cng thc tp cng ngh ln men 2012

cng thc tp cng ngh ln men

Sinh tng hp ethanol bng phng php bt ng t bo
I/. Chng ging: Saccharomyces cerevisiae 1. V tr - Gii nm (Fungi) - Ngnh nm ti (Ascomycota) - H Saccharomycetaceae Cn gi l men bnh m, men bia, men ru 2. Ngun gc - Bnh men ru - V hoa qu (vd nho) - t, nc 3. c im Nm men (Yeast) n bo Cu to t bo nhn tht Sinh sn: ny chi (ch yu), nhn i a. Hnh thi Hnh cu / trng / oval, c th to thnh chui hay m nh chm nho Kch thc nh ~ 5 14 m (KHV: quan st vt knh 10 or 40) b. Sinh l + Ngun dinh dng: Carbohydrat: ng n (vd Glu, Fruc, Gal); ng i (vd Sac, Mal) Nit: hu c (vd cao nm men); v c (vd amoni sulfat) Vi lng: Mg, P + k nui cy: to: ti u 28 30oC pH: 6.5 7.0 Cp kh: hiu kh (nu thu sinh khi) hoc k kh (nu thu sn phm chuyn ha nh EtOH) + Chuyn ha: 2 con ng Hiu kh: Glu CO2 + ATP K kh: Glu EtOH + CO2 + ATP + sp ph (glycerol, a.succinic) 4. ng dng + Sn xut EtOH, ru vang, bia: qt ln men k kh + Sn xut a.a: l ngun nguyn liu giu protein Page 1

 cng thc tp cng ngh ln men 2012

+ TB biu hin gen: l Eucaryote n bo, sinh sn nhanh, c nghin cu chi tit v di truyn nghin cu di truyn sinh vt nhn tht + Probiotic (tr sinh) cho ngi: b sung saccharase cho TE tiu ha km saccharose + Thc phm b sung trong chn nui gia sc, nui trng thy sn II/. Phng php bt ng E & TB 1. M E t do: Thng khng bn vi to, pH Ch sd c 1 ln Tinh ch tn km Khc phc: k thut c nh E & TB K thut c nh E (TB) + K/n: E (TB) c nh v / KG x | bo ton c hot tnh SH + M: Sd nhiu ln D tch khi sp C th ngng p/ khi cn 2. YC a. E tinh sch Gi c hot tnh b. TB Cha Eni bo hot tnh mnh Khng c E can thip vo p/ E chnh, to ra p/ ph Sp ca p/ E i qua c mng TB c. Cht mang Bn vi to, pH, VSV, tc nhn ha hc PP c nh n gin bn ph hp, gi c hot tnh ca h trong t di Khng c vi E, TB, c th sng Gi thnh ph hp Phn loi + V c: bentonite, silica, thy tinh + Hu c: polysaccharide nh cellulose, agar, alginate; protein nh collagen, albumin; polymer tng hp nh polyacrylamide, polystyrene 3. PP c nh Page 2

 cng thc tp cng ngh ln men 2012

Nhc + k phc tp + C th nh hng E lm gim hot + Khng ti sd cht mang By bao + E (TB) c nht / + Hu nh khng + C cht PTL ln gi mng li polymer (thin nh hng cu trc khng qua mng nhin or tng hp) E (TB) c + E (TB) c gi / vi + Mt li nh s + Kch thc mt nang c mng bn thm gi cht E (TB) li kh /chnh ng nht, E c th thot ra 1 phn + 1 s gc t do to thnh / qt trng hp c th km hm E + Khng ti sd cht mang Hp ph + E (TB) c hp ph ln + t nh hng cu Lk lng lo, d b BM cht mang R nh lc trc E (TB) ra tri hp ph, ion, to phc + D thc hin, H + Cht mang: C hot, cao bentonite, d/c ca + Ti sd c cht cellulose mang To lk cho + Gn E vi E = lk ng + t b ra tri + Lng E c nh ha tr i phn t + Tng kh nng t khng tan chng chu cc yu + C th gim hot + Tc nhn gn E: hay t bin tnh tnh E dng glutaraldehyde + t Tn To lk ng ha tr M t + E to lk ng ha tr vi cc nhm chc / cht mang khng tan / H2O 4. u nhc chung a. u E (TB) n nh Chuyn sang p/ lin tc: c th t ng ha D tch sp Ch ng ngng p/ khi cn Page 3 u + Bn vng + Cht mang a dng + E d t/x c cht

 cng thc tp cng ngh ln men 2012

C th ti sd E (TB) C th pht trin h thng p/ nhiu E (TB) b. Nhc Hot tnh < Etan C cht kh tx E (TB) C th cnh tranh c cht gia E & cht mang Duy tr s sng TB c nh kh hn TB t do 5. d a. Sx nguyn liu BTH lactam + C nh E: penicillinamidase + C nh TB: E.coli, Bacillus megatherium b. Sx a.a C nh E amino acylase / DEAE Sephadex: chuyn h2 racemic L a.a c. Sx fructose C nh E glucose isomerase d. Sx EtOH C nh TB: Saccharomyces cerevisiae e. Sx amylase C nh TB: Bacillus cereus f. Sx a. citric C nh TB: Aspergillus niger etc 6. ND thc tp + Bt ng TB Saccharomyces cerevisiae + PP bt ng: by TB / gel Ca alginate Alginate (a.alginic) + Ngun gc: rong bin, to nu + Cu to: a.polyuronic, gm cc /v a. D manuronic (M) & L guluronic (G) kt hp vi nhau = lk 1,4 M dng thng tnh mm do G dng gp khc tnh vng chc + C ch to gel alginate: Cha c ion ha tr II: alginate tn ti dng b ri 3 dng block: MMMM GGGG MGGM Page 4

 cng thc tp cng ngh ln men 2012

C ion ha tr II (vd Ca2+): Ca2+ gi cho cc mch xp // di dng v trng

Ca2+

Ca2+

Ca2+

TB + C ch by TB: trng hp gel Ca alginate / DD cha TB Na alginat tan / H2O trn c vi dch nui cy TB thm tng git vo DD CaCl2 to ht gel Ca alginat khng tan cha TB (TB s c cha trong cc ngn ca v trng - xem hnh v) III/. Cc yu t nh hng n hiu sut qu trnh ln men to EtOH bng PP bt ng TB Saccharomyces cerevisiae Cc yu t lin quan sinh l t bo Chng ging Dinh dng to pH Cp kh tln men Cc yu t thuc v k thut Ht Ct vdng tln men

1. Chng ging + Cht lng ging + Lng men ging 2. Dinh dng + TB nm men c th ng ha trc tip ng n v ng i + Nu sd ngun carbohydrat khc: cn x l Vd - Cellulose: thy phn bng a. v c - Tinh bt: thy phn bng a. v c hoc E amylase (dng Etinh khit hoc sd cc VK, nm mc sinh amylase) + Cng cng cn c nghin cu | TB c dinh dng m Pthm thu khng nh hng xu n TB 3. Nhit Page 5

 cng thc tp cng ngh ln men 2012

Nm men pht trin tt trong khong 28 32oC Nhit ti u 30oC Ngoi khong ti u: to c th lm gim hot tnh E ca TB pH Nm men pht trin tt k pH 6.5 7.0 Cp kh + Giai on nhn ging: cn k hiu kh to nhiu sinh khi + Giai on ln men to EtOH: cn k k kh sinh nhiu EtOH tln men Nn kt thc trc khi t bo nm men bc vo thi k suy tn Ht + S lng + Cht lng (gi c bao nhiu t bo, t bo c b thot ra khi ht khng, c cht v sn phm c ra vo c khng) Ct a. Chiu di ct + Ct qu di: cc TB bn di d thiu dinh dng, b EtOH c ch cht + Ct qu ngn: lng ht nhi ct gim b. ng knh ct + qu nh: dch ln men kh chy, lng ht nhi ct gim + qu ln: tn dch ln men, Slp t Tc dng + Qu nhanh: TB cha kp thu nhn dinh dng + Qu chm: gim nng sut; cc TB bn di c th b thiu dinh dng

4. 5.

6. 7.

8.

9.

IV/. Mt PP nh lng ethanol, glucose 1. nh lng ethanol Xem SGT c cht hc a. PP vt l + Lng + Cn k b. PP ha hc + OXH K2CrO7 / H2SO4 c KMnO4 / H2SO4 K2CrO7 / HNO3 + Nitroso ha to phm mu azo o quang

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 cng thc tp cng ngh ln men 2012

+ Alkyl iodid: to C2H5I (dn CO2 qua DD EtOH si v DD HI 70%), cho td vi Br2 / acid c ri vi KI d GP I2. c. PP sinh hc Sd men alcol dehydrogenase (ADH) ADH C2H5OH + DPN CH3CHO + DPNH2 semicarbazid o quang 2. nh lng glucose a. PP Schoorl Regenbogen + TT Schoorl A (CuSO4) + TT Schoorl B (KNa tartrat) phc Cu2+ xanh + Phc Cu2+ + aldehyd or ceton / kim Cu+ + Cu2+ d + KI tha / acid I2 nh lng I2 bng Na2S2O3, ch th h tinh bt b. o I2 Glucose b OXH bi I2 / Na2CO3 Chun tha tr c. nh lng Cu2O + Cu2+ + aldehyd or ceton / kim Cu2O + Cu2O c lc, ra ri ha tan trong Fe2(SO4)3: Cu+ Cu2+, Fe3+ Fe2+ nh lng Fe2+ bng KMnO4 / acid d. o gc quay cc e. o f. o quang: lm p/ mu vi acid dinitrosalicylic (DNS) g. nh lng bng enzym glucosidase V/. Bo co thc tp ND - Nguyn tc thc tp - S qui trnh, s ct ln men - Kt qu: sinh khi, nh lng ng, nh lng EtOH - Bn lun Nhn xt kt qu nh lng u nhc im ca ln men lin tc so vi ln men m Gp v qui trnh thc tp

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