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Nc mm cao m ngn ngy t c nc

PGS.TS. L VN VIT MN

PH LC
Tiu Trang 1. Nguyn liu ................................................................................................................... 5 1.1 C nc........................................................................................................................ 5 1.1.1 c im............................................................................................................... 5 1.1.2 Phn phi v mi trng sng ........................................................................... 6 1.1.3 Thnh phn ha hc v tiu chun ca c nc................................................. 6 1.2 Mui......................................................................................................................... 14 2. Vi sinh vt v enzyme trong sn xut nc mm ................................................... 16 2.1 H vi sinh vt trong sn xut nc mm ............................................................ 16 2.2 Nm mc Aspergillus.oryzae ................................................................................ 17 2.3 c im protease ca nm mc ......................................................................... 20 2.4 Cc h enzyme trong nc mm ......................................................................... 22 3. S ............................................................................................................................ 23 4. Thuyt minh................................................................................................................ 24 4.1 Ra .......................................................................................................................... 24 4.2 Ln mem.................................................................................................................. 25 4.3 Lc .......................................................................................................................... 28 4.4 C c ..................................................................................................................... 29 4.5 B sung hng ....................................................................................................... 31 4.6 ng chai ................................................................................................................ 33 5. Sn phm .................................................................................................................... 34 5.1 Tiu chun nc mm thnh phm .................................................................... 34 5.2 Gi tr dinh dng ca nc mm ...................................................................... 35 5.3 Cc cht bay hi .................................................................................................... 35 5.4 Cc cht khc ......................................................................................................... 35 6. Thnh tu cng ngh ................................................................................................. 35 6.1 Ch phm enyme protease t vi sinh vt ............................................................. 36 6.2 Phn lp chng vi sinh vt to hng .................................................................. 36 7.Ti liu tham kho ...................................................................................................... 36

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Nc mm cao m ngn ngy t c nc

PGS.TS. L VN VIT MN

PH LC BNG
Bng Trang

Bng 1 : Thnh phn khi lng ca mt s loi c kinh t .......................................... 6 Bng 2 : Thnh phn ha hc ca mt s loi c bin .................................................... 7 Bng 3: Hm lng mt s acid amin tnh theo % lng protid ca da,vy,vy ca mt s loi c ............................................................................................................... 7 Bng 4 : Thnh phn cc amino acid ch yu trong c so vi tng lng protin v trong cc ng vt khc (%prtin) ........................................................................................... 7 Bng 5 : Thnh phn lipid ca cc loi c .................................................................... 11 Bng 6: Phm vi hot ng ca mt s loi enzyme ..................................................... 13 Bng 7: Ch tiu ha ha l ca cc loi mui n.......................................................... 16 Bng 8: Mt s tnh cht ca protease VSV .................................................................. 18 Bng 9: Mt s nm mc c kh nng tng hp mnh Protease (P) ........................... 19 Bng 10: c im protease nm si ............................................................................ 20 Bng 11 : Cc ch tiu cm quan ca chp .................................................................. 32 Bng 13 : ch tiu ha ha hc phn loi nc mm ................................................... 32

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Nc mm cao m ngn ngy t c nc

PGS.TS. L VN VIT MN

PH LC HNH
Hnh Trang

Hnh 1 : C nc................................................................................................................... 5 Hnh 2: My ra bng chuyn ........................................................................................ 25

Hnh 3 : Thit b ln men ............................................................................................... 27


Hnh 4: My lc tinh vi kiu ng ................................................................................... 29 Hnh 5: Thit b c c chn khng ............................................................................... 30 Hnh 6: Thit b chit rt t ng ................................................................................... 33 Hnh 7: Thit b ng np t ng ................................................................................. 34

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Nc mm cao m ngn ngy t c nc

PGS.TS. L VN VIT MN

LI M U
Nc mm l sn phn ln men t cc loi c , l sn phm truyn thng ca dn tc Vit Nam . Nc mm c sn xut rt lu , cho n nay cha c ti liu no xc nh c thi dim chnh xc v ai l ngi Vit Nam u tin a ra qui trnh sn xut sn phm ny. Ch bit rng , nc mm gn lin vi i sng hng ngy v l mt bn sc vn ha rt ring ca dn tc Vit Nam . Cng trnh nghin cu nc mm u tin l do bc s Rode vo nm 1914. Sau nh nghin cu ngi Php khc nh Matxna, Krem, Bots, Ghibec. Kt qu nghin cu v thnh phn ha hc v cng ngh sn xut nc mm c cc nh khoa hc a ra cc kt lun sau : 1. Nc mm l hn hp cc acid amin. Cc acid amin ny c to thnh do s thy phn ca protease. Cc protease ny l do vi sinh vt tng hp nn. 2. Mun c tc dng c ch vi sinhvt gy thi. T l mui thch hp l 20 25% 3. Tc dng lm ngu v to hng ngoi protease ca vi sinh vt cn do cc enzyme tiu ha c trong ni tng c. 4. Nhit c tc dng rt ln n hot ng ca cc enzyme trong qu trnh sn xut nc mm. Nhit thch hp l 36 440C 5. Trong qu trnh thy phn acid tng. Ban u ca qu trnh lm nc mm mi trng kim c tc dng rt tt.

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Nc mm cao m ngn ngy t c nc

PGS.TS. L VN VIT MN

1. NGUYN LIU 1.1 C nc : - Ngnh:Chordata - Lp: Lp ph - B: Perciformes - Gia nh: Carangidae - Chi: Decapterus

Hnh 1 : C nc 1.1.1 c im : Thn hnh thoi, dp bn. Chiu di thn bng 4,0 - 4,5 ln chiu cao thn, bng 3,3 - 3,7 ln chiu di u. Mp sau xng np mang trn, gc trn xng np mang lm. Mm di, nhn. Ming ln, chch, hm di di hn hm trn.Rng nh, nhn, hm trn v hm di u c mt hng. Ton thn, m v np mang ph vy trn, nh. ng bn hon ton, vy lng ph kn c on thng.Vy lng di, thp. Vy ngc di, mt vy t n hoc qu l hu mn.Phn lng mu xanh xm, bng mu trng. nh vy lng th hai c mu trng. C nc c mu xanh trn u v mu trng di bng . Mang c c mt im nh mu en. C nc c chn gai trn vy lng v 30-34 tia mm. Vy hu mn ca c nc ch c ba gai v 26-29 tia mm . C nc thng c mt sc mu vng chy t u n cung ui, con c nc di nht th chiu di ghi nhn c l 30cm, khng xa vi chiu di ngi ln trung bnh c c tnh l 12 inch (30,48 cm). Thnh phn chiu di: C nh bt c c chiu di dao ng t 60 n 239 mm, ch yu l nhm chiu di 120 n 189 mm. Thnh phn chiu di ca c nh bt c trong ma gi ma ng Bc v Ty Nam hi khc nhau. Thnh phn chiu di ca c Nc s nh bt c bng cc loi li khc gn nh nhau. iu ny c th c gii thch bi s di c thng ng theo ngy m ca loi ny .

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Nc mm cao m ngn ngy t c nc 1.1.2 Phn phi v mi trng sng

PGS.TS. L VN VIT MN

- C nc c bit n t i dng i Ty Dng, c th loi ny n t Nova Scotia pha bc n Rio de Janeiro pha nam, bao gm c vng bin Carib v Vnh Mexico pha ty. V pha ng, chng c bit n t Maroc pha Bc n Nam Phi pha nam, bao gm c nhng hn o Madeira, qun o Canary, Cape Verde, o Ascension v St Helena. - C nc trng quanh nm, cng ti vng bin xa b , trng ca chng tri ni trong vng nc ni trc khi nui. - C nc s thuc ging c nc (Decapterus). Ging Decapterus Vit Nam cho n nay bt gp 4 loi l : D.maruadsi, D.lajang, D.kuroides v D.russelli. Trong 4 loai ny, c Nc s (D.maruadsi) c sn lng cao nht - C Nc s phn b rng bin Vit Nam, bt gp vng bin ven b t Vnh Bc B ti Vnh Thi Lan, ch yu su 30 60m. 1.1.3 Thnh phn ha hc v tnh cht ca c Thnh phn ha hc tht ca c c th bin i trong phm vi: Protid chim t 15.8 24%, hu ht cc loi c t protid t 16-21% c rt t loi trn 22 v di 16% Lipid chim t 0.3 9.25% Nc chim t 37.7 81% Hm lng tro trong cng mt loi v gia cc loi u bin i rt t, chim t 0.9 1.68% Bng 1 : Thnh phn khi lng ca mt s loi c kinh t Tn c Tht phi-l (%) Nc s Thu 55.80 75.10 u c (%) 22.90 11.20 Xng c (%) 11.08 7.19 Vy, vy c (%) 3.82 1.92 Ni tng (%) 5.60 3.50

