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Ti liu tham kho Cng ngh vi sinh vt tp 2, 3. PGS-TS Nguyn c Lng Cng ngh vi sinh ng dng, PGS-TS Trn Minh Tm Cng ngh ln men ng dng trong CNTP, Bi i Cng ngh sn xut malt v bia, Hong nh Ha Cng ngh sn xut v kim tra cn etylic, PGS TS Nguyn nh Thng, Nguyn Th Thanh Hng Enzyme vi sinh vt, PGS TS L Ngc T Food microbiology, William C.Frazier Applications of biotechnology to traditional fermented foods (http://www.nap.edu/catalog/1993.html) ..
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Ti liu tham kho (tt) Bamforth C.W. Food, Fermentation and Micro-organisms, Blackwell Publishing, USA, 2005. Hutkins R.W. Microbiology and Technology of Fermented Foods, Blackwell Publishing, USA, 2006. Elmer H. Marth, Applied dairy microbiology, Second edition Springer, Wine microbiology practical and procedures, 2007 Springer, Modern techniques in the microbial ecology of fermented foods, 2008 Elsevier, Handbook of culture media for food microbiology, 2003 The microbiology of safe food, Blackwell Publishing, USA, 2000 Practical food microbiology, 3rd edition, Blackwell Publishing, USA, 2003
NH GI KT QU HC TP CA SV
1. IM QU TRNH (30%)
1.1. KIM TRA 15 PHT (15%) 1.2. BO CO SEMINAR (15%): chn 1 trong 3 phng n sau
1.2.1. Chn 1 bi bo ting Anh, dch sch lin quan ti mn hc c hiu v thuyt trnh power point (4sv/nhm) 1.2.2. Chn 1 bi bo ting Vit lin quan ti mn hc c hiu v thuyt trnh power point (2sv/nhm) 1.2.3. Chn 1 ti c thc hin lin quan ti mn hc c hiu v thuyt trnh power point (4sv/nhm).
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NI DUNG CHI TIT HC PHN 2 Phn 1. M u www.gbd.edu.vn Phn 2. K thut ln men Phn 3: Cng ngh ln men ng dng
3.1. Ln men ethanol v cc ng dng 3.2. Cng ngh sn xut acid hu c thc phm 3.3. Cng ngh sn xut acid amin 3.4. Cng ngh sn xut sinh khi vi sinh vt 3.5. Cng ngh sn xut polysaccharide t VSV 3.6. Cng ngh enzyme 3.7. Cng ngh sx cc sp ln men truyn thng
PHN 1. M U 1.1. CNG NGH LN MEN 1.1.1. Khi nim v ln men (fermentation)
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1.1.2. Khi nim v CNLM (fermentation technology) 1.2. PHN LOI CC SN PHM LN MEN
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1.1.1. Khi nim v ln men (tt) First Fermentation concept, or Pasteur concept in 1857, Fermentation is the transformation process of the sugar to alcohol in presence of "la vie sans l'air" (means life without air).
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1.1.1. Khi nim v ln men (tt) Conclusions of Pasteur from its study of wines: The alcoholic fermentation of grape juice occur only in presence of yeasts. The wine acidification occur in presence of bacteria. When the grape juice is heated the fermentation do not take place. When the wine (the sugar) is heated the acidification do not occur.
Fermentation is the change of the substrate (sugars) by the action of the ferments
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The fermentation technology is the combined application of the knowledge of process engineering, biochemistry and microbiology for designing or evaluation a fermentation process.
Sn phm
Acid foods (citric, lactic) Alcohol Amino acids Antibiotics Enzymes Pharmaceutical proteins Sn lng (tn/nm) 100,000 1,000,000 10 100,000 10 30,000 0.1 3,000 0.001 - 3
Useful Applications
ENVIRONMENT
PHARMACY FOOD
Bioprocess Bioinformatics
Biosensors
Process analysis
Biochemistry Microbiology
SCIENTIFIC KNOWLEDGE
Physiology Statistic
Molecular Biology
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C cht
C cht
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Cost distribution
High volume and Low value product High investment cost (ph u t cao) High raw material cost (ph nguyn liu cao) High conversion efficiency (hiu qu chuyn i thp) Low recovery cost (Ph quay vng thp) Low margin profit (li nhun thp) Only few regulatory problems (phi theo quy nh php lut) Required not much R&D expenditures (khng tn nhiu tin cho RD Low qualification of the labor force is acceptable (Ngi lao ng c th ko cn trnh chuyn mn cao) Low volume and High value product High investment cost Cost are distributed in all production stages High recovery of the product in the fermentation stage High purification cost High margin profit Too much regulatory and legyslation restrictions, and high plant hygiene Required too much R&D Cost Very high qualification of the labor force is nedeed
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VSV TRONG CNG NGH LN MEN TP TP ln men vi s tham gia ca VSV trong t nhin (TP ln men truyn thng) TP ln men vi s tham gia ca VSV thun khit (TP ln men cng nghip)
Sx th cng, quy m nh, Sx quy m ln, nng sut cao nng sut khng cao Ch ng cy 1 lng VSV Khng kim sot c vo nguyn liu qu trnh, cht lng cha Kim sot c qu trnh ln n nh v cha ng u. men, cht lng n nh & Mang bn sc m thc, ng u kinh nghim, vn ha ca mi dn tc
2.1.1. Yu cu ging VSV trong CNLM Phi c tc sinh trng v pht trin mnh, thun Phi to ra sn phm c nng sut sinh tng hp cao, cht lng tt Phi c tnh thch nghi nhanh trong iu kin sx CN Phi c kh nng chng chu li VSV tp nhim Phi c kch thc ln, thun tin cho qu trnh lng, lc, tinh ch sau ny. Sn phm sinh khi d tch ra khi mi trng nui cy Chng VSV c bo qun d dng, tn ti cc c tnh trong sut thi gian s dng C kh nng thay i cc c tnh bng k thut di truyn ci thin, nng cao nng sut Khng hoc t to thnh sn phm khng mong mun
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Nguyn tc
PL trong c c cht, c VSV ngun VSV phong Tn thi gian, t nhin phn gii c cht ph a dng hot lc VSV cn thp PL trong Tnh thch nghi cao Hiu qu cao, VSV / sx CN quen vi iu kin sx cng nghip To Tip hp, ti t hp To c ging Yu cu trnh VSV = VSV c c tnh cao, thit KTDT mong mun b hin i
Cc trung tm lu tr ging trn th gii v Vit Nam ABBOTT: Abbott Lab, North Chicago, III.60064, USA ATCC: America Type Culture Collector, 12301, Parklaw Drive Rockvill Md20852, USA HIR: Food and Fermentation Divisio, Hokkatdo Profectural Industrial Research Institute Saporo, Japan FERM: Fermentation Research Institute, Agency of Industrial Science and Technology Ministry of Industrial Trade and Industry, Chiba, Japan VTCC, IMBT, National University, HaNoi, VietNam vtcc@vnu.edu.vn; www.biotechvnu.edu.vn (Ngn hng ging quc gia, HN) Bo tng ging: www.bacteriummuseum.org
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2.1.3. K thut nhn ging Nhn ging trong PTN Nhn ging trong quy m sx ln
Cc yu t nh hng n qu trnh nhn ging Thnh phn cc cht trong MT nhn ging (khng cha cht khng sinh, cht c ch). Penicillin, chloramphenycol c ch sinh trng ca VK lactic. Nng cc cht trong MT nhn ging. Nng ng hoc mui cao tng p thm thu c ch VSV pH: chn pH ca MT nhn ging = pHop ca VSV. pH nh hng trc tip ln b mt t bo tch in khc nhau lm cho hot cc loi enzyme VSV thay i. pH nh hng n s phn ly ca cc cht dinh dng c trong MT Nhit : chn To nhn ging = Toop ca VSV
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Ging VSV b tp nhim, thoi ha phi phn lp li hoc thay ging khc
Kim tra thun ca ging thng xuyn (b nhim t ng gc) Kh trng MT dinh dng; vi cc MT c bo t cn kh trng trit hn. V d dit bo t Bac. subtilis 1800C/60 - 90 (B nhim trong qt nhn ging) Ging VSV b thoi ha: do tc ng ca mi trng bn trong v tc ng ca nhng sn phm TC do chnh VSV tit ra
Khun lc nm men trn Hansen Agar c hm lng ng 26, 28% (phng php hun luyn ging)
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S lng khun lc/ nng khng sinh trong MT nui cy 10-7 (khng b 0,01mg/ml 0,1mg/ml 0,5mg/ml sung khng sinh) Nhm 1 2 3 4 34 58 107 Nhim 8 5 7 Nhim 2 3 5 5 Khng xut hin khun lc
TN tin hnh vi vk E.coli. Nui cy E.coli nng pha long 10 -1 trong cc mi trng b sung khng sinh
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Cch thc hin lai ging nm men 1. Cy 2 ging men vo mi trng y M1(mi trng lng). 24h. 2. Cy ging hn hp vo mi trng thch ti thiu M2 v M1 (thch bng). t nht 5 ngy 3. Hai ging nm men lai vi nhau nu trn M2 xut hin khun lc 4. Kim tra ging thu nhn c phi l lng bi th bng cch: Cy khun lc trn mi trng M2 vo mi trng Thch acetat (M3). 48h 5. Lm tiu bn, quan st di knh hin vi, nu thy nang cha 4 bo t (t bo lng bi th) chng t lai thnh cng. 6. Kim tra tnh trng cn quan tm.
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Sng bn gen
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2.1.6.1 Bo qun ging trong thch nghing (cy truyn nh k) Nguyn tc: lun i mi t bo, khng gy ra bt thng. Bin php: mi trng ti thiu, nu mi trng giu dinh dng, VSV pht trin nhanh s thoi ha nhanh. u: n gin, d thc hin, t tn km, thch hp quy m nh Nhc im: tn thi gian, thi gian BQ ngn (1- 2 thng) V tnh hun luyn VSV sng iu kin lnh, lm bin i ging VSV ban u. 2.1.6.2. Bo qun ging trong lp du khong Nguyn tc: c ch qu trnh h hp VSV trong c VSV y m kh v hiu kh, hn ch tip xc vi oxy, ngn hin tng mt nc ca mi trng v VSV. Bin php: 1 lp du khong hoc parafin lng u im: n gin, hiu qu cao, thi gian BQ di (1 nm) Nhc im: C ln du
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VSV c nui mi trng thch. ct, t v trng vo ng nghim, lc u, sau rt qua ng nghim khc. Hn kn ming ng u im: thch hp BQ cc ging VSV trong x l mi trng, trong nng nghip (phn bn) khng i hi mc tinh khit cao. S dng bo qun ging VSV to bo t. Thi gian bo qun di (2 nm) Nhc im: khng dng trong sn xut cng ngh thc phm
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2.1.6.5 Bo qun ging trong giy lc Nguyn tc: p dng vi VSV c bo t. Ngoi giy lc, c th s dng B.C (bacterium cellulose) Cch thc hin: 1. Chun b giy lc v trng:Ct giy lc 1 3 cm. Cho giy lc ct vo ng nghim, y nt bng, sy 1600 C/ 2h hoc kh trng 1210 C/30. 2. Nui VSV trong mi trng lng n khi to thnh bo t 3. Dng pi pet v trng ht 1 git Vi khun vo giy lc. Sy 400 C dn khi thy ming giy lc kh th chuyn giy lc vo ng nghim. Thi gian BQ: 5 nm
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2.1.6.7 Bo qun ging bng phng php lnh ng Nguyn tc: S pht trin ca VSV s b c ch nhit lnh su. Cn s dng cht bo v VSV (glycerin 15%, saccharose 10% + gelantin 10%... Gip VSV khng b cht nhit lnh su Thi gian BQ: -300 C: 9 thng - 400 C: 1 nm - 700 C: 10 nm
2.1.6.8 BQ ging bng pp ng kh (c s dng trong cc ngn hng ging, c quan nghin cu ln)
Nguyn tc: sy nhit thp trong iu kin chn khng, khng nh hng n cht lng ging v c th to ra cc ng ging theo quy m cng nghip. Cch thc hin: ging, nhn ging trong mi trng lng, ki m tra ging (c im sinh ha, sinh l), nu t tiu chun, my ng kh 24h, hn np li. Thi gian bo qun: 20 nm u im: cht lng ging khng i, bo qun nhit thng, thi gian bo qun di Nhc im: chi ph ln
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2.2. NG HC QU TRNH LN MEN (the kinetics of the fermentation processes) 2.2.1. PHNG TRNH MONOD 2.2.2. CC YU T NH HNG N HIU SUT LM
Mc ch nghin cu ng hc Thit k 1 qu trnh ln men mi nh gi hiu qu ca qt ln men ang thc hin Ci tin qu trnh (cht lng, nng sut, gim chi ph) Nng cp hoc gim quy m sx
How to describe a fermentation process? biomass Conversion process of nutrients to products of the microbial metabolism
nutrients cells CO2 H2O Products
CHmOl + aNH3 + bO2 cCHpOnNq + dCHrOsNt + eCO2 + fH2O (1) Substrate Metabolic by
(source of N& O2) (biomass) Rest of the nutrients products or Interest product
Yield is an indicator of the efficiency of the process. From chemical engineering: the Yield is stoichiometry of the reaction.
