn liu ny rt phong ph, a dng, r tin, d kim nh: r ng, khi thu phn g tp, rm r, b ma do vy gi thnh ca sn phm s thp. ng thi s dng nguyn liu ny s gp phn gii quyt vn nhim mi trng do cht thi v nc thi. Nhc im Tuy vy, ngun Protein thu nhn c t vi sinh vt cn c nhng hn ch: - Hm lng cc acid amin cha lu hunh thp. - Kh nng tiu ha ca protein: c phn b hn ch bi thnh phn phi protein, nh acid nucleic, peptid ca t bo, hn na chnh thnh v v t bo VSV kh cho enzyme i qua.
Trang 3
Chng 2
NGUYN LIU V GING VI SINH VT S DNG

I. NGUYN LIU Nguyn liu c dng trong sn xut nm men gia sc l b ru - ph phm ca cc nh my ru, c b sung thm r ng - ph phm cc nh my sn xut ng. Dch b ru l ngun dinh dng nui cy nm men rt tt. Trong dch ny c khong 7.5-10% cht kh ho tan rt giu vitamin B v cc acid amin. Khi nui nm men, dng dch ny pha thm r ng mi trng lun lun c khong 2-3% ng, ta c th thu c t 10- 15 kg men kh t 1 m3 dch b ru. Nguyn liu b ru (Hm) Trong sn xut cn t mt r, b thi ra mi trng l b ru. B ru cha nm men, cht ha tan v c lng cn st. C hai loi b ru : loi b ca cc nh my ru vi ngun nguyn liu t ng cc, sn (cc loi cha tinh bt) v ca cc nh my ru r ng. B ru sau khi chng ct cn l mt loi nguyn liu tt dng nui cy nm men. Bng 2. Thnh phn ha hc ca b ru t r ng Hm lng (% cht kh) Hp cht hu c 70-80 17-27 Protein Nit 3-5 tng 0.4-1.0 protein 0.3-0.6 amin 0.1-0.3 NH3 Acid amin 6-10 Trong b ru c cha cc vitamin: Acid nicotinic (PP) Riboflavin (B2) Priridoxin (B6) Acid pentotenic (B3) Biotin (B7) Acid folic http://www.ebook.edu.vn Trang 4 Vt cht Vt cht Cc acid hu c Trong c acid bay hi Glyxerin Vt cht kh Tro tnh ra K2O Na2O CaO Vi lng Hm lng (% cht kh) 5-27 3-12 6-13 3-7 17-24 7-8 0.5-3 0.5-3
Trong s cc cht ha tan ca b ru c ng (12,5%), cc hp cht nit, cc vitamin nhm B. Ngoi ra, trong b ru cn c cc nguyn t khong, cc nguyn t vi lng... nh vy trong b ru c t ng, nhng rt phong ph cc cht sinh trng. V vy khi dng b ru nui cy nm men ngi ta lc ly phn dch trong ri b sung t 12% r ng, thm supephosphate tng ngun phospho v (NH4)2SO4 hoc urea lm ngun nit. Lng b ru chim khong 0.36% so vi lng mt r em vo sn xut. Trong dch b ru tinh bt sau khi lc loi b, b th dng cho thc n chn nui c 7-8% cht kh v dch b ru r ng thng c 7.5-10% cht kh, nhiu cc hp cht N, nhiu vitamin v khong cht. Trong qu trnh sy nm men chuyn 50-55% cht kh ca r ng vo dch b. Do , dch l mi trng rt giu cc cht sinh trng. Dch b r ng cha 7.5-10% cht kh, trong c ti 3% l cc hp cht v c. Dng kh 0.2-0.5%. glyxerin 0.6-0.9%, cc axit hu c 1.5-2.5%, cc axit amin, cc loi ru, glucoside, cc hp cht v c v hu c, cc mui P, K, Fe, vitamin v cc nguyn t vi lng. Cc hp cht ny men c th hp thu c. Bn cnh , v mt kinh t, b ru cn c mt s u im so vi cc nguyn liu khc: Gi thnh r. Khi lng ln v di do. S dng tin li. Ngun cung cp kh ph bin. Nguyn liu r ng: R ng l ph liu cha ng nhiu ng khng kt tinh trong sn xut ng t ma hoc c ci ng. Yu cu ca r ng dng trong sn xut nm men gia sc: Hm lng cht kh khng thp hn 75% ng 4050% Hm lng cht tro khng thp hn7,5% Tng nit khng thp hn 1,4%
S lng cc vi sinh vt khng qu 15000 t bo trong 1g r ng. Ngun Nit Cc hp cht cha nit ca r ng (a. asparaginic, a.glutamic, lxin, isolxin, tyrosin, cc mui nitrat) c th c nm men s dng n 30-40%. Ch c betain l khng c s dng.
Trang 5
thay th ngi ta dng cc ngun nit v c nh dd NH3, cc mui nitrat, urea nh ngun cha nit, diamoni phosphate (DAP) nh ngun cha nit v photpho. Ngoi DAP v urea, ngi ta c th s dng cc ngun cung cp nit v phospho khc nh: (NH4)2SO4, NH4OH, H3PO4, Ca(H2PO4) nui cy nm men. Tuy nhin, hai ngun nit urea v diamoni phosphate (DAP) l nhng loi phn v c c s dng nhiu trong nng nghip, d mua v r hn rt nhiu so vi cc cht khc nn chng c s dng nhiu trong sn xut sinh khi nm men. Supephosphat v amoni sulfat c s dng nhng khng nh mong mun v chng to ra canxi amoni sulfat, to CaSO4 bm vo mt trong thit b v ng dn, lm cho thit b bm nhiu cu cn gy cn tr cho thanh trng v truyn nhit. Ngun khong Lng DAP c th s dng trong khong 0,15-0,3%, urea c s dng t hn. Nu dng nhiu ure, phi tng cng lng biotin (di dng tin cht l desthibiotin) trong dch ln men v enzyme phn gii ure ca nm men-ure amidoliaza c th i hi mt lng ln biotin. B sung P di dng mt loi mui thch hp hay dng acid orthophosphoric. Ngun Kali v Magie: Trong sn xut sinh khi nm men, ngi ta s dng K2CO3 v KCl nh nhng ngun kali v MgSO4.7H2O hoc MgCl2 nh ngun cung cp magie. Nc Nc s dng trong sn xut sinh khi nm men l nc s dng trong sinh hot (nc my). Nu s dng nc ging phi x l chng cht lng loi nc ny t cht lng nh nc my dng trong sinh hot. Nc c coi nh nguyn liu chnh dng trong sn xut v y l cng ngh ln men hiu kh. Cc yu cu v nc : C cng t 4 8o (1o tng ng 10 mg CaO/l) Khng mu, khng mi, khng v. Cc cht sau khng c qu mc cho php (mg/l): Cl- < 0,5; SO4-2 < 80; As < 0,05; Zn < 5; Cu < 3; FeO < 3. Tng s vi khun hiu kh <10 cfu/L (37oC), khng cha Coliforms, khng cha Faecal streptococci v cc vi khun clostridia kh sulphit.
II. GING VI SINH VT Trong sn xut men chn nui trn mi trng b ru, ging VSV c s dng l cc loi nm men Candida utilis (hay c tn gi khc l: Torula utilis, Torulopsis utilis) hay Candida Tropicalis. Candida utilis Candida tropicalis http://www.ebook.edu.vn Trang 6
SX sinh khi VSV giu protein cho gia sc Gii: Nm Ngnh: Ascomycota Di ngnh: Saccharomycotina Lp: Saccharomycetes B: Saccharomycetales H: Saccharomycetaceae Ging: Candida Loi: Candida utilis 1. Candida tropicalis
GVHD: PGS.TS L Vn Vit Mn Gii: Nm Ngnh: Ascomycota Di ngnh: Saccharomycotina Lp: Saccharomycetes B: Saccharomycetales H: Saccharomycetaceae Ging: Candida Loi: Candida tropicalis
c im hnh thi T bo nm men c hnh ovan, hoc hnh trn, kh ln, kch thc trung bnh ca nm men thng l (5-10) x (4-8) m, phn ln cc t bo kt thnh nhnh, him khi ng ring r. H si gi pht trin tt t nhng si gi ko di, phn nhnh thnh chui. Khng to bo t ti. Trong t bo gi tch t nhiu ht cht bo. Tnh cht nui cy Qua ngy m 36C trong nc malt (4Be) to thnh cn khng nhiu v qua mt thng to thnh mng dy nhn nheo. Khun lc mc trn thch malt hnh trn, mu kem trng. Ra khun lc b chia ct theo hnh rng ca hoc c ta (him kh phng nhn). Men ny l loi d hnh thi : mt chng c khi mc thnh khun lc dng R (nhn nheo) hay dng S( nhn). Tnh cht ho sinh Ln men tt cc dch ng glucose, galactose, sacarose, maltose, treharose, rafinose, melixitose, inulin, sucxinic, citric. Khng hp thu c sovbiose, xentobiose, lactose, milibiose, dulxit, inozit v aicd xalysalic. Cn mt s vitamin lm cht sinh trng : acid pantotenic, paraminbenzoic, tiamin, inozit... c tnh cng ngh Hiu sut thu hi sinh khi ca nm men Candida tropicalis t khong 3846% trong mi trng nui cy. Tc sinh trng ring l 0.15-0.2/h. Nu cho thm vo mi trng cao men (0.5%). Nng sut s tng n 48-50% v tc sinh trng l 0.25-0.28/h. Nhit nui cy thch hp l 36-37C, pH = 4.2-4.5 2. Candida utilis c im hnh thi T bo di c kch thc 4x8.3m, ng ring r hoc i khi kt thnh chui ngn, phn nhnh, khng thy sinh h si hay gi si, khng sinh ra bo t ti. Tnh cht nui cy http://www.ebook.edu.vn Trang 7
Trong mi trng lng to thnh vng v cn lng c. Khun lc mc trn mi trng thch malt c mu vng a, ng nh nh, phng nhn, ra hi b chia ct, thnh thong gia khng lng bng m hi gn nhn nheo Tnh cht ho sinh Ln men c glucose, sacarose, 1/3 rafinose ng ho bng cch oxi ho glucose, sacarose, maltose, rafinose, xylose; yu vi galactose v arabinose. C th hp thu c cc ngun nit nh KNO3, amonisunfat,ure, peptone. c tnh cng ngh Nng sut thu hi sinh khi l 40% so vi cht kh trong mi trng. Bng 4. Thnh phn ha hc ca t bo nm men candida utilis Thng s % cht kh m 5-7 protein (N x 6,25) 50 - 54 Tro 6.5-7.0 Cht bo 3.0-5.0 carbohydrates 25-30 iu kin sinh trng Nhit sinh trng: i vi sinh trng ca a s nm men th nhit sinh trng ti thch vo khong 28- 32oC. Oxi ha tan v hiu kh hiu kh ca mi trng c th hin bng lng oxi ha tan trong mi trng. S c mt ca oxi to iu kin cho nm men h hp v sinh sn. Lng oxi ha tan trong mi trng ph thuc vo nhit , nhit cng cao th oxy ha tan cng km. Bnh thng lng oxy ha tan ti a trong nc l 9 mg/l. Khi nng ny gim xung 1mg/l th nm men s ngng sinh sn. pH ca mi trng Mi loi vi sinh vt ni chung u sinh trng v pht trin ti thch mt gi tr pH nht nh. pH thch hp cho sinh trng ca nm men l 4,0- 4,5. Hm lng ng Hm lng ng (glucose, fructose, galactose, maltose, saccharose, ...) cng cao th nm men sinh trng cng tt. Tuy nhin, nu hm lng ng qu cao s to ra p lc thm thu ln, t c ch nm men sinh trng. Cn lu vi glucose, hm lng cao c th c ch h hp (hiu ng Captree). Tiu chun la chn ging: Nm men gm c nhiu ni (chng) khc nhau, mi ni c mt vi c tnh ring bit, ni chung nm men dng trong sn xut sinh khi phi m bo nhng yu cu sau: http://www.ebook.edu.vn Trang 8
SX sinh khi VSV giu protein cho gia sc -
Nm men Candida utilis c kh nng sinh sn rt nhanh chng: kh nng tch ly sinh khi nm men l 0.2 g/h. Trong iu kin nui nm men c sc kh, mi trng c nng cht kh l 8% 30oC trong 6h. Hm lng protein v sinh t cao, d tiu ha nn c dng sn xut thc n cho ngi v ng vt. Thnh phn ha hc p ng c nhu cu v dinh dng v khng cha cc cht c c tnh i vi ng vt. Khng gy bnh cho ng vt v ngi. ng ha c cc cht dinh dng c trong mi trng vi h s kinh t cao. Thch nghi vi mi trng b ru. C sc chng chu vi tp khun v nhng cht km hm sinh trng T bo nm men c kch thc ln, ng u c th d tch bng separator (my li tm tch)
Chng 3
QUY TRNH CNG NGH

