B GIO DC V O TO TRNG H K THUT CNG NGH TP.

HCM KHOA MI TRNG V CNG NGH SINH HC

BI BO CO

THC HNH CNG NGH LN MEN

GVHD: T.S. NGUYN HOI HNG
NHM SVTH:

1. Nguyn Th Nh Yn 2. Nguyn Th ThanhTrang

0851110308 0851110262 0851110263 0851110303 0851110168 0851110213

0851110304

08DSH4 08DSH4 08DSH4 08DSH4 08DSH4 08DSH4

08DSH4

3. Nguyn Th Thanh Trang 4. Cao Tun V

5. L Hong Thy Oanh 6. Bi Th Anh Th
7. L Minh V

TP.HCM, 12/2011

THC HNH CNG NGH LN MEN

MC LC
Bi 1: Ln men bia ......................................................................................................... 3 Bi 2: Sn xut nc tng ln men ............................................................................ 18 Bi 3: Ln men yoghurt ................................................................................................ 28

NHM 2 _ 08DSH4

THC HNH CNG NGH LN MEN

Bi 1: LN MEN BIA.
PHN 1: L THUYT. 1.1. Gii thiu. Bia l loi ung c cn v c gas c t c xa, thm ngon v b dng. Cng ngh sn xut bia. xut hin trn th gii hang ngn nm v du nhp vo Vit Nam hn mt trm nm nay. c bt ngun t Ai Cp. cn trong bai thp 3-8%, nh c CO2 khi rt ra to nhiu bt gy sng khoi khi ung. Gi tr dinh dng kh cao, cha nhiu vitamin nhm B v cc nguyn t vi lng. V vy cng ngh sn xut bia khng ngng pht trin v hon thin trn th gii. 1.2. Nguyn liu. 1.2.1. Malt (i mch ny mm). C th sn xut bia thun ty t malt, hoc gim gi thnh c th b sung th liu nh go, i mch Khi nu malt nhit thch hp, tinh bt malt s c h enzyme amylase (alpha v beta amylase) gi chung diastase thy phn thnh ng maltose.

1.2.2. Hoa bia (houblon). Nguyn liu c bn ng th 2 sau malt. Hoa bia gip cho bia c v ng du, hng thm c trng v tng kh nng to v gi bt. Thnh phn chnh ca hoa bia: acid ng, nha ng, tannin v tinh du. Trong cng ngh sn xut bia ngi ta c th s dng trc tip hoa bia hoc ch phm t hoa bia: bt hoa hay cao hoa.

1.2.3. Nm men.

NHM 2 _ 08DSH4

THC HNH CNG NGH LN MEN Saccharomyces cerevisiae v Saccharomyces carlsbegensis l 2 loi nm men thng s dng trong ln men bia. Cc loi nm men ln men bia thuc loi ym kh ty tin c oxy tng sinh khi nh h hp t bo, khng c oxy th ln men to ethanol v CO2. ng + Nito amine t do + Nm men + Oxy 150g/l + 150mg/l 1g/l 25mg/l Ethanol + CO2 + Nm men. 45g/l 42g/l 5g/l

Saccharomyces cerevisiae

Saccharomyces carlsbegensis

c tnh ln men ca 2 loi nm men ny c nhiu im khc nhau dng ln men cc loi bia khc nhau.

Trong bi thc hnh ny chng ta s dng chng nm men S. carlsbergensis v ln men chm. 1.2.4. Ph gia. - Lm gim cng ca nc: Al2(SO4)3, CaSO4. - iu chnh pH= H2SO4, acid lactic, CaCl2.
4

NHM 2 _ 08DSH4

THC HNH CNG NGH LN MEN - Cht chng oxy ha: acid ascorbic. - Cht to mu: caramen. 1.3. Yu cu cht lng (TCVN7042:2002). 1.3.1. Bng yu cu cm quan ca bia hi.

1.3.2. Bng yu cu v ch tiu ha l.

Tn ch tiu 1. acid, s ml NaOH 1N trung ha ht 100ml bia hi ui ht CO2. 2. Hm lng diacetyl, mg/l 3. Hm lng ethanol % V/V 4. Hm lng cht ha tan ban u.

Mc <=1.8

<=2 Theo tiu chun c cng b ca nh sn xut.

1.3.3. Ch tiu v kim loi nng v vi sinh vt (TCVN7042:2002). 1.4. S cng ngh ln men bia malt thun ty. 1.4.1. S cng ngh

NHM 2 _ 08DSH4

THC HNH CNG NGH LN MEN

Malt

Nu malt

Lc v ra b Hoa houblon

Nu hoa

ng

Lng trong Cn Dch Men ging Lm lnh nhanh

Nhn ging

Ln men chnh Nm men

Men

Ln men ph Thu hi CO2 Lc trong bia X l Bo ha CO2

X l li

CO2 tinh khit

Thc n gia sc

1.4.2. Thuyt minh quy trnh. 1.4.2.1. Nu bia. Tinh bt di tc dng ca enzyme trong malt (hot lc diastase). Khi nu bia c th b sung enzyme tng vn tc ng ha. Enzyme thng s dng l Termamyl c ngung gc t vi khun chu nhit. Gin nu l th hng dn qu trinh nu s thay i ca nhit theo thi gian.
6

NHM 2 _ 08DSH4

THC HNH CNG NGH LN MEN

Cc giai on trong qu trnh nu bia. - Malt: nc = 1:4 , a nhit ln 38-40oC trong 30 pht enzyme hemicellulase, glucanase, protease thy phn glucan hoc protein phc tp bao quanh ht tinh bt , to iu kin cho enzyme amylase thy phn tih bt. - 52oC 30 pht: enzyme protease thy phn protein thnh amino acid v peptide to dinh dng cho nm men. To hng v m v gi bt cho bia. Chim 5-7%. - 65oC 30 pht: enzyme beta amylase hot ng thnh cc ng c kh nng ln men. - 72oC 20 pht: enzyme alpha amylase hot ng. - 78oC 30 pht: gim nht gip qu trnh lc d dng hn. iu kin hot ng ca enzyme trong malt.

ng ha kt thc khi phn ng gia iod v tinh bt cho thy mu iod khng b thay i. 1.4.2.2. Lc ra b. Ra b bng nc nng 76oC 3 ln. m bo lng nc trong dch ng. Ra n nng cht ha tan trong nc ra b cn 0,3-0,5% th ngng.
7

NHM 2 _ 08DSH4

THC HNH CNG NGH LN MEN 1.4.2.3. Nu hoa. Nng nhit ln 76-78oC gi 10 pht ng ha ht phn tinh bt cn li. un si dch ng 10 pht th cho hoa bia vo to v ng. Trc khi kt thc 10 pht cho hoa bia vo to hng.

Mc ch.
- Bt hot enzyme trong malt. - Thanh trng dch ng. - Trch ly hoa bia. - ng t protein trong dch ng. - Hnh thnh hng v v mu. - Hnh thnh cc cht kh chng oxy ha trong giai on tip theo. - Gim pH dch ng. 1.4.2.4. Lm lnh nhanh. a nhanh chng dch ng sau khi nu hoa v nhit ln men. 1.4.2.5. Nhn ging nm men. Cn b sung Zn v N. Mi trng ging khi ng cng ging mi trng ln men chnh cng tt. Sc kh v lc ng vai tr quan trng. Mi trng nhn ging l YEPG: yeast extract 5g/l, peptone 10g/l, glucose 20g/l.

S nhn ging nm men.

NHM 2 _ 08DSH4

THC HNH CNG NGH LN MEN

1.4.2.6. Cy ging. T l cy ging ti u l 6-10.106 t bo/ml men ni. 10-14.106 t bo/ml men chm. Dch ng cng m c t l cy ging cng cao. 1.4.2.7. Ln men chnh. Phng php thnh hnh hin nay l ln men theo m. Cc thng s cn theo di trong qu trnh ln men chnh. - Hm lng ng Brix, pH. - pH u l 5,2-5,6. - pH cui ln men chnh l 4,4- 4,5 do H2CO3 sinh ra. - Nng ethanol. + Nng ethanol 2% nm men pht trin bnh thng. + Nng ethanol 2-5% nm men gim tng trng. + Nng ethanol >5% nm men chm dt tng trng nhng vn ln men. + Nng ethanol = 12% nm men chm dt ln men. - Thi gian ln men 7-10 ngy ph thuc vo cn v pH. Kt thc qu trnh thu c bia non, mi v nng, cm quan cha t.
9

NHM 2 _ 08DSH4

THC HNH CNG NGH LN MEN 1.4.2.8. Ln men ph. y l qu trnh - Cc phn ng enzyme v phi enzyme din ra dn n n nh cht lng bia. - Cc hp cht to hng quan trng hnh thnh. - Diacetyl b phn hy xung i mc quy nh 0,2 mg/l, ch tiu quyt nh thi gian ln men ph. - Bo ha bia bng CO2 tng v v kh nng to bt, c ch s pht trin ca vi sinh vt c hi.

Tin hnh.
Sau khi tch men, h nhit xung 0-1oC. Duy tr p sut 0,9-1,2mg/cm2 ngng thu hi CO2 khi qu trnh ln men ph din ra 15 ngy. Kt thc qu trnh ln men ph nng ng cn li khong 1,9 Brix, tch nt phn nm men cn li. Bia sau ln men c bm qua thit b lc.

PHN 2: THC HNH. 2.1. Nguyn liu. - 200g malt. - Cao hoa: 0,02%. - Nm men: Saccharomyces carlbergensis. 2.2. Tin hnh. Xc nh m ca malt: cn cc v np sau cho 3g malt ta c khi lng trc sy l (m1) vo em sy nhit 1050C, cn li ta c khi lng (m2). Lu trong lc sy th m np lc cn th y np v sy n khi no khi lng khng i th ngng sy. m c xc nh theo cng thc:

Xc nh mt t bo nm men v xy dng ng chun t bo: Ging t mi trng gi ging Hansen c cy sang erlen cha 50ml mi trng nhn ging YEPG.

10

NHM 2 _ 08DSH4

THC HNH CNG NGH LN MEN - Tin hnh xc inh mt t bo: ht 1ml dch huyn ph t mi trng nhn ging YEPG, tin hnh pha long trong nc mui sinh l 0,85% n pha long 10-7, 10-8, 10-9. Ht 0,1ml t cc nng pha long 10-7, 10-8, 10-9 cho vo a petri cha mi trng Hansen v tin hnh cy trang, mi pha long lp li 3 ln, qu trnh ny tin hnh trong iu kin v trng. em cc a petri i 300C trong 72h . Sau m khun lc v xc nh mt t bo. - Xy dng ng chun t bo: Ht 1ml huyn ph t bnh erlen nhn ging, cho vo eppendorf, em i ly tm 10000v/pht x 10 pht. Sau b phn dch ly phn cn v ht 1ml nc mui sinh l cho vo, lc u. Tin hnh pha long n khi nng OD t gi tr l 0,4 bc sng 600nm. Qu trnh nu v ln men bia: - Nghin 200g malt sau phi trn vi nc theo t l 1:4 iu chnh v pH 5,5. - Tin hnh nu malt theo gin nu bia thun malt.

Gin nu bia thun malt. - Qu trnh nu malt l qu trnh ng ha, qu trnh ng ha kt thc khi tinh bt b thy ht. kim tra ta dng dugn dch lugol, mt im i chng th ch cha lugol, im th nghim l dch ng ha + dung dch lugol, nu dung dch lugol im th nghim khng c s khc bit so vi im i chng th ta kt thc qu trnh ng ha. + Xc nh ha tan ca malt: Cn 55g malt v nghin trn u, ly ra mt phn nh xc nh m.
11

NHM 2 _ 08DSH4

THC HNH CNG NGH LN MEN t nhit bnh iu nhit. Cn 50g malt nghin cho vo cc bit trong lng, sau thm 200ml nc ct nhit 460C, dng a thy tinh khuy trnh vn cc v ta bt. Gi cc nhit 450C trong 30 pht, khuy lin tc. Tng nhit ln 700C vi tc 10C/ pht. Nhit ca cc t 700C th rt thm vo 100ml nc ct 700C. 5 pht th ly mu mt ln th dch ng ha bng cch ha mt git dung dch iod 0,02N vi mu n khi hn hp c mu vng rm (khng lm mt mu dung dch iod) coi nh ng ha kt thc. Thi gian t khi hn hp trong cc t 700C n khi khng lm mt mu dung dch iod c gi l thi gian ng ha. Nu qu trnh ng ha khng kt thc sau 1h th nghim th ngng li hoc lm li hoc thay i lng malt th nghim. Gi nhit 700C trong vng 1h (tnh t khi t 700C), ly cc ra lm ngui n nhit phng khong 10-15 pht. Ra a khuy bng mt cht nc ct. ct (ch khng k trng lng cc). Khuy u ri ton b lng dch lc vo phu lc, lc li 100ml dch lc ban u ri mi bt u tnh thi gian lc. Ly mt phn dch lc xc nh mu. Qu trnh lc coi nh kt thc khi b mt b kh hoc qu trnh lc chm li (ko di qu 2h). Xc nh t trng dch ng: Phng php cn bnh t trng: Chun b: Bnh t trng sch kh, cn trng lng bnh k c nt (mo); nc ct; dch cn xc nh t trng c lm lnh n 200C. Mu trng: trng bnh t trng bng nc ct lm lnh 200C, nc ct n vch nh mc ca bnh sau y nt li ri t bnh vo bnh n nhit 200C trong vng 30 pht. Sau 30 pht s ly bnh ra v lau kh bn ngoi v yn trong 5 pht, sau ta cn bnh v ghi kt qu (m1). Mu thc: ht nc ra khi bnh t trng trng bnh vi ln bng dung dch phn tch lm lnh 200C. dch phn tch n vch nh mc ca bnh, y nt li v t bnh vo bnh n nhit 200C, 30 pht. Sau 30 pht ly ra, lau kh bn ngoi, yn trong 5 pht, cn ghi kt qu (m2). T trng ca dch lc c tnh theo cng thc sau: (g/g)
12

Lau kh bn

ngoi cc v em cn li sau cho trng lng trong cc t 450g bng cch b sung thm nc

NHM 2 _ 08DSH4

THC HNH CNG NGH LN MEN

Hoc t trng c o bng cch s dng ph k (t trng k). Tnh kt qu: hm lng cht ha tan ca dch ng c xc nh t t trng bng ph lc 1 Bng tra hm lng cht kh ha tan theo t trng ca dch - Goldiner v Klemann. Hm lng cht ha tan ca malt tnh bng cng thc sau: E1(%) = e.(W + 800)/(100 e) ha tan ca malt tnh theo cht kh tuyt i: E1(%) = e.(W + 800)/(100 e) Trong : E1: hm lng cht ha tan ca malt (%) theo khi lng. E2: hm lng cht ha tan ca malt kh tuyt i (%) theo khi lng. e: hm lng cht tan trong dch ng. + Nng lc ng ha. Chun b mu: Cn mu: nu malt vng nht 21g, malt mu sm 41g, malt enzyme 11g, em nghin mn. Cn chnh xc 20g malt vng nht, 40g malt mu sm, cho 10g malt enzyme cho vo cc nu. Thu dch chit: un bp cch thy ti nhit 40oC. Rt 480 ml nc lnh vo mu malt. Khuy u v tnh vn cc. t cc vo bp cch thy 40oC khuy lin tc trong 1 gi c th chnh lch 2 pht. Lm ngui dch chit ti nhit phng. Ra a khuy bng mt t nc. Lau kh pha ngoi cc, iu chnh khi lng cc ti 520, 540, hoc 510g c th chnh lch 0,2g tng ng vi lng malt nu trn. Lc: khuy tht u dch chit, ngay ton b hn hp vo phiu l, lc b 200ml dch lc u, ly 50 ml dch lc sau phn tch. Thy phn dung dch tinh bt. Mu th nghim: dng pipet ly 100 ml dung dch tinh bt vo bnh nh mc 250 ml, them vo 6,5 ml dung dch m acetate, t vo bnh cch thy 20oC, yn 20 pht. Cho tip 6,5 ml dch chit malt lc vo hn hp trn, lc u ri tip 20oC ng 30 pht tnh t
13

NHM 2 _ 08DSH4

THC HNH CNG NGH LN MEN lc bt u them dch chit malt. Sau cho 5ml NaOH ngng hot ng ca enzyme. Thm nc ti ngn nh mc v lc u kim tra kim bng cch nh mt git phenolphthalein, mu ca dung dch phi l mu hng. Mu kim chng: ly 100ml dung dch tinh bt vo bnh 250 ml thm 5 ml dung dch NaOH lc k. Thm 6,5 ml dch chit malt ri thm nc ti ngm bnh v lc u. Xc nh ng kh theo phng php iod: ht 50 ml dung dch trn vo bnh tam gic 250 ml, them 25 ml dung dch iod v 3 ml dung dich NaOH v lc u. y nt bnh trnh tn tht iod v yn trong 15 pht. Them 4,5 ml H2SO4 v chun phn iod d khng phn ng bng dung dch Na2S2O3 ti khi mt mu xanh. Lng iod phn ng nn trong khong 6-12 ml, nu ngoi khong gii hn ny th phi lm li th nghim vi lng malt nhiu hn hoc t hn. - Tin hnh lc v ra b bng nc 760C, ta thu c dch nha. - Nu hoa: ta b cao hoa di dng dch vo dch nha, nu trong 10 pht. Tin hnh o Brix nhit 200C, chnh Brix v 120 Brix. - Np vo bnh ln men 450ml, ngui v tin hnh cy ging , t l cy ging 10%. Sau cho ln men trong t lnh 7-80C. Qua trnh ln men s to ra CO2 v vy ta cn phi bch ming chai bng bong bng.

Sn phm bia cha lc

Kim tra bia thnh phm: kim tra nng ru v cht ha tan ban u. - Tin hnh: Lc bia bng phu lc chn khng, sau ht 100ml bia em i ui CO2 bng cch lc. Sau chng ct, cho 100ml bia ui CO2 vo bnh ct. Lp vo b ct cn.
14

NHM 2 _ 08DSH4

THC HNH CNG NGH LN MEN Ch u ra ca ng ngng phi di khong 5ml nc ct trong bnh thu dch ct. Bt bp bt u chng ct cn. sau khi ta thu c khong 100ml cn th ngng qu trnh chng ct. Trong qu trnh chng ct cn b sung nc lm lnh. - Lm ngui phn dch cn li sau khi chng ct. S dng Brix k o Brix v cn k o cn.

PHN 3: KT QU. - m ca malt: Mu (cc +np) 36,687 36,74 37,158 = 110,3 Msy (cc + np + nguyn liu) 39,454 39,580 39,458 = 118,88

W1 = W2 =

3,0138 2,76 100 8,4 3,0138 3,0574 2,84 100 7,1 3,0574

W% = 7,75% x - Xc nh mt t bo: + V qu trnh thao tc c s sut lm cc mu cy trn a petri u b nhim mc - Mt khun lc: S0 = N/ (n1.V1.F1 + n2.V2.F2 + ) + Ta s dng s liu chun sau: S0= 91010 cfu/ml - Xc nh ng chun t bo: pha long l 17 ln (S1). S1 4ml 3,5ml 3ml Nc mui sinh l (ml) 0ml 0,5ml 1ml S1(cfu/ml) 0,5291010 0,51010 0,4741010 Log S (cfu/ml) 9,72 9,7 9,68
15

OD 600nm 0,4 0,37 0,34 NHM 2 _ 08DSH4

THC HNH CNG NGH LN MEN 2,5ml 2ml 1,5ml 1ml 0,5ml 1,5ml 2ml 2,5ml 3ml 3,5ml 0,451010 0,4291010 0,4091010 0,3911010 0,3751010 9,65 9,63 9,61 9,59 9,57 0,32 0,28 0,26 0,23 0,2

0.45 0.4 0.35 0.3 0.25 0.2 0.15 0.1 0.05 0 9.55 9.6 9.65 logS(cfu/ml) 9.7 9.75

OD 600nm

y = 1.2958x - 12.196 R2 = 0.9929

Biu : ng chun t bo nm men S.carlbergensis. ha tan: Theo Brix = 120 Brix d2020 = 1,05 e = 12,39 E1 = e.(W + 800)/(100 e) = 12,39.(7,75 + 800)/(100 12,39) = 114,23% E2 = (E1.100)/(100 W) = (114,23.100)/(100 7,75) = 123,83% Nhn xt: ha tan ca malt kh tuyt i l 123,83% >100% ( trong qu trnh tin hnh cn v sy c sai s ln nn cho kt qu sai ) Nng lc ng ha: DP1 = F.(VB VT) = 34,2(0,5 0,3) = 6,48 (WK). DP2 = (DP1.100)/(100 W) = (6,48.100)/(100 7,75) = 7,41 (WK). Nhn xt:

16

NHM 2 _ 08DSH4

THC HNH CNG NGH LN MEN - Thng th nng lc ng ha phi t 290-320 (WK) nhng trong th nghim ny nng lc ng ha ch t 7,41 (WK) Qu thp khng t yu cu. - Gii thch: + C th do qu trnh pha ha cht b sai. + Khu chun b dch chit malt lm bt hot enzyme trong malt. + Do khu chun b mu th nghim th cho NaOH vo sm hn thi gian quy nh nn lm enzyme trong malt cha kp thy phn tinh bt th n b bt hot. + Do qu trnh tn tht iod trong lc chun . + Do thao tc chun sai. Kim tra cht lng bia thnh phm: + Brix: 50Brix + cn: 90. ln biu kin = [(Brix0-Brix)/Brix0] .100= [(12 - 5)/12].100 = 58,33%. ln men thc = ln men biu kin .0,81= 58,33 x 0,81 = 47,25% .

17

NHM 2 _ 08DSH4

THC HNH CNG NGH LN MEN

i 2 : SN UT N C TNG
PHN 1: L THUYT. 1.1. Gii thiu:

N MEN

- Nc tng c ngun gc thc vt l dch thy phn ngun m c trong nguyn liu. Tc nhn thy phn c th l enzyme do vi sinh vt tit ra, c th l acid nh HCl (nc chm ha gii). - Nc chm ln men thun ty l nc tng c truyn cc nc Chu c sn xut Vit Nam l nc chm ha gii. - Gn y, ngi ta pht hin trong nc tng ha gii bng HCl c cha nhiu 3-MCPD (3-monochloropropane-1,2diol) c kh nng gy ung th. V vy c xu hng quay tr v cng ngh ln men truyn thng. 1.2. N u n liu: Kh bnh du u nnh (u phng) hm lng b o cng thp cng tt <5%. Nm si Aspergillus oryzae: - Asp.oryzae sinh trng dng h si, bao gm cc si rt mnh chiu ngang 5-7 m, phn nhnh nhiu, c vch ngn chia si thnh nhiu t bo. - Asp.oryzae c khun lc, mu sc bo t v hnh thi rt ging Asp.flavus, mt loi nm tng hp aflatoxin. Asp. oryzae Asp.flavus . Thuy nhin thy phn k o di hng thng, gi thnh cao. V vy trong mt thi gian di hu ht nc tng

1.3. Y u cu cht ln : h ti cm an:

18

NHM 2 _ 08DSH4

THC HNH CNG NGH LN MEN Tn trong Mu V Loi c bit Loi 1 Khng vn c Mu nu m hay mu nu cnh gin Ngt m, sau khi nm khng Du ngt, sau khi nm khng c cm gic kh c, khng c cm gic kh c, khng ng chua hay c cc v ng, chua hay c cc v khng thch hp khc Mi Tn ch tiu m tng s (g/l) m formon/m tng s m NH3 (g/l) acid tng ng acid acetic (g/l) pH NaCl (g/l) 5.5 6.7 230-250 3 5-7 3 5-7 3 5-7 h ti khng thch hp khc. Thm ngon, khng kh t, hay c cc mi khng thch hp khc h al Loi c bit 20 55 Loi 1 16 55 Loi 2 12 55 Loi 2

Quan trng : khng cha aflatoxin (m tnh test aflatoxin) 1.4. Qu t nh c n n h:

19

NHM 2 _ 08DSH4

THC HNH CNG NGH LN MEN

Ln men cho n khi mc chuyn sang mu vng (t=45h)

Kh du

nghin

Thy phn bng enzyme nm mc 550C , > 72h

Phi liu v trn nc

Trch ly

iu v Hp chn (thanh trng) (1atm, 30 pht) Thanh trng Lm ngui 0 v t=40 C ng chai Cy bo t sp.oryzae

PHN 2: THC HNH. 2.1. l n u n liu: - Mc ch : to cu trc ht nhm to mi trng thong kh khi ln men (nui mc). - Nghin bnh du (BD): 1500g = 1.5 kg 2.2. o m D: Cn 3 cc s c np, ta cn cc + np bit trng lng ca cc + np. Sau ta cn 3g BD cho vo mi cc, lc ny ta c khi lng mu trc khi sy. M np, t cc v cc vo t sy nhit 1050C. Sy trong 3h, y np, ly cc ra lm ngui trong bnh ht m v nhit phng trong 20 pht ri cn. Sy li trong vng 2h na v cn li, thc hin ging thao tc trn cho n khi khi lng sau khi sy khng thay i.
20

NHM 2 _ 08DSH4

THC HNH CNG NGH LN MEN Kt qu o c: STT Cc I Cc II Cc III KL cc KL sau khi sy (cc +BD) 38,2 38,1 36,8 40,9 40,82 39,47

- m (W %) ca BD c tnh theo cng thc sau: W= Trong : m1: Khi lng BD trc khi sy m2 : khi lng BD sau khi sy - m cc I : W=
3 2,7 100% = 10% 3

= (%)

- m cc II: W=
3 2.72 100% = 9,33% 3 3 2,67 11% 3

- m cc III: W=

WBD = 10.11% 2.3. Chun b D v nh n ing: Nhn ging Aspergillus Oryzae: - Cn BD (W=10.11%) vo 4 bnh tam gic mi bnh 20g c b nc t m 45% nui mc tng. - Th tch nc cn b sung vo 20g BD t 45% m: = 0.45

21

NHM 2 _ 08DSH4

THC HNH CNG NGH LN MEN - Nc thm =

(0,45 20 ) (20 0,1011 ) = 12,67 (ml) 0,55

- Bnh du b sung thm 12,6ml nc t m 45%, em hp tit trng, sau cy ging t ng thch nghing sang, em nhit phng n khi ra bo t. Hp thanh trng BD : - Cn 200g BD (W=10.3%) phi trn vi nc to m cho BD nui mc tng. Ta tin hnh thc hin 3 m (45%, 55%, 65%) nhm xc nh ra m thch hp nht cho nm mc tt nht. - Th tch nc cn b sung vo 200g BD t 45% m: = 0.45 Nc thm =
(0,45 200 ) (200 0,1011 ) = 126,87 (ml) 0,55

- Th tch nc cn b sung vo 200g BD t 55% m:

= 0.55
Nc thm =
(0,55 200 ) (200 0,1011 ) = 199,51 (ml) 0,45

- Th tch nc cn b sung vo 200g BD t 65% m: = 0.65 Nc thm =

(0,65 200 ) (200 0,1011 ) =313,657 (ml) 0,35

- em hp 1210C, 30 pht, 1atm. 2.4. C in v l n m n: S dng cc khay nha c l c lt bo pha di khay, ta cho 200g BD c m khc nhau (45, 55 ,65%) vo trong khay. T l cy ging l 0,1% .(200g BD th c t l cy l 0,2 g bo t nm mc). Sau ly bo ph nh trn khay. - Nhit phng (30 320C): trong sn xut th cng cn lt mc duy tr nhit bn trong khi nguyn liu ln men 30 320C. Trnh nhit qu cao v hi nc ngng t lm hng sinh khi nm .
22

NHM 2 _ 08DSH4

THC HNH CNG NGH LN MEN - m khng kh 95%. - Thi gian ln men: cho n khi thy bo t c mu vng hoa cau trong thi gian 42h.

m tt nht ca nm mc khi m 65%. V ma ny l ma kh nn

m s cao hn. Cn cc m khc thu c sinh khi mc t hn.

Mc tn c m u v n hoa cao 2.5. Th ph n: - Mc ch: thy phn m trong nguyn liu bng enzyme do nm tng hp. Cc thng s k thut: - Lng nc, nng nc mui b sung (c ch tng trng sinh khi enzyme bt u thy phn) - Nhit : 54 580C - Thi gian: 64 -72h (do hm lng m formon quyt nh). - Lng nc b sung c tnh nh sau: 100g mc c m 65% cn b sung khi lng nc W sao cho lng nc trong mc gp 1,2 ln khi lng cht kh trong mc. Lng nc b sung = 1.2 khi lng mc Lng nc b sung vo mc = 1.2 100g = 120 ml - Nng mui ha tan trong nc b sung Nng mui 15% 20% 25%
23

Khi lng mui trong 120 ml nc 18 g 24 g 30 g NHM 2 _ 08DSH4

THC HNH CNG NGH LN MEN

Vi nng mui ha tan trong nc b sung nh trn tin hnh cho lng nc mui b sung vo sao cho ngp ht b mt khi nguyn liu.

Thy phn 2.6. Ph n t ch tn .

Nc tn th nh phm
24

NHM 2 _ 08DSH4

THC HNH CNG NGH LN MEN Tin hnh: Ht 2ml mu nc tng cho vo bnh t mu. Cho 0.2g xc tc (K2SO4:CuSO4 = 3:1) + 5ml H2SO4 m c. Tin hnh t cho n khi thy dung dch c mu trng trong th ngng. Sau tin hnh chng ct m Mu nh mc n 100ml chia lm 2 phn mi phn 50 ml dng bnh Kjeldal 50 ml nc ct + 3 git Tasiro. Sau cho 50 ml mu vo + 25 ml NaOH 45%. Chun b bnh hng dng erlen: cho 20ml H2SO4 0.1N + 3 git Tasiro sau cho u ng ngp trong acid H2SO4 . Trong bnh KJeldal, NH3 bay ra gp hi nc to thnh NH4OH, NH4OH tc dng H2SO4 d to thnh (NH4)2SO4 + H2SO4 d. em chun lng H2SO4 d ny s xc nh c lng H2SO4 tc dng vi NH4OH t tm c lng NH4OH. Tnh theo cng thc
(V1 V2 ) 0,00142 1000 100 (g/l) Vmau 10

N=

V1: s ml dung dch H2SO4 0.1N cho vo bnh cha V2: s ml dung dch NaOH 0.1N dng chun s acid d 0.00142 s g nit tng ng vi 1 ml NaOH 0.1N 1000: h s i ra g/l 100: dung tch bnh nh mc ml Vmu: s ml mu ly v c ha 10: s ml mu pha long cho vo my ct Chnh lch kt qu gia 2 ln xc nh song song khng c qu 0,25 g/l Nng mui 15% Th tch NaOH chun 16,5 ml Nito tng s N = (20-16,5)1,42 = 4,97 (g/l)

25

NHM 2 _ 08DSH4

THC HNH CNG NGH LN MEN 20% 17,2 ml N = (20-17,2) 1,42= 3,976 (g/l) N =(20-18) 1,42= 2,84 (g/l)

25%

18 ml

Nhn .

t:

nng mui 15% cho hm lng nito tng s l cao nht (4,97g/l)

nng ny cho mu tng p v m hn cc nng khc. Tuy nhin, trong sn Tin hnh

xut, sau khi thy phn ngi ta phi b sung thm mui t mn cn thit.

Ht 10 ml nc tng cho vo bnh nh mc 250 ml thm nc ct vo nh mc n vch, lc u. Gi dung dch ny xc nh hm lng mui. Dng pipet ht 10 ml dung dch trn + 15 git ch th hn hp sau cho vo tng NaOH 0.1N n khi dung dch c mu pht xanh. Cho tip 5 ml formol trung tnh, chun bng dung dch NaOH 0.05N cho n khi dung dch chuyn t mu vng sang mu tm. Tin hnh xc nh mu trng, thay 10 ml nc tng pha long bng 10 ml nc. Kt qu tnh hm lng nito amin theo cng thc Namin = 0,0014210001(V2-V1) 0,1 (g/l)

V1, V2: th tch NaOH cn dng khi chun mu trng v mu th nghim 0,00142: s g Nito tng ng vi 1 ml NaOH 0.1N 1000: h s i ra g/l 0,1: th tch mu nguyn 1: th tch mu pha long cho vo my ct Chnh lch kt qu gia 2 ln xc nh song song khng c qu 0.25g/l Nng mui Th tch NaOH chun 15% 2,4 ml Namin = (2,4-0) 1,42 = 3,408 (g/l)

Nito amin

26

NHM 2 _ 08DSH4

THC HNH CNG NGH LN MEN 20% 2,15 ml Namin = (2,15-0) 1,42 =3,053 (g/l) Namin = (1,325-0) 1,42 = 1,882 (g/l)

25%

1,325 ml

Nng mui 15% 20% 25%

nng mui 15% cho hm lng Nito amin l cao nht 3,408 (g/l) Nito tng s 4,92 (g/l) 3,976 (g/l) 2,84 (g/l) T s
3,408 0,693 4,92 3,053 0,768 3,976 1,882 0,663 2,84

T s Nito formol / Nito tng Nito formol 3,408 (g/l) 3,053 (g/l) 1,882 (g/l)

Nhn -

t: nng mui 15% trch li nhiu m nht. y l nng mui thch hp cho

hot ng enzyme thy phn protein ca bnh du. - Nu hm lng mui qu cao c ch hot ng enzyme v qu trnh trch ly, s nh hng n cht lng ca nc tng sau ny.

27

NHM 2 _ 08DSH4

THC HNH CNG NGH LN MEN

Bi 3: LN MEN YOGHURT
PHN 1: L THUYT. Yoghurt l mt dng sa ln men. 1.1. Nguyn liu. - Sa: Sa b trung bnh cha 3, 5% cht bo; 3,4% protein, 4,8% lactose. - Ging vi khun lactic: + Vi khun ln men lactic: trong ch yu l Lactobacillus delbrueckii subsp. bulgaricus (LB) v Streptococcus thermophilus (ST) hoc Lactobacillus acidophilus (LA) + Vi khun Gram dng hnh que (Lactobacillus) hoc lin cu (Streptococcus), khng sinh bo t,khng di ng, k kh khng bt buc, chu acid.

Lactobacillus delbrueckii subsp. Bulgaricus

Streptococcus thermophilus

Lactobacillus acidophilus

- Sn phm trao i cht ca vi khun lactic trong qu trnh ln men + Acid lactic (CH3-CHOH-COOH, Mw = 90)
28

NHM 2 _ 08DSH4

THC HNH CNG NGH LN MEN Lm ng t casein (protein sa) pH 4,6-4,7, do to nn cu trc gel. To v c trng cho yoghurt (chua). c ch vi sinh vt khng mong mun . + Polysaccharide ngoi bo (exopolysaccahride EPS) lm tng nht ca yoghurt. + Cc hp cht to hng Acetaldehyde Diacetyl 1.2. Yu cu cht lng yoghurt: Bng 1 Cc ch tiu cm quan ca sa chua

Bng 2 Cc ch tiu l - ho ca sa chua

29

NHM 2 _ 08DSH4

THC HNH CNG NGH LN MEN

1.3. Quy trnh cng ngh.

30

NHM 2 _ 08DSH4

THC HNH CNG NGH LN MEN

X l s b

ng ha

X l nhit

Nhn ging

Lm ngui n t0C ln men

nhit 450C, thi gian 4h

Lm lnh nhanh

X l sa s b: Lc sa: Lm sch sa khi cc vt ln trong sa hoc t bo ng vt. Chun ha nguyn liu: - Chun ha hm lng cht bo: ty theo yu cu hm lng cht bo trong sn phm, loi b cht bo bng ly tm tch bo hoc b sung sa nguyn kem hoc kem. - Chun ha hm lng cht kh: tng cht kh tng s ln 14-16g/100g t trng thi mn v c st nh yu cu cht lng. ng ha (quy m cng nghip) p sut cao 2000-2500 psi. Mc ch lm n nh cu trc ht nh sa, tng thi gian bo qun. X l nhit: Thanh trng 950C trong 10 min. Lm ngui n 450C (nu s dng mt chng Lactobacillus) v 430C nu s dng hn hp LB v ST. Cy ging: Nhn ging:
31

NHM 2 _ 08DSH4

THC HNH CNG NGH LN MEN - ng thch nghing. - Mi trng: sa gy hon nguyn hm lng cht rn khc bo 10-12g/100g tit trng 1210C, 15 min. - T l cy ging 2% 430C trong 3-4 gi (t 0.85 g lactic acid/100g) Cy ging theo t l 2%. Ln men: 450C nu s dng mt chng v 430C nu s dng 2 chng (ST hot ng ti u 390C v LB 450C) trong vng 5 gi( kim tra pH xung n 4.3 hoc acid tng l 0.850.9%) Lm lnh nhanh v 50C PHN 2. THC HNH. Chun b sa cho qu trnh ln men v nhn ging: - Sa ti 1L - Sa bt gy: 100g - Ging: 2 h sa chua Vinamilk v 2 h sa chua Well yo ca Kinh

Pha sa: + Nhm 1,2: pha 240ml sa ti => thanh trng + Nhm 3,4: pha 240ml sa ti +10% sa bt gy => thanh trng + Nhm 5,6: pha 240ml sa ti +15% sa bt gy => thanh trng - Mi nhm 2 h nha, mi h ht 60ml sa pha v cy 10ml ging. - Cy ging: s dng hai ging LB v ST (phn lp t sa chua Vinamilk) cho nhm 1,3,5 v LA (phn lp t sa chua Well yo) cho nhm 2,4,6.
32

NHM 2 _ 08DSH4

THC HNH CNG NGH LN MEN Kho st: 370C, o cc ch tiu qua cc mc thi gian: 0, 2, 4, 6h .ng thi tin hnh chun nguyn liu sa trc khi cy ging + Trng thi. + Mi. + pH: o bng giy o pH. + chua (chun acid): cho 10ml sa qua cc mc thi gian vo erlen 250ml, thm vo 20ml nc ct v vi git phenoltalein, chun bng NaOH 0,1N n khi xut hin mu hng ghi nhn s ml NaOH s dng chun theo cc mc thi gian.

PHN 3: KT QU. Kt qu cm quan kt cu, mi, o pH v chun bng NaOH: acid: (oT) = VNaOH 5 Nhm 1 Ch tiu Nguyn liu Mi trng sa pH acid 2 Trng thi Dng lng Mi Mi trng sa pH acid 3 5-6 15,25 Mi trng Dng lng Dng lng Dng lng Hi t Hi r hn 6 64,25 Dng lng c Thong 6 66,5 Hi t Hi c c c Mi ca trng sa 5-6 35,5 Dng lng c Mi ca trng c Mi c Thong 0h Dng lng 2h Dng lng c Mi 4h Dng lng c Thong 6h Hi t Hi r hn c

Trng thi Dng lng Mi

c Mi ca trng sa

ca trng ca cht chua sa

mi chua

ca trng ca cht chua sa 5-6 38,75 Dng lng c Mi

mi chua

Trng thi Dng lng Mi

ca trng ca cht chua

33

mi chua NHM 2 _ 08DSH4

THC HNH CNG NGH LN MEN sa pH acid 4 Trng thi Dng lng Mi Mi trng sa pH acid 5 Trng thi Dng lng Mi Mi trng sa pH acid 6 31,5 Mi trng sa pH acid 36 Dng lng c Mi ca trng sa 57,5 Dng lng c Mi 83,5 Dng lng c Thong 95,5 Hi t Hi r hn c Dng lng Dng lng Dng lng Hi t Hi r hn c c Mi ca trng sa c Mi c Thong Dng lng Dng lng Dng lng Hi t Hi r hn c c Mi ca trng sa c Mi c Thong sa sa r hn

ca trng ca cht chua sa

mi chua

ca trng ca cht chua sa

mi chua

Trng thi Dng lng Mi

ca trng ca cht chua sa

mi chua

Nhn xt: - Trng thi v mi ca cc t l cy ging khc nhau hu nh l ging nhau, lc u u c mi c trng ca sa, sau qu trnh ln men th to ra mi c trng ca sa chua. - Theo l thuyt th pH s cng ngy cng gim do acid lactic sinh ra trong qu trnh ln men. Kt qu th nghim th pH ca cc nhm vn ln hn pH ca sa chua l 4,6. L do c th l bi v trong qu trnh th nghim ch s dng giy o pH o pH nn vic xc nh khong pH khng c chnh xc.
34

NHM 2 _ 08DSH4

THC HNH CNG NGH LN MEN - acid: Ta thy kt qu acid ca nhm 1,2 u khng t so vi ch tiu (75140),cn cc nhm 3,4,5,6 th acid cao hn t ch tiu (75- 140)v tng theo t l hm lng sa bt gy => do b sung sa bt gy gip tng lng ng lactose, thy phn nhiu ng lactose hn to acid lactic -> tng acid. Nhng cc nhm u c acid tng dn theo thi gian chng t l lng acid lactic to ra cng v sau th cng nhiu. - Qua kt qu th nghim cho thy t l cy ging m khng phi trn l th tt hn l cc ging phi trn. pH cc t l cy ging khng phi trn c gim nhanh theo thi gian.

35

NHM 2 _ 08DSH4

THC HNH CNG NGH LN MEN

36

NHM 2 _ 08DSH4

THC HNH CNG NGH LN MEN

37

NHM 2 _ 08DSH4

THC HNH CNG NGH LN MEN

38

NHM 2 _ 08DSH4