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Nc mm cao m ngn ngy t c nc chm Ng 59.00 19.10 7.76

PGS.TS. L VN VIT MN

0.81

Bng 2 : Thnh phn ha hc ca mt s loi c bin Tn c Nc Nc s Mi thng Trch xng Cm 1.1.3.1 Protein c cu to t cc acid amin, cc acid amin khng thay th quyt nh gi tr dinh dng ca thc phm. Bng 3: Hm lng mt s acid amin tnh theo % lng protid ca da,vy,vy ca mt s loi c Loi c C nc C thu C trch ln Glycin 4.5 7.5 Acid aspatic 8.7 9.0 9.0 Acid glutamic 13 12 13 75.14 11.25 2.10 0.90 75.9 21.76 3.15 1.70 76.8 77.5 S gam trong 100g tht c Protid 21.75 19.26 Lipid 0.85 0.85 Mui 1.32 1.8

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Nc mm cao m ngn ngy t c nc

PGS.TS. L VN VIT MN

Bng 4 : Thnh phn cc amino acid ch yu trong c so vi tng lng protin v trong cc ng vt khc (%prtin) Amino acid Lysine Trytophan Histidine Phenylalanine Leucine Isoleucine Threonine Methioninecystine Valine 6.0 7.6 ( Ha thy sn) Protein ca c l ngun giu cc acid amin c cha lu hunh v lysine. Protein ca c chia lm 3 phn: Protein cu trc (protein t c) Gm cc si myosin, actin, actomyosin v tropomyosin chim khong 65 75% tng hm lng protein trong c. Cc protein cu trc ny c chc nng co rt m nhn cc hot ng ca c. Myosin v actin l cc protein tham gia trc tip vo qu trnh co dui c. Protein cu trc c kh nng ha tan trong dung dch mui trung tnh c nng ion kh cao (> 0,5M). Protein cht c (protein tng c) Gm myoglobin, myoalbumin, globulin v cc enzyme, chim khong 25 30% hm lng protein trong c. Cc protein ny ha tan trong nc, trong dung dch mui trung tnh c nng ion thp (< 0,15%). Hu ht protein cht c b ng t khi un nng trong nc nhit trn 500C. Trang 8 5.0 8.1 C 8.8 1.0 2.0 3.9 8.4 6.0 4.6 4.0 Sa 8.1 1.6 2.6 5.3 10.2 7.2 4.4 4.3 Tht b 9.3 1.1 2.8 4.5 8.2 5.2 4.2 2.9 Trng 6.8 1.9 3.2 5.4 8.4 7.1 5.5 2.3

Nc mm cao m ngn ngy t c nc

PGS.TS. L VN VIT MN

Trong qu trnh ch bin v bo qun, myoglobin d b oxy ha thnh metmyoglobin, nh hng n mu sc ca sn phm. Protein m lin kt Bao gm cc si collagen, elastin. Hm lng collagen c tht c thp hn ng vt c v, thng khong 1 10% tng lng protein v 0,2 2,2% trng lng ca c tht. Chim khong 3% c xng v khong 10% c sn. C trong mng li ngoi bo, khng tan trong nc, dung dch kim hoc dung dch mui c nng ion cao im ng in pI ca protein c vo khong pH = 4,5 5,5. Ti gi tr ny protein c ha tan thp nht. Cu trc hnh thi ca protein c d b bin i do mi trng vt l thay i. Tnh tan ca protein trong si c thay i sau khi ng kh. Vic x l vi nng mui cao hoc x l bng nhit c th dn n s bin tnh sau cu trc protein b thay i khng hi phc c. Protein tng c c kh nng ha tan cao trong nc, lm mt gi tr dinh dng do mt lng protein ng k thot ra khi ra, p mui, tan gi,... V vy cn ch duy tr gi tr dinh dng v mi v ca sn phm. Protein m lin kt da c, bong bng c, vch c khc nhau. Cc si collagen cc m ca c to nn cu trc mng li mng vi mc phc tp khc nhau. Tuy nhin collagen ca c km bn nhit v t c cc lin kt cho hn nhng nhy cm hn collagen ng vt mu nng c xng sng. 1.1.3.2 Thnh phn trch ly cha nit phi protein Cht phi protein l thnh phn ha tan trong nc, c khi lng phn t thp v chim khong 9 18% tng hm lng protein c c xng, khong 33 38% cc loi c sn. Thnh phn chnh ca hp cht ny bao gm cc cht bay hi (amoniac, trimethylamin), trimethylamineoxide (TMAO), dimethylamineoxid (DMAO), cc acide amin t do, ure (c nhiu trong c sn), nucleotide, ... Thnh phn cht trch ly cha nit phi protein khc nhau ph thuc vo loi, kch c, ma v, phn c ly mu, ...

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Nc mm cao m ngn ngy t c nc

PGS.TS. L VN VIT MN

Cc cht trch ly cha nit phi protein rt quan trng i vi cc nh ch bin thy sn bi v chng nh hng n mi tnh cht ca thc phm nh: mu sc, mi v, trng thi cu trc, dinh dng, s an ton v s h hng sau thu hoch. Trimethylamineoxid (TMAO) TMAO l thnh phn c trng v quan trng ca nhm cht cha nit phi protein. TMAO c ch yu trong cc loi c nc mn v t c tm thy trong cc loi c nc ngt. Hm lng TMAO trong cc loi c khc nhau ty theo loi, iu kin sinh sng, kch c. C hot ng bi li nhiu, kch c ln cha nhiu TMAO hn c nh v t bi li trong nc. Hm lng TMAO cha cao nht trong cc loi c sn (c nhm), mc,........ Theo Tokunaga (1970), hm lng TMAO trong c ni nh c trch, c thu, c ng , c nc tp trung cao nht trong c tht sm, trong khi cc loi c y tht trng c hm lng TMAO cao hn nhiu trong c tht mu sng. TMAO c vai tr iu ha p sut thm thu ca c, v vy gip c chng li p sut thm thu gy ra do s chnh lch nng mui trong nc bin. Cc acid amin t do Cc acid amine t do chim khong 0,5 2% trng lng c tht, chng gp phn to nn mi v c trng ca nguyn liu. Hm lng acid amin t do cng nhiu th vi khun gy h hng pht trin cng nhanh v sinh ra mi amoniac. Cc loi c c c tht sm v thng vn ng nh c ng, c thu c hm lng histidine cao. C tht sm cha histidine cao hn tht trng. Trong thi gian bo qun histidine b vi sinh vt kh nhm carboxyl hnh thnh c t histamine. Ure Ure c ph bin trong tt c c tht c, nh ni chung c t hn 0,05% trong c tht ca c xng, cc loi c sn bin cha mt lng ln ure (1 2,5%). Trong qu trnh bo qun ure phn hy thnh NH3 v CO2 di tc dng ca enzyme urease ca vi sinh vt. Do ure ha tan trong nc v thm qua mng t bo nn n d c tch ra. Amoniac Amoniac c mi c trng (mi khai). Trong c tht ca c ti c lng nh amoniac. Trong c xng lng amoniac thp nhng khi b h hng do vi sinh vt th lng Trang 10

Nc mm cao m ngn ngy t c nc

PGS.TS. L VN VIT MN

amoniac tng nhanh. Khi s h hng tin trin, pH ca c tht chuyn sang mi trng kim do lng amoniac tng ln v to nn mi n thi ca c. Creatine L thnh phn chnh ca hp cht phi protein. C trng thi ngh ngi creatine tn ti di dng mch vng phospho v cung cp nng lng cho qu trnh co c. 1.1.3.3 Lipid C s dng cht bo nh ngun nng lng d tr duy tr s sng trong nhng thng ma ng, khi ngun thc n khan him. Hm lng lipid dao ng nhiu (0,1 30%). C c phn loi theo hm lng cht bo nh sau: + C gy (<1% cht bo) nh c tuyt, c tuyt sc en, ... + C bo va (<10% cht bo) nh c mp, c bn li nga, c nhng. + C bo (>10% cht bo) nh c hi, c trch, c thu,... Bng 5 : Thnh phn lipid ca cc loi c Loi c C nc s C tuyt C bn C sao C herring C thu S phn cht bo trong c Cht bo ca c bo thng tp trung trong m bng v y l v tr c t c ng nht khi bi li trong nc. M m cn tp trung m lin kt, nm gia cc si c. Vi c gy, hm lng cht bo trong c d tr ch yu trong gan. Hm lng cht bo (%) 0.85 0,1 0,9 0,5 9,6 1,1 3,6 0,4 30 1 - 35