I. S KHI QUY TRNH CNG NGH
B ru
Lc
B Mui dinh dng
Mt r
To mi trng nui cy Thanh trng
Men ging
Lm ngui
Nhn ging
Ln men
Oxi
Ly Trang 9tm
II. THUYT MINH QUY TRNH CNG NGH 1. Qu trnh lc a) Mc ch: Chun b Trong sn xut ru c mt lng dch b rt ln : c 100 lt cn thi ra ti gn 3 12m . Qu trnh lc b nhm thu dch lc lm mi trng nui cy nm men, phn b cn li c tn dng lm thc n gia sc. b) Cc bin i: Vt l: loi c mt s tp cht trong b Ha l: nht gim c) Phng php thc hin : Do b ru c nht cao, tng i kh lc nn ta chn phng php lc p lc nhit cao v c s dng cht tr lc. y, b ru sau khi chng ct (nhit 55-60oC ) c em i lc ngay nn khng cn phi gia nhit b ru trc khi lc. Cht tr lc Cht tr lc l mt loi bt mn c a vo h tr cho qu trnh lc. Cht tr lc c nhim v lm cho l mao qun nh v to thnh trn b mt lc mt lp b b sung lm tng kh nng gi pha rn v gim tr lc ca pha lng. Cc yu cu cn thit ca bt tr lc To c trn b mt lc lp b c xp ln ( = 0,85-0,9), nhng kch thc l xp b.
Trang 10
B mt ring ca bt tr lc khng ln lm (v b mt ring ln th kch thc ht b v tr lc ln). Gii hn thnh phn c ht ca bt tr lc trong phm vi hp (kch thc c ht tng i ng nht). nn p di p sut khng ln lm.
Khng ha tan v c phn ng ha hc vi pha lng ca huyn ph. Ngi ta thng c hai cch s dng bt tr lc + Ha bt tr lc vo huyn ph (khong 0.01- 4 % huyn ph em lc). + Ph lp bt tr lc ln b mt (thng dng cho thit b lc gin on) vi chiu dy khong 0.8 - 2.5mm (tng ng vi khi lng 0.1 - 0.75 kg.m2). trong cc thit b lc lin tc ngi ta thng pha bt tr lc vo huyn ph ri tin hnh lc vi tc ln. Trong sn xut thng s dng nhiu loi bt tr lc khc nhau nh diatomit, amiang, cellulose, mn ca, than g, than hot tnh d) Thit b lc Lc bng phng php ly tm th tt nhng thit b ti tn, gi thnh t. Cho nn ngi ta thng dng thit b lc p khung bn. Qu trnh lc nhm tch ht cn trong b ru thu c dch chit trong. Thit b lc p khung bn lm vic di p lc 4-5 kp/cm2( ti a 15 kp/cm2). Cc khung lc p lc c lm t g, kim loi, hp kim nhm, thu tinh hay compozit. Gia cc tm lc c vch lc (thng bng vi) to thnh bng cha b. Cu to My lc khung bn gm mt dy cc khung 1 v bn 2 c cng kch thc, xp lin nhau. Khung v bn c tay ta trn hai thanh nm ngang 7, gia khung v bn c vi lc 3. Gii hn hai u gm tm c nh 5, u kia l tm di ng 6, dch chuyn c nh tay quay 8.
Hnh 1. My lc khung bn 1. Khung 2. Bn 3. Vi lc 4. Chn 5. Tm y khng chuyn ng 6. Tm y chuyn ng 7. Thanh nm ngang 8. Tay quay 9. Mng tho http://www.ebook.edu.vn Trang 11
Hnh 2. Khung v bn a) Khung b) Bn 1. Khung 2. Bn 3. Rnh huyn ph 4. Rnh nc ra 5,6. Rnh nm ngang 7. Rnh n van Nguyn tc hot ng: Ph lp bt tr lc ln b mt (thng dng cho thit b lc gin on) vi chiu dy khong 0.8 - 2.5mm (tng ng vi khi lng 0.1 - 0.75 kg.m2). Huyn ph c a vo rnh 3. Kh ra, nc ra a vo rnh. Trn b mt ca bn, ngi ta x cc rnh thng ng song song nhau v hai rnh nm ngang hai u. Rnh nm ngang bn di c thng vi van tho nc lc v nc ra. Khung rng to thnh cc phng cha cn. Huyn ph di tc dng ca p sut c a qua rnh 3 ri vo khong rng ca khung cht lng chui qua vi lc sang cc rnh ca bn ri theo van ra ngoi, cn b b gi li trong khung. ra b, ngng cho huyn ph v cho nc ra vo. Nc ra chui qua lp vi lc, qua ton b b dy lp b ko theo cht lng cn li trong b qua lp vi lc th hai sang bn bn cnh ri theo ng ra ngoi. Do khi ra b c mt van ng v mt van m.
Trang 12
Hnh 3. S lm vic ca my lc khung bn a) Qu trnh ra b) Qu trnh lc 1,3. Bn 2. Khung Khi ra xong ngi ta m tay quay, khung v bn tch xa nhau, b s ri xung mng di ri ly ra ngoi. u im : B mt lc trn mt n v din tch sn xut ln; ng lc qu trnh lc (hiu s p sut) ln; c th kim tra qu trnh lm vic c v c th dng khng cho mt vi bn lm vic ( khi thy nc lc chy qua van ca bn no b c th ta ng van li). Nhc im : Thao tc bng tay nhiu, ra b cha tht tt, vi lc chng b rch. Thng s cng ngh: - p lc lc: 5at (c to ra bi bm nhp liu) - Thi gian lc: 1-2h - Nhit : 55-60oC - B dy lp bt tr lc diatomit trn b mt vi: 0.8 - 2.5mm 2. Chun b mi trng nui cy a) Mc ch: chun b Thnh phn v nng cc cu t ca mi trng dinh dng nui cy nm men nh hng rt nhiu n tc sinh sn, thnh phn t bo v hiu sut thu hi sinh khi ca nm men. Cc ngun ng n v ng i, acid amin, glycerin l ngun cacbon dinh dng ca nm men. B ru qua cc k thut sn xut hu nh khng cn cha cc cht sinh trng cho nm men, nn vic b sung cc cht sinh trng vo b ru l cn thit. Trong b thng thiu cn i v t l C:N:P. V vy, mun nm men s dng ht ngun cacbon ta phi thm ngun N v P. b) Phng php thc hin: Ta trn dch b ru vi 1-2 % r ng (tnh theo th tch dung dch) B sung vo dch nui cy vi 1 m3 b ru thng b sung vi 1,3 kg DAP v 0,5 kg ure ri h nhit xung 30oC. Nui nm men : nhit 28-30o, pH 4.0-4.2 , thi gian nui 16-18h. Nu dch b c nng cht kh trn 8% th pha long ti nng 6.8-7.2% Tin hnh pha dung dch DAP : Urea d ho tan tuy nhin s lm gim nhit ca nc, DAP rt kh ho tan trong nc do khi sn xut n c cha mt s cht ph gia khng tan.V vy, trc khi s dng phi ho tan vo nc nng theo t l 1:5-6 (nc) trc. Dung dch c trn ln vi lng acid phosphoric tnh ton, pha long bng nc theo t l 1: 5 ri cho qua b phn o lng. Chun b kh v trng : Phng php vi lc gm 3 giai an
Trang 13
- X l s b : tch bi v hp cht c hc khc ( = 5-10m). Vt liu membrane polymer, phi kim loi - X l th: tch phn ln (98%) cc vi sinh vt trong khng kh ( = 1-1.5m). Vt liu membrane polyamide, polyester - X l tinh: tch cc vi sinh vt c kch thc nh ( = 0.3m), tinh sch 99.99%. Vt liu lc: membrane ceramic,cellulose acetate 3. Qu trnh thanh trng a) Mc ch: chun b Qu trnh ny s tiu dit mt s vi sinh vt c trong dch b ru chun b cho qu trnh nui cy sau ny. b) Cc bin i trong qu trnh: Vt l: nhit tng. Ha hc: mt i mt s cu t mn cm vi nhit cao, c th xy ra phn ng Maillard. Ha l: nht dung dch gim. Ha sinh: cc enzym b bin tnh bt thun nghch. Sinh hc: h vi sinh vt trong mi trng b tiu dit mt phn. c) Thit b thanh trng bn mng:
Hnh 4. Thit b thanh trng bn mng Cu to B phn chnh ca thit b l nhng tm bn hnh ch nht vi dy rt mng v c lm bng thp khng g. Mi tm bn s c 4 l ti 4 gc v h thng cc ng rnh trn khp b mt to s chy ri v tng din tch truyn nhit. Nguyn tc hot ng Mi trng nui cy c bm vo thit b trao i nhit bn mng cng vi dng nng. d) Thng s cng ngh: Nhit thanh trng: 95-98 oC Thi gian thanh trng: 30 pht 4. Qu trnh lm ngui a) Mc ch: chun b http://www.ebook.edu.vn Trang 14
H nhit ca canh trng chun b cho qu trnh nui cy. b) Cc bin i trong qu trnh: Vt l: nhit gim Ha l: nht dung dch tng. c) Thit b trao i nhit bn mng: Nguyn tc hot ng Mi trng nui cy c bm vo thit b trao i nhit bn mng cng vi dng hi lnh. d) Thng s cng ngh: Nhit mi trng dinh dng sau qu trnh lm lnh: 28-32oC 5. Qu trnh nui cy men ging a) Mc ch: chun b y l giai on chun b cho qu trnh nui cy m rng nhm lm tng dch nm men ging sau mi chu k nui cho n khi lng ging cn thit cho qu trnh sn xut. b) Bin i: Sinh hc: sinh khi vi sinh vt tng. c) Phng php thc hin: Nui cy nhn ging u tin c thc hin trong phng th nghim: Ging c nui cy trn ng nghim thch nghing ri cy chuyn vo ng nghim 20ml cha 10ml mi trng nui cy c v trng. Tin hnh nui 2832oC trong vng 16-20 gi. Sau khong thi gian trn th ta ln lt cy vo bnh tam gic v trong cc thit b ln hn, t l ging chuyn cp l 1:10 cho n khi c 100l th ta chuyn sang giai on nhn ging ch phn xng. Nhn ging trong giai on phn xng: Tin hnh nhn ging trong thit b c th tch 3-4 m3 cha 1m3 dch nui cy cng nhit v thi gian nh trn, sau ta tip tc nhn ging vo nhng thit b ln hn vi t l mi cp chuyn ging l 1:10 cho n khi c 100 m3 . Qu trnh nhn ging trong phng th nghim v nhn ging trong sn xut cn to iu kin cho ging thun chng sinh sn nhanh, khng b tp nhim vi cc dng c v thit b, phc v cho nui cy v khun (k c lc kh phc v cho sc kh). Khi nui cy u cn phi m bo cc bin php trnh nhim tp nh: un nng hoc lc cc thnh phn mi trng: Mi trng nhn ging v khi tin hnh nhn ging cng phi m bo v trng. Kh trng thit b nui cy Cc qu trnh nui cy cn thng kh mnh cho nm men h hp v pht trin sinh khi.
Trang 15
Nhit sinh ra trong qu trnh nhn ging phi c loi b nh mt h thng lm lnh. d) Thng s cng ngh: pH trong qu trnh nhn ging v nui ging: 4-4.2 Nhit trong qu trnh nhn ging v nui ging: 28-32 oC Thi gian nui mi ln chuyn cp: 16-20 gi. 6. Ln men a) Mc ch: khai thc Qu trnh nui cy men m rng nhm mc ch tng sinh khi t bo nm men n mc nh mong mun. C s khoa hc Khi nui cy nm men ru trong mi trng giu ng nu oxy chng s sinh trng, tng sinh khi v sng trng thi h hp, nu thiu oxy hoc khng c oxy chng chuyn sang ln men (h hp k kh). Nu cung cp oxy lin tc vo mi trng th t bo nm men s chuyn t trng thi ln men sang h hp. Qu trnh hp thu glucose trong t bo ang h hp xy ra chm hn so vi cc t bo ang ln men. Hin tng ny c gii thch nh sau: qu trnh ln men sinh ra t nng lng hn qu trnh h hp. V vy, c nng lng, trong iu kin ln men cn phi phn gii nhiu glucose hn, t bo ln men hp thu nhiu glucose hn V nguyn tc, sinh khi nm men c th c thu nhn bng 2 cch: nui cy k kh v nui cy hiu kh. Trong iu kin nui cy hiu kh sn phm ch yu l sinh khi, cn CO2 l sn phm th cp. Trong nui cy k kh thu c sinh khi t hn (ch khong so vi hiu kh), cn li l CO2 v mt s sn phm trao i cht ca t bo nm men, quan trng nht l ethanol. V th, trong sn xut sinh khi nm men, t c hiu sut thu hi sinh khi cao cn phi to iu kin hiu kh m bo cho nm men sinh trng v tng sinh khi tt ng thi trnh tiu hao nhiu cht dinh dng cn thit trong qu trnh nui cy b) Cc bin i xy ra trong qu trnh nui cy Cc bin i sinh hc Cc giai on sinh trng ca nm men giai on tim pht, nm men cn phi c thi gian thch nghi vi iu kin mi. giai on ny vic to thnh cc acid amin, cc peptid v acid nucleic xy ra nhanh hn cc qu trnh khc. Thi gian c th ko di cng c th ngn tu thuc vo tng chng nm men. Giai on ny thng khong 0,5-1 gi v ch cn cung cp lng khng kh khong 50 m3/h/m3 mi trng l . giai on tng trng, nm men tin hnh cc qu trnh trao i cht rt mnh, khi lng nm men trong thit b tng ln r rt. Vic tng hp cc enzyme http://www.ebook.edu.vn Trang 16
c xc tin nhanh. Hm lng RNA c tng hp nhiu nht. Giai on ny thng ko di khong 7-14 gi v cng l giai on nm men cn nhiu oxy nht, do lng oxi cung cp phi gp i giai on u tin, khong 80-100 m3/gi/m3 mi trng. Giai on k tip l giai on cn bng. Giai on ny c nhiu thay i quan trng. Lng t bo mi sinh gn bng lng t bo cht. Cc qu trnh trao i cht gim mnh, do nhu cu oxy khng nhiu. Giai on ny ko di khong 1-2 gi v ta phi gim lng khng kh cp vo mi trng. Lng khng kh cp vo mi trng ny t 20-50 m3/ gi/m3 mi trng. Cc bin i ha sinh Cc bin i ha sinh phn ln xy ra bn trong cng vi s trao i cht ca t bo nm men. Bin i u tin quan trng nht l s chuyn ha ng saccharose thnh D-glucose v D-fructose di xc tc ca enzyme invertase c tng hp bi nm men. Sau , glucose tip tc tham gia cc chui phn ng ha sinh trong cc chu trnh sinh ha tng hp vt cht t bo v nng lng cho nm men sinh trng. Cc bin i vt l Trong qu trnh nui cy, qu trnh trao i cht ca nm men xy ra lin tc lm cho nhit canh trng tng ln. Cc bin i ha hc Hm lng cht kh (ch yu l ng saccharose): gim dn theo thi gian do qu trnh h hp hiu kh v qu trnh sinh tng hp vt cht t bo ca nm men. pH: s thay i theo 2 c ch chnh: S sinh tng hp cc acid hu c (cc sn phm ph ca qu trnh trao i cht) C ch ng vn chuyn ion H+ trong v ngoi t bo nm men trong qu trnh trao i cht. Cc bin i ha l S ha tan oxy V nm men ch c th s dng oxy dng ho tan nn tc ho tan ca oxy vo dung dch bng tc s dng oxy ca t bo nm men th sinh khi t bo nm men t cc i. ha tan ca oxy ph thuc vo nhiu yu t nh nhit , tc khuy trn, lu lng sc kh, s c mt ca cc cht hot ng b mt, cc cht ph bt. S hnh thnh bt Trong qu trnh nui cy, do sc kh lin tc v nm men h hp gii phng mt lng ng k CO2 nn lm tng th tch bn nui cy, gy hin tng tro bt trn b mt. V vy dch nui cy rt d b tp nhim. Do , dung tch s dng thit http://www.ebook.edu.vn Trang 17
b phi nm trong khong 0,7-0,8 ng thi c s dng cht ph bt (keo a nc, m c, hn hp x phng ha, acid oleic, du silicone...). c) Cc yu t nh hng n qu trnh nui cy Thnh phn mi trng nui cy Mi trng nui cy phi n nh v thnh phn cht dinh dng phi n nh cht lng nm men ng u tt c cc m nui trong sut qu trnh sn xut v mi trng nui cy phi tuyt i v trng. Thnh phn mi trng nui cy nh hng n cht lng ca qu trnh nui cy nm men nh sau: nh hng ca hm lng ng Lng ng trong mi trng khong 22% (nui cy gin on) v khong 2-3% (nui cy bn lin tc) khng nn nhiu hn v cng khng nn t hn v: Nu hm lng ng cao s va gy lng ph (v trong qu trnh tng sinh khi trong iu kin hiu kh nm s dng ng rt t) va to ra nhng sn phm trao i cht khc, s gy c ch ngc n qu trnh to sinh khi. C th l, iu kin hm lng ng thp v c mt ca oxi, nm men s tin hnh qu trnh trao i cht v nng lng theo chu trnh Crebs (t 1 phn t ng hexose to thnh 38 phn t ATP). Con ng ny gip s dng t c cht (tit kim c cht) nhng nng lng to ra rt nhiu. Trong khi , nu hm lng ng cao th mt lng ng d tha s tham gia cc con ng trao i cht khng cn thit (nh con ng hnh thnh ethanol t acid pyruvic), d cho qu trnh nui cy din ra trong iu kin hiu kh. Mc khc, hm lng ng qu cao c th to nn mt p sut thm thu cao trong mi trng nui cy c th kim hm s trao i cht v sinh trng ca nm men. Hm lng ng cao cn c th to ra dung dch c nht ln nh hng n s ha tan v khuch tn oxi cng nh kh nng tip xc ca nm men vi thc n. Kt qu l lm gim hiu sut sinh tng hp sinh khi nm men v lm ko di thi gian nui cy. Nu hm lng ng qu nh s khng ngun carbon cn thit cho qu trnh to sinh khi, t lm cho ko di thi gian thu nhn sinh khi. Cn lu rng, nu trong mi trng c nng ng glucose cao cng c kh nng c ch h hp ca nm men (hiu ng Captree). Khi qu trnh h hp b c ch th qu trnh ln men s din ra thm ch ngay c khi c mt ca oxy lm tn tht ng. p H pH trong mi trng khong 4.0-4.2. Khng nn h thp pH ca mi trng xung qu gi tr ny v c th lm cho nm men kh pht trin. Nu pH cao qu va c ch nm men pht trin va to iu kin thun li cho vi khun nhim vo pht trin, cnh tranh cht dinh dng, lm gim cht lng nm men thnh phm. C ch c ch sinh trng ca nm men pH thp nh sau: pH c trng cho kh nng phn ly H+ ca cc acid hu c hoc v c. pH cng thp H+ cng nhiu, khi H+ s tham gia vo qu trnh vn chuyn cht trong v ngoi mng t http://www.ebook.edu.vn Trang 18
bo, c bit l lm ri lon cc phn ng sinh ho c enzyme xc tc. iu kin nui cy Nhit Nhit nui cy phi c kim sot cht ch v iu chnh n nhit ti thch cho nm men pht trin. Nhit qu cao hay qu thp u nh hng qu trnh sinh trng v trao i cht do nh hng n hot tnh ca cc enzyme trc tip tham gia vo cc chui phn ng sinh ho ca c th. Mc khc, nhit cn nh hng n ho tan ca oxy trong mi trng. Nhit cng thp th ho tan ca oxy cng cao. Nhng nu nhit qu thp (di 28oC) s lm chm qu trnh sinh trng ca nm men ko di thi gian nui cy. V vy, trong nui cy nm men ngi ta thng iu chnh nhit mi trng nui cy thng l 28-32oC. Oxy Oxy l mt trong nhng yu t quan trng nh hng n sinh trng v pht trin ca nm men Cc qu trnh ln men thong kh i hi phi cung cp O2. O2 cn dng lm cht nhn hydrogen v in t cui cng, ngoi ra O2 cn c dng trong cc qu trnh oxy ha do oxygenase xc tc. Vi sinh vt ch s dng O2 ha tan, nhng ha tan ca O2 trong nc l rt nh. iu kin p sut kh quyn v nhit 30oC, trong cc dung dch dinh dng c thng kh c ha tan khong 4 - 5 ml O2/l. Tri li trong cc qu trnh sn xut, vi sinh vt i hi lng O2 mc t 500-5000 ml O2/l.h. Nhu cu O2 c quyt nh bi hot tnh trao i cht ca vi sinh vt. V d tng hp c nguyn liu t bo ca 1g cht kh nm men t ng th cn khong 500 ml O2. V trong qu trnh ln men loi ny mt t bo l 10g/l, mt khc s tng gp i khi lng t bo nm men trn glucose ko di trong 2h nn tr s v nhu cu O2 l 2500 ml O2/lt dch dinh dng.h. Cc tr s ny ph thuc vo mc oxy ha c cht. Ngoi ra nhu cu O2 cn ph thuc vo iu kin nui cy v tc sinh trng ca vi sinh vt, khi sinh trng chm nhu cu ny cng gim i. m bo vic cung cp O2 mt cch c hiu qu v ti u l mt trong nhng chc nng chnh ca ni ln men. Nh cc h thng khuy theo cc kiu khc nhau c tc dng khuy trn v ph v cc bng khng kh, hoc nh cc h thng bm a khng kh vo dung dch m c th tng cng s xm nhp ca O2. Vic cung cp oxy cho qu trnh to sinh khi l rt cn thit. Tuy nhin, nhu cu cung cp oxy cho qu trnh pht trin ca nm men khng phi lc no cng ging nhau. Do phi thay i mc cung cp oxy theo ng nhu cu thc ca nm men. Ngi ta thng cung cp oxi cho mi trng nh khng kh. Khng kh trc khi a vo thit b ln men, phi c lc v trng lm sch khi vi sinh v tp cht. Nm men ch c th s dng oxy dng ha tan trong mi trng lng. http://www.ebook.edu.vn Trang 19