YX/S: biomass-substrate yield = c Mr(Biomass) / Mr(Substrate) YP/S: product-substrate yield, is calculated through d YP/X: product-biomass yield, is calculated through d/c
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Direct methods
Direct estimation of cell mass: Cell dry or wet weight Optical density
Other Nitrogen + required source nutrients
O 2+
Cells
Measure cell components: Protein DNA and RNA Carbohydrates & Lipids Enzymes
+ Products + CO2 + Heat + H2O Monitor CO2 production: IR analysis Mass spectrometer
Monitor the consumption of the nitrogen source: Chemical analysis NH3 electrode pH-stat Nitrate electrode
Monitor product formation: Chemical analyses Mass spectrometer pH and viscosity Chromatography (gas and HPLC) Enzyme electrode
Indirect methods
Monitor the consumption of essential nutrients: Chemical analysis Ion selective electrodes
P: sn phm (g/L)
=
1 dX X dt
S: c cht (g/L) nh hng ca loi c cht ti Loi c cht Glucose Maltose Maltotriose max (h-1) 0,28 0,22 0,18
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= max
= max
S Ks + S
S S2 Ks + S + KI
VSV E. coli
tng sinh khi trong 1 n v thi gian (ngy, gi) t 1 n v sinh khi X). = dX/XdT (1/ngy; 1/gi) max: hng s tc sinh trng max [S]: nng c cht
C cht Ks (mg/L) 25 4 20 5 4,5 0,042 0,45 0,7 0,4 120 3
Saccharomyces sp. Glucose Glucose Lactose Aspergillus sp. Candida sp. Pseudomonas sp. Hansenula sp. Glucose Glycerol Oxygen Methanol Methane Methanol Ribose
KS: hng s Monod, l nng c cht khi tc to thnh sinh khi = max/2. KS c x bng thc nghim. KS = f(chng VSV v c cht) .
2.2.1.Phng trnh Monod (tt) Phng trnh Monod: mi lin h gia nng c cht v tc sinh trng ca VSV ngha thc tin ca pt Monod: Tnh c, d on c tc to sinh khi ring () nng c cht (S) nht nh. Hn ch: - Pt Monod khng ng khi [S] qu cao. Khi [S] qu cao th gim do [S] qu cao: to p sut thm thu, c ch VSV. - Pt Monod ch ng trong mt khong gii hn ca [S]
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2.2.2. Cc yu t nh hng n hiu sut ln men 2.2.2.1. nh hng ca ging VSV 2.2.2.2. nh hng ca nng c cht 2.2.2.3. nh hng ca nhit Khi nhit tng, hng s tc phn ng tng, tc phn ng tng. Khi gim nhit , tc phn ng gim, nhng c tnh thun nghch. Nu ta tng nhit tr li vng ti u th hot ng ca enzyme v tc phn ng s tng tr li 2.2.2.4. nh hng ca pH enzyme VSV ch hot ng tt pH nht nh. pH ti u ca mi VSV l khc nhau 2.2.2.5. nh hng ca mt s yu t khc: cht c ch
IN
Internal
Cells
OUT
External
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2.3.2. Tit trng trong cng nghip ln men 2.3.3. Cc phng php ln men 2.3.4. Cc thit b ln men
2.3.1. Chun b mi trng Nguyn tc thit lp mi trng: - cht v lng -Da trn nhu cu dinh dng ca VSV v: Cc nguyn t c bn (Macroelements: 90-95% dry mass): C, H, N, H, O (Conc: > 10-4 mol/L) Cc nguyn t khong (Microelements): Ca, Mg, Fe, Zn, Mn, Fe, Cu, B, Cr, Mo Yu t sinh trng (Growth factors): vitamin B, Amino acids, hp cht khc (acid bo, acid nucleic) Mi trng lng: cht kh, pH Mi trng rn: m, cht n (tru, rm)
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Cc bc chun b mi trng
Tit trng khng kh: phng php vi lc, Tit trng/ thanh trng mi trng: -Phng php vt l: nhit , nhit + p sut, vi lc -Phng php ha hc: iu chnh pH, b sung SO2 -Tit trng thit b, h thng ng ng, nh xng: U.V, xng formol, ha cht ty ra
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2.3.3. PHNG PHP LN MEN Phn loi phng php ln men: Theo s pht trin ca VSV trong mi trng: Ln men chm: LM trong cc fermentor vi mi trng lng Ln men b mt: LM trong cc khay vi mi trng lng hay mi trng c c cht rn hay xp Theo nguyn l hot ng ca thit b (Operation mode) Ln men tnh (ln men theo m) (batch culture) Ln men tnh c b sung c cht (fed batch culture) Ln men lin tc (continuous culture) iu kin ln men Ln men hiu kh: thng kh bng h thng trc khuy c sc kh (sx biomass) Ln men k kh (sn xut ru, bia)
Ln men tnh c coi l 1 h khp kn do khng Air c tc ng ca cc yu t bn ngoi (tr kh) Harvesting
Cell growth can be expressed as: Dry cell weight (g dry biomass /L culture) Wet cell weight (g wet biomass /L culture) Optical density (O.D) Cell counts on Petri dishes Other parameters used for the characterization of batch cultures Number of generations or duplications Duplication time Yields: Product/Substrate (YP/S); Biomass / substrate (YX/S) Productivity: Volumetric productivity (P); Specific productivity (qP)
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LnX
time
Nng sut =
Pha Lag: di t vi pht ti vi gi. Tlag = f (k mi trng: pH, To, MT nui cy v VSV). Tlag khng to ra sp, nh hng ti tng nng sut ca qt LM T lag
Culture
Unload
Mass Balance in batch culture Supposition: Perfect mixing = all parameters in the fermentor are similar at a time in any point For biomass: Biomass in + Biomass grow = Biomass out + Cumulative Biomass In a Batch fermentor: grow = cumulative For substrate: Substrate in = Substrate out + Metabolized substrate + Cumulative substrate In Batch fermentor: 0 = metabolized + cumulative
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n gin, hn ch c s ly nhim. Thch hp bt u nghin cu 1 sn phm. Cho php chn cc thng s v nghin cu c s tng tc ca chng. X c MT dinh dng, , tc ti u to thnh sp
Nng sut thp. Nng c cht cao c ch VSV. Khng kim sot c tc sinh trng ca VSV. Tnh ng u ca SP khng cao so vi cc PP khc. Chi ph ln (h thng tit trng bng hi nc, nc v sinh thit b, in, din tch, sc lao ng
Khc phc nhc im ca Batch culture bng Fed- Batch Culture v Continuous Culture
Preparation of the starting culture medium Preparation of the feed culture medium
Fermentor sterilization
Fermentor inoculation
Feed sterilization
Feed addition
Fermentation
Air Harvesting
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Tng nng sut ca qu trnh do gim 1 s cng on (ra thit b, tit trng, loading v unloading). Sn phm c tnh ng nht cao
Fermentor sterilization
Fermentor inoculation
Air
1 Fermentation
Feed sterilization
2 Feed addition
Harvesting
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Ln men lin tc - Continuous culture u im Nhc im Dch ln men lun c b sung Nguy c vy nhim cao nn cc cht c ch b pha long Kh nng xut hin t Tng nng sut ca qu trnh do bin cao YX/S, YP/S khng i (= constant) Chi ph thit b t tin iu khin v kim sot c tc So vi cc pp khc, pp c cht chnh vo thit b nn ny cha c nhiu kinh nghim p dng quy duy tr YP/S ti u Tng nng sut ca qu trnh do m cng nghip gim 1 s cng on (ra thit b, tit trng, loading v unloading). Sn phm c tnh ng nht cao
2.3.4. Thit b ln men PHN LOI THIT B LN MEN Theo nguyn l hot ng: fermenter hot ng lin tc, gin on, bn lin tc Theo cu trc thit b: fermenter c cnh khuy (Shaking Tank), khng c cnh khuy Theo hnh dng thit b: Thng (H/ 3); Thp (H/ > 3); ng (dng thp nm ngang, t s dng) Theo nng sut: nh, va, ln Fermenter Types: Shaking Tank, Bubble Column, Airlift, Packed Bed, Fluidized Bed
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Anchor
Rake anchor
Baffles
Propeller
Rushton Turbine
Palette
Coiled helix
Working volume = (60-70) % of V tank Height Diameter ratio H/D =1; H/D > 1 Heat transfer **
Airlift
CYTOLIFT V= 580 ml H= 38 cm D= 10 cm
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a) Continuous jacket b) External coiled jacket c) Internal coiled jacket d) Coiled baffles e) With external heat interchanger
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600 BC
Spontaneous, natural processes
XIX century
XX century
Submerged Aerobic Culture
Cn x nhu cu oxy
CHmOl + aNH3 + bO2 ycCHpOnNq + zCHrOsNt + dCO2 + cH2O
The oxygen demand of the cell and the low solubility of oxygen in water, required a continuous air supply into the fermentor and a continuous transfer of oxygen from the gas phase into the liquid phase
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Oxygen Mass Balance: O2 in - O2 out = O2 metabolized + O2 cumulative O2 in = yO2 inQAir Air O2 out = yO2 outQEA EA O2 metabolized = qO2XV O2 cumulative = d(CO2V)/dt = VdCO2/dt Assuming: QAir Air = QAE AE
P
manometer
O2 probe
pO2
Flow meter
But the oxygen transfer in a fermentor finds resistances before to reach the cells
Mass transfer takes place by two basic processes: Convection and Diffusion Therefore, a full treatment of mass transfer requires a fully known flow field. However, a simplified treatment in which the overall mass transfer is schematically divided into different transfer steps is used with good results
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How we can improve the YP/X? Strain development approach - Fermentation process optimization
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3.2. Cng ngh sn xut acid hu c thc phm 3.3. Cng ngh sn xut acid amin 3.4. Cng ngh sn xut sinh khi vi sinh vt 3.5. Cng ngh sn xut polysaccharide t VSV 3.6. Cng ngh enzyme 3.7. Cng ngh sx cc sp ln men truyn thng
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Cc hnh nh kho c v vic lm ru t khi nn nng nghip xut hin (ngi Xume, 4000 B.C) Tm thy ru trong lng m ca ngi Ai Cp CTHH: C2H5OH, M= 46 Cht lng, trong sut, khng mu, mi v c trng T trng d2020 = 0,79067; d204 = 0,78927 Nhit si: 78,35oC c i vi ngi v VSV
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nh hng ca ETHANOL vi VSV Ethanol xm nhp vo mng t bo VSV, gy ng t mt s protein, nh hng n qu trnh TC ca t bo. VSV b cht hoc b c ch
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Brasil: sx 0,9-1 triu lt ethanol/ ngy (ng u th gii) 2007, sn lng ethanol ton cu: 55,7 t lt 2015, sn lng ethanol ton cu: 162 t lt (Theo FAO, y ban Kinh t M Latinh (CEPAL) v Ngn hng pht trin Braxin (BNDES)
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TNH HNH SN XUT ETHANOL VN 1898, sn xut cng nghip ru VN. 1962-1965, xut khu ru sang ng u, ch yu l Lin X 1975-1978, nh my ru H Ni sn xut >8 triu lt cn/ nm, 12 triu lt ru mi/ nm (xut khu 6 triu lt/ nm). 1970-1980, xy dng cc x nghip ru a phng (qui m 100 - 500 nghn lt cn/ nm). C nc c gn 300 nh my ru ln nh, tng cng sut 40-80 triu lt/ nm (cha k lng ru dn nu t do). 1997, sn lng ru quc doanh t 17 triu lt/ 1997. Nu k c ca dn th t 264 triu lt). Bnh qun 3,4 lt/ ngi/ nm. 2005, c khong 180 -200 triu lt ru cc loi, trong ru cn t tinh bt chim 30 - 40%.