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Nc mm cao m ngn ngy t c nc Dng t nhin trong cht bo

PGS.TS. L VN VIT MN

Lipid trong ca c c chia thnh 2 nhm chnh: phospholipid v tryglycerit. Phospholipid to nn cu trc mng t bo, v vy chng c gi l lipid cu trc. Triglycerit l lipid d tr nng lng c trong cc ni d tr cht bo, thng trong cc t bo m c bit c bao quanh bng mt mng phospholipid v mng li collagen mng hn. Triglycerit thng c gi l lipid d tr. Mt s loi c c cha cc este dng sp nh mt phn ca cc lipid d tr. Thnh phn cht bo trong c khc xa so vi cc loi ng vt khc. im khc nhau ch yu l chng bao gm cc acid cha bo ha cao. Hm lng acid bo cha bo ha trong c bin (88%) cao hn trong c nc ngt (70%). Cht bo trong c cha nhiu acid bo cha bo ha do rt d b oxy ha sinh ra cc sn phm cp thp nh aldehyde, ceton, skaton. Tuy nhin lipid trong c rt c li cho sc kho ngi tiu dng. Cc hp cht c li trong lipid c l cc acid bo khng no cao, c bit l: acide eicosapentaenoic (EPA 20.5), v acide docosahexaenoic (DHA 22.6). 1.1.3.4 Glucid Hm lng glucid trong c tht c rt thp, thng di 0,5% tn ti di dng nng lng d tr glycogen. Tuy nhin hm lng glycogen c chim khong 3%. C va trng lng glucid d tr rt thp. Sau khi cht, glycogen c tht chuyn thnh acid lactic, lm gim pH ca c tht, mt kh nng gi nc ca c tht. S bin i ca pH c tht sau khi cht ca c c ngha cng ngh rt ln trong c tht. 1.1.3.5 Enzym Enzym l protein, chng hot ng xc tc cho cc phn ng ha hc trong ni tng v trong c tht .Enzyme tham gia vo qu trnh trao i cht t bo, qu trnh tiu ha thc n v tham gia vo qu trnh t cng. Sau khi cht, enzyme vn cn hot ng, v th gy nn qu trnh t phn gii ca c lm nh hng n mi v, trng thi cu trc v hnh dng ca chng. Sn phm ca qu trnh phn gii do enzyme l ngun dinh dng cho vi sinh vt, lm tng tc n hng. Trong nguyn liu c nhiu enzyme khc nhau. Cc nhm enzyme chnh nh hng n cht lng nguyn liu l: - Enzyme thy phn : protease ( pepsinase, trypsinase, peptidase) , esterase, carbonhydrase( amylase. Cellulase, lactase,) Trang 12

Nc mm cao m ngn ngy t c nc

PGS.TS. L VN VIT MN

- Enzyme oxy ha kh: catalase, phenoloxydase, Bng 6: Phm vi hot ng ca mt s loi enzyme Enzyme Pepsin Trypsin Cathepsin Erepsin Ni tn ti Dch d dy Ty tng C tht, gan Dch rut pH hot ng 14 610 2.56.5 6.59.5 pH ti thch 23 89 4 8

Nhiu loi protease c tch chit t c tht c v c tc dng phn gii lm mm m c. S mm ha m c gy kh khn trong ch bin. Cc enzyme thy phn protein quan trng trong nguyn liu gm: Cathepsin, protease kim tnh, collagenase, pepsin trypsin, chimotripsin Cc enzyme thy phn lipid quan trng gm: lipase, phospholipase. Chng thng c trong cc c quan ni tng v trong c tht. Enzyme thy phn lipid rt quan trng i vi c ng lnh, cc loi c ny lipid c th b thy phn khi hot ng ca nc thp. Qu trnh bo qun lnh ng cc acid bo t do c sinh ra t phospholipid v triglycerit, c nh hng xu n cht lng ca c. Acid bo t do gy ra mi v xu, nh hng n cu trc v kh nng gi nc ca protein c tht Cc enzyme oxy ha bao gm: phenoloxydase, lipoxygenase, peroxydase. Polyphenoloxydase c bit quan trng trong tm v chng l nguyn nhn gy nn m en cho nguyn liu sau thu hoch 1.1.3.6. Cc loi vitamin v khong cht: C l ngun cung cp chnh vitamine nhm B (thiamin, riboflavin v B12), vitamine A v D c ch yu trong cc loi c bo. Vitamine A v D tch ly ch yu trong gan, Vitamine nhm B c ch yu trong c tht c. Vitamine rt nhy cm vi oxy, nh sng, nhit . Ngoi ra, trong qu trnh ch bin nh hng ln n thnh phn vitamine. V vy cn phi ch trnh tn tht vitamine trong qu trnh ch bin

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Nc mm cao m ngn ngy t c nc

PGS.TS. L VN VIT MN

Cht khong ca c phn b ch yu trong m xng, c bit trong xng sng. Canxi v phospho l 2 nguyn t chim nhiu nht trong xng c. Tht c l ngun giu st, ng, lu hunh v iod. Ngoi ra cn c niken, coban, ch, asen, km. Hm lng cht st trong tht c nhiu hn ng vt trn cn, c bin nhiu hn c nc ngt, c tht c mu sm nhiu hn tht c mu trng. Sunfua c ph bin trong tht cc loi hi sn, chim khong 1% cht kh ca tht. Sunfua trong tht c phn ln tn ti dng hp cht hu c sunfua ha tan. Hm lng sunfua nhiu hay t c nh hng ln n mu sc sn phm. Hm lng ng trong c t hn so vi ng vt thy sn khng xng sng. Hm lng iod trong tht c t hn so vi ng vt hi sn khng xng sng. C bin c hm lng iod cao hn c nc ngt. Hm lng iod ca hi sn ni chung nhiu hn gp 10 50 ln so vi ng vt trn cn. Tht c c nhiu m th hm lng iod c xu hng tng ln. 1.2 Mui: 1.2.1 Thnh phn ca mui n: Thnh phn ca mui ch yu l NaCl, ngoi ra t nhiu u c tp cht. Nhng tp cht c th chia lm hai loi: + Nhng cht khng c hot tnh ha hc nh nc v cc cht khng ha tan nh si, ct ,v.v. + Nhng cht c hot tnh ha hc nh: hp cht Clor ca Ca2+, Mg2+, Fe2+, Fe3+ v mui ca gc sulfat. Mui n tinh khit l cht kt tinh, khng mu. Khi lng ring d = 2.161, im nng chy: 8030C, im si 14390C. Mui n c mu trng, tan trong nc, glycerin, hi tan trong cn, d ht m, c v mn. Lng nc trong mui lun thay i. Nu trong khng kh c m trn 75% mui s ht nhiu nc. Nu m khng kh di 75% th mui s kh rt nhanh. Ngoi ra lng nc trong mui cn ph thuc vo to nh ca ht mui, ht mui to c hm lng nc t hn ht mui c tinh th nh. 1.2.2 Tc dng ca mui n: 1.2.2.1 Tc dng phng thi ca mui n Mui n thm thu vo nguyn liu lm cho nc thot ra, vi khun thiu nc khng th pht trin c. NaCl khi ha tan s cho ra ion Cl-, Cl- kt hp vi protide mi peptide lm cho cc men phn hy protide ca vi sinh vt khng cn kh nng ph v protide ly cht dinh dng sinh sng. Cng c th do ion Cl- c c tnh lm cho vi khun trng c. Nng mui cng ln th p sut thm thu cng mnh, v vy c th lm rch mng t bo vi khun, gy cho chng st thng. Do c mui nn oxy t ha tan trong mi trng p mui. V vy nhm vi sinh vt hiu kh khng c iu kin pht trin. Gii hn pht trin ca khun ln men nng mui 15% v ca khun ln mc l 20 30%.