Thng thng, lng oxy ho tan trong nc rt t. Trong qu trnh pht trin, nm men s nhn oxy ha tan v nh vy lng oxy ha tan s gim, do cn phi cung cp oxi t bn ngoi thit b. Nu ch thiu oxy trong thi gian ngn, ngay lp tc chng chuyn qu trnh ln men hiu kh sang qu trnh ln men k kh. Nh vy lng sinh khi to thnh s rt t v ng s c chuyn ha theo cc chu trnh ng phn c cc chu trnh khc cui cng to ra sn phm trao i cht bc hai. Do qu trnh nui cy nm men thu nhn sinh khi bt buc phi c cung cp oxy lin tc. S tng hm lng oxy ha tan khng t l thun vi s tng sinh khi. S khuy trn v thi kh Trong thit b ln men ngi ta phi lp t h thng phn phi kh v h thng cnh khuy. H thng phn phi kh trong nui cy nm men thng c lp t di y thit b. Vic cung cp oxy cho nhng thit b nh trong cc phng th nghim thng rt kh khn. Do ngi ta thng s dng my lc tng s ha tan ca oxy vo mi trng. Khi s vng quay ca cnh khuy v tc thi kh tng th ho tan ca oxy vo dung dch mi trng s tng theo. ng thi lm tng s xo trn mi trng v thc y qu trnh truyn nhit v truyn khi tt hn, t lm tng nhanh qu trnh trao i cht v sinh sn ca nm men. Tuy nhin, vic tng tc khuy v tc thi kh ch n mt gii hn nht nh. Nu tc ca hai yu t ny qu cao s lm tng nhanh hin tng t phn ca t bo v bt s to ra nhiu. Nu trong qu trnh ln men, nhit mi trng tng nhanh (do qu trnh h hp ca vi sinh vt) th ngay lp tc phi thi kh mnh lm gim nhit ca mi trng. Nu khng lm h nhit, ho tan ca oxy s gim v t nng sut sinh khi cng gim. S c mt ca cht hot ng b mt hay cht ph bt Trong trng hp phi dng nhng cht hot ng b mt, cht ph bt hoc mt s cht chng nhim trng, ha tan ca oxy cng gim. Dch men trong qu trinh nui nu c thi kh s to thnh bt. Khi nui men t yu cu, dch men tho ra bnh cha l mt th dch bt vi nng 0,25 g/cm3, trong bt chim 2/3 th tch. Trc khi tch men cn phi ph bt bng bin php c hc v ha hc. Cc cht ph bt l cc hat cht b mt: cc cht keo a nc, m c voi, c mp, hn hp x phng ha, acid oleic, du silicon. tng tc dng ph bt, cc cht ny c s dng dng nh ha vi nc, vi t l 1:6. Ph bt bng c hc da vo kt cu ca thit b. Trong sn xut sinh khi nm men gia sc, ngi ta thng kt thc qu trnh nui nm men gn cui giai on hai tc l giai on tng trng, bi v mt trong nhng yu cu quan trng l s lng t bo nm men sng phi chim i a s nn khng th i n giai on cn bng mi tin hnh thu nhn sinh khi, lm nh vy nm men thu c s cha rt nhiu t bo gi. http://www.ebook.edu.vn Trang 20
d) Cc phng php nui cy: Trong sn xut cng nghip ngi ta thng dng phng php nui cy theo dng lin tc. Hm lng oxy ha tan l nhn t gii hn trong thnh phn ti u ca mi trng v iu kin thun li cho nui cy. Tc s dng oxy ca nm men ch ph thuc vo hat tnh ca chng v thnh phn ca mi trng. Hiu sut ca thit b nui cy ph thuc vo h s hp ph ca oxy, mc s dng khng kh, ph thuc vo nhu cu ring v oxy tng hp 1 n v sinh khi t tng lai nguyn liu v ph thuc vo h s tng hp. xc nh ch lm vic ti u ca mt thit b, ngi ta xc nh mc s dng oxy v theo s chn c tc pha long, thnh phn tng ng ca mi trng v tng cao nng sut sinh khi. Cc t bo cng c gi lu trong thit b th hot tnh ca ging cng gim v nng sut sinh khi cng thp. V vy khi tin hnh nui ln men theo phng php bn lin tc th s cho hiu qu kinh t cao: khi t lng sinh khi c trong dch nui cy s ly dn ra ri cho thm mi trng mi vo ni ln men c hm lng ng khong 1-2 %. S dng ging nm men c hat tnh thp s lm gim hiu sut thu hi sinh khi.. e) Thit b ln men trao i khi mnh Cu to:
Trang 21
Hnh 5 Thit b ln men trao i khi mnh Thit b l mt dung lng xilanh 17, bn trong lp xilanh hng 2. Hai on ng 4 v 18 nh v cho xilanh hng trong dung lng. on ng 18 c lp cht n y v chia dung lng ra lm hai phng: phng 19 dng nui cy canh trng, cn trong phng 20 tn dng b sung nguyn liu ban u. on ng 4 lp cch mt y ca dung lng. Bn trong xilanh hng 2 v trong khng gian gia tng dung lng v on ng 4 c b tr cc ng gp 16. Cc ng gp c lp cht bi cc ng t l 21. Trong khng gian gia xilanh hng 2 v cc on ng 18 v 4 c cc b trao i nhit 1. np khng kh n cc ng gp trong phng 19 dng ng gp phn phi 7, cn trong phng 20 dng ng gp 5. phn trn ca dung lng c ng gp 14 thu nhn v lm kh bt, bn trong c lp cc a hnh nn 15. Khng kh thot ra t phng 19 qua b tch kh 9. My kh bt bng c hc 12 vi b dn hng c lp t trn cc a 15. Mi trng dinh dng c y vo thit b qua khp 3. Sinh khi c tho ra khi thit b qua khp 11, cn khng kh - qua khp ni 8 v 13. http://www.ebook.edu.vn Trang 22
Nguyn tc hot ng Np hn hp dinh dng ban u vo phng 19 qua khp ni 3, cn khng kh - vo thit b qua khp ni 6. Trong phng 19 xy ra nui cy sinh khi. Tun hon v o trn cht lng c thc hin bi thit b bm dng bng kh nn. T phng nui cy, cht lng canh trng chy qua on ng 12 vo phng 20, ti y b sung nguyn liu.Bn trong phng 19 v 20 dung dch canh trng c thi kh nh cc ng c c nhiu l. Khi khuy o v sc kh mnh lin tc trong ni ln men s to ra bt, n c khuynh hng tro ra khi ni ln men v gy nhim tp mi trng ln men, ngoi ra bt kh cn cn tr s tip xc gia vi sinh vt v mi trng dinh dng. Do vy, trong qu trnh ln men ngi ta cn kim sot lng bt to thnh v tm cch ph hu chng Sinh khi tho ra khi phng cng vi pha bt c to thnh phn trn ca phng. Mt phn bt ni ln theo cc ng rnh gia cc a nn 15, c tch khi cht lng v c c li. Bt c c bng b kh bt c hc 12 v tho ra qua khp ni 11. Thi khng kh khi phng 19 qua khp ni 8 nh b tch kh 9, cn khi phng 20 qua khp ni 13. Khi nui men t yu cu, dch men tho ra bnh cha l mt th dch bt vi nng 0,25 g/cm3, trong phn bt cn li chim 2/3 th tch. C th s dng cc cht ph bt l cc hot cht b mt: cc cht keo a nc, m c voi, c mp, hn hp x phng ha, acid oleic, du silicon. tng tc dng ph bt, cc cht ny c s dng dng nh ha vi nc, vi t l 1:6. c tnh k thut ca thit b: 200 m3 Th tch ca thit b ln men: H s cha y: 0,6 p sut lm vic: 0,02- 0,03 Mpa Nhit hot ng: 28-32oC Mi trng pH: 4,0-4.2 7. Qu trnh ly tm a) Mc ch cng ngh: khai thc v chun b Qu trnh ny nhm mc ch phn tch thu nhn t bo nm men t canh trng nui cy c thc hin ngay sau khi kt thc qu trnh ln men. b) Phng php thc hin tch nm men khi dung dch ln men, ta dng phng php ly tm. Nm men thng c t trng ln hn dung dch ln men (t trng ca nm men thng nm trong khong 1,13-1,14 cn dch nui cy khng cha nm men l 1,011,03). Do vy, t bo nm men s chu lc ly tm ln hn v c tch ra khi dung dch nui cy. c) Thit b http://www.ebook.edu.vn Trang 23
Cu to My phn ly gm khung 1 vi c cu dn ng, trng quay 2 vi cc a v trc, b phn cha cht c nm men 4 v on ng tho cht lng canh trng x l 3. Dn ng my phn ly c thc hin t ng c ring bit qua khp ni ly hp ma st v b truyn trc vt bnh vt c tc cao. Trng quay c lp t t do trn trc con v c lp vo cc rnh x ca trc bng thanh ging, nh m bo vic t iu chnh tm ca trng quay. Bn trong trng c lp cc a hnh nn c cc g trn b mt ngoi, khong cch gia cc a bng 0,8 mm. Gia c cc ti trong ng quay nm trong b gi a. phn di ca my theo vng trn phn b cc rnh xuyn qua c t cc ng tho cht c nm men.
1- C cu dn ng 3- ng tho sn phm Nguyn tc hot ng Huyn ph nm men np qua ng phn phi vo khoang trong ca b gi a, ti y nh cc g ca n m chuyn ng quay c truyn n. Huyn ph chy qua gia b gi a v y trng. Di tc ng ca lc ly tm, cc t bo nm men ln hn bn vo ngoi vi ca trng quay. Huyn ph t khoang cha nm men vo ti ca cc a hnh nn v trong ch chy tng th b trn ra thnh lp mng u nhau. Di tc ng ca lc ly tm cc t bo nm men, khi c t trng ln so vi pha lng, lng trn b mt trong ca cc a v c chuyn o theo b mt vo khng gian cha bn ca trng. Cht c nm men qua ming phun ngoi vo thng cha. http://www.ebook.edu.vn Trang 24
Hnh 6 :Tthit b ly tm 2- Trng quay 4- B phn cha dung dch nm men
C th iu chnh nng nm men trong huyn ph c c bng phng php thay i ng knh cc l trong ming phun, tuy nhin t s cc ng knh l trong v bn ngoi ca ming phun ln hn 1:1/7. Khi gim lng ming phun th mc c huyn ph nm men tng ln lm cho nng sut ca my gim. Cht lng c phn ly khi qua ti ca cc a, chy ngc ln dc theo b mt ngoi ca b gi a v chy vo khoang ri c tho ra ngoi nh a p lc. Mc c huyn ph ph thuc vo nng nm men trong huyn ph ban u. Khi nng ca nm men c 75% nc, bng 20-30 g/l, mc c l 8-10%, cn khi nng 80-120 g/l l 5-6%. C huyn ph n hm lng nm men 550-600 g/l c tin hnh trong khong 2-3 mc phn ly lin tc. Cht lng cn phi t do cho n khng th chy ra khi vi phun, khng bt vi v khng to vm bn trong rto. Lin quan vi iu vic lc s b c nh hng tt ti hot ng ca my phn ly. Loc s b qua b lc li loi cc tp cht c hc, lm bn khong khng gian gia cc a v lm bn cc l trong ming phun. S dng cc my phn ly trn thu nhn cc cht cn c m nh nht l khng cn thit c tnh k thut: Nng huyn ph sau ly tm: 20-25% cht kh c im sinh khi nm men sau ly tm Sinh khi nm men thu c dng st c 75-80 % nc, 20-25 % cht kh trong cacbon chim 40-50 %, nit 7-10 % tng ng vi 40-60 % protein, hydro 5-7 %, oxy 25-30 %, cc nguyn t v c 5-10 % ( phospho v kali chim 95-97 % tng lng tro, s cn li l canxi, magie, nhm, lu hunh, clo, st, mt lng rt nh cc nguyn t mangan, km, molipden, bo, coban), Ngoi ra,trong t bo nm men cn ch hu ht cc cht cn thit cho s sng nh glucid, lipid, enzym, cc acid nucleic 8. C c a) Mc ch: chun b Chun b cho qu trnh sy d dng hn bng cch tng nng cht kh, lm gim chi ph v thi gian sy. Git men v tt c cc vi sinh vt tp nhim hn ch cc mm bnh c th nhim t vi sinh vt Ph v t bo nm men nhm tng h s hp thu nm men cho ng vt . b) Cc bin i Vt l: Tnh cht dung dch thay i, h s dn nhit, nhit dung gim, khi lng ring, nhit si tng. Ha l: nht ca huyn ph tng ln Ha sinh: mt s enzym b bin tnh. Sinh hc: h vi sinh vt tn ti trong nm men b tiu dit. c) Phng php thc hin: http://www.ebook.edu.vn Trang 25
C th tin hnh phng php bng cc thit b c c bng thit b dng mng hoc tm bn. Nng cui cng ca nm mn ln n 40%. Phng php thng c s dng vi sn phm sy phun. d) Thit b
Hnh 7. Thit b bc hi dng mng ri Cht lng c phn phi mt cch u n mt trong ca ng, chy xung to thnh mt mng mng. Qu trnh bc hi s din ra ti y nh nhit c cung cp bi hi nc. Hi nc ngng t v chy xung mt ngoi ca ng. C nhiu ng c lp t cnh nhau to thnh mt chm ng. Hai u ca chm ng c c nh bi 2 v ng v ton b chng c bao bc bi mt lp v o. B phn nh th c gi l calandria. Hi nc c a vo bn trong lp v o. Khong khng gian gia cc ng gi l vng gia nhit, mt trong ca ng c gi l vng bc hi. Cht lng c c c v hi nc ra khi calandria ti phn y. Ti , phn ln cht lng c c s c tho ra. Phn c gi li s i vo b phn tch hi cng vi hi nc. Sau khi tch hi, phn cht lng ny cng c tho ra (thng s dng bm ging nh phn chnh ca dch c c tho ra t calandria). Cn hi s ri khi b phn tch hi ti nh. Hi gia nhit ngng t ti mt ngoi ca ng v c tp trung di dng nc ngng ti y ca vng gia nhit v cng c tho ra bng bm. V hi th to ra t dung dch c c cn cha rt nhiu nng lng, cho nn ngi ta tn dng n lm tc nhn gia nhit. iu ny c thc hin bng cch thm mt calandria khc vo thit b bc hi. Calandria mi ny c nhit bc hi thp hn, lm vic nh l mt bnh ngng t hi th t calandria th nht. http://www.ebook.edu.vn Trang 26
t c s khc nhau v nhit gia sn phm v hi nc trong calandria th hai, vng bc hi trong calandria c vn hnh iu kin chn khng cao hn tng ng vi nhit thp hn . d. Thng s cng ngh: Huyn ph sau ly tm a qua thit b c c chn khng. Nhit : 75oC. Ap sut : 360mmHg., Nng cht kh sau c c 40% khi lng cht kh. 9. Sy a) Mc ch:ch bin, bo qun Huyn ph sinh khi nm men sau qu trnh c c c nng cht kh 40% . Qu trnh sy nhm mc ch tch m ra khi men, a m ca chng v di 8% ko di thi gian bo qun ca nm men. ng thi, qu trnh sy cng lm a dng ha sn phm nhm thun tin cho qu trnh vn chuyn v s dng b) Cc bin i trong qu trnh sy. Vt l: hm m gim nhanh chng. Ha l: s bay hi nc v cc cht d bay hi di tc ng ca nhit cao. C s chuyn pha t dng lng ( dch ln men ) sang dng rn. Ha sinh: mt s enzym b bin tnh. Sinh hoc: t bo nm men v mt s vi khun b tiu dit. Tuy nhin do thi gian sy rt ngn nn bin i v ha sinh v sinh hc khng ln lm. c) Thit b sy phun Thit b sy phun y hnh nn Cu to v nguyn tc hot ng
Trang 27
Hnh 8. My sy phun y hnh nn Sy phun gm mt bung sy hnh tr, u di hnh nn (ng knh bung hnh tr 8-10 m, phn tr cao 5,5-7 m, phn nn cao 6,6-8,7 m). Phn bn trong trn nh bung sy lp h thng phun. Hn hp khng kh nng theo ng trung tm bung pha di a phun lm nng bung sy. Kh tha s theo xyclon lc kh ra ngoi. Thnh phm dng bt c y ra di tc ng ca lc ly tm. Dung dch y vo sy b phun ra nh c cu ly tm (13) c a (10). a phun (10) quay vi tc 10000 vng/pht t ng c qua hp gim tc. bi trn c cu phun, phn trn ca thit b c lp c cu c hc v b lc m (14). V lng in (15) dng nng c cu phun. Tc nhn sy a vo phn trn ca thit b theo ng dn (7). cui ng dn (7) lp c cu phun hnh nn (8). Nh c cu (8), to ra dng xoy ca kh a vo. Cc git sn phm c phun bng a b bao ph bi dng khng kh v chuyn xung di. m c bc hi, cc phn t bt nh cn li lng xung y hnh nn v tho n c cu (1) chuyn sn phm vo h bng ti kh ng hc. Lp my rung (17) ty sch cc tiu phn ca sn phm bm trn tng,.
Trang 28
Tc nhn sy b thi c mang theo cc tiu phn nh ca sn phm ra khi thit b sy qua ng dn (2) vo xyclon tch bt. V tr (9) c y hnh nn tho bt kh. trnh chy sn phm trong my sy, ngi ta t cc c cu bo him 3 v 18. kho st bn trong, c xe nng (4), ngun chiu sng (6) v ca (5). Tm ngn my sy (11) c cc van bo him dng cc a chng nhau v dng ng ng (12) x kh sy khi tng p sut ng k.
. Hnh 8. H thng sy phun t hp. B sy gm thng cha dung dch cht lng canh trng 2, cc bm ly tm 3 v 9, thit b lc kh 1, phng sy 4, c cu tho d y bt kh vo bng ti kh ng 10, cc b lc vi khun 7, qut hai chiu 6, calorife 8, thng cha sn phm kh 12, cc b lng bng xyclon 11, b tho d xyclon 13, b lc khng kh 5 y vo calorife 8. Cc thng s. Tc a phun 10000 vng/pht Nhit vo ca tc nhn sy: 180-200oC. Nhit ra ca ra bung sy : 85-95oC. Nng nguyn liu vo: 40% cht kh m sau khi sy: 8% Nhit nguyn liu trong qu trnh sy 60-70 oC Thi gian tip xc vi tc nhn sy ngn, ch khong vi giy. Nm men c a nng ln khng qu 95oC lm cho cht lng ca cc cht thnh phn trong nm men nh protein, vitamin, mu sc v cu trc khng b bin i, c hon thin hn cng nh d tiu ha hn. http://www.ebook.edu.vn Trang 29
10. Qu trnh bao gi a) Mc ch: hon thin sn phm b) Phng php tin hnh: Sn phm sau khi sy kh c bc trong cc gi bng giy v bng polietylen theo tng l t 0,3 n 1,6 kg. Cng on bao gi sn phm c tin hnh trn dy chuyn t ng B6-BPA, dy chuyn kho st kh nng bin i kch thc ca hp theo chiu cao t 150 n 300 mm vi ng knh khng i bng 242 mm, v nh lng sn phm trong gii hn 0,4-0,5 kg. Dy chuyn c s dng hot ng trong phn xng chia gi nhit t 18 30oC v m tng i ca khng kh n 60%. c) Thit b Cu to
Hnh 9 S ca dy chuyn t ng nh lng phn chia bao gi 1. B nh lng sn phm t ng 2. C cu cp liu mng mng 3. B to ng 4. My hn mi dc ca ng 5. C cu cng ng 6. My hn y v np gi 7. C cu ct ti 8. Cu chuyn ti hp rng 9. C cu t gi thnh phm vo hp 10, 11 C cu nn i cc ti vo cc hp 12. My t ng ghp np 13. B o hp 14. My dn nhn Nguyn tc hot ng http://www.ebook.edu.vn Trang 30
Np sn phm vo ng lm bng mng polyetylen c hn t b nh lng 1. Sau khi kt thc hn m cp dc nh ra. ng c hn cng sn phm h xung di nh cc bng ti ko ca c cu h ng 5 xung mt khong bng chiu di ca gi, sau hn gi, ct gi di, np sn phm cho gi tip theo. Gi ng y sn phm ri xung hp kim loi qua phu nhn nm trong bng ti xung ca c cu xp. Np cc hp kim loi rng ti bng ti xung c tin hnh bng phung php gt hp qua cu chuyn. T bng ti xung ca c cu xp hp, cc gi c chuyn n bng ti kiu tm ca my ghp m t ng ghp y v chuyn n my dn nhn qua my lt hp. Hp c a vo my dn nhn v tr nm ngang ri dn vng trn v ti hp ti mng nghing ca my dn nhn. Sau hp theo bng ti vo kho thnh phm. c tnh k thut ca dy chuyn t ng nh lng phn chia bao gi Nng sut: 480 gi/h Khi lng mt ln nh lng: 0,4 0,5 kg Phng php nh lng: Cn chnh xc nh lng: 1% so vi liu lng nh mc Cng sut thit k ca ng c: 9,16 kW Kch thc c bn: 6820x2370x3210 Khi lng: 4850 kg
Trang 31
Chng 4
SN PHM SINH KHI NM MEN