9/2007, Cty Petrosetco k tha thun hp tc thnh lp lin doanh xy dng nh my sx Ethanol vi Tp on Itochu Nht Bn. Nh my d nh t ti KCN Hip Phc - TP.HCM vi cng sut 100 triu lt Ethanol/ nm t sn lt. Vi 1,2 triu tn sn lt xut khu hng nm, Vit Nam c th sn xut c t nht 400 triu lt Ethanol/ nm. Ethanol s dng pha vo xng (p ng 50% nhu cu ca th trng xng Vit Nam)
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3.1.1.2. NGUYN LIU SN XUT ETHANOL NGUYN LIU sx Ethanol theo quy m CN Nguyn liu cha ng: mt r Nguyn liu cha tinh bt: go, khoai m (sn), bp (ng), nc thi cha tinh bt (b khoai m) Nguyn liu cha cellulose (rm, tru, b ma Whey (lactoserum)
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C2H5OH
C6H12O6
VSV
2C2H5OH + 2CO2
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Ch tiu
Thnh phn ho hc ca mt s nguyn liu cha tinh bt trong sx ethanol (%) Khoai ty Ng Sn Go t Tm 12 - 21 68,4 74,74 69,2 42,0 1,2 3,2 8,3 0,205 7,3 5,3 0,5 1,5 4,1 1,11 0,5 22,5 0,1 0,3 5,1 0,41 1,2 2,0 72 -80 12,5 13,12 11 11,5
VK lactic Nguyn + mc+ liu cha men tinh bt Mycomalt Ch phm mc + men LM CN
i hi k thut cao Quy m sx khi cy VSV vo CN bnh ln men Tn din tch, cng sc Quy m sx va & CN
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1,5 2,5%
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Sn xut ethanol bng pp ln men (tt) Phng php mycomalt: g ng ha v ru ha din ra 2 thit b.
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Sn xut ethanol bng pp ln men (tt) Phng php amylose: g ng ha v ru ho xy ra gn nh ng thi, trong cng thit b
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THIT B NGHIN NGUYN LIU (tinh bt) My nghin ba kiu git nc Nghin trc
Nghin bt mn
Nghin cao p
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NU NGUYN LIU H HA - GELATINISATION Mc ch Ph v mng t bo ca ht tinh bt Chuyn ho tinh bt sng thnh tinh bt chn Chuyn tinh bt thnh dng ha tan Thng s k thut Nhit nu = f (nguyn liu v kch thc ca ht tinh bt) Phng php Gin on Bn lin tuc Lin tc
DCH HA
NG HA
Chuyn tbt thnh ng, cung cp c cht cho qt ln men Quyt nh phn ln hiu sut thu hi ru.
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LN MEN ETHANOL T TB THNG S K THUT CA MI TRNG LN MEN Nng cht kh: 16 18% Hm lng ng kh: 90 -100 g/L Khong (Nit, Mg, P) Chnh pH: pHop 4,5 - 5 Dch ln men 12oP 16oP 18oP Dch ln men t sn Dch LM t r ng Yu cu Nit trong dch ln men (nit hu c, v c) 140 -150 mg/L 200 mg/L > 280 mg/L 300 mg/L 150 200 mg/L
THNG S K THUT TI CTY RU BNH TY Trnh t cng vic Xung bt: 1080 L nc + 300kg go (xay 1mm) + 445g CaCl2 + 283mL Termamyl Nng nhit Gi nhit Nng nhit Gi nhit, cho vo 120mL H2SO4 To (oC) 32 32 83 83 83 95 95 pht 10 40 50 15 30 30
Bm v h nhit dch nu bng corbin sang 95 58 thng ng ha, m bo To thng ng ha l 58oC Cho 32g San super, o trn, gi nhit dch khng lm mt mu iod, h nhit Bm sang thng ln men, + 50g ure 56 58 56 30
60 40
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THNG S K THUT TI CTY RU BNH TY Trnh t cng vic Ln men: t l men 8 10% Kim tra mu sau 4 ngy ln men: Bal, chua Kt thc ln men: Bal <0; chua < 2,5mg/L; ru > 10%v/v Lc bng my ly tm Chng ct (600mm Hg) Lc lnh ru thnh phm 5oC To (oC) pht
68 - 72
Ch tiu Nng lng th (MJ/kg VCK) VCK (%) ng kh (%) Saccharose (g/100g VCK) Glucose (g/100g VCK) Fructose (g/100g VCK) Nit (g/100g VCK) Protein th (%) Khong (g/100g VCK) Ca (g/100g VCK) Mg (g/100g VCK) K (g/100g VCK) P (mg/100g VCK) Na (mg/kg VCK) S (g/100g VCK) Cu (g/kg VCK) Fe (mg/kg VCK) Mn (mg/kg VCK) Zn (mg/kg VCK)
Mt r sx cng nghip c 9,47 75,9 34,6 23,3 5,1 6,2 0,46 2,87 11,7 0,48 0,32 1,94 47 661 0,3 1,58 97 52 3,8 VN 9,87 76,7 39 21,2 8,7 8,47 0,29 1,80 6,2 0,44 0,18 0,92 30 75 0,33 3,01 417 52 12,6
THNH PHN HA HC CA MT R
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LN MEN ETHANOL T MT R
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LN MEN ETHANOL T MT R
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CELLULOSE
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Tc nhn thy phn Acid Ch phm nm mc Ch phm enzyne cellulase (thy phn lin kt 1,4beta-D-glycosidic trong cellulose, lichenin v beta-Dglucans) Kt hp acid v enzyme
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3.1.1.5. CHNG CT (DISTILLATION) & TINH CH (RETIFICATION) ETHANOL SAU LN MEN Qu trnh thu ethanol v tp cht d bay hi t dch ln men
Chng ct: thu sn phm cn th (raw spirit) 82 87% Tinh ch: thu sn phm cn tinh luyn 94 98% Kt hp chng ct & tinh ch: s dng nhiu thp chng ct
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Sn phm: C2H5OH Aldehydes: by oxidation of alcohol Acids: by oxidation of aldehydes Fusel Oil: by conversion of free amino acids in water to higher alcohols Easters: by esterification of alcohols and acids Volatile sulphur compounds: by combination of sulphate and sulphur with amino acids
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CH TIU K THUT CA ETHANOL Nng ru 15oC Acid hu c (tnh theo mg acetic/l ) Furfurol Metanol Aldehyde tng s (mg/L) Ru cao phn t (mg/L) Este (mg/L) Hm lng kim loi ng ( Cu mg/l ) Xyanhydric ( HCN mg/l cn 100o ) (vi cn sx t khoai m > 96,2 < 15 0 0 <4 <4 < 30 < 30 <50
HNG S DNG MEN Men bnh m SPC (Thc n cho gia sc) Cao nm men (yeast extract)
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CC SN PHM PH CA QU TRNH LN MEN ETHANOL HM RU Thnh phn Cht kh (%) Protein th (% tng cht kh) Lipid Cellulose Cht chit phi nit Tro Dch hm t nguyn liu Khoai ty 6 27 2,7 8,1 49,9 12,6 Khoai m 3,3 14,2 1,8 2,8 74,2 6,7
HNG S DNG HM RU Thc n cho gia sc Nguyn liu lm phn vi sinh Nguyn liu sx vitamin bng VSV
Sn phm
Oxi ha Tch nc
Nn ha lng
Lm ngui
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Trung Quc Cy la min Philippines Thi Lan Brasil Ma, Cy la min Ma, sn Ma
This is the simplest of all the processes for producing bioethanol by fermentation. Sugar cane has an energy production per hectare that is substantially higher than the other feedstocks considered here, but the process conversion efficiency is only about 0.35-0.40 GJ bioethanol per GJ feedstock. The sugar cane residue or bagasse can be burned to generate electricity, producing about 0.08 GJ electricity per GJ feedstock.
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Wet milling, several types of residues are produced; dry milling produces only one type of animal feed product. The starches were broken down into C6 sugars. The current conversion efficiency of both process routes is about 0.55 GJ of ethanol per GJ wheat (USDA 2002). The processes are well established but there is some limited scope for efficiency improvements.
The process is similar and very slow. The wheat is first crushed or milled. In its passive form, malting is a process by which under controlled conditions of temperature and humidity, enzymes present in the wheat break down starches into C6 sugars. These sugars are washed out of the wheat with water, whilst the leftover residue can be sold for animal feed. Fermentation at 32-35oC; pH 5.2. Ethanol is produced at 10-15% concentration and the solution is distilled to produce ethanol at higher concentrations.
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It requires the production of ethanol from both C5 and C6 sugars. This process is technically feasible but is complex and expensive and there are few industrial examples. Ongoing research and development in the US aims to address cost issues and develop a more efficient process. This is thought by many to be a step on the way to the eventual goal of an enzyme hydrolysis process
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9 FUEL ICONS Unleaded Petrol Super Unleaded High Octane Diesel Bio-Ethanol Bio-Diesel CNG LPG Leaded Four Star.
www.gbd.edu.vn
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Husks All the discarded husks of the sugar cane. The first step in making rum is to crush the cane to get out all the juiceyou can see they make a lot of rum here at River's.
SX RU RUM T MA
After all the sugar cane juice is squeezed out, the next step in the process of making rum is to boil it down in giant kettles (ex: copper kettles) - like the ones you see here.
SX RU RUM T MA
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After the sugar cane juice is boiled down in the coppers it has to ferment for a couple of days.
SX RU RUM T MA
Condenser The final step is the condenser, where the alcohol gets seperated out thru coils and a cooling process. What comes out is the final product- pure white rum.
SX RU RUM T MT R
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SX RU RUM T MA
Whisky: The difference between rum and whisky manufacture begins with the starting point. Whisky starts with barley that must be malted before it is converted from a starch to a sugar that can be fermented. After fermentation, it is also distilled usually using a pot still or single column. Cognac: The sugar that is fermented to make cognac, come from grapes. As is the French style, only grapes grown in a particular area in France can be used. First the grapes are fermented into wine and this is distilled in special stills to yield cognac. Brandy is made the same way but because the grapes arent grown in the Cognac region, it can not be called cognac. All spirits are aged in a special barrel prior to blending and bottling.