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Nc mm cao m ngn ngy t c nc

PGS.TS. L VN VIT MN

1.2.2.2 nh hng ca cc thnh phn khc trong mui n: Trong mui n, ngoi NaCl cn c cc mui khc nh CaCl2, MgCl2... Cc mui ny u lm tr ngi tc thm thu ca NaCl vo c th c, v ha tan ca chng ln hn. Ngoi ra, cn lm cho mu sc, mi v v vng chc ca sn phm km i. 1.2.2.3 Tnh thm thu v khuch tn: Do tnh ht nc ca mui nn p c vi mui, nc trong c thot ra lm tan mui (gi l qu trnh khuch tn). Song song l qu trnh mui thm vo c (gi l thm thu). Sau cng nc t c khng thot ra na nhng mui trong dung dch mui c (nc bi) vn tip tc ngm vo c th c cho n khi bo ha mui. bo ha mui c thng thp hn nc bi. C cng ti th tc thm mui cng nhanh v nhiu hn c n Mui cng ln nhiu tp cht th mu cng ng, i khi c mu en. Nc trong mui n l mt thnh phn rt d thay i v mui n c c tnh hp th nhiu nc trong khng kh , lng nc trong mui cn ph thuc vo to nh ca ht mui.V cht ca mui l do trong mui c MgCl2, CaCl2, theo ti liu nghin cu ca S Ng Nghip Hi Dng Lin X, nu nng ca Ca2+, Mg2+ trong dung dch nc mui nu t 0,6% s c v cht. 1.2.3 Tiu chun mui n

Yu cu ca mui trong sn xut nuc mm phi l loi mui n, cng tinh khit cng tt, kt tinh ht nh c rn cao, mu trng ng nh ( khng vn cc, m t, v ng cht ). Ch tiu cm quan ca mui : o Mu sc: trng trong, trng nh xm, trng nh vng, trng nh hng. o Mi v: khng mi, dung dch mui 5% c v mn thun khit, khng c v l. o Dng bn ngoi kh ro, sch. Ch tiu ho l: Bng 7: Ch tiu ha ha l ca cc loi mui n Ch tiu Hm lng NaCl, tnh theo % khi lng cht kh khng nh hn m tnh theo phn trm khng ln hn Hm lng cht khng tan trong nc tnh theo % khi lng cht kh, khng ln hn Hm lng cc ion tnh theo Ion Ca2+ % khi lng cht kh Ion Mg2+ khng ln hn Ion SO42Thng hng 98 5 0.25 0.15 0.1 0.3 Hng 1 96.5 6 0.3 0.2 0.15 0.7 Hng 2 95 8 0.5 0.4 0.4 1.1

Mui l nguyn liu quan trng trong ch bin nc mm, khng c mui th khng lm nc mm c. Mui c nhiu loi khc nhau v c tc dng khc nhau trong ch bin nc mm.

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Nc mm cao m ngn ngy t c nc

PGS.TS. L VN VIT MN

Mui dng trong sn xut nc mm v p mui c l loi mui n cng tinh khit cng tt. Tt nht l loi mui kt tinh ht nh, c rn cao, mu trng ng, khng vn cc, khng m t, khng c v cht, ng. Trong mui n nu c ln cc mui khc nh :CaCl2, MgCl2, KCl s nh hng n ha tan v tnh thm thu ca mui vo c th c, c bit lm cho sn phm c v ng, cht. Trong ch bin nc mm phi s dng mui c, mui c bo qun cng lu cng tt. Trong qu trnh ch bin chp c s di chuyn ca mui vo c th c v nc t c th c ra bn ngoi to thnh nc bi. NaCl l mt mui trung tnh, nu lng mui cho vo t n bo ha sm (358g/lit 20oC) th protein trong c th c kh c ha tan v nhng protein dng ha tan trong dung dch cng c th b kt ta nh vy lng m theo l thuyt s gim nhng thc t th li tng. Nguyn nhn l do s thy phn protein di tc dng ca enzyme to sn phm: peptone, polypeptide v acid amin t do. Cc cht ny khng b kt ta bi NaCl bo ha nn mc d protein trong c th c khng ha tan vo dung dch nhng li c sn phm thy phn ca n ha tan. Trong mui n, ngoi NaCl ra cn c cc mui khc nh CaCl2, MgCl2 cc mui ny u lm tr ngi tc thm thu ca NaCl vo c th c, v ha tan ca chng ln hn. Ngoi nh hng n cn lm cho mu sc, mi v, cht lng sn phm km hn 2. VI SINH VT V H ENZYM TRONG SN XUT NC MM: 2.1 H vi sinh vt trong sn xut nc mm: 2.1.1 Ngun gc: Vi sinh vt c mt t u ca qu trnh ch bin do tn ti trong nguyn liu hoc t ngoi mi trng ngoi nhim vo. Do nng mui trong qu trnh sn xut nc mm qu cao nn c ch rt nhiu hot ng ca vi sinh vt. Tuy nhin, trong mt giai on ngn u tin, khi mui cha kp tc dng, c mt s vi sinh vt gy thi hot ng. Sau vi s hnh thnh nc bi, mn tng dn v khi nng mui t t 12% tr ln th cc vi sinh vt gy thi hu nh ngng hot ng v cc vi sinh vt khc cng b c ch cao . Nh vy, trong qu trnh ch bin nc mm, vi sinh vt tham gia vi hai chc nng: Tham gia vo qu trnh thy phn protein nhng rt yu v b c ch bi nng mui cao. Tham gia tch cc vo vic hnh thnh hng v ca nc mm, ch yu l cc vi sinh vt k kh c kh nng sinh hng. Cho enzyme protease vo rt ngn thi gian ln men : dng ch phm enzyme protease ca nm mc Aspergillus oryzae thy phn. Nh men ca nm mc m qu trnh phn gii s xy ra nhanh hn rt ngn c thi gian ln men. 2.1.2 Phn loi: - H vi sinh vt tn ti trong sn xut nc mm c chia thnh 2 loi: Nhm vi sinh vt a mui: c th pht trin c trong mi trng c nng mui trn 10% Nhm vi sinh vt khng a mui: ch pht trin c nng mui di 10% - Ngoi ra , ta cng c th chia h vinh vt thnh hai nhm hiu kh v k kh theo nhu cu s dng oxy. giai on u ca qu trnh ch bin nc mm vi sinh vt hiu kh pht trin c v c tham gia vo qu trnh thy phn c. Sau mt thi gian, mui ngm vo c gy c Trang 16

Nc mm cao m ngn ngy t c nc

PGS.TS. L VN VIT MN

ch hot ng ca nhm vi sinh vt ny lm hot ng ca chng gim dn. Cn nhm vi sinh vt k kh th hot ng c giai on sau ca qu trnh d bn thn chng cng khng a mui nhng do c s thch nghi dn vi mn ca mui nn pht huy c tc dng. 2.2 Nm mc Aspergillus oryzae: - A.oryzae l mt loi nm vi th thuc b Plectascales, lp Ascomycetes (nang khun). C th sinh trng ca n l mt h si bao gm nhng si rt mnh, chiu ngang 5-7 m, phn nhnh rt nhiu v c vch ngang, chia si thnh nhiu bao t bo (nm a bo). T nhng si nm ngang ny hnh thnh nhng si thng ng gi l cung nh bo t, c c quan sinh sn v tnh. Cung nh bo t ca Asp.oryzae thng di 1-2 mm. Pha u cung nh bo t ca Asp.oryzae phng ln gi l bng t bng ny phn chia thnh nhng t bo nh, thun di, gi l nhng t bo hnh chai. u cc t bo hnh chai phn chia thnh nhng bo t nh vo nhau, nn gi l nh bo t. - nh bo t ca Asp.oryzae c mu vng lc hay mu vng hoa cau. c im ca ging Asp.oryzae giu cc enzyme thu phn ni bo v ngoi bo (protease, amylase, pectinase). - Trong nm mc Asp.oryzae c kh nng tng hp 3 loi enzyme protease: axit, kim, trung tnh, nh: innulase, ivertase, asparaginaza, glutaminase, proteolytic. Nhng enzyme ny kh nng thu phn protein thnh axit amin iu kin nhit v pH thch hp l: nhit 37- 41 o C; pH = 6-8. Bng 8: Mt s tnh cht ca protease VSV Nhm P Xerin Ngun Enzyme Bac.subtilis Bac.pumilus Str.griseus Str.fradiae Art B22 Asp.oryzae Asp.flavus Asp.sojae E.coli P-tiol Streplococcus Clostridium histoly-ticum P kim Loi Bac.subtilus Indoaxeta-mit Ps.cloromercurbenzoat EDTA++ Kim loi ha tr Trung tnh Trang 17 -SH 7,5 7,0 hrobacter DFP+ Xerin Kim Cht km hm c imTTH pH ti thch

Nc mm cao m ngn ngy t c nc Bac.subtilus NRRL B3411 Bac. Subtilisamy Losaccarilicis Bac. Megaterium Psuedomonasa eruginoa Atreptomeces naraensis Asp. Oryzae Acremonium kiliense Clostridiumhist tolytium P Acid Asp.niger Asp.Awamori Sailoi,penicilli um Janthinellum Rhizopus chinensis Mucor pucillus Endothia parasilica Dizoaxetil Dlnorloxinmetil este COOH 1,10octafenantrolin hai

PGS.TS. L VN VIT MN

Acid

Bng 9: Mt s nm mc c kh nng tng hp mnh Protease (P) Nm mc Asp. Oryzae Asp. Satoi P-trung tnh, P-axit, P-kim. P acid, P trung tnh, P kim. P acid (aspergilopepti- dase A) Asp. Awamori Asp. Niegr Asp. Shirousami Asp. Fumigatus P acid Hai P acid P acid Hai P: Pacid, v P kim Trang 18

Nc mm cao m ngn ngy t c nc Asp. Tericola

PGS.TS. L VN VIT MN P acid c tc dng lm ng sa P trung tnh P kim Hai P: P kim v P trung tnh P acid P kim P acid c tc dng lm ng sa c dng Nht thay th cho rennin.