I. SN PHM SINH KHI NM MEN
II. YU CU CHT LNG CA MEN THNG PHM Tiu chun ca sinh khi nm men thng phm dng cho chn nui nh sau: Ch tiu cm quan: Dng bt Mu sc: xm, trng, vng, nu. Mi v: c trng ca nm men, khng c c mi v l. Kch thc: hiu sut qua ry 3mm trn 95% Ch tiu ha hc: m: khng qu 8 % Protein: khng nh hn 45% (tnh theo cht kh), vi men lai kh l 56,0 % Lizin, metionin v tryptophan tng ng khng di 0,5; 1,4; 1,1 % ca protein kh tiu ha ca protein khng di 75- 80 % Gi tr sinh hc ca protein kh khng di 55 % Cc vitamin B1, B2, B5 tng ng khng di 10, 30 v 300 mg/kg. Hm lng tro: i vi men ru t r ng khng qu 14% men kh tuyt i, cn men ru t b ru ng cc th khng qu 10 %. Tp cht kim loi sau khi tch st c th cn c trong ch phm men dng cc mu vy nh l kim loi bt t hoc khng bt t. Nhng tp cht kim loi l th mnh kim loi khng bt t phi c kch thc mnh, ming kim loi khng qu 2mm. Hm lng kim loi mnh c kch thc<2 mm (mg/ 1kg men kh):< 20 Cc kim loi t tnh: khng qu 0,003% (ch v asen khng qu 5 mg/kg) Ch tiu vi sinh: - Tng vi sinh vt hiu kh khng qu 7500 cfu/kg men kh - Vi khun thng hn: khng c c - Nm mc: khng qu 50 cfu/kg men kh
Chng 4
THNH TU CNG NGH