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3.1.3.2. NGUYN LIU SN XUT BIA 3.1.3.3. VSV TRONG QU TRNH LN MEN BIA 3.1.3.4. CNG NGH LN MEN BIA SN PHM
3.1.3.1. GII THIU V BIA - LCH S PHT TRIN 7000 B.C, Ngi Babylon (a ra vn t lut php quy nh v vic bn ru bia) pha ch c 20 loi bia. 3000 B.C, ngi Ai Cp c i pht minh ra ng ht ung bia cn ln cn v la m 500 400 B.C, bia l sn phm dnh cho ngi lao ng 768: Baviere s dng hoa houblon (trc kia dng tho mc to v ng cho bia) 1516: lut sx bia u tin c p dng (bia nu t malt i mch, hoa houblon v nc 1857: Pasteur nghin cu qu trnh ln men ethanol, thanh trng bia 63oC/ 20 30 1883: Hansen v ging men thun khit
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GII THIU V BIA Sn lng bia Sn lng bia trn th gii: >120 t lt/nm Lng bia tiu th: CHLB c 170L/ngi/nm VN: 5 10 L/ngi/nm
Nguyn liu
c: 100% malt i mch Php: s dng 30% th liu VN: 2030% go
Ph gia
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Anherue busch (M) Heineken (H Lan) Sn lng bia trn th gii: Miller (M) >120 t lt/nm Kirin (Nht) Lng bia tiu th: Fosters (c) CHLB c: 170L/ngi/nm Danone (Php) Brahma (Brasile) VN: 5 10 L/ngi/nm Guiness (Anh) (bia en) SAB (Nam Phi) Pilsen (Tip) Carlsberg (an Mch) (men chm)
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Weizen
Bia l sn phm ca qu trnh ln men ethanol t dch nha, khng qua chng ct
Bia en Munich
Nguyn liu chnh: malt ai mch Th liu: nguyn liu giu glucid Hoa Houblon Nc
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H enzyme trong ht malt s lm nhim v ct cc hp cht cao phn t trong ni nh ca ht thnh cc cht c trng lng phn t thp hn (dextrin, acid amin, pepton, ng n) ho tan vo nc tr thnh cht chit ca dch ng.
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NGUYN LIU SN XUT BIA - TH LIU TH LIU CHA TINH BT: ht i mch, go, tm (VN), bp tch phi (chu u), khoai m, khoai ty, tinh bt
nguyn liu giu glucid, thay th mt phn hoc ton b malt i mch trong sx bia
U IM H gi thnh sn phm Ci thin mt vi tnh cht ca sn phm (tng bn ca keo) To ra chng loi bia c cc mc cht lng khc nhau
NHC IM nh hng cho qu trnh nu (do th liu c t hoc ko c enzyme) nh hng ti qu trnh ln men (Protein ha tan trong dch nha gim, khng cn i thnh phn dinh dng)
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NGUYN LIU SN XUT BIA - TH LIU TH LIU S DNG TI VN - GO cu trc ht tinh bt cht, nhit h ha cao, kh ng ha Bia Heineken: 100% malt i mch Bia Tiger: 15% go Bia Si Gn: 20% go TH LIU S DNG TI CHU U BP Tch phi loi cht bo Cht bo ph hy h bt, nh hng n cht lng cm quan
NGUYN LIU SN XUT BIA - HOA HOUBLON Nguyn liu khng th thay th trong cng ngh sx bia To v ng du, hng th m c trng Tng kh nng to v gi bt Tng bn keo v tnh n nh sinh hc ca sn phm. Trong sx bia ch s dng hoa ci cha th phn (c ht lupulin) do hoa c v hoa ci th phn c hm lng cht mi v cht to v thp (khng c ht lupulin)
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NGUYN LIU SN XUT BIA - HOA HOUBLON HOUBLON Houblon to hng 3,85 5,14% acid Ch tiu Mi v Alpha acid m Hoa vin Thm c trng, khng mi l 8 0,3 <10 Houblon to v ng 8,62 9,31% acid Hoa cao Thm c trng, khng mi l 30 2
NGUYN LIU SN XUT BIA - HOA HOUBLON Thnh phn ha hc c trong hoa houblon (% khi lng cht kh) m 10 -11 Alpha acid 2 -12 Beta acid 2 - 10 Tinh du (> 200 cht nh terpene, ester, 0,5 2 ketone, hp cht cha lu hunh) Polyphenol 2 -5 Protein 12 18 Lipide 2-4 Cellulose 40 50 Pectin 12 Tro 1-2
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nh gi cht lng hoa houplon Ch s Mu hoa Mu ht lupulin Mi Loi 1 Vng ng nh Thm d chu, c trng Loi 2 Vng, vng sm Thm, khng c mi tp cht Cnh hoa c th b rch (1,5cm), c chm c ph <3% <1 % >12 % <11 % <3 % Loi 3 Vng xanh Vng sm Hi nng Rch nhiu, chm c ph nhiu <9% <5 % >10 % >12 % <13 % Vng vng ng Vng lc
Cnh hoa To u chc, khng rch Tp cht Hoa bnh Hoa ng Tro m <1,75% 0% <10 % <15 % <13 %
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Hoa kh (W 4 -6%) Nghin Ry (2-8mm) Bt hoa kh Bao gi (chn khng, CO2, N2) Bo qun Hoa dng ht, vin: W 4 6%, bt hoa kh (c th thm 20% bentonote SiO2 ri to thnh vin Hoa cao trch ly; dng st c hm lng polyphenol thp, lm gim bn keo
NGUYN LIU SN XUT BIA NC Quyt nh n cht lng bia Chim 90 95% trong bia YU CU V NC TRONG SN XUT BIA YU CU CM QUAN: trong, khng c mi v l YU CU VI SINH: khng c E.coli v Streptococci/100 ml nc. YU CU HO L pH 7.0 cng 2-4mg/L kim <50mg CaC03/L Cht kh <1500mg/L Sulphate < 500/L Clorute 250 300mg/L Nitrate <30 mg/L
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Mnh, xy ra trn b Mnh, xy ra trong mt mi trng lng mt 33% Cc tb kt chm, chui to thnh lp dy ni ln b mt mt, bia trong chm 100% Cc tb kt chm, chui lng xung y thng ln men rt nhanh, bia t trong nhanh hn
Tap chat
Nghien
Nc
Loc
Ba malt
Houblon
Tach ba houblon
Ba houblon
vo trung
Lam lanh
, bao hoa O
Nam men
Nhan giong
Can
Can
CO
Bao hoa CO
, tang tr
Chai
/ lon
Chiet chai
/ lon
Thanh trung
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3.1.3.4.1. Sx bia NGHIN Mc ch NGHIN Tng b mt tip xc vi nc Nc xm nhp vo cc thnh phn trong ni nh nhanh hn Cc yu t nh hng n qu trnh nghin m Cu trc ht tinh bt Thit b Phng php nghin Nghin th liu: go Nguyn tc: nghin go cng mn cng tt
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Sn phm nghin malt: v tru, ht tm ln, tm nh, bt. Mc nghin MALT Nghin th: 20 25% (280mL/100g malt) Nghin mn: 50 60% bt (210mL/100g malt) Nghin rt mn: 85 90% bt (200mL/100g malt) Mc nghin nh hng ti trch ly, thi gian lc b Yu cu v mc nghin Ch tiu Nghin trc (lc bng ni lc) Tru 15 18 Ht tm ln 12 15 Ht tm nh 30 35 Bt 25 - 35 malt (% khi lng) Nghin ba (lc bng thit b lc khung bn) 9 12 12 15 30 35 40 - 45
3.1.3.4.2. Sx bia NU DCH NHA Trch ly cc cht chit c Mthp (ng, acid amin, vitamine, khong) t malt i mch v th liu vo nc Thy phn mt s cht c Mln thnh cc cht c Mnh (c cht cho nm men) Cc phn ng xy ra trong qu trnh nu dch nha Thy phn Protein: to thnh polypeptide, peptide (endoenzyme), acid amin (exoenzyme) Thy phn tinh bt: to thnh dextrin, oligosaccharide (alpha amilase), maltose (beta amilase) Thy phn hemicellulose: to thnh beta glucan mch ngn Thy phn mt s cht hu c c cha acid phosphoric: to thnh acid phosphoric
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3.1.3.4.2. Sx bia NU DCH NHA (tt) THNG S K THUT T l malt: nc = 1:4 1:5 T l th liu: nc = 1:1,5 2 pH: 5,4 5,6 (chnh pH 1 ln) Thc hin cc im dng trong qu trnh nu Thit b nu c cnh khuy
CNG NGH SX BIA Nh my Bia LIDA 500kg nguyn liu/m thu c 3100 3300L dch ln men. T l malt: go = 3/7 (130 kg go: 370 kg malt). Ni go: 130 kg go + 520- 650 lit nc + 200g CaCl2. Dng H3PO4 chnh pH 5,3 -5,4. 40oC/15. Trong 15 tng t 40oC n 72oC, 72oC/20. Trong 10 tng n 83oC, 83oC/5. Trong 5, h xung 72oC, 72oC/20. Trong 20 tng ln 100oC, 100oC/15. Trong 5, h xung 65 67oC Ni malt: 370 kg malt + 1480 1850 lit nc + 300g CaCl2. Dng H3PO4 chnh pH 5,3 -5,4. Khi nu go c 60 th nu malt. 40oC/20. Trong 15, Tng n 50oC, 50oC/20. Trong 10 tng n 65 67oC .
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Ni malt v go u 65 67oC. Trn 2 ni lm mt. Duy tr 67oC/30. Trong 15, tng ti 72oC, 72oC/15. Trong 15, tng ti 76oC, 76oC/15. Tng thi gian nu go v malt l 210. Sau lc dch v ra b (40 50). Dch lc c un si trong 120. Trong thi gian un si, cho hoa houblon 3 ln. Thi gian cho hoa: ln 1 sau 10 un, ln 2 sau 30, ln 3 sau 75. B sung 250mL caramen chnh mu dch ng. + 2g ZnCl2 b sung nguyn t vi lng cho men. Sau 120 un, chuyn qua lng 25 tch cn, cc cht khng ha tan nh xc hoa, cht kt ta Lm lnh nhanh. Chuyn dch vo tank cy men trc
3.1.3.4.3. Sx bia Lc dch nha Mc ch Tch b b malt Ra b malt Phng php Ra b malt bng nc nng Toop = 78 80oC
Cc yu t nh hng n qu trnh lc Mc nghin malt Nhit nht Thit b lc tch b malt: thng lc hnh tr y bng (thng s dng) Cu to thit b: phc tp (1 y gi l mt sng v h thng cnh o trn, dao gt b, h thng tho b malt)
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3.1.3.4.4. Sx bia un si dch nha vi hoa Houplon MC CH Chit cht ng t hoa houplon (alpha acid) vo dch nha, ng phn ha mt s cht ng V hot enzyme, ng t Protein km bn nhit iu chnh nng cht kh trong dch nha To thnh mt s cht c li cho sn phm Tch mt s hp cht d bay hi nh hng xu n cht lng sn phm
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3.1.3.4.4. Sx bia un si dch nha vi hoa Houplon (tt) LNG HOA HOUPLON B SUNG (Tnh theo acid) 7 20 g acid/hL THI GIAN UN SI: 60 120 PHNG PHP B SUNG HOA Mt ln: sau khi dch nha si Hai ln: sau khi dch nha si + 30 trc khi kt thc qu trnh nu Ba ln: sau khi dch nha si + 30 trc khi kt thc qu trnh nu + 15 trc khi kt thc qu trnh nu
Nguyn l: Bm dch nha vo thit b hnh tr ng, dung dch chuyn ng theo ng xon c. Dng lc ly tm lng (lng xoy tm)
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Thnh phn dch nha pH Nng cht kh Glucose Fructose Saccharose Maltose Trisaccharide (maltotriose, raffinose) Dextrin Beta glucan v gum Nit tng Nit amin t do (FAN) Cc cht khc (acid nucleic, vitamine, khong, polyphenol) 5,2 5,4 10 13oPt 5 15 g/L 0,5 2 g/L 2 6g/L 45 65g/L / / 200 800mg/L 500 1.100mg/L 100 250mg/L /
3.1.3.4.6. Sx bia Lm ngui v np oxy vo dch nha MC CH H nhit dch nha, chun b cho ln men Cung cp oxy cn thit cho sinh trng ca men YU CU Lm ngui nhanh, thit b kn Np Oxy (khng kh) v trng, lng oxy ha tan (6 8mg oxy/lt dch nha, P = 1atm, To = To ln men
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3.1.3.4.7. Sx bia NHN GING MEN & LN MEN Nhn ging 2 giai on PTN: thch nghing, cy vo ng 10ml nc nha+ pepton 12h Cy qua bnh tam gic 150 170mL dch, 12h. Cy qua bnh tam gic 1,5L, 24h Chuyn vo tank 1 (V tank 1: 200L, V dch ng: 150L), c cung cp oxi trong tank, Nui 48h. Chuyn tank 1 vo tank 2 (V tank 2: 1000L, V dch 800L, nui 48h, 20 25oC Bm men t tank 2 vo Tank cha dch ln men (V 12.000L)
3.1.3.4.7. Sx bia NHN GING MEN & LN MEN (tt) MC CH LN MEN: Chuyn dch nha thnh bia Ln men chnh Chuyn ha ng thnh ethanol, CO2 v cc sn phm ph, sp trung gian Tc ln men din ra nhanh, cc cht cn (protein) v t bo men cn st li lng dn xung y, bia trong dn Ln men ng cn li, bo ha CO2 cho bia ti, n nh hng v, cht lng, trong cho sn phm, tng gi tr cm quan
Ln men ph
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3.1.3.4.7. Sx bia NHN GING MEN & LN MEN (tt) Ln men chnh Primary fermentation THNG S K THUT 0,5L men/hL dch nha hay 10 25.106 tb/mL dch nha To= 13 15oC Thi gian: 7 9 ngy Nng cht kh cn 1,15 0,2% kt thc ln men chnh CC YU T NH HNG N LN MEN CHNH Mi trng ln men (dch nha): pH, nng cht kh, thnh phn ha hc Nm men iu kin ln men: To,
3.1.3.4.7. Sx bia NHN GING MEN & LN MEN (tt) Ln men Ln men ph Secondary fermentation THNG S K THUT LM PH 4 8.106 t bo men/mL dch nha To= 0 5oC Thi gian: 7 30 ngy (ph thuc vo tng loi bia) Thi gian ln men ph quyt nh hm lng diacetyl trong bia, cng nh mc chn ca bia. Lin tc tch nm men cht v cc kt ta ra khi thit b trong qu trnh ln men ph
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3.1.3.4.8. Sx bia Lm trong bia MC CH Tch trit cc phn t rn lng, khuych tn trong bia Tng trong, Lm n nh thnh phn c hc Tng bn keo v bn sinh hc cho sn phm YU CU Nhit lm trong: 1 2oC THIT B LC Thit b lc khung bn: r, d vn hnh, bi v VSV d xm nhp Thit b lc ly tm, lc da, lc nn.