Asp. Candidus

Asp. Ochraceus

Asp. Sojae Asp. Flavus Pen. Janthinellum Pen. Chrysogenum Pen. Cyaneo fulvum Mucor. Pusillus Rh. Chinensis Ph. Delemar Ph. Niveus Ph. Nodous Ph. Pseudokinensis Ph. Peka Phymatorrichum omnivorum

Bng 10: c im protease nm si STT Vi sinh vt Khong pH phn hy Hemoglobin 1 2 3 Aspergillus saitoi Aspergillus Oryzae Aspergillus Oryzae (Protease trung tnh) 4 Aspergillus Oryzae (Protease kim) 5 6 7 Paccilomyces varoiti Rhizopus chemninsis Mucor pusillus 3.5 5.5 5.0 3.5 4.5 3.0 2.9 3.3 5.6 3.0 5.0 3.8 6.5 3.0 -6.0 6.0 -9.5 6.5 10.0 7.0 8.0 DIFD 3.0 4.5 3.0 4.0 5.5 7.5 Cazein 2.5 3.0 2.5 3.0 2.0 5.0 5.0 7.0 EDTA n nh pH Cht c ch NBS

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Nc mm cao m ngn ngy t c nc 2.3 c im ca protease:

PGS.TS. L VN VIT MN

- Protease l cc enzyme xc tc cho s thy phn lin kt peptid (CO-NH) trong phn t v cc cht tng t, to thnh cc peptide phn t thp, axit amin. - Protease c phn chia thnh hai loi: endopeptidase v exopeptidase. Endopeptidase(proteinase): phn gii cc lin kt peptid gia chui mch polypeptide (pepsine, tripsine, chymotripsine...). Da vo ng hc ca c ch xc tc, endopeptidase c chia thnh bn nhm: + Serin proteinase: l nhng proteinase cha nhm OH ca gc serine trong trung tm hot ng v c vai tr c bit quan trng i vi hot ng xc tc ca enzyme. Nhm ny bao gm hai nhm nh: chymotrypsin v subtilisin. Nhm chymotrypsin bao gm cc enzyme ng vt nh chymotrypsin, trypsin, elastase. Nhm subtilisin bao gm hai loi enzyme vi khun nh subtilisin Carlsberg, subtilisin BPN. Cc serine proteinase thng hot ng mnh vng kim tnh v th hin tnh c hiu c cht tng i rng. + Cysteine proteinase: Cc proteinase cha nhm SH trong trung tm hot ng. Cystein proteinase bao gm cc proteinase thc vt nh papain, bromelin, mt vi protein ng vt v proteinase k sinh trng. Cc cystein proteinase thng hot ng vng pH trung tnh, c tnh c hiu c cht rng. + Aspartic proteinase (protease acid): Hu ht cc aspartic proteinase thuc nhm pepsin. Nhm pepsin bao gm cc enzyme tiu ha nh: pepsin, chymosin, cathepsin, renin. Cc aspartic proteinase c cha nhm carboxyl trong trung tm hot ng v thng hot ng mnh pH trung tnh. + Metallo proteinase: Metallo proteinase l nhm proteinase c tm thy vi khun, nm mc cng nh cc vi sinh vt bc cao hn. Cc metallo proteinase thng hot ng vng pH trung tnh v hot gim mnh di tc dng ca EDTA. Exopeptidase (peptidase): thy phn cc lin kt u mt ca chui mch. Da vo v tr tc ng trn mch polypeptide, exopeptidase c phn chia thnh hai loi: Aminopeptidase: xc tc thy phn lin kt peptide u N t do ca chui polypeptide gii phng ra mt amino acid, mt dipeptide hoc mt tripeptide. Carboxypeptidase: xc tc thy phn lin kt peptide u C ca chui polypeptide v gii phng ra mt amino acid hoc mt dipeptide.

Ngoi ra, protease c phn loi mt cch n gin hn thnh ba nhm Protease acid : pH 2-4 Protease trung tnh: pH 7-8 Protease kim: pH 9 -11 Protease t vi sinh vt Protease vi sinh vt l mt h thng rt phc tp bao gm nhiu enzyme rt ging nhau v cu trc, khi lng v hnh dng phn t nn rt kh tch ra di dng tinh th ng nht. Cng do l phc h gm nhiu enzyme khc nhau nn Protease vi sinh vt thng c tnh c hiu rng ri cho sn phm thy phn trit v a dng. Nhiu vi sinh vt c kh nng tng hp mnh protease. Cc cng trnh nghin cu protease vi sinh vt ngy cng nhiu. Cc kt qu nghin cu cho thy ngay c cc protease ca cng mt loi vi sinh vt cng c th khc nhau v Trang 20

Nc mm cao m ngn ngy t c nc

PGS.TS. L VN VIT MN

nhiu tnh cht. Cn c vo c cht phn ng, PH hot ng thch hp, cc nh khoa hc phn loi cc protease vi sinh vt thnh 4 nhm nh sau: Protease-xerin; Protease-thiol; Protease-kim loi; Protease-acid. Mt s tc gi chia protease ra 3 nhm, da vo PH hot ng ca chng bao gm: Protease-acid, Protease trung tnh, Protease kim. Trong bn protease k trn, cc Protease-xerin v Protease-thiol c kh nng phn gii lin kt este v lin kt amid ca cc dn xut acid ca amino acid. Ngc li protease kim loi v protease acid thng khng c hat tnh esterase v amidase i vi ca cc dn xut ca amino acid. Trong TTH ca Protease vi sinh vt ngoi gc acid amin c trng cho tng nhm cn c mt s gc acid amin khc. Cc kt qu nghin cu chung v TTH ca mt s Protease vi sinh vt cho php rt ra mt s nhn xt chung nh sau: TTH ca Protease ln v bao gm mt s gc acid amin v trong mt s trng hp cn c c cofact kim loi.

i vi cc Protease khng cha cysteine, TTH ca chng c tnh mm do hn v cu trc khng gian ca chng khng c gi vng bi cc cu disulphide 2.4 Cc h enzym trong sn xut nc mm Gm 3 h enzym ln 2.4.1 H enzym Metalo-protease (Aminodipeptidase) H enzym ny tn ti trong ni tng ca c v chu c nng mui cao nn ngay t u n hot ng mnh, gim dn t thng th 3 tr v sau. Loi enzym ny c hot tnh kh mnh, c kh nng thy phn rng ri i vi cc loi peptid. y l nhm thy phn enzym trung tnh, pH ti thch t 5-7, pI = 4-5, n n nh vi ion Mg2+, Ca2+ v mt hot tnh vi Zn2+, Ni2+, Pb2+, Hg2+.. 2.4.2 H enzym serin-protease in hnh l enzym tripsin, tn ti nhiu trong ni tng ca c. giai on u ca qu trnh sn xut nc mm hot ng ca n yu n thng th 2 v pht trin dn t gi tr cc i thng t 3 ri gim dn n khi chp chn (protein phn gii gn nh hon ton khng cn dng peptone). H enzym ny lun b c ch bi chui acid amin trong cu trc ca enzym. tho g chui ny phi nh n hot ng ca men cathepsin B nhng men cathepsin B d b c ch bi nng mui cao. V vy men cathepsin B hot ng c ngi ta thc hin phng Trang 21

Nc mm cao m ngn ngy t c nc

PGS.TS. L VN VIT MN

php cho mui nhiu ln. Enzym serin-protease hot ng mnh pH t 5-10, mnh nht pH=9. 2.4.2.3 H enzym acid-protease C trong tht v ni tng c, in hnh l enzym cathepsin D. H enzym ny d b c ch bi nng mui khong 15% nn thng n ch tn ti mt thi gian ngn u thi k ca qu trnh thy phn. Loi men ny ng vai tr th yu trong qu trnh sn xut nc mm. 3. S QUI TRNH CNG NGH

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Nc mm cao m ngn ngy t c nc


c

PGS.TS. L VN VIT MN

Ra

tp cht

Mui Protease t A.oryaze Nc


Ln men

lc

c c chp c hng tt

b sung hng

Chai v trng

ng chai

Nc mm ng chai

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Nc mm cao m ngn ngy t c nc

PGS.TS. L VN VIT MN

4. THUYT MINH QUI TRNH CNG NGH: 4.1 Ra: 4.1.1 Mc ch: chun b Qu trnh ra s lm sch v loi tr cc tp cht c hc nh t, ct, bi v lm gim lng VSV. 4.1.2 Cc bin i ca nguyn liu: Sinh hc: gim lng VSV Vt l: Loi tp cht nh ct t bi bm ln b mt con c Cc bin i khc khng ng k.