http://www.ebook.edu.vn Trang 32
I. SN XUT PROTEIN VI SINH VT T DU M V KH T 1. S lc T lu ngi ta bit nhiu vi sinh vt sng da vo du m v kh t, cc b cha du, cc mt ng nha Nm 1925, con ngi pht hin thy kh nng oxy ho hidrocacbua ca mt s vi sinh vt. Nm 1940, nhiu cng trnh nghin cu s dng vi sinh vt vo vic tm kim du m v kh thin nhin gn mt t c tin hnh.Ti nm 1962, phng php sn xut protein t du m v trin vng ca n c trnh by ti hi ngh du m quc t ln th VI. Sau , mt s nc trn th gii xy dng nh my sn xut sinh khi nm men m sn phm cha ti 70% protein nhm mc ch thu protein. Sn phm thu c khi nui vi sinh vt ( ch yu l nm men ) trn du m rt giu dinh dng c th coi l 1 loi protein-vitamin m c, khng mang mi v g ca nguyn liu v khng c c t. Cc sn phm ny rt giu protein nhm B cng nh ecgostenn (tin vitamin D2).Chnh v vy chng c dng rng ri vo chn nui v mt phn c tch protein tinh khit lm thc n cho ngi. Sn phm ca Php c tn l Toprina cha ti 70% protein, ca Lin X l BVK c trn 52% protein, ca Anh l BP cha khong 40% protein. 2. Vi sinh vt s dng Hydrocacbua dng kh thng c vi khun Mycobacterium v Pseudomonas ng ho. Kh nng ny cn thy mt s vi khun khc v x khun: Streptomyces, Flavobacterium, Chromobaterium, Acremonium, Corynebacterium, Micrococus, Staphylococus, Methylococus. ng ho tt cc hydrocacbua lng l cc chng thuc ging Candida: C.tropicalis, C.maltosa, C.lypolitica, C.rubusta, C.pelliculosa, C.scotti, C.rugosa, C.olophila. Ngoi ra cn c cc ging khc cng ng ho c hydrocacbua du m nh: Torulopsis, Rhodotorula T.colliculosa, T.sake, T.dattila, T.famata, Rh.glutinis, Rh.gracilis. Mc tt trn mi trng ny cn c nm men Lodderomyces enlongisporus, vi khun Pseudomonas ovalis. Vi khun c kh nng sinh trng trn nhiu loi hydro cacbua hn nm men v nm mc. Nm men ch pht trin trn n-alkan v alken. Nm mc pht trin trn nalkan, cn trn alkan mch nhnh sinh trng km hn.Vi khun pht trin tt trn cc dy alkan mch thng, mch nhnh, trn cc hydro cacbua thm v kh thin nhin. Qu trnh thm nhp ca hydrocacbua vo t bo cho ti nay cng cha c lm sng t y . C gi thuyt cho rng, qu trnh thc hin nh c s tham gia ca lipit mng t bo. C ch phn gii hydrocacbua tuy c nhiu gi thuyt v c nghin cu nhiu nhng cng cha hon ton r rng cc i tng khc nhau. Vn chn la cc chng vi sinh vt c hot lc sinh tng hp cao dng trong sn xut rt c ngha quan trng. Trong cng nghip sn xut protein t kh t v du m phi chn cc chun sao cho p ng cc nhu cu sau: http://www.ebook.edu.vn Trang 33
C kh nng s dng tt ngun hidrocacbua dng lm nguyn liu sn xut. Sinh trng nhanh chng, cho sn lng cao trong thi gian ngn, khng i hi cc yu t sinh trng b sung. C thnh phn ha hc v iu kin nui cy n nh c hm lng protein cao, cha y cc acid amin cn thit, khng c c t v phi c ng vt ng ha tt. Hin nay ngi ta ang y mnh nghin cu sn xut sinh khi vi khun giu protein, tuy nhin mc p dng rng ri th cha c v vi khun c nhng u v nhc im sau: - Tc sinh trng nhanh. - Dng c nhiu loi c cht. - pH cn c gi 5-7, nu khng s c nguy c nhim cc vi khun ly - bnh. - Thu hi bng li tm kh. - Hm lng protein th c th rt cao (ti 80%) song hm lng bnh thng ca cc axit nucleic, c bit l ARN cng cao (ti 20%) v cn phi c loi b - 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 sinh sn c t ca chng. Phn ln cc chng nm men c sn lng cao trn c cht l hidrocacbua c phc lp t nhng mu t v bn nhng ni c m du hoc quanh cc nh my ch bin du.Vin sinh tng hp cc hp cht protein Lin X nm 1967 chn hn 500 chng nm men phn lp t cc mu trn c kh nng ng ha c hidrocacbua trong c chng Candida cho sn lng cao nht. T kt qu nghin cu phng th nghim cho thy: nui cy chng trn mi trng l n-parafin s cho hiu sut sinh khi kh t 85-100% trng lng men kh so vi trng lng parafin c dng , hm lng protein trong sinh khi th khong trn di 50%. V d: Tn nm men Hiu sut nm men Hm lng protein % kh % cht kh Candida tropicalis 94.4 58.8 Candida intermedia 87.1 51.0 Qu trnh ng ho cacbon t du m v kh t c th ra dng tng qut nh sau:
Trang 34
SX sinh khi VSV giu protein cho gia sc (1) Hydrocacbua ru bc 1 hay ru bc 2
GVHD: PGS.TS L Vn Vit Mn aldehyt axit bo.
(2) i vi n-alkan c th l:
(3)i vi cc hp cht khng no ( th d: loi 1 olefin), ngi ta cho rng qu trnh oxy ho nh vi sinh vt c th i theo con ng sau:
Cc vi sinh vt c kh nng phn gii kh metan thnh CO2 v H+ hot ng. Sau chng s dng H+ ny kh CO2 v to thnh cc hp cht hu c:
Cc qu trnh sinh ho trong t bo vi khun vi metan i th nh sau:
Trang 35
Cc axit bo c li cun vo qu trnh ng phn ho tip theo oxy ho n axetyl CoA ri i vo vng Krebs. Mt phn cc axit amin hu c c to thnh s kt hp vi NH3 cho cc aminoaxit. Nh cc dy chuyn amin m s loi amin ngy cng phong ph v cui cng di s iu khin ca AND trong t bo vi sinh vt cc axit amin ny s c t hp vi nhau hnh thnh cc phn t protein. 3. Quy trnh sn xut sinh khi vi sinh vt t hidrocacbua:
Du th
X l Men ging To mi trng Mui dinh dng Oxi
Nhn ging
Ln men Ly tm Ra nc Lm kh X l bng dung mi Ra nc sch Lm kh ng gi
Men thnh phm
Trang 36