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3.1.3.4.8. Sx bia Lm trong bia (tt) Cc cht lm trong Cho thm mt hay nhiu cht lm trong vo bia (khng b bt buc phi cng b nh l mt thnh phn ca bia) Vit Nam, thng s dng thit b lc khung bn, b sung cht tr lc diatomid
Lc khung bn
3.1.3.4.9. Sx bia Bo ha CO2 Hm lng CO2 trong bia non: 4-4,5g/l. Nu khng t tiu chun: b sung thm CO2 MC CH Chun ha hm lng CO2 cho bia chai v bia lon YU CU Nhit : 0 1oC P = 0,15 -0,2 MPa
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3.1.3.4.10. Sx bia Chit v thanh trng bia MC CH Ko di thi gian BQ sn phm D vn chuyn v s dng YU CU Nhit : 0 1oC P = 0,15 -0,2 MPa
3.1.3.4.10. Sx bia Cht lng sn phm Ch tiu cm quan Mu sc trong bt Mi V Yu cu Vng rm sng Trong sut Bt trng, mn lu tan Thm c trng ng du, m , c hu, c trng
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Ch tiu ha l Cht ha tan ban u (% khi lng) ng st (%) cn pH chua (mL NaOH/100mL bia) mu (EBC) CO2 (g/L) bn ca bt (giy) Oxi ng Diacetyl
Yu cu 10,2 0,2 1,8 2,2 4,3 0,2 4,0 4,2 1,6 0,2 7,0 7,2 4,0 4,5 150 250 < 3ppm 20oBU < 0,1ppm
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www.gbd.edu.vn
3.2. Cng ngh sn xut acid hu c thc phm 3.2.1. Sx acid lactic 3.2.2. Sx acid acetic 3.2.3. Sx acid citric
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3.2.1.1. GII THIU V ACID LACTIC 1780, Carl Wilhelm Scheele (nh ho hc Thy in) tch acid lactic t sa b chua. 1881, sx theo quy m cng nghip nh Frmy (nh khoa hc Php) sx acid lactic bng pp ln men C3H6O3 CH3-CHOH-COOH M= 98,08; khng mu, mi nh, ht m cao, To si = 122oC, tan 17oC 2 dng ng phn D- v L+. Dng ng phn sinh hc L+ c trong c th ngi Ph thuc vo tinh sch ca acid lactic p dng vo cc lnh vc khc nhau
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D-acid lactic
L-acid lactic
T l D:L = 50:50 (Hn hp raxemic (k hiu l DL-acid lactic) Raxemic c dng lng, tan trong nc, cn, khng tan trong CHCl3, To si = 122oC, nhit nng chy 16,8oC Raxemic ch c c khi tin hnh tng hp hu c
(dng L c trong c th ngi)
LACTIC TRONG LNH VC THC PHM LACTIC TRONG LNH VC PHI THC PHM Sn lng acid lactic c tnh khong 50 ngn tn/nm, khong 2/3 sn lng c sn xut bng pp ln men
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3.2.1.2. VAI TR CA ACID LACTIC (tt) trong thc phm Tht gia sc, gia cm, c (Meat, Poultry & Fish) ung (Beverages) Rau ngm gim (Pickled vegetables) Salad v nc st (Salads & dressings) Bnh ko (Confectionery) Cc sn phm t sa (Dairy) Baked Goods Savory Flavors
3.2.1.2. VAI TR CA ACID LACTIC (tt) LNH VC PHI THC PHM Dc phm (Pharmaceutical), m phm (Comestics) Vt liu sinh hc (Biomaterials) Mt s ngnh k thut (Technical) Cht ty ra (Detergents) Thc n cho gia sc (Animal Feed) Cht do t phn hy (biodegradable plastics) Nha PLA (Poly lactic acid) chu c 175oC, thch hp lm chai rt nng, khay s dng trong l vi ba, cc loi si v thit b in t chu nhit
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Glucose, sucrose, lactose Mt r ng Huyt thanh sa Nguyn liu cha tinh bt (ng, khoai ty, nc thi cha tinh bt)
X L MT R
H2O
5% H2SO4 (w/w)
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3.2.1.3. C CHT SN XUT ACID LACTIC (tt) X L NGUYN LIU CHA TINH BT
Nguyn liu cha tinh bt Thy phn Nguyn liu cha cc ng n gin
Nit CaCO3
3.2.1.4. VSV LN MEN ACID LACTIC 1857, Pasteur kt lun VK khun lactic ln men sa 1878, Joseph Lister phn lp c VK lactic t tn l Bacterium lactic nay gi l Streptococcus lactic. VK lactic thuc h Lactobacterium, khc nhau v hnh dng, sinh l v kh nng ln men VK lactic thuc nhm k kh khng bt buc VK lactic chia lm 2 nhm: -VK lactic ng hnh (cu khun v trc khun): sn sinh 85-95% acid lactic. -VK lactic d hnh: sn sinh 50% acid lactic v ru. Trong cng nghip ln men lactic s dng VK lactic ng hnh
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Cc VSV chnh s dng trong cng nghip sx acid lactic hin nay: Lactobacillus casei. Streptococcus thermophilus. Lactobacillus acidophilus. Lactobacillus bulgaricus. Lactobacillus delbrueckii Chng c th s dng tinh bt: L. amylophilus, L. amylovorus
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Strep. thermophilus
Lactobacillus bulgaricus
Lb. acidophilus Lb. delbrueckii (2 (2 7 0,4 0,4 0,8m) Dng k thut bc x t ngoi to Lac. delbrueckii t bin. Chng c tc pht trin mnh hn, pha lag ngn hn, nng sut v hiu sut sinh acid lactic ln hn
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3.2.1.4. VSV LN MEN ACID LACTIC (tt) Nhu cu dinh dng ca VK lactic - Carbon: cu trc t bo v cung cp nng lng - Nit: nit c trong MT (cc acid amin), b sung thm peptone, cao tht, cao nm men, casein - Vitamine: VK lactic t c kh nng tng hp vitamine. Chng cn cc vitamine nh riboflavin, thiamin, acid pantotenoic, nicotinic, biotine -Cc cht hu c: + Base cha nit: adenine, guanine, uraxine, + Acid hu c: acid citric, acid acetic, + Acid amin: L- asparagine, L-glutamine -Khong: P, S, Mg, Ca, Mn, Fe, K, Na Mn2+, Mg2+, Fe2+ c tc ng tch cc ln s pht trin v sinh ra acid lactic ca VK Lactobacillus
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Commercial lactic acid is produced naturally by fermentation of carbohydrates such as glucose, sucrose, or lactose.
Sn xut L-Lactic Acid (lactide) bng PP ln men trc tip khoai ty Hokkaido
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Sago Industry: PLA (Poly Lactic Acid) production incorporated with Sago
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GALACTIC, sx L(+) lactic theo pp Nh my B&G, sx lactic ln nht LM ln th 2 trn TG, xut khu > Chu TQ, cung cp cho th 90% sn phm cho > 50 nc. trng thc n gia sc v sx PLA (Poly-Lactic Acid)
3.2.1.5. CNG NGH SN XUT ACID LACTIC (tt) THNG S K THUT Mi trng ln men 10 15oBx pH = f (pHop ca VK s dng ln men) VSV S dng chng VK lactic ng hnh T l ging 3-5% (v/v), mt t bo 106 tb/ml iu kin ln men To = f (Top ca VK s dng ln men) Fermenter c cnh khuy v thi kh Duy tr pHop trong sut qu trnh ln men
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3.2.1.6. THU NHN V TINH SCH LACTIC Cng ngh truyn thng Cng ngh hin i
PP thm tch in &trao i ion PP tch pha Dch sau ln men (lactate canci) Lactic un dch n 8080-90oC
CaCO3
Lc khung bn
7070-80oC
Dch lc
sinh khi
Lc
Na2CO3
sinh khi
Dch trong
Tb C c chn khng Thm tch in trch ly Dung dch acid lactic Ct trao i ion
PP thm tch in & trao i ion
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SN PHM ACID LACTIC Nng acid lactic 95% Chloride 0.1% Cyanide 5mg/kg Kim loi nng (ch) 10mg/kg St 10mg/kg. Cn 0.1% Sulfate 0.25%
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3.2.2.1. GII THIU V ACID ACETIC (ETHANOIC ACID) C2H4O2 CH3-COOH L carboxylic acid n gin nht v cu to M= 60,05 To to tinh th = 16,7oC n mn nhiu kim loi, gy bng da Gim n (acetic 4 8%)
ACETIC TRONG LNH VC THC PHM ACETIC TRONG LNH VC PHI THC PHM Nhu cu v acetic acid khong 6.5 triu tn/nm (Mt/a). Khong 4 Mt/a c sn xut t cng nghip ha du hoc t cc ngun sinh hc. Tnh ti 2006, VN nhp khu acid acetic 100%
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3.2.2.2. VAI TR CA ACID ACETIC (tt) VAI TR CA ACETIC TRONG THC PHM S dng lm cht ph gia thc phm (E260) Gim n
Nhiu nc quy nh, acetic acid dng lm gim phi c sx bng pp ln men. Mi v v ca gim ph thuc vo nguyn liu v cch sx.
3.2.2.2. VAI TR CA ACID ACETIC (tt) TRONG LNH VC PHI THC PHM CN ha cht: s dng nhiu trong thuc n Sx polyethylene terephthalate (s dng trong nc gi i kht ng chai) Sx cellulose acetate: s dng trong phim chp hnh Sx polyvinyl acetate: dng trong keo dn g S dng lm dung mi pha ha cht Sx vinyl acetate monomer dng trong sn Cng nghip ch bin cao su Tng hp cht do t si Trong gia nh: acetic long lm sch cn bn.