4.1.3 Thit b: s dng thit b ngm ra xi ti , dng bng ti. Cu to : - My c cu to gm mt bng ti bng thp khng r v thng cha nc ra c th tch tng i ln. Bng ti c chia lm 3 phn, phn nm ngang ngp trong nc, phn nghing c cc ng phun nc mnh v mt phn nm ngang pha cao. Bn di bng ti phn ngp trong nc c b tr cc ng thi kh nhn khng kh t mt qut t bn ngoi. Hot ng : - Trong giai on ngm, nguyn liu trn phn bng nm ngang ngp trong nc, cc cn bn bm trn ngoi b mt nguyn liu b bong ra. Bng ti di chuyn s mang nguyn liu i dn v pha phn bng nghing. Hiu qu ca qu trnh ngm c tng cng nh thi kh lm xo trn nc v nguyn liu trn mt bng, lm tng din tch tip xc ca nguyn liu v nc nn thi gian ngm c rt ngn. Khi nguyn liu di chuyn n phn nghing ca bng, cc vi phun nc vi p sut cao n 2-3 at s ra sch cn bn. cui qu trnh ra, nguyn liu di chuyn n phn nm ngang pha trn c lm ro nc. - Ty thuc loi nguyn liu v mc bn, c th iu chnh tc di chuyn ca bng chuyn cho ph hp. Nu nguyn liu qu bn, cho bng chuyn i chm li, lm tng thi gian ra. Ngc li, nu cn bn bm trn ngoi nguyn liu t, c th cho bng chuyn i nhanh hn nhm tng nng sut qu trnh. Nc sch t vi phun vo thng ngm s b sung nc cho h thng, cn cn bn c tho ra lin tc qua van x v nc tha theo mng chy trn ra ngoi.

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Nc mm cao m ngn ngy t c nc

PGS.TS. L VN VIT MN

Hnh 2: My ra bng chuyn 4.2. Ln men: 4.2.1. Mc ch cng ngh: ch bin,bo qun. - Ch bin: Thy phn protein thnh cc acid amin nh h enzyme trong bn thn nguyn liu c v ch phm enzyme protease b sung rt ngn thi gian ln men. - Bo qun:di tc ng cu mui lm c ch VSV gy thi , ko di thi gian bo qun sn phm 4.2.2. Cc bin i ca nguyn liu: Vt l: - Nng mui trong c tng ln. Do s dch chuyn ca cc tiu phn mui vo trong c v s khuch tn ca nc vo trong tht c. - Trong qu trnh thy phn mt s tnh cht vt l ca mu thy phn s b thay i: nhit tng, th tch khi chp thay i, t trng khi chp cng thay i, thay i v mu sc : - S bin i v mu sc l do 2 nguyn nhn: Ha l: - Qu trnh thy phn cng lm nn mt s bin i v pha, nh t tht c rn chuyn thnh dng huyn ph. Khi qu trnh ln men kt thc ta thu c khi chp l dng huyn ph. Trang 25 Do sc t trong nguyn liu to nn. Nc mm c nc th c mu cnh gin (vng m). Do phn ng ca cc acid amin gy nn nh phn ng to melanoidin, phn ng oxy hato mu sm cho sn phm.

Nc mm cao m ngn ngy t c nc Ha hc

PGS.TS. L VN VIT MN

- Trong qu trnh ch bin nc mm hm lng protein trong c gim xung theo thi gian cn hm lng acid amin, peptide v cc m bay hi khc trong nc chp u tng ln r rt. - Trong qu trnh ch bin nc mm c hin tng sinh tng hp mt s acid amin ca vi sinh vt trong chp. Mt s acid amin c chiu hng tng ln nhng cng c mt s acid amin c chiu hng gim i. Nguyn nhn c th do chng tham gia vo cc phn ng melanoidin, polyphenolamin, oxy ha cn khi b phn hy thnh cc sn vt thi ra nn hm lng gim i kh nhiu. Ha sinh: - Do tc dng ca h enzyme protease tc ng vo thy phn protein tht c qua cc dng trung gian nh peptone, polypeptide, peptide v cui cng l acid amin. Protein Peptone Polypeptid Peptid

acid amin

- Vi sinh vt gy thi th c tc dng lm ra nt tht c c khi ngay giai on u hay trong qu trnh ch bin, hoc nu khng khng ch kp thi sau khi to thnh nc mm cng d b thi do vi sinh vt gy nn. - Qu trnh phn gii tht c chuyn t protein n acid amin l mt qu trnh rt phc tp vi s tham gia ca rt nhiu enzyme. Mi enzyme c tnh c hiu ring i vi c cht hay v tr tc dng. V d nh peptidase ch tc dng ln mi lin kt peptide thy phn mi lin kt ny.

-CO-NHSinh hc:

nc , peptidase

-COOH + -NH2 +

- Bin i quan trng l s trao i cht v sinh trng ca vi sinh vt - Do nng mui cao vi khun gy thi hu nh ngng hot ng v cc vi khun khc cng b c ch cao . - Mt khc mt s vi sinh vt c kh nng tng hp acid amin nn c mt s acid amin tng ln trong qu trnh ch bin: acid glutamic, alanin, serin, prolin, cystin, cystein 4.2.3. Thit b: ni hai v c o iu nhit c cnh khuy.

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Nc mm cao m ngn ngy t c nc

PGS.TS. L VN VIT MN

Hnh 3 : Thit b ln men Cu to:


- Ni hai v c o nhit nng nhit khi chp lun t 450C 2 0 C v gi nhit trong qu trnh thy phn . Dng ni dung tch 500 1000l. - Thit b hnh tr , np v y elipse c gn vo thn nh 2 mi ghp bch . Mt trong thit b c lp cao su chu acid, chu p lc cao. Trn thn thit b c ng dn hi trc tip vo dch thy phn . Trn np thit b c ca nhp nhp liu v y thit b c ca tho liu . Bn thnh thit b cn c ca d dng v sinh v sa cha thit b .

4.2.4 Thng s cng ngh : - Ch phm enzyme : 3 5 % so vi c


- Nc : 4 6% so vi c - Thi gian : 10 20 ngy - Nhit : Nhit ti thch cho enzyme hot ng l t 30 - 470C. Nhit cng cao qu trnh phn gii tht c cng nhanh. Nhit 700C tr ln hu ht cc h enzyme trong c mt hot tnh. Trong qu trnh thy phn tin hnh nng nhit ln t t ln 50-550C, nhit ny c tc dng c ch s hot ng ca vi khun gy thi v lng mui cho vo lc u rt hn ch. Ch nhit khng qu 600C . Sau khuy o, khi lng mui cho vo nhiu hn th tin hnh h nhit v gi nhit 450C. - PH: Enzyme rt nhy cm vi pH ca mi trng, mi enzyme ch hot ng mnh pH xc nh. Trong qu trnh phn gii c rt nhiu enzyme tham gia cho nn ta phi xem loi enzyme no c nhiu nht v ng vai tr ch yu trong qu trnh thy phn to mi trng thch hp cho n hot ng. Enzyme protease trong bn thn nguyn liu: Pepsin v tripsin l 2 enzyme ch yu. Pepsin hot ng tt mi trng acid c pH = 1.5 - 2.2 . Mi trng mui mn cng nh mi trng c mui MgSO4, NH4Cl mi trng kim c ch s hot ng ca enzyme ny.