Parafin
X l
Men ging
To mi trng nui cy
Mui dinh dng
Nhn ging
Ln men
Oxi
Ly tm
Ra nc
Lm kh
ng gi
Men thnh phm
4. Mt s vn cn lu Khi chun b mi trng dinh dng, ngi ta b sung octofosforic hoc supephosphat, NaCl, MgSO4, vo hn hp hidrocacbua vi H2O, ngun nit thng dng l nc amomiac c 20-25% NH3 v 1 lng nh (NH4)2SO4 acid ha mi trng ban u. Nc amoniac c dng vi 2 mc ch va l ngun dinh dng nit va l cht iu chnh pH trong thi gian nui cy. Nguyn liu ban u l hidrocacbua khng c cc nguyn t vi lng, v vy phi cho vo mi trng cht dinh dng cc mui sau: MnSO4, ZnSO4.7H2O, CuSO4.5H2O, KI, NaMoO.H2O, FeCl3.6H2O.
Trang 37
Cng vi n-parafin, du m th cng c dng lm nguyn liu sn xut protein t nm men, phng php dng du th cha tch parafin tng i d, nhng i hi quy trnh cng ngh phc tp hn. Nu dng parafin th khi tch nm men c th b bt khu ty ra dung mi hu c v thc t parafin c nm men s dng hon ton. n-parafin 12.5 kg Supephosphat 2.7 kg Amonsunphat 0.45 kg kg Nc NH3 25% 4.0 KCl 0.56 kg 0.28 kg MgSO4 Nc 1000 ml II. SN XUT SINH KHI VI SINH VT T NGUYN LIU CHA TINH BT V CELLULOSE. 1) S lc Cellulose l cht hu c thng gp nht trn tri t v hng nm c ti to vi mt s lng khng l. Cc loi rm r cha ti 30-40% cellulose. Cellulose cng gp nhiu trong nc thi ca cng nghip g, cng nghip dt , b ma v cc cht thi ca ngnh cng nghip thc phm. Sn xut protein n bo t c cht cellulose l mt hng c nhiu trin vng song 2 vn cn t ra l s ph thuc vo ma v v nhng kh khn trong vn chuyn. 2) Vi sinh vt s dng Cc vi sinh vt thch hp cho vic s dng cellulose l x khun a nhit, vi khun t d c ca ng vt nhai li, cc loi Cellulomonas, Alcaligenes faecalis, Tricderma reesei v nhiu nm si khc nh Myrothecium verrucaria, Humicula grisea, Sporotrichum thermophilum, Chaetomium thermophilum, nm si chi nhit Chaetomium cellulolyticum k c khi c nui hn hp vi Trichoderma reesei v nhiu loi khc. Ngoi nm men v nm si, ngi ta cn s dng rng ri vi khun sn xut protein t nguyn liu cellulose v protein vi khun c hm lng axt 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 nh nghin cu thnh cng rc r trong vic nui vi khun to protein t cy c, rm r. Srunivaan v Han (1969) phn lp c hai loi vi khun c kh nng cng sinh rt l th l Cellulonas v Alcaliges. Trong mi trng cellulosea, nu ch ring mt mnh Alcaligens th hu nh vi khun khng pht trin c, nu ch mt mnh Cellulomonas th vi khun pht trin rt km. Nhng nu nui cy cng mt lc c hai vi khun ny th sinh khi tng vt ln. http://www.ebook.edu.vn Trang 38
Cc nh bc hc M trng i hc Luisiana phn lp t b ma mt loi vi khun phn hy mch cellulosea ca ngun nguyn liu ny. Cng trnh nghin cu ny ang c ng dng c kt qu M v Cuba v c 113-136 kg b ma, ngi ta c th sn xut c 18-23 kg protein. y l mt thnh tu c nhiu ngha thc tin, n cho php s dng b ma, li ng, rm r sn xut protein mt cch trc tip m khng phi qua khu thu gii bng H2SO4. Hai nh bc hc ngi Austrayllia l Roper v Moss a ra mt phng php sn xut protein vi khun t rm, c, 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. t bit, protein do Roper v Moss thu c t rm r c cht lng tng ng vi lng trng g. Gio s Macmilan, ngi lnh o phong tro chng i Austraylia gi cng trnh ca hai nh pht minh ny l mt ting n k du trong cuc chin u vi nn i protein ca th gii . 3) Quy trnh cng ngh sn xut sinh khi vi sinh vt t cellulose Nguyn liu
Thy phn Mui dinh dng Oxi
Men ging
To mi trng nui cy Ln men
Nhn ging
Ly tm
Ra nc
Sy kh
ng gi
Men thnh phm
Trang 39
4) Mt s lu khi sn xut sinh khi vi sinh vt t cellulose Trong t nhin t gp cellulose thun khit m n thng nm dng lin kt vi cc polime khc nhau nh linhin, pectin, hemicellulose linhin l 1 polime 3 chiu c to nn nh s ngng t ca cc gc ru coniferil. Linhin bao quanh cc si cellulose bng 1 mng li 3 chiu v nh vy ngn cn s phn gii cellulose nh enzym. Ring vic lm gim ln ca ht cho php tng ng k s phn gii cellulose. Dung dch kim sunfit l cc ph phm ca cng nghip cellulose: g c nu vi canxi bisunfit. y lignin c ho tan vo dung dch dng mui canxi ca axit ligninsunforic; hemicellulosea thu phn thnh ng. Trong dung dch kim sunfit c c ng pentoza v hexoza. Phn ch yu ca dung dch kim sunfit l mui canxi ca axit ligninsunforic (khong 60% cc hp cht hu c) khng c nm men s dng. Tt c cc ng c trong dch kim sunfit v cc axit axetic c nm men Candida s dng vi hiu sut cao: sinh khi to thnh (tnh theo vt cht kh) ti 50% so vi ngun cacbon c trong mi trng. Ngi ta tnh thy rng, khong 5 tn bt cellulosea dng sn xut giy s thi ra mt lng kim cha ti 180 kg ng. Dch ny hp ph nhiu oxy cho nn khi nui cy nm men c th gim mc cung cp oxy ti 60% so vi bnh thng. Mun s dng c dung dch kim sunfit nui nm men th khi dung dch cn nng phi thi kh mnh ui S02, furfurol ra khi dch (cc cht ny km hm sinh trng ca nm men). Dng bt ng cc lm ngun nguyn liu sn xut nm men rt tt. Bt hoc tinh bt cc loi dng vo mc ch ny trc tin phi tin hnh thu phn bng axit hay enzim ca mm m hoc enzim vi sinh vt bin cc polysaccarit thnh cc dng ng m nm men ng ho c. III. SN XUT SINH KHI TO Ngoi ra vi mi trng nui cy dng b ngoi tri phi c iu kin v nh sng v din tch cu b th vic nui cy to l chim phn ln hn c. Cc ging to c ch nhiu nht vn l: Chlorella, Scenedesmus v Spirulina. Vi 3 phng php: sinh dng quang hp, ho tng hp hay l d dng, trong phng php sinh dng quang hp l c p dng rng ri hn c. To tuy trong thnh phn ca n ngoi hm lng protein cao cn c cha nhiu nguyn t vi lng v rt giu vitamin B, cng cha c p dng nhiu v trong sn xut to cn c nhng yu cu v k thut cao hn. 1. c im chung ca to. s dng to vo iu kin cho thch hp hn, ngi ta i su vo nghin cu v pht hin ra chng c mt s c im ni bt. t bit l khi s dng chng vo sn xut qui m ln cn phi c s tuyn chn cc chng ging c nhng c im thch hp cho vic nui trng cng nh cho nng sut to sinh khi cao. V khi s dng chng vo sn xut th chng c mt s c im sau: http://www.ebook.edu.vn Trang 40
Tc sinh trng nhanh Nng sut quang hp cao. C sc chng chu tt vi s dao ng ca iu kin ngoi cnh nh: nhit , chiu sng, nng mui cao, mt s bnh tt - Sinh khi c thnh phn ha hc thch hp, khng cha c t, d tiu ha, hm lng protein rt cao. - C cht dip lc, cht ng mt vai tr quan trng trong vic hp th nh sng mt tri ca to, chnh v vy to c kh nng quang hp m hu ht gii vi sinh vt khng c. - Chu k sinh sn ngn, trong iu kin ti u c th thu hoch to trong mt ngy v trong iu kin bn t nhin khong t 3-5 ngy ty theo thi tit, v vy c th thu hoch quanh nm. - To sinh trng trong mi trng nc, cho php ta d dng to c cc iu kin nui cy ti u thu c sinh khi m c. V cng ngh - T bo lun trng thi huyn ph, khng dnh kt vo thnh b nui hoc lng xung y. - D tch bng cc cch vt, lc, li tm. - Cng ngh n gin, d thc hin. 2. Gi tr dinh dng ca to. Hm lng protein ca to ni chung khong 40-55%, ring to Spirunila c cha ti 70%. Hm lng cc axit amin ca nhng protein ny gn vi quy nh ca protein tiu chun, c bit hm lng lizin trong protein ca to cao hn hn so vi protein la mch. iu ng ch l tng s cc axit amin khng thay th trong protein rt cao, c khi ln ti 42%. Gi tr dinh dng ca to cn th hin rt r cht lng v s lng cc vitamin c trong . To Cholorella c nhiu vitamin A, vitamin B, trong t bo ti c rt nhiu vitamin C. Ngoi ra, ngi ta cn thy vitamin B, K, axit nicotinic, axit pantoneic, biotin, lencophorintrong cc loi to. To Spirullina cha vitamin B12 nhiu hn hn to Chorella. Khng nhng th, Spirulina cn cha nhiu xantophyl, nhiu loi khng sinh chng vi khun v cc loi nm, v vy m ct gi to dng kh rt lu m vn khng b mc. Trc y ngi ta sn xut nhiu Chorella, nhng dn dn, do nhng yu im ni bt to Spirulina chim v tr ch yu. V km theo l hm lng vitamin ca spirulina rt a dng: Vitamin. Hm lng(mg/kg khi lng kh). Bta Carotene( tin vit. A) 1700 Cyanocobalamine (vit.B12) 1,6 D-Ca pantothenate 11,0 Acid folic 350,0 Inositol 118,0 http://www.ebook.edu.vn Trang 41
Niacine( vit.B3) 118,0 Pyridoxine( vit. B6) 3,0 Thiamine( vit.B1) 55,0 Tocopherol( vit. E) 190,0 To Spirulina sinh sng nhng iu kin v a phng khc nhau c khc nhau v hnh thi, song nhng c im v cht lng ni chung tng t nhau: - Hm lng protein cao (65-70% trng lng kh), hm lng cc axit amin khng thay th trong protein ny bng hoc cao hn tiu chun do FAO quy nh, tr xistein. - Giu cc loi vitamin nht l vitamin B12. - Cc axit nucleic chim khong 4% cht kh. - Tng s cht bo khong 6-7%. Trong cc axit bo v cc cht khng x phng ho chim t l: 83% v 17%, Nhn chung cc axit bo ca Spirulina l palmetic v C18 khng no. Khng c cc axit bo vi mch cacbon l. - Kt qu th nghim trn chut cho thy: s dng protein thun (NPU) =6163; hu hiu (PER) =2,3 (PER ca cazein l 2,5). H s tiu ho l 84%, h s ny rt cao s vi ging to khc, v d to n bo Chlorella c h s tiu ho ch trn 50% do v t bo cng bn vng i vi enzim tiu ho. V c tnh, th nghim 90 ngy trn chut khng thy cc biu hin ca c t. 3. Phng php nui cy. Trong vic pht trin nui to th cc to lc n bo hoc to c mt s t bo, nh Chlorella, Scendesmus v c ch hn c l Spirulina. Sn lng protein to trn mt hecta c th t c 10-15 tn nm, cao hn nhiu ln so vi cy nng nghip. Mun t c sn lng ny cn c nhiu iu kin thun li, trc ht l mt thi k c nh sng mt tri mnh v ko di c nng lng nh sng. Ngoi ra vic nui to i hi nhng thit b nui c bit. Thng thng ngi ta dng nhng b phng (b trn) hoc nhng mng phng un khc. Nhng thit b ny c cc h thng lt o nhm hn ch s lng ca t bo v a t bo lun lun tr li b mt c chiu sng. t c nng CO2 ti u khong 4-5% th vic cung cp CO2 l cn thit. Nhm mc ch ny c th s dng cc kh thi cng nghip. lm gim s tiu hao nng lng cn cho vic ly tm t bo t nhng dch huyn ph khng m c lm (5-10 mg/l) ngi ta ang nghin cu tch t bo bng phng php tuyn ni v phng php kt bng t bo. V c nhu cu cao v nh sng nn vic nui to hin nay ch c th c thc hin vi hiu qu kinh t nhng ni c ngun nh sng mt tri mnh v ko di. Vic nui bng ni ln men c chiu sng nhn to l qu t v qu phin phc v mt k thut. Vic nui d dng hay tp dng trong ni ln men (di nh sng yu) vi axetat hay glucoza lm ngun C c tin hnh quy m cng nghip, tuy nhin ch c hiu qu kinh t i vi phm vi s dng c bit trong cng nghip dc. http://www.ebook.edu.vn Trang 42
Trn th gii tu hon cnh ca tng nc ngi ta t chc sn xut to theo nhiu phng thc khc nhau. C th sn xut th cng cc ao h t nhin. Cch nui th cng n gin Nht Bn nh sau: trn phn, nc tiu sc vt vi b c, thm photphat, canxi, t, ri y tht kn, t 10-20 ngy. Sau gn ly nc trong, pha thm 20-30 ln nc cho long lm mi trng nui cy, c khi thm t st, lu hunh. Nui Chlorella cng nghip c thc hin u tin Hoa K. Ti y ngi ta nui to trong cc ng trong sut bng cht do hnh ch U di 21m, ng knh 1,2m vi cao ti a ca mi trng trong ng l 6,25cm. Kh CO2 c bm vo mi trng. Khi mi trng lun lun c tun hon nh mt bm khc. Nng lng mt tri c bin thnh nhit nng trong ng duy tr nhit 26oC. Phng php ny, lc thi tit tt c th t nng sut 11g/m /ngy. CHLB c, to Scenedesmus c nui trong nhng b trn ngoi tri khuy trn mi trng bng c kh. CH Sec cng nui loi to Scenedesmus vng Trabon trn din tch 900 m2. Nng sut t 15 g/ m2/ ngy. Nm 1967, Vin du m Php kt hp vi Vin du m Angieri cng bt u xy dng nhng c s sn xut to Spirulina kh ln (din tch 5000 m2) theo kiu Sosa Texcoco. 4. Mt s lu khi sn xut sinh khi to Lm c s b Lc bng trng lc v chn khng. Ph v t bo Sy kh Nghin (i vi sn phm to sy kh bng my sy tng quay hnh tr) ng gi. Nhng khu cng ngh trn y thuc lnh vc ho cng ngh c in. Tuy nhin, trong quy trnh sn xut to Spirulina c nhiu c im ring. V d, lm c s b cn phi c mt mng li c bit chn lc, lm m c mt sinh khi t 0,1g/ l n 10g/l, phc v cho mc ch ny ngi ta chn phng php trng trng, y l mt phng php kinh t hn c (tn km nng lng t hn phng php li tm). Khu ph v t bo c ngha quan trng, nh m dnh ca sn phm gim i, to thnh mt dng sinh khi c th vn chuyn d dng qua nhng bm tng thng. Cng on sy c thc hin bng my sy trng quay hnh tr. IV. MT S NGHIN CU MI 1. Nghin cu kh nng s dng nc chit ci bp lm c cht sn xut sinh khi vi sinh vt giu protein. Ngi ta tin hnh th nghim vi 4 loi nm men: Candida utilis, Pichia stipitis, Kluyveromyces marxianus, v Saccharomyces cerevisiae. Khi c nui cy trn mi trng nc ci bp th tr loi Candida utilis ra 3 loi cn li u cho lng protein tng nhiu hn so vi khi c nui trn mi trng l nc luc tht cha cng mt lng ng. Trong mi trng c lc bng membrane nm men pht trin tt hn trong mi trng c tit trng. Ch cn b sung thm ngun nit v c http://www.ebook.edu.vn Trang 43
l ammonium sulfate vo mi trng nc ci bp nng nit 0.5g/l. B sung bt bp nh ngun nit hu c lm tng sinh khi t bo khong 11%. B sung thm cht vi lng l 0.5 mM km. Tm li nc ci bp sau khi x l nhit s b vi iu kin x l thp hn tit trng th c th dng lm mi trng trong sn xut sinh khi vi sinh vt giu protein m khng cn b sung thm c cht. 2. Sn xut sinh khi vi sinh vt t b khoai ty Ngi ta tin hnh nghin cu sn xut sinh khi vi sinh vt giu protein da trn c cht l b khoai ty. Ging vi sinh vt s dng y l nm koji c ln men trn mi trng rn. Tin hnh: Trn hn hp gm 35 kg b khoai ty (75-78% m), 5.9 kg bt khoai ty (b khoai ty c lm kh, 10-15% m), 0.4 kg ammonium phosphate, calcium carbonate. To iu kin hiu kh thch hp. Sc hi nc p sut 0.8 kg/cm2 vo hn hp trong 10 pht. Sau khi lm ngui trn thm 1.2 kg bt khoai ty, 0.4 kg ure v 0.18 kg ging koji vo hn hp. Hn hp ng trong cc khay koji t trong phng ln men v c 30oC trong 2 ngy. Hm lng protein trung bnh t 15%. Hm lng lysine methionine and isoleucine gn nh trong u nnh v tht. tiu ha ca protein thc l 56.8. Cht dinh dng tng 60.9% cht kh v nng lng cung cp khong 2.72 Mcal/kg 3. Nghin cu sn xut sinh khi nm giu protein t nc thi ca nh my ru bng cch s dng vi nm. Ba ging nm s dng l Trichoderma viride WEBL0702, Aspergillus niger WEBL0901 and Aspergillus oryzae WEBL0401. S dng cc ging ny ngoi kh nng to sinh khi cao cn gip lm gim COD trong nc thi. Trong , ging T. viride c hiu sut to sinh khi cao nht v cn ngun nit t nht. C th to ra 5g/l sinh khi nm, nu s dng T. viride th khng cn b sung c cht cn nu dng A. oryzae v A. niger th phi b sung (NH4)2SO4 0.5 1.0 g/l. Nu dng T.viride th sinh khi nm thu c c hm lng protein l 19.8%, cn nu dng 2 loi cn li th hm lng protein khong 36%. T.viride cho lng sinh khi nhiu nht sau 24h cn A. oryzae v A. niger th sau 48h 4. Sn xut sinh khi vi sinh vt giu protein v enzym - Amylaza t nc thi nh my sn xut tinh bt s dng nm mc Aspergillus oryzae. S dng thit b ln men c thi kh air lift bioreactor, iu khin qu trnh ln men iu kin ti u cho vi sinh vt: pH=5.0, to = 35oC. Nng sut qu trnh:sau 1 m (thi gian nui 12h) thu c 6-10g sinh khi/1 lt nc thi, trong cha 38% protein v 55 EU/ml emzym - Amylaza
Trang 44
Sinh khi thu c em di x l loi b 95% COD, 93% BOD v 98% cc tp cht rn l lng. Sinh khi vi sinh vt ny c s dng cho nong nghip nn qu trnh x l nguyn liu khng cn thanh trng v cng khng cn thit b sung thm cht dinh dng trong qu trnh nui cy. Sn phm sinh khi protein t nm men ny cha ti hn 45% protein vi hm lng ng k cc acid amin thit yu v hy vng n s tr thnh thc n giu cht dinh dng m gia sc c th s dng c. Mt khc, ta cn loi b hn 95% COD v BOD, 75% cc hp cht nit v pht pho v cc huyn ph cn li trong nc thi ca quy trnh sn xut tinh bt. Nc thi c ci to v ph hp c th ti tiu trong nng nghip. Nu phng thc mi ny ra i v mang tnh kh thi th n s mang li li ch rt ln cho ngnh cng nghip thc phm, ngnh nng nghip v c th bn c di dng mt sn phm sinh hc gi tr gia tng. 5. Sn xut SCP bng cch ln men dch qu chanh Nui cy h nm mc Aspergillus niger v Trichoderma viride trong cc bnh ln men, nguyn liu l dch lc t qu chanh c em nghin. 6. Sn xut SCP t cht thi giu cellulose : S dng h 2 vi sinh vt Cellulomonas flavigena v Xanthomonas sp c nui cy kt hp vi nhau. Tn dng li th ca tng loi vi sinh vt to ra c sinh khi c cht lng cao t cht thi giu cellulose. 7. Phn lp, tuyn chn v nghin cu cc chng nm men c kh nng phn gii cellulose nhm ng dng trong x l b thi hoa qu lm thc n chn nui 7.1 M u Hng nm cng nghip ch bin hoa qu nc ta thi ra hng trm ngn tn b thi. Lng b thi ny hin nay vn cha c x l ring r m vn c chung vi ngun rc thi thnh ph va lng ph va gy nhim mi trng. Nu lng b thi ny c x l lm thc n gia sc hoc phn bn th s l mt ngun li ln. Mt kh khn ca vic x l b thi hoa qu l pH ca chng rt thp (3-5). Ly v d b thi da trong cng nghip sn xut ru vang, hp, b thi ny va c pH thp va cha mt lng ln cellulose. pH ny thng thng nhm vi khun v x khun khng hoc km sinh trng v pht trin nhng nhm nm men li hon ton c th. c mt s nghin cu s dng nm si trong x l b thi hoa qu lm thc n gia sc vi mc ch b sung protein n bo. Nhm nm si cng c kh nng chu pH nhng vi c tnh d to thnh bo t nu chng c dng trong ch bin thc n gia sc c th s gy ra cc bnh v ng h hp. Nm men phn gii cellulose khi pht trin trn ngun c cht b thi da c pH thp v giu cellulose s chuyn ho cellulose thnh ngun protein n bo, nu http://www.ebook.edu.vn Trang 45
c dng lm thc n gia sc s rt tt. Tuy nhin trong thc t, s lng cc loi nm men phn gii cellulose li t hn nhiu so vi nm si, vi khun, x khun c cng chc nng. Nghin cu ny tp trung vo phn lp, tuyn chn cc chng nm men c kh nng phn gii cellulose vi hy vng cc chng c la chn s c trin vng ng dng trong vic x l b thi hoa qu lm thc n gia sc. 7.2 Nguyn liu v phng php nghin cu Vi sinh vt : 343 chng nm men bao gm cc chng phn lp c t cc mu bn, g ang phn hu, t bnh men ru xung quanh H ni v b nm men phn lp t trc thuc Bo tng ging chun vi sinh vt. Cc phng php nghin cu: - Phng php xc nh kh nng phn gii mt s ngun cacbon bng phng php khuch tn trn thch - Cc phng php xc nh c tnh nui cy ca cc chng nm men - Phng php ln men xp xc nh hot tnh phn gii cellulose ca cc chng nm men 7.3 Kt qu v tho lun a/ Tuyn chn cc chng nm men phn gii cellulose: Cc chng nm men phn lp c t cc mu bn, g ang phn hu, t bnh men ru xung quanh H ni v b ging nm men thuc Bo tng ging chun vi sinh vt c phn lp t trc v s tuyn bng phng php cy vch trn mi trng Hansen vi ngun cacbon l CMC(carboxymethylcellulose) (10g/l). Hot tnh phn gii CMC ca cc chng c da vo kh nng to vng phn gii xung quanh vch sau 2 ngy nui cy khi th vi Lugol Nhng chng nm men c vng phn gii CMC tip tc c kim tra kh nng phn gii CMC. Trong th nghim ny, tin hnh ng thi nui cy cc chng nm men trn mi trng Hansen c ngun ng saccharose v trn mi trng ngun ng c thay th bng CMC chit dch enzym cellulase. Mt s cng trnh nghin cu cho thy cellulase l mt enzym cm ng, mt s ngun cacbon nh glucose, saccharose, axetat, succinat li chnh l tc nhn c ch qu trnh tng hp enzym ny. Mc tiu tuyn chn tip theo l chn ra cc chng va sinh trng trn ngun ng saccharose va c kh nng sinh enzym phn gii cellulose. iu ny gn lin vi mc tiu chn ra cc chng va c kh nng sinh trng trn b thi hoa qu vi hm lng ng cn kh cao va c kh nng phn gii cellulose cha trong cc b thi ny. Hot tnh cellulase c xc nh bng phng php c l, khuch tn trn thc Kt qu cho thy r rng c nhng chng c hot tnh cellulase cao khi nui cy trn mi trng c cha CMC nhng hot tnh ny li rt thp hoc thm ch khng c hot tnh khi nui trn mi trng c cha saccharose. iu ny ph hp vi mt s kt qu nghin cu trc y. http://www.ebook.edu.vn Trang 46
Nhng chng c hot tnh cellulase khi nui cy trn 2 loi mi trng trn v 7 chng khc khng c hot tnh cellulase khi nui trn mi trng c cha saccharosea nhng li c hot tnh cao khi nui trn mi trng c cha CMC tip tc c th kh nng phn gii mt s ngun cacbon khc: Avicel, cellulose Nhng chng nm men c hot tnh phn gii Avicel v cellulose cao nht c chn tip tc cc th nghim sau ny. b/ Nghin cu mt s c im nui cy ca cc chng nm men c la chn Cc th nghim ny phc v cho mc ch nghin cu to dng ging khi ng (starter culture) cho qu trnh x l b thi hoa qu giu cellulose t cc chng nm men c la chn nh hng ca pH nui cy ban u Cc chng nm men c nui cy trn mi trng cha CMC vi cc thang pH t 3 n 6. Nui cy lc 220 vng/pht, 300C. Sau 3 ngy nui cy, xc nh hot tnh CMC-aza ca cc chng nm men Kt qu cho thy cc chng u c kh nng tng hp enzym cellulase trong di pH mi trng nui cy l 4,0 - 6,0 nhng pH ti u nht l 5,0. nh hng ca nhit nui cy Cc chng nm men c nui cy lc trong mi trng cha CMC, pH 5,0; 220 vng/pht cc thang nhit t 200C - 450C. Sau 3 ngy xc nh hot tnh CMC-aza. men. Kt qu cho thy cc chng nm men c chn nghin cu u khng phi l cc chng nm men a nhit. Chng pht trin thch hp nht nhit 30 - 350C ty tng chng. nhit cao (40 - 450C), hot tnh ca cc chng u gim. nh hng ca thi gian nui cy Vi chng c nui cy lc 300C ,vi chng khc c nui 350C; pH 5,0. Sau mi ngy xc nh hot tnh phn gii CMC, ng thi xc nh sinh khi t bo thng qua hm lng protein Sau ngy nui cy u tin, cc chng c nui 350C t c hot tnh ln nht. Cc chng c nui 300C t c hot tnh cao nht bt u t ngy nui cy th hai. Qua ngy th ba, hot tnh ca tt c cc chng u gim dn c th do sinh trng ca nm men gim dn, ngun dinh dng cn dn hoc do cellulase b thy phn. Nh vy thi gian nui cy c nh hng r rt n kh nng tng hp enzym cellulase. Cc kt qu nghin cu trn y ln lt tr li c cc cu hi: iu kin nhit , pH no, thi gian nui cy bao lu hot tnh phn gii cellulose ca cc chng nm men cao nht ng thi t c sinh khi t bo cao nht. Cc kt qu ny rt quan trng trong vic sn xut ging khi ng, l tp hp ca cc chng nm men c tuyn chn, dng trong ch bin thc n gia sc t ph thi ca cc nh my ch bin hoa qu, c bit l nh my ch bin da. http://www.ebook.edu.vn Trang 47
c/ Kh nng phn gii cellulose t nhin Thm d kh nng phn gii cellulose trong b thi da ca cc chng nm men Cc chng nm men c nui cy trn mi trng c ngun cellulose t nhin l b thi da. Sau 3 ngy nui cy, kt qu thu c nh sau: Hot tnh phn gii ngun cellulose t nhin ca cc chng nm men
Trong thc t, rt nhiu chng vi sinh vt c hot tnh phn gii cellulose ha tan (CMC) nhng li khng c kh nng phn gii ngun cellulose t nhin hoc phn gii rt km nhng kt qu nghin cu cho thy c 5 chng nm men c tuyn chn u c kh nng phn gii ngun c cht t nhin l b da tuy hot tnh khng cao nh khi nui cy trn mi trng c cha cellulose ha tan. Mc d vy, y l mt kt qu c ngha bi n gip khng nh rng cc chng nm men c tuyn chn c th c ng dng trong x l b thi da giu cellulose lm thc n chn nui. Kh nng tch ly sinh khi v hot tnh cellulase ca cc chng nm men khi nui trn ngun c cht b da Ging khi ng l hn hp ca cc chng nm men trn c to ra di dng bt kh. S dng dng bt kh ny cy vo b thi da vi cc t l 1/100, 1/1000, 1/10.000 nhm tm ra t l ging cy thch hp sao cho va t c sinh khi t bo ln nht va c hot tnh phn gii cellulose trong b da cao nht. Kt qu s bin i sinh khi t bo theo thi gian trong cc mu b da c khi ng vi cc t l khc nhau
ng dng cc chng nm men ny trong thc t x l b thi da lm thc n chn nui cn cn tin hnh rt nhiu nghin cu su hn na quy m trong v ngoi phng th nghim.
Trang 48
4. Kt lun T 343 chng nm men phn lp c kt hp vi b ging nm men ca Bo tng ging chun vi sinh vt, 5 chng nm men c hot tnh phn gii cellulose cao nht vi k hiu 9B1, 30B1, 97m, VTCC 2.0243, 4B2 c tuyn chn. xc nh c cc iu kin nui cy thch hp nh pH, nhit , thi gian nui cy cho cc chng nm men c tuyn chn Bc u th nghim nui cc chng nm men trn ngun c cht t nhin l b da cho thy cc chng u c hot tnh cellulase v sinh trng mnh trn ngun c cht ny to ra lng sinh khi t bo ng k. Kt qu ny m ra mt hng nghin cu tip, l ng dng cc chng nm men trong ch bin b thi hoa qu giu cellulose lm thc n chn nui. 8. Nghin cu cng ngh v thit b tn thu, lm sch v ch bin sinh khi nm men bia trong qu trnh sn xut bia Nm men bia l mt ph phm trong qu trnh sn xut bia - mt ch phm sinh hc giu cht dinh dng, c gi tr kinh t cao v c s dng rng ri trong ch bin thc phm, thc n gia sc, dc phm. nhiu nc trn th gii, sinh khi nm men c tn thu mt cch trit , ch bin thnh cc ch phm c gi tr kinh t cao. Mt s nc c ngnh cng nghip pht trin cn u t cc nh my sn xut sinh khi nm men n bo vi cng sut rt ln ch bin thnh cc sn phm phc v cho nhu cu khc nhau ca nn kinh t quc dn. Hin nay ngnh sn xut bia Vit Nam ang ngy cng pht trin. Vi sn lng 800 - 900 triu lt hin nay v 2,1 t lt n nm 2020. Trung bnh thu c 12 - 23 kg men st/ 1000 lt bia (tng ng khong 1,5 - 2,8 kg men kh). Nh vy hng nm ta c th thu c hng ngn tn sinh khi nm men kh. Hin nay, ti cc nh my bia phn ln lng nm men ny c x cng nc thi gy nhim mi trng. Tn thu sinh khi nm men trong qu trnh sn xut bia, nghin cu ch bin thnh cc sn phm c gi tr kinh t cao l mt vic lm cn thit tn dng u th ca ngun li ny, khp kn dy chuyn sn xut, trnh nhim mi trng. Nguyn liu v phng php nghin cu 1. Nguyn liu - Chng nm men: Saccharomyces carlsbergensis - Ho cht: H2SO4, NaCl, ... dng tinh khit. 2. Cc phng php nghin cu Xc nh hnh dng v kch thc ca t bo nm men, xc nh sch c hc, xc nh s lng t bo nm men, xc nh t l t bo men cht, xc nh glycogen, volutin trong t bo nm men bng mt s phng php nghin cu vi sinh vt thng p dng ti cc phng th nghim Xc nh hm lng protein theo phng php Kjeldal http://www.ebook.edu.vn Trang 49
3.Kt qu v tho lun 3.1 Phng php lm sch nm men 3.1.1 nh hng ca nhit nc ra n kh nng lng kt ca t bo nm men - Nhit nc ra qu cao hay qu thp ch s OD800 cao, tc l kh nng kt lng ca t bo nm men km. - Nhit nc ra nm men thch hp l: 40C. 3.1.2 nh hng ca thi gian n s kt lng ca nm men T cc kt qu bng 2 cho thy thi gian nm men kt lng tt v c th chp nhn c l 40 pht. 3.1.3 nh hng ca t l nc/nm men n kh nng kt lng ca t bo nm men Kt qu th nghim cho thy nu t l gia nc ra v nm men qu cao trn 6/1 hay nh hn 4/1 th kh nng kt lng ca nm men km. T l nc ra thch hp nht l: 5 - 6lt nc/1lt men st, vi t l ny ch s OD thp nht. 3.1.4 nh hng ca s ln ra n ch s OD800 T kt qu cho thy: sau 5-6 ln ra sinh khi nm men c sch cao nht, OD800 thp nht. Vy s ln ra nm men thch hp l 5 - 6 ln. 3.2. Nghin cu s dng dung dch axit H2SO4 v NaCl x l nc ra, nng cao cht lng nm men * Nghin cu s dng dung dch axit H2SO4 x l nc ra nm men Nu dng dung dch axit H2SO4 1% x l th mc tp nhim ca sinh khi nm men gim t 5% xung 1,3%. T l t bo cht so vi mu i chng tng, gi tr OD800 cc mu th nghim ln hn mu i chng. Thi gian t n mc ln men ti hn ca mu i chng ngn hn cc mu th nghim. Nh vy s dng H2SO4 1% x l c tc dng gim mc tp nhim v lm tng t l t bo sng. * Nghin cu s dng dung dch NaCl x l nc ra nm men Th nghim c tin hnh vi cc nng NaCl thay i t: 0,2; 0,25; 0,3; 0,35; 0,4; 0,45; 0,5; 0,55 %; kt qu c trnh by bng 6 T cc kt qu th nghim cho thy cc mu c s dng NaCl trong x l sinh khi nm men so vi mu i chng c t l t bo sng tng ln r rt v ch s OD800 thp. Nng mui NaCl qu thp 0,2 - 0,25%, hay qu cao 0,5% t l t bo sng tng khng cao. Nng mui NaCl thch hp nht x l t bo nm men l: 0,40%. Bo qun nm men Trong qu trnh bo qun c nhiu yu t nh hng n cht lng sinh khi nm men. Trong 2 yu t quan trng nht l: nhit v thi gian bo qun. Nhit bo qun thch hp: l 2 - 40C v thi gian bo qun khng qu 6 ngy. Vi nhit v thi gian ny t l t bo nm men cht t nht, ch s AP thay i khng ng k. http://www.ebook.edu.vn Trang 50
T cc kt qu trn cho thy s lng t bo nm men cht gim mnh sau khi ra t 8% xung 3%, v tng ln 15% sau 5 ngy bo qun. Hm lng glycogen, volutin gim hn sau khi ra, v tiu hao gn ht sau 5 ngy bo qun. tp nhim sinh hc gim mnh sau khi ra v c chiu hng tng ln trong qu trnh bo qun. Kt lun T cc kt qu th nghim ngi ta rt ra mt s kt lun sau: - Xc nh c phng php lm sch sinh khi nm men vi nhit nc ra : 2 40C, thi gian kt lng: 40', t l nc ra v sinh khi nm men: 5nc/ 1nm men, s ln ra: 5 - 6 . - nng cao cht lng sinh khi nm men trong qu trnh ra c th s dng dung dch H2SO4 1% v dung dch NaCl 0,4% - bo qun sinh khi nm men: nhit bo qun thch hp 2 - 40C, thi gian bo qun khng qu 6 ngy.
Trang 51
TI LIU THAM KHO

[1] Kiu Hu nh, Gio trnh vi sinh vt hc cng nghip, Nh xut bn Khoa hc v K thut [2] L Xun Phng, Vi sinh vt hc cng nghip, Nh xut bn xy dng H Ni, 2001. [3] Lng c Phm, Nm men cng nghip, Nh xut bn Khoa hc v K thut, 2005, 331 trang. [4] M. T. ENSIKOV, Tn dng ph liu ca cng nghip thc phm, Nh xut bn Khoa hc v K thut, H Ni, 1977, 249 trang. [5] Nguyn c Lng, Cng ngh vi sinh ( Tp 2 )
[6] Nguyn c Lng, Vi sinh vt hc cng nghip, Nh xut bn i hc Quc gia Tp.HCM, 2002, 371 trang. [7] PGS.TSKH L vn Hong, Cc qu trnh v thit b cng ngh sinh hc trong cng nghip, Nh xut bn Khoa hc v K thut, H Ni, 2004, 355 trang. PGS.TS Trn Minh TmCng ngh vi sinh vt ng dng [8] Min Ho Choi and Yun Hee Park Production of yeast biomass using waste Chinese cabbage Department of Molecular Science and Technology, Graduate School, Ajou University, 5 Wonchon-dong, Suwon 442-749, South Korea, 2002 Hidenori ABE Microbial Biomass Protein Production from Potato Waste by Solid State Fermentation of Koji Fungi and This Nutritive Value in Sheep Hokkaido Animal Research Center, 2002. Zhan Ying Zhanga, Bo Jina, b, , , Zhi Hui Baia, c and Xiao Yi Wanga Production of fungal biomass protein using microfungi from winery wastewater treatment a School of Earth and Environmental Sciences, The University of Adelaide, Adelaide, SA 5005, Australia b Australian Water Quality Centre, Bolivar, SA 5095, Australia c Research Centre for Eco-Environmental Sciences, Beijing 100085, China 2006
[9]
[10]
Trang 52

Sinh Khoi Nam Men

Uploaded by

Document Information

Original Title

Copyright

Available Formats

Share this document

Share or Embed Document

Sharing Options

Did you find this document useful?

Is this content inappropriate?

Copyright:

Available Formats

Sinh Khoi Nam Men

Uploaded by

Copyright:

Available Formats

SX sinh khi VSV giu protein cho gia sc

GVHD: PGS.TS L Vn Vit Mn

SX sinh khi VSV giu protein cho gia sc

GVHD: PGS.TS L Vn Vit Mn

SX sinh khi VSV giu protein cho gia sc

GVHD: PGS.TS L Vn Vit Mn

SX sinh khi VSV giu protein cho gia sc

GVHD: PGS.TS L Vn Vit Mn

NGUYN LIU V GING VI SINH VT S DNG

SX sinh khi VSV giu protein cho gia sc

GVHD: PGS.TS L Vn Vit Mn

SX sinh khi VSV giu protein cho gia sc

GVHD: PGS.TS L Vn Vit Mn

SX sinh khi VSV giu protein cho gia sc

GVHD: PGS.TS L Vn Vit Mn

SX sinh khi VSV giu protein cho gia sc -

GVHD: PGS.TS L Vn Vit Mn

QUY TRNH CNG NGH

B Mui dinh dng

To mi trng nui cy Thanh trng

SX sinh khi VSV giu protein cho gia sc

GVHD: PGS.TS L Vn Vit Mn

SX sinh khi VSV giu protein cho gia sc -

GVHD: PGS.TS L Vn Vit Mn

SX sinh khi VSV giu protein cho gia sc

GVHD: PGS.TS L Vn Vit Mn

SX sinh khi VSV giu protein cho gia sc

GVHD: PGS.TS L Vn Vit Mn

SX sinh khi VSV giu protein cho gia sc

GVHD: PGS.TS L Vn Vit Mn

SX sinh khi VSV giu protein cho gia sc

GVHD: PGS.TS L Vn Vit Mn

SX sinh khi VSV giu protein cho gia sc -

GVHD: PGS.TS L Vn Vit Mn

SX sinh khi VSV giu protein cho gia sc

GVHD: PGS.TS L Vn Vit Mn

SX sinh khi VSV giu protein cho gia sc

GVHD: PGS.TS L Vn Vit Mn

SX sinh khi VSV giu protein cho gia sc

GVHD: PGS.TS L Vn Vit Mn

SX sinh khi VSV giu protein cho gia sc

GVHD: PGS.TS L Vn Vit Mn

SX sinh khi VSV giu protein cho gia sc

GVHD: PGS.TS L Vn Vit Mn

SX sinh khi VSV giu protein cho gia sc

GVHD: PGS.TS L Vn Vit Mn

SX sinh khi VSV giu protein cho gia sc

GVHD: PGS.TS L Vn Vit Mn

SX sinh khi VSV giu protein cho gia sc

GVHD: PGS.TS L Vn Vit Mn

Hnh 6 :Tthit b ly tm 2- Trng quay 4- B phn cha dung dch nm men

SX sinh khi VSV giu protein cho gia sc

GVHD: PGS.TS L Vn Vit Mn

SX sinh khi VSV giu protein cho gia sc

GVHD: PGS.TS L Vn Vit Mn

SX sinh khi VSV giu protein cho gia sc

GVHD: PGS.TS L Vn Vit Mn

SX sinh khi VSV giu protein cho gia sc

GVHD: PGS.TS L Vn Vit Mn

SX sinh khi VSV giu protein cho gia sc

GVHD: PGS.TS L Vn Vit Mn

SX sinh khi VSV giu protein cho gia sc

GVHD: PGS.TS L Vn Vit Mn