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Nguyn liu cha tinh bt Cellulose (s dng Clostridium lentocellum SG6 nghin cu ca H Osmania, n ) Nguyn liu cha ng Cn, ru vang
VK hiu kh Sau 12h, sinh khi tng 17 triu ln Chuyn ha ru thnh acid acetic
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3.2.2.5. CNG NGH SN XUT ACID ACETIC PP HA HC (3) Methanol carbonization Butane oxidation Acetaldehyde oxidation PP SINH HC
3.2.2.5. CNG NGH SN XUT ACID ACETIC (tt) PP HA HC Cho ti 1960s, acetic acid c sx ch yu bng pp oxi ha cc ngun hydrocarbon. 1960s, PP sx acetic quy m cng nghip (methanol carbonization). 1970s, Monsanto ci tin pp ny, gim chi ph ph sx. Hin nay, s dng pp methanol carbonization, kt hp methanol v carbon monoxide to thnh acetic acid. Lng acetic sx t pp ny chim 80% tng sn lng
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CATIVA acetic acid plants in Hull, UK (left), and Chongqing, China (right)
At the heart of this success is BP's proprietary and trademarked CATIVA process for acetic acid manufacture, which, with a market share of more than 25 per cent of the 10 million tones of acetic acid produced globally each year, can justifiably claim to be leader of the pack.
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3.2.2.5. CNG NGH SN XUT ACID ACETIC (tt) PP SINH HC Pasteur pht hin ra VK Acetobacter ln men gim Ngi Php a ra cng ngh sx gim (pp ln men chm) Thi k chin tranh th gii th 2, ngi c ci tin v a ra cng ngh ln men nhanh.
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Gim n Cc yu t nh hng
Mi trng dinh dng Gim: B sung vo mi trng ban u acid ho mi trng, ngn chn s pht trin vi sinh vt c hi, b sung lng t bo nht nh. Ru ethylic: c s dng nh l mt c cht, t 2 10%V, > 10% s lm gim nng sut ln men. Cc cht C, N, P v cc nguyn t vi lng: cn b sung thm mt s mui khong nh: K2HPO4, KH2PO4, MgSO4.7H2O, FeCl3.6H2O, (NH4)2SO4, (NH4)2HPO4 ty theo nhu cu c th ca tng loi. B sung thm vitamin v cc cht kch thch sinh trng nh: glucoza, cao nm men, pepton...
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Gim n Cc yu t nh hng
nh hng ca oxy Cn lng khng kh ln cho qu trnh ln men gim. oxy ho ht 1 kg ru khan cn 2,3 m3 khng kh (theo l thuyt), hay oxy ho ht 1 mol ru cn 1mol O2. Qua thc t nhiu tc gi cho thy lng oxy cn thit phi ln gp i lng oxy ho theo l thuyt.
Gim n Cc yu t nh hng
nh hng ca nhit Acetobacter thuc nhm a m, Toop: 25 32oC nhit thp: qu trnh ln men gim xy ra chm. nhit cao: c ch s hot ng v n mc no s nh ch s sinh sn ca t bo lm tng tn tht cho vi khun v lm gim hiu sut qu trnh ln men do s bay hi acid acetic v ru etylic. nh hng ca tc nhn sinh hc (Ln gim, b gim, rui gim
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Cc h hng thng gp (Gim b c, gim chua) Do oxy ho ho hc (oxy ho ru thnh CO2 v H2O) Do oxy ho sinh hc: tc nhn l nm men Comdida mycoderma, Acetobacter xylium, Acetobacter aceti, ln gim (2), b gim, rui gim (3) gy ra. Khc phc (1,3): v sinh thit b ln men sch, thanh trng Pasteur dch nhit 60 -70oC ri cho ln men li, cho thm vo K2S2O5 vi liu lng 5 15gam/100 lt gim kt hp vi thanh trng Pasteur tng bn ca gim, y kn thit b Khc phc (2): cho 1 2% NaCl vo gim thnh phm. Sau 1 2 ngy ln gim cht, lc hoc a nhit ln n 40 500C ri lc loi b ln gim.
3.5.2.5.1. PP LN MEN CHM (PHP - PP Orleans) Xut x: t 1670 vng trng nho ni ting ti Php (dch vang nho trong nhng thng g h np dn dn b c, c mng trng ph trn b mt cha VK Acetobacter orleaneuse v tr nn chua). Hin nay ch cn mt vi nc chm pht trin cn s dng phng php ny, trong c VN
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3.5.2.5.1. PP LN MEN CHM (PHP - PP Orleans) (tt) Nguyn liu: dch nho Ln men trong thng g si 250 300 lit V dch ln men: 1/2 - 2/3 V thng Mi trng ln men: 12 14oBx VSV: Ace. orleaneuse Thi gian ln men: 10 20 ngy 5 tun Hm lng acid acetic t c: 5 -6%
3.5.2.5.1. PP LN MEN CHM (PHP - PP Orleans) (tt) Nhc im Thi gian ln men di Hm lng acid acetic thp Nng sut thp, kh c gii ha, hin i ha Tn din tch Sau 20 ngy, acetic b oxi ha (ha hc v sinh hc)
Khc phc s oxi ha acetic Trong dd acetic phi cn 5% cn Thanh trng Pasteur B sung 0,2% NaCl tiu dit ln dm
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3.5.2.5.2. PP LN MEN NHANH (C) Xut x: Shiizenbachi tm ra nm 1923. Nguyn tc: to ra b mt tip xc ln hn gia VK Acetobacter vi khng kh oxy ha ru bng cch s dng vt liu xp (v bo, li ng xp trong thit b hnh tr, cn, nn ct) lm cht mang. Yu cu cht mang: c b mt tip xc ring ln, th tch t do ln, khi lng ring b, bn c hc cao, r tin, d kim, phi c nhm, xp vi sinh vt bm cht. u im: Thi gian ln men nhanh 5 6 ngy. Nhc im: Hiu sut kinh t cha cao (ru v acid l cht d bay hi), thit b phc tp, kh iu chnh thit b thng kh.
3.5.2.5.2. PP LN MEN NHANH (C) (tt) B1.Thanh trng cht mang B2. Gn VSV vo cht mang B3. Cho dch ln men tip xc vi VSV c nh
u im Thi gian ln men nhanh Hm lng acid acetic cao Thit b n gin t tn din tch
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3.5.2.5.2. PP LN MEN NHANH (C) (tt) Thng ln men g, hnh tr, H=2,5 6m; =1,2 3m; H= 2 C h thng thi kh t di ln trn MT ln men: Cn, 0,1 0,5% ng, P, N, K VSV: Ace. xylinum gn vo cht mang (mt ca, li bp) Tgian LM: 8 - 10 ngy Hm lng acid acetic t c: 8 -10% Thit b ln men gim (Vinegar Generator)
3.5.2.5.3. PP LN MEN CHM (Submerged Process) H thng ln men acetic c gi l acetator. Cho dung dch ln men vo thit b v thi kh rt mnh. Khi trong dung dch ln men s to ra th huyn ph v dung dch ln men. Hai th ny lun trn ln vo nhau v nh vy qu trnh oxy ho xy ra rt mnh lit. u im: hiu sut chuyn ho c khi t c 98 99%.
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3.5.2.5.3. PP LN MEN KT HP
Cu to h thng ln men: phn trn l lp m (lp cht mang cha VSV), lp gia l 1 thng cha dung dch sau khi ln men phn trn chy xung, di cng l h thng thi kh mnh, kh s c thi qua phn dung dch ny ri chuyn ngc ln phn trn (ging thit b ln men chm). u im: hiu sut ln men rt cao, nng acid acetic thu c thng nm trong khong 10 -12%. Nhc im: o thit b yu cu qu phc tp, cng knh. o ging vi khun phi ph hp
Bnh ln men hi lu, c sc kh ti PTN Vi Sinh H NL. VK Acetobacter aceti c c nh trn cht mang l b ma. C cht l dung dch nc xoi
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NNG CAO NNG ACETIC TRONG PP LN MEN PP CHNG CT BNG NHIT PP CHNG CT BNG MUI S dung mui c kh nng phn ly mnh (CaCl2 hoc CH3COONa). Hiu sut thu c s cao PP CHNG CT TRCH LY Chn cu t phn ly c bay hi nh hn cu t c trong hn hp cn chng ct. cu t phn ly + cu t c trong dung dch to thnh hn hp kh bay hi v thot ra di y thp chng ct PP CHNG CT CHN KHNG
S-107 S-108
Water 1
S-106 S-103 S-102 PFLN-101 / PF-101 S-109 Plate & Frame Filtration HSRL-101 / ST-101 Heat Sterilization S-117 S-113 AFLN-102 / AF-102 S-119 Air Filtration
Biomass
Nutrients
S-112
S-111 S-121
Water 2
STRG-101 / V-103 S-110 HSRL-102 / ST-102 Heat Sterilization BLND-103 / V-104 Blending FRMN-101 / F-101 Fermentation Storage Tank
S-101
Air
S-115 AFLN-101 / AF-101 S-118 Air Filtration CMPS-101 / G-101 CF Compressor
Sulfuric Acid
S-125
Gypsum
S-122
S-131
Product
S-139 S-126
Lime
S-124 RXN-101 / R-101 Product Precipitation
S-134 RVFL-103 / RVF-103 CCRN-101 / R-103 Cont. Crystallization Rotary Vacuum Filtration RXN-102 / R-102 Gypsum Formation S-130
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$/ao 3 949 000 10 866 000 2 668 000 19 000 400 000 1 290 000 3 003 000 22 195 000
17,8%
Costo de produccin
3.3. Cng ngh sn xut acid amin 3.3.1. Sx Lysine 3.3.2. Sx Glutamic
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3.3. Cng ngh sn xut acid amin NG DNG CA ACID AMIN Thc phm Ha hc Y h c Cc lnh vc khc CC PP SX ACID AMIN THY PHN PROTEIN TNG HP HA HC CHUYN HA SINH HC LN MEN
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3.3.1. Sx Lysine
3.3.1.1. GII THIU V LYSINE 3.3.1.2. C CHT SN XUT LYSINE 3.3.1.3. VSV LN MEN LYSINE 3.3.1.4. LN MEN LYSINE 3.3.1.5. THU NHN V TINH SCH LYSINE
3.3.1.1. GII THIU V LYSINE L mt trong 8 aa khng thay th C nhiu trong tht, trng, sa, u nnh D b phn hy nhit cao C vai tr trong tng hp enzyme, hocmon, khng th, gip tng sc khng Yu cu: 1g/ngy/ngi ng dng: thm vo khu phn n ca tr em gip tr n ngon ming, hp th dinh dng tt, pht trin chiu cao; cht ph gia thc n; thc phm chc nng, dch truyn amin, b sung vo thc n gia sc
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3.3.1.1. GII THIU V LYSINE (tt) LYSINE (2,6-diaminohexanoic acid) ( amino acid) C6H14N2O2, M= 146.188, cu hnh L v D CH2 CH2 CH2 CH2 CH COOH | | NH2 NH2 1985, Nht Bn sx lysine qui m ln bng Corynebacterium glutamicum Tng sn lng trn TG: 1triu tn/nm
3.3.1.2. C CHT SN XUT LYSINE C cht: mt r, methanol, glucose. MT ln men Glucose: 50g (10 12%) tinh bt ng ( loi bo): 20 g Ngun nit: ure, cc loi mui amon hoc nc amoniac: (NH4)2SO4: 25g; Urea: 1 g Khong: mui Photpho 0,008 0,02mg/l; KH2PO4: 2.5g MgSO4.7H2O: 0.75g (0,03 0,5%) biotin 7,5g/l; treonin 40mg/l duy tr pH = 7.0 bng NaOH 2M Nhit : 28 300C.
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3.3.1.4. VSV LN MEN LYSINE Micrococcus glutamicum Brevibacterium flavum Brevibacterium lactofermentum Corynebacterium acetophilum Corynebacterium glutamicum Gleocladium sp Ustilago maydis Cc vi khun tham gia tng hp lysine u c kh nng ng ho glucose, fructose, maltose, saccharose. Khng c kh nng ng ho lactose, rafinose, pentose
3.3.1.4. VSV LN MEN LYSINE (tt) VSV Cyronebacterium glutacium b-6 C.glutacium H-8241 C.glutamicum C.lactofermentum AJ 12592 Brevibacterium lactofermentum AJ 12937 B.lactofermetum Bacillus methanolicus C cht R ng 32oC/48h Y (g/L) 100
10%Sucrose 32oC/72h 48 18%Glucose 27oC/70h 60 Glucose 31,5oC/48h 11,8 Glucose 31,5oC/58h 120,5
Glucose Methanol
32oC/72h 32oC/48h
48,8
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SX Lysine t Methanol bng Bacillus methanolicus MG3: cht t bin Thi kh: 5,5L/pht Duy tr pH 7,1 (b sung NH4OH 8N) To: 50oC 1M KH2PO4: 0,1M MgCl2: 0,01M CaCl2 = 100: 10: 1 Duy tr nng Methanol trong bnh phn ng: 100mM
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3.3.1.4. LN MEN LYSINE Cng ngh sinh hc LM LLysine c ci thin khng ngng v s tin b v k thut to ra s lng ln lysine p ng nhu cu ngy nay Thit b ln men c cnh khuy v thi kh lin tc Qu trnh gm hai pha: 18 24 h u (Pha to thnh sinh khi) + 40 60h sau (Pha to thnh lysine)
Lysine C. glutamicum
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Asp: Aspactat Asp-P: beta Aspactyl phosphate ASA: Aspactate beta Sinialdehyde DAP: Dihydropicolinasyntetase AK: aspartokinase Lys: Lysine
Hom: Homogerin Thr: Threonin Ile: Ilexin Met: Methionine HSD: homoserine dehydrogenase
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Pilot-Scale Batch Production of High Lysine Content Animal Feed Suppliment (20h/m)
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Variant pathways for the synthesis of m-DAP/lysine in plants and bacteria. m-DAP/lysine biosynthesis genes present in chlamydiae are boxed
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3.3.1.6. THU NHN V TINH SCH LYSINE ly tm dch ln men, loi sinh khi v cht khng tan Dch sau khi ly tm qua h thng trao i ion cationit Ra ct nhiu ln cho ht mu v nh hp ph bng 0,5 5 % dung dch amoniac un nng dch cha lysine loi amoniac H pH 4,9 5 bng HCl, (to monochlohydrat lysine) C chn khng nng cht kh t 30 50% Kt tinh 10 120C(tinh th mu vng nht) Ly tm thu nhn tinh th Dch sau khi ly tm s ti kt tinh thu nhn trit lysine c trong dch ln men Ha tan trong cn, kt tinh li ( tinh sch cao) Sn phm: sch 97%; m: 0,7%; tro 0,3%
3.3.2. Sx Glutamic
3.3.2.1. GII THIU V GLUTAMIC 3.3.2.2. C CHT SN XUT GLUTAMIC 3.3.2.3. VSV LN MEN GLUTAMIC 3.3.2.4. LN MEN GLUTAMIC 3.3.2.5. CC YU T NH HNG 3.3.2.5. THU NHN V TINH SCH GLUTAMIC
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3.3.2.1. GII THIU V GLUTAMIC M = 147,13 To nng chy: 247 249oC Tan hon ton trong nc Khng tan trong cn, eter v mt s dung mi
3.3.2.1. GII THIU V GLUTAMIC (tt) VAI TR CA ACID GLUTAMIC Trong y hc v dc hc: nng thp, acid glutamic l cht an thn, thuc tr nhc u, c tc dng b no Trong thc phm: mui ca acid glutamic (glutamate) l cht iu v l tng M chnh (bt ngt) c s dng ti hu ht cc nc trn TG Sn lng khong 1,6 triu tn/nm. Cc dng sn phm: cht iu v 621, ht nm, bt ngt cnh to, bt ngt xay nh Hnh thc s dng: thc phm ch bin tm p bt ngt, b sung trc tip vo thc phm
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3.3.2.1. GII THIU V GLUTAMIC (tt) 1860, Ritthaussen, c, x thnh phn cc pr ng vt, cc a.a trong c a glutamic Woff, c, x trng lng phn t, cu trc v cc hng s vt l ca cc a.a 1889, Kikunae Ikeda (Nht) tch a. glutamic v iu ch natri glutamate t rong bin 21/4/1909, ng ng k patent s 9440 ti Anh: sx cht to v 1920, ngi Trung Quc cng tm c cng ngh sx glutamate
3.3.2.1. GII THIU V GLUTAMIC (tt) M chnh t nhin To Formate Ch xanh Mc C chua S Ng Khoai ty To Mg/100g 2240 1206 668 146 140 132 130 102 102 M chnh t nhin Bp ci N m u tng Khoai lang Tm Hn C rt Sa m Sa b Mg/100g 100 67 66 60 43 41 33 22 2
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Bt ngt - cht iu v E621. Bt ngt c v Umami - l mt trong 5 v chnh m con ngi vn cm nhn trong ba n hng ngy cng vi chua, ngt, mn, ng. "iu v: c php s dng trong thc phm gip iu ha, lm tng v ngon ca mn n, khng cung cp dinh dng Cht iu v c php s dng: E621, E627 (disodium inosinate) v E631 (disodium guanylate) 627 v 631 ("siu bt ngt) c "ngt" cao hn nhiu so vi 621.
3.3.2.1. GII THIU V GLUTAMIC (tt) 1970s, JECFA - y ban hn hp v Ph gia thc phm ca T chc Y t th gii (WHO) v T chc Lng nng (FAO) nh gi tnh an ton ca m chnh v a ra quy nh v lng s dng hng ngy: 0-120mg/kg th trng 1987, Hi ngh ln th 31 ca JECFA nh gi li an ton ca m chnh v a ra kt lun liu dng khng xc nh Ro cn ngn Glutamate xm nhp vo no tr: Ro cn rut Ro cn nhau thai Ro cn no Tr em tiu ha m chnh ging ngi ln nn khng tm thy mi nguy hi no i vi tr em
n bt ngt c hi khng?
Tr em n bt ngt c hi khng?
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3.3.2.2. C CHT SN XUT GLUTAMIC Nguyn liu cha tinh bt: sn, ng, go Nguyn liu cha ng: mt r, dch tinh bt thy phn. Khi s dng r ng phi s dng cht khng biotin kim sot sinh trng ca VSV
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3.3.2.2. C CHT SN XUT GLUTAMIC (tt) MT 1 Mt r: 20%; Urea: 2%; K2HPO4: 0,05%; MgSO4.7H2O: 0,03%; CaCO3: 1%; pH: 6,8 7,8 MT2 Saccharose: 8,5 - 10% Urea: 0,5% K2HPO4: 0,1% MgSO4.7H2O: 0,1% ZnSO4: 0,1% Dch chit bp: 0,25% (nhn t sinh trng) pH 7 - 8
3.3.2.3. VSV LN MEN GLUTAMIC Brevibacterium Micrococcus Arthrobacter Brevibacterium flavum Corynebacterium glutamicum Corynebacterium glutamicum Ging t nhin: nng sut 0,2g/L Ging chuyn gen: nng sut 135 g/L (nm 1960) Ajinomoto: s dng ging siu tng hp: 170 180 g/L
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3.3.2.4.1. PP HA HC: (thy phn protein t bt m, 3.3.2.4.2. PP SINH HC: (ln men)
3.3.2.4.1. PP HA HC
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3.3.2.4.2. PP SINH HC: (ln men) PHNG PHP LN MEN Ln men tnh Ln men tnh c b sung c cht (khng thng dng) Ln men tnh (V = 100m3) T l ging: 5 6% To: 28 30oC (c iu nhit) pH 7 - 8 Sc kh: 40 85mg O2/L.min. nu khng sc kh th sn phm to ra lactate. Thi gian: 40 60g Sn phm: 100 150 g/L
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FBP: Fructose 1-6 Biphosphate GAP: Glyceraldehyde 3 phosphate PEP: phospho enolpyruvate AcCoA: acetyl coenzyme A IsoCit: IsoCitrate KG: alpha ketoglutanate Glu: glutamate Suc: Succinate Fum: Fumarate Oxa: oxaloacetate
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pHop = 6,8 8 iu chnh pH bng NH4+ (ure, nc NH3, kh NH3, NH4Cl) Toop: 26 37oC Thc t: Giai on u: 30-32oC Giai on sau: 36-37oC
Thiu oxy: sn phm ch yu l acid lactic Tha oxy: sn phm ch yu l acid alpha xetoglutavic PP cung cp oxy Sc khng kh v trng kt hp vi khuy trn (150 vng/pht) S dng cht ph bt trong qu trnh ln men
3.3.2.5.YU T NH HNG TRONG PP LN MEN (tt) Cht kch thch sinh trng biotin (2 5g/l) Ngun cung cp biotin: cao ng, mt r Khng nh hng n kh nng tng hp glutamic nh hng n tnh th m thu ca mng t bo, gip glutamic thot ra khi tb khuych tn vo mi trng Tha biotin: sinh tng hp acid glutamic t Tha biotin + sc kh t: sn phm ch yu l alanin v acid lactic Khc phc: b sung 0,15% Tween 60, penicillin 5UI/mL, km hm s pht trin ca VK trong mt giu biotin ng thi tng kh nng tng hp acid glutamic
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Cc phng php kt tinh glutamic t mi trng nui cy PP im ng in PP to hydrochloxit ca acid glutamic PP dng dung mi hu c PP trao i ion PP chuyn acid glutamic thnh cc mui kim loi
HCl
Kt tinh glutamic
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Dch MT sau ln men Thit b t tin Ct trao i cation (gi li tp cht) Ct trao i anion (gi li glutamic) Dung dch m Ra ct anion Hiu sut thu nhn glutamic cao
3.3.2.6. THU NHN V TINH SCH GLUTAMIC (tt) M chnh non (t) Tinh th glutamic
Nc
Ly tm Ty mu Kt tinh
Mm
Na2CO3/ NaOH
Trung ha HCl
Na2S + C2H2O4
C c n 30,2 30,5 Be
65oC
pH 6,8
Kh Fe2+, Ca2+
Ty mu
60oC
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Sn phm bt ngt: Hiu sut thu hi: 70% - 80% khi lng glutamic Cc ch tiu cht lng ca bt ngt: Tinh th ln, ng u, m Lng gluNa: 98 -99% m: <0,5% NaCl <0,5% Cc tp cht cn li khng cha asen, kim loi v hp cht canxi
Tinh sch
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3.4. CNG NGH SN XUT SINH KHI VSV 3.4.1. k c bn sx sinh khi VSV 3.4.2. Sn xut sinh khi nm men 3.4.3. Sn xut SPC single protein cell
Ging VSV
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Chn phng php thch hp cho qu trnh nui cy Nguyn liu cho sn xut Thit b v phng php thu nhn, tinh sch ph hp
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3.4.2.1. Nguyn liu sx sinh khi nm men 3.4.2.2. Cng ngh sn xut sinh khi nm men 3.4.2.3. Cc sn phm sinh khi t nm men
3.4.2. SN XUT SINH KHI NM MEN 3.4.2.1. Nguyn liu sx sinh khi nm men
Mt r (r ng) Nc: t tiu chun nc cp thnh ph Nguyn liu cha nit: ure, D.A.P Nguyn liu cha photpho: P2O5
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MT R
SACCHAROSE BIOTIN MU SM H KEO
Cn tr s ha tan ca oxy
X L MU V H KEO TRONG MT R
PP NNG Mt r 80oBx+ H2SO4 (1 tn mt r : 3,5 kg H2SO4 ) Khuy lin tc 85oC/6h Khuy lin tc 28 32oC /24h PP LNH
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Nhn ging
Men paste
TP cho ngi
Men kh
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CC THNG S TRONG THIT B LN MEN Fermenter c dng thng: H = 2 Fermenter cn c cnh khuy v h thng cung cp kh. Tc khuy: 50 300 vng/pht Kch thc l thng kh = 10 ln kch thc t bo u khng kh vo phi gn mng lc kh. u khng kh ra phi cho kh qua dung dch KI 10% ri lc bng bng thy tinh trnh kh ra c hi du diezen khi chy bng my nn. T l Khi lng khng kh: Khi lng dung dch: Thi gian = 1:1:1 (m3 oxy: m3 th tch: 1 pht)
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3.4.2.3. Cc sn phm sinh khi t nm men MEN BNH M - STARTER Men kem (Men lng) Hot tnh mnh Thi gian BQ ngn BQ lnh Thng mi kh Men p: m =75 % Hot tnh gn bng men kem Thi gian BQ: 10 20 ngy BQ lnh D vn chuyn, thng mi ha c Men kh: m < 10% Thi gian BQ lu. D vn chuyn D thng mi ha Hot tnh khng mnh bng men kem, men p. Men d cht do tc ng nhit khi sy
Sinh khi nm men Sinh khi nm men lm thc n gia sc lm men bnh m Quy trnh cng ngh Ging nhau Mc v sinh Yu cu cao Khng yu cu cao Thi im thu nhn u phase cn bng Cui phase cn bng sinh khi
Ch tiu
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3.4.3.1. Thut ng SCP 3.4.3.2. L do sx SCP 3.4.3.3. Nguyn liu sx SCP 3.4.3.4. SCP t vi khun 3.4.3.5. SCP t nm men 3.4.3.6. SCP t nm mc 3.4.3.7. SCP t nm qu th 3.4.3.8. SCP t to
3.4.3.1. Thut ng SCP Thut ng SCP c dng ln u tin ni v cc dng Pr thu nhn t nm men (c th VSV n bo) Protein Protein a bo Protein V Protein TV Protein n bo Protein c ngun gc t VSV
- Nm men - Spirulina
- Casein - Globulin
SCP
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3.4.3.2. L do sx SCP Nhu cu Protein: 70 100g Pr/ ngi/ngy Thiu Protein HNG GII QUYT (5) 1. Nng nghip 2. Chn nui 3. Khai thc bin 4. Tng hp ha hc 5. Sn xut v s dng SPC
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Sn xut Pr t nm men: Sac. cerevisiae, Candidas utilis, Tropicalis utilis Sn xut Pr t to: Chlorella, Spirulina platensis, Slenedemus Quy m sx: quy m ln, c li ch kinh t cao
3.4.3.2. L do sx SCP (tt) U IM CA SCP Hm lng Pr: SCP cao hn Pr V v Pr TV Cht lng Pr: SCP tng ng Pr V, cao hn Pr TV VSV dng nguyn liu r tin Sx SCP khng ph thuc vo kh hu, iu kin a l C th sn xut theo quy m cng nghip NHC IM SCP Khng ph hp vi 1 s tnh cht thc phm: mu, mi, c l C mt s t bo VSV c th ngi khng th hp thu
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3.4.3.3. Nguyn liu sx SCP whey, sulphate waste liquors, hydrocarbon waste from the pet petroleum industry, and the vats used to produce alcoholic beverages. The production process involves growth of the organisms in large fermenting tanks with forced aeration for vigorous cell-growth. Manufacture process used by British Petroleum Industry for single-cell protein from hydrocarbons is represented in Fig. 17.7. :
VSV VK Hygrogenomonas, Cellulomonas, Pseudomonas Candida spp., Saccharomyces fragilis, Torula sp., Rhodotorula spp Aspergillus terreus, Trichoderma reesei, Penicillium spp. Scenedesmus acutues, Scenedesmus acutues, Spirulina, Chlorella Nguyn liu Hydrocarbon wastes from petroleum industry. Whey, ethyl alcohol, starches, n-paraffins, sulphite waste, etc. Paper-pulp, starches, coffee wastes, straw, bassage, etc. culture ponds
Nm men
Nm mc To
3.4.3.4. SCP t vi khun Cc ging thng c s dng u c kh nng ng ha cc alkan (C6 C18) Phi tinh sch v ch bin lm thc n cho ngi
C cht Metan Hydocacbua (du t m) VSV Methylomonas methanica Methylococens capsulatus Pseudomonas, Corynebacterium Mycobacterium Brevibacterium
Nui Mycobacterium trn 1kg metan thu c 1kg sinh khi c hm lng Pr 60 70%; Gl 20 40%; L 2 4%; khong 2-6%, cc vitamin, c bit l B12
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Candidas utilis
Trong chin tranh th gii 2, c sx Candidas utilis nh 1 ngun thc phm thay th cho Pr. SCP t Can. utilis c FDA cho php s dng. c b sung trong thc phm, nhiu nc trn th gii s dng n trong CBTP
3.4.3.5.2. PP thu nhn cc sn phm trong t bo nm men lm thc phm cho ngi PP ha hc PP vt l PP sinh hc PP kt hp
PP HA HC Thy phn bng acid hoc kim Ph hy a.a trong t bo, gi m gi tr dinh dng Sn phm thu c khi s dng kim: 50% a.a dng L + 50% a.a dng D (c th ngi khng hp thu c) Thit b phi chu c acid v kim c hi
PP VT L Sock nhit: ng lnh sinh kh i VSV -20oC -40oC/2-4h ri a ngay v 40 45oC. Lm sock nhit 3 ln lin tip s ph v c 100% t bo S dng nng lng: l viba, hng ngoi, t ngoi Nghin: t s dng khi sn xut ln
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3.4.3.5.2. PP thu nhn cc sn phm trong t bo nm men lm thc phm cho ngi (tt)
PP SINH HC Thy phn bng enzyme ngoi bo v ni bo C ch s dng enzyme ngoi bo: glucanase hoc dng hn hp cellulase ph v thnh t bo C ch s dng enzyme ni bo (t phn): to k cho enzyme c sn trong t bo t phn hy tb bng cch iu chnh to, pH Thng s k thut trong qu trnh t phn T l nc: sinh khi men paste= 1,8 -2 lit : 1kg Duy tr pH 4,5 5 Duy tr To = 48 52oC Thi gian: 10 -12h Sau qu trnh t phn, sy thu bt Pr m c (60% Pr). B sung bt Pr vi cc sn phm thc phm khc
3.4.3.6. SCP t nm mc
www.gbd.edu.vn Nm si thng sinh trng chm hn nm men.
Thi gian th h: khong 10h Hm lng Pr trong nm si thp: khong 30% Cc loi nm dng sn xut SPC trong thc phm: - Kt hp Trichoderma viridea v Sac. cerevisiae - Kt hp Trichoderma viridea v Candida utilis -Kt hp Endomycopsis v Candida utilis D tch sinh khi, hng v c bit
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3.4.3.7. SCP t nm c qu th Cc loi nm c qu th: nm m, nm mo, nm rm, Morchella (Mor. Crassique, Mor. esculata, Mor. Hortesis nui Morchella rt tn km v d b tp nhim) Cung cp 30 50% Pr Kh cng nghip ha Mi trng nui cy r tin: rm, r, cy mc, mt ca, ph thi, v chui, v c ph, r ng SPC t nm ln sx nhiu : c 3000 tn/nm, i Loan, Indonexia
Mor. esculata
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VSV t dng quang nng Cn cung cp nh sng, mi trng nui su 20 -30cm. T tng hp cht hu c Tng sinh khi nhanh. Kch thc ln, thun li cho vic thu nhn Khng b VSV khc tn cng.
3.4.3.8. SCP t to iu kin sinh trng, c im SPC t to pht trin ca to Hm lng Pr cao: 70 75%
cht kh Pr to l Pr hon ho, cht lng cao Khng c c t nguy him tn ti trong sinh khi to
To Spirulina
1963, Brandily tm thy ti h Tchad (Chu Phi) 1968, hi ngh quc t (Thy in) bn v li ch v cch nghin cu sn xut to Spirulina lm thc n cho ngi, c bit l tr s sinh, vn ng vin 1980, cng b rng ri cc gi tr dinh dng v chc nng sinh hc ca Spirulina Php, M, Nht, Canada, Mehico, i Loan 1985, Spirulina c a vo VN.
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To Chlorella
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Thnh t bo Hnh thi TB Pr (%) Glucid (%) Lipid (%) Tro (%) Khong (Ca, P, Fe, Na, Cl, Mg, Mn, K, Vitamin (B3, B12, B6, B1, E, caroten, acid folic) Cc acid amin thit yu Net protein utilization - N.P.U
Spirulina Mng 5m *2mm, xon 60 - 73 4,7 - 6 7-8 4-5 6 Nhiu Nhiu 80-85%
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Spirulina Chlorella Ni trn b mt Khng khuy s chm pHop = 8,5 9 To op = 35 t b chi phi n gin pHop = 5,5 7
To op = 35
B chi phi Phc tp
TNH HNH SX V S DNG TO TRN TG Cc b tc Chu Phi v Chu M la tinh s dng Spirulina lm nc chm, nu cho, ch bin thc n. B sung Pr cho ngi v gia sc, thc phm chc nng X l mi trng Cc nc sn xut to theo quy m cng nghip: M, Mehico, i Loan, Nga, Tip, c, Bungari, Nht, Myanma, Thi Lan, n , TQ.
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TNH HNH SX V S DNG TO TI VN 1985-1995, TS. Nguyn Hu Thc nghin cu ti "Cng nghip nui trng v s dng to Spirulina". Bs Nguyn Th Kim Hng (HCM) "Nghin cu sx v s dng thc n c to Spirulina trong dinh dng iu tr Mt s a phng nui trng to Spirulilla: Vnh Ho (Bnh Thun), Chu Ct, Lng Sng (Thun Hi), Su i Ngh (ng Nai), c Min (c Lc). Cng ngh: nui to trn cc b nng xi mng s dng kh CO2, ngun CO2 ly t cc nh my bia, cn, runn ha lng vo bnh cha.
SPIRULINA FARM: Production, harvesting, drying and conditioning of micro algae spirulina, located in Guanacaste, the only one in C.A., 5 hectares land or 12.35 acres, culture size is 2000m2, production of around 3.5 tons per year with capacity to 5 tons, 8 productions pools of 250m2 each covered with greenhouses, full working equipments, independent home of 130m2, 60 meters deep well and more. International clients from Germany / Switzerland / France, classification as one of the best production and product over the world. USD $954,000
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D n SEVAS nui to n
Spirulina Packets: (Powder) 100 gms - Rs. 200 ($ 5) One Kilo - Rs. 2000 ($ 50) Spirulina Tablets: 100 Gms - Rs. 250 ($ 6.5) One Kilo - Rs. 2500 ($ 65) 25 % of total amount will go to SAVAS Spirulina project
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B sung sn phm to Chlorella vo sn phm to Spirulina lm tng kh nng o thi kim loi nng khi c th.
186.000 VND/ hp
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3.6. Cng ngh enzyme 3.6.1. Cc khi nim c bn 3.6.2. Ngun thu nhn ch phm enzyme 3.6.3. Sn xut enz t VSV 3.6.4. C nh enz
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3.6.1. Cc khi nim c bn Enzyme: cht xc tc sinh hc v n c tt c cc t bo sinh vt Tnh cht enzyme - c hiu - Kh nng xc tc cao - Ch hot ng Top, pHop Ch phm enzyme: sp thng mi hay hng ha, sp c hot tnh enz
Pht hin thy > 2000 enz. 200 ch phm enz c s dng nhiu Cc sn phm c ng dng r rng km theo khuyn co v cch s dng. Cc ch phm ph bin: ch phm enz thy phn: protease, amylase, pectinase.