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Nc mm cao m ngn ngy t c nc

PGS.TS. L VN VIT MN

Tripsin gi vai tr quan trng nht hot ng mnh mi trng pH = 8-9, nng mui NaCl cao tripsin cng hot ng c. Enzyme tripsin pht trin tt khi gp mi trng c cht keo nh albumin, peptone, gelatin nhit thch hp. Do trong qu trnh ch bin cn phi to iu kin tt cho enzyme ny pht trin mnh giai on u. Trong iu kin t nhin th tripsin hot ng mnh giai on gia ca qu trnh ch bin. Chp ch bin bng phng php t nhin c mi trng pH = 5.5-6.5. Khi dng ha cht acid hay baz iu chnh pH mi trng th thy cht lng nc mm khng bng so vi ch bin chp t nhin. Qua th nghim pht hin c nu iu chnh pH=1.5-2 thy mu sc ca chp p nhng tc thy phn chm v mi v so vi chp ch bin bng phng php t nhin th km hn. Cn nu gi pH=7.5-8.5 (thch hp cho enzyme tripsin hot ng) thy tc thy phn nhanh hn nhng m thi nhiu hn v mu sc khng bng so vi chp ch bin bng phng php t nhin. mi trng t nhin pH = 5.5-6.5 tuy khng u tin pht trin loi enzyme no, nhng c hai loi enzyme chnh l pepsin v tripsin u pht trin c. Mt khc li c tc dng c ch mt phn s hot ng ca vi sinh vt. Enzyme protease trong nm mc: hot ng tt nht pH= 7 Vy pH t nhin ca mi trng l thch hp cho cc enzyme trong c v vi sinh vt hot ng. - Mui : Lng mui cho vo c chia lm nhiu ln. Lc u cho mt lng mui hn ch khng nh hng n hot ng ca enzyme v c ch vi khun gy thi. Sau lng mui cho c tng dn ln khng ch lng vi khun gy thi v h enzyme vn hot ng tt . Khi cho lng mui cn thit th h nhit xung 40- 45 0C. Ch cho mui vo nh sau : + Sau 12-18 gi cho 7% mui so vi c . + Sau 30- 48 gi cho 3% mui so vi c + Sau 3 ngy na cho thm 15 - 17 % lng mui tng cng l 25 27 % so vi c . Hm lng m (m ton phn , m formon v m amin) bt u n nh t ngy th 7 . Sau 10 15 ngy ko rt c nc mm. 4.3. Lc : 4.3.1. Mc ch: khai thc cu t ha tan trong chp chn. 4.3.2. Bin i ca nguyn liu : Vt l: sau khi lc ta thu c dch lc v b lc. Mt s ch tiu vt l ca dch lc s thay i so vi huyn ph ban u nh : t trng , trong. Ha hc: qu trnh lc khng gy ra nhng bin i ha hc trong huyn ph . Tuy nhin trong iu kin tip xc vi khng kh th cc cu t trong nguyn liu c th b bin i hoc tng tc vi nhau to ra mt s hp cht ha hc mi nhng khng ng k. Ha l: phn ring hai pha mt l rn (b chp) v lng ( dch thy phn). Mt s cu t d bay hi nh cht mi trong dch lc c th b tn tht

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Nc mm cao m ngn ngy t c nc

PGS.TS. L VN VIT MN

Sinh hc : qu trnh lc khng gy ra bin i sinh hc. Nhng nu thi gian lc huyn ph ko di th h vi sinh vt c sn trong huyn ph hoc trong mi trng sn xut b nhim vo huyn ph s pht trin. V vy cn phi lc nhanh v trong iu kin kn. Ha sinh : khng xy ra. 4.3.3. Thit b: Cu to : My lc kiu ng tinh vi loi PG l do nhiu l cng cp kt cu thnh thit b lc, n v lc c th t n micrmt, cc vt liu c cht dnh t m khong 0.5m-100m ch cn qua mt ln lc, t c hiu qu lc cht lng. c im ca my lc tinh vi kiu ng: - tinh cht lc cao, phm vi rng, c th tha mn c yu cu cc tinh cht lc khc nhau. - Chu c acid, kim, mui v cc dung mi hu c, khng c, khng mi, khng trc di vt. - Kt cu theo hnh lp ng, chim din tch t my nh. - D dng thao tc, lng x l ln, gi thnh u t thp. - Dng my nn kh hoc hi nc ti sinh bi cn ra tr li, cng c th ti sinh li kha hc, lm cho tui th s dng ca tm lc di hn.

My lc kiu ng tinh vi PG

ng polyetylen (PE) l micrmt

Hnh 4: My lc tinh vi kiu ng 4.3.4 Thng s cng ngh: - Kch thc l lc: 40-80 m - p sut lc P = 3at - Thi gian lc t = 40 ph/m

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Nc mm cao m ngn ngy t c nc 4.4 C c chn khng: 4.4.1. Mc ch: khai thc, bo qun

PGS.TS. L VN VIT MN

- Khai thc: qu trnh c c lm tch bt nc ra khi nc mm do lm tng m ca nc mm thnh phm. - Bo qun: qu trnh c c lm bay hi nc t nc mm, do lm gim hm lng nc v lm tng hm lng cht kh trong sn phm do hot ca nc trong nc mm thnh phm cng gim i, y l yu t gy c ch h vi sinh vt trong nc mm, gip ko di thi gian bo qun. 4.4.2. Cc bin i ca nguyn liu: Vt l: hm lng cht kh tng, nht, khi lng ring tng tng khi lng v th tch nc mm gim, hot ca nc trong nc mm gim. Ha hc: c c chn khng nhit thp nn hn ch c s bin i, mt mt acid amin do cc phn ng caramen, maillard. Ha l: s chuyn pha ca nc, nc tn ti trng thi lng trong nc mm trc c c s chuyn sang trng thi hi v thot ra khi nc mm. Sinh hc: mt s vi sinh vt b c ch. 4.4.3. Thit b: thit bi c c chn khng. - Cu to: Thit b bao gm cc phn: Ni c c, b ngng t s b dng tch kh v nc. B ngng th hai dng lm mt v thu hi thnh phm. Phn tip xc nguyn liu c ch to bng thp khng g, my hon ton ph hp cho vic v sinh, my khng b g, n mn. - Nguyn tc hot ng: nguyn liu c a vo khoang un nng. Hi c cp v khoang un nng v xy ra hin tng bc hi. Phn hi nc s chuyn sang khoang ngng, sau c lm mt v ngng t. Qu trnh bc hi s lm cho nguyn liu dn dn c c c.

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Nc mm cao m ngn ngy t c nc

PGS.TS. L VN VIT MN

Hnh 5: Thit b c c chn khng

4.4.4 thng s cng ngh - Dung tch: ty chn: 200-500-700-1000-2000 (lt) - Nng sut bay hi tng ng: 60-100-130-150-350 (kg/h) -p lc hi: <0.1MPa - chn khng: 0.06 MPa 4.5 B sung hng: Nc mm sn xut theo phng php ngn ngy cho hng v khng tt, khc phc iu ny pha vi nc mm c hng v tt. 4.5.1. Mc ch: hon thin. to hng v c trng cho nc mm thnh phm. 4.5.2. Phng php thc hin: s dng chp chn c lm t c nc, c hng thm, rt nc mm ct. Ly nc mm ct( ngn ngy) em trn trong b. Khi thy nc l chy ra t trong th bt u ly nc th nht, ri tip tc ly nc chan khc ly nc th 2 v th 3. Lu rng nc chan ny c i qua nhiu b chp chn lin hon nhm mc ch nng cao thm m cho sn phm, ng thi khi rt mi b phi lu li mt lng nc, khng nn rt cn. 4.5.3. Cc bin i trn nguyn liu: Ha hc: nng cu t hng trong nc mm tng ln. Ha l: trch ly mt s thnh phn ha tan trong b chp to hng vo trong nc mm, s khuch tn ca cc phn t cht tan t trong b chp tt vo nc mm Cc bin i khc khng ng k Trang 31

Nc mm cao m ngn ngy t c nc 4.5.4. Thit b v thng s cng ngh

PGS.TS. L VN VIT MN

- S ko rt c thc hin trong thng chp c hng tt c ch bin t c nc. C nc c ln men thnh chp chn c hng v tt , sau rt nc mm thnh phm ra th k t y thng chp ny c th c s dng gy hng to v cho nc mm cha c hng v tt. - Thi gian nc mm ngn ngy c ngm trn trong b chp : 2 3 ngy Ch tiu nh gi chp chn Ch tiu cm quan : Mu sc : mu nu ti , nu xm , hoc xm . Ring nc ct c mu vng rm n cnh gin. Mi : mi thm c trng , khng c mi chua , mi l . Trng thi : + i vi chp gi nn : c cn nguyn con , nu s ra tht c tch khi xng , nu khuy tht s nt vn. + i vi chp cnh khuy: c nt nhuyn , ci chp sng , khi nh khuy khng c hin tng si bt . Bng 11 : Cc ch tiu cm quan ca chp Ch tiu M i Chp loi A C mi thm khng c mi tanh Ngt m, du, khng kh c Nc cn mu vng rm hoc lu . B c mu Chp loi B C mi thm nc mm khng c mi tanh, chua, ng Ngt m, khng c v mn cht, chua hoc v khc Nc c mu cnh gin hoc mn chn. B hoc mu trng ng Lng mt dn, khng ln vn bt c Chp loi C C mi nc mm, cn c mi tanh , chua. v hi hi ngt, hoc cht Nc c mu nc ch. B xm hoc en B ln vi nc sn st , khng trng snh

Mu sc

Tnh trng chp Lng mt dn, khng ln vn bt c

Ch tiu ha hc: T l nit amin trn m ton phn ca nc ct >45% Bng 12 : Ch tiu thnh phn ha hc ca chp Thnh phn Chp loi A Chp loi B 14 -22 Chp loi C 13 -17 Trang 32

Nit ton phn trong 20-30

Nc mm cao m ngn ngy t c nc nc ct (g/l) Nit ton phn trong 20 -22 chp (g/Kg) T l Nit 13 -19
27%

PGS.TS. L VN VIT MN

9 -13
30%

amoniac 25% ton phn


>40%

T l Nit

a min ton phn

40%

<35%

Mt vi phng php khc : Phng php phi nng hoc sy 500C , nu nc mm khng c bin i g so vi mu i chng l nc mm chn . Nu t mu vng rm hoc cnh gin chuyn sang vng nht mt hng v c trng vn c th chp cha chn Phng php lng ng: ngi ta s dng phng php c hc lc mnh mu nc mm , lc 30 40 ln sau yn 20 pht . Nu mu nc mm khng c bin i g so vi mu i chng l chp chn

4.6. ng chai: trong iu kin v trng. Sn phm s c chit rt theo chai t ng bng my. Bao b s dng l chai nha, c v trng trc khi i vo quy trnh ng chai. S dng np nha kn. 4.6.1. Mc ch: hon thin. 4.6.2. Bin i: khng xy ra bin i ng k. Yu cu chit rt v ng chai trong iu kin kn hn ch s xm nhp ca vi sinh vt v oxy ca khng kh vo sn phm. 4.6.3. Thit b thng s cng ngh: H thng chit rt t ng. c im ca my: Mm xoay cp chai t ng. Bng ti bng inox hoc bng nha Nguyn tc chit: chn khng, bm hi lu. My chit rt c cho nhiu loi chai c hnh dng v kch c khc nhau. D v sinh.

Thng s cng ngh: p lc kh nn: 5kf. ng knh sn phm: (16-30) / (20-60) mm. Dung tch chai : (200-10000) ml. Trang 33

Nc mm cao m ngn ngy t c nc Tc bng ti : (10-20) m/ph

PGS.TS. L VN VIT MN

Hnh 6: Thit b chit rt t ng

Hnh 7: Thit b ng np t ng. 5. SN PHM: 5.1 Tiu chun ca nc mm thnh phm Ch tiu cm quan - Mu sc: nc mm c mu vng rm n cnh gin. - Mi thm c trng, khng c mi chua mi l. Ch tiu nh gi: (TCVN 5107:2003) Bng 13 : ch tiu ha ha hc phn loi nc mm

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Nc mm cao m ngn ngy t c nc Loi m c bit > 30oN Thng hng >25oN

PGS.TS. L VN VIT MN Hng 1 >15oN Hng 2 >10oN

Ch tiu vi sinh vt: - Vi khun hiu kh: <10 (khun lc/ml) - Staphylococcus aureus, E.coli: khng c c - Nm men, nm mc: <10 (khun lc/ml) - Coliforms: <100 (khun lc/ml) - Clostridium perfgingens: <10 (khun lc/ml) 5.2. Gi tr dinh dng ca nc mm Cc cht m Chim ch yu v quyt nh gi tr dinh dng ca nc mm. Gm 3 loi m: - m tng s: l tng lng nit c trong nc mm (g/l), quyt nh phn hng ca nc mm. - m amin: l tng lng m nm di dng acid amin (g/l), quyt nh gi tr dinh dng ca nc mm - m amon: cng nhiu nc mm cng km cht lng Ngoi ra trong nc mm cn cha y cc acid amin, c bit l cc acid amin khng thay th: valin, leucin, methionin, isoleucin, phenylalanin, alanin.v.v .. Cc thnh phn khc c kch thc ln nh tripeptid, peptol, dipeptid. Chnh nhng thnh phn trung gian ny lm cho nc mm d b h hng do hot ng ca vi sinh vt. Thnh phn dinh dng ca nc mm ph thuc vo nguyn liu em i ch bin. 5.3. Cc cht bay hi Rt phc tp v quyt nh hng v ca nc mm. Hm lng cc cht bay hi trong nc mm mg/100g nc mm - Cc cht cacbonyl bay hi: 407-512 (formaldehyde) - Cc acid bay hi: 404-533 (propionic) - Cc amin bay hi: 9,5-11,3 (izopropylamin) - Cc cht trung tnh bay hi: 5,1-13,2 (acetaldehyde) Mi trong nc mm c hnh thnh ch yu do hot ng ca vi sinh vt ym kh trong qu trnh sn xut nc mm to ra. 5.4. Cc cht khc - Cc cht v c: NaCl chim 250-280g/l v mt s cc cht khong nh: S, Ca, Mg, P, I, Br. - Vitamin: B1, B12, B2, PP. Trang 35

Nc mm cao m ngn ngy t c nc

PGS.TS. L VN VIT MN

6.

THNH TU CNG NGH 6.1 Ch phm enzyme protease t vi sinh vt:

ti ng dng cng ngh sinh hc ln men nc mm c trch do Ph Gio sTin s Nguyn Vn B, Ph Gim c Vin Nghin cu v Pht trin cng ngh sinh hc, Trng i hc Cn Th ch nhim, c nghim thu trong thng 9-2005 l mt bc t ph ln trong vn b sung ch phm enzyme protease vo chp nhm rt ngn thi gian thy phn. Nghin cu b sung Enzyme nm si Aspergillus oryzae v vi khun Bacillus subtilis lm cht xc tc y nhanh tc thy phn protein tht c v kt hp vi tin hnh p mui nhiu ln tng hot ng ca enzyme protease, rt ngn qu trnh thy phn. Kt qu nghin cu cho thy lng m tng s tng 14,8% (25,82g/l), m amin tng 40,1% (13,58g/l) trong vng 6 thng. Cng cng thi gian ny, mu i chng c sn xut theo phng php dn gian, m tng s ch t 22,5g/l v m amin l 8,13g/l 6.2 Phn lp chng vi sinh vt to hng: T vng ni ln trn trong qu trnh ch bin nc mm, cc nh nghin cu phn lp c hai chng vi sinh vt to hng nh sau: - Chng 1 : Staphylococcus intermediu, thuc ging Staphylococcus,h Micrococcaceae - Chng 2 : Vibrio costicola, thuc ging Vibrio, h Vibrionaceae 7. TI LIU THAM KHO 1. Tin s Nguyn Trng Cn, Cng ngh ch bin thc phm thy sn tp 1, NXB Nng Nghip, H Ni, 1990. 2. Tin s Lng Hu ng, K thut sn xut nc mm, NXB Khoa hc v K thut, H Ni, 1975. 3. PGS.TS L Vn Vit Mn, Cng ngh ch bin thc phm, NXB i Hc Quc Gia TPHCM, Thnh Ph H Ch Minh, 2009. 4. PGS.TS Nguyn c Lng, Cng ngh vi sinh,tp 2, c s vi sinh vt cng nghip, NXB i Hc Quc Gia TP HCM 5. PGS.TS Nguyn c Lng, cng ngh vi sinh,tp3, thc phm ln men truyn thng,NXB i Hc Quc Gia TP HCM 6. www.maymocthietbithanhphat.com.vn 7. www.mayruaca.com 8. www.fermentedmachine.com 9. www.industry.vn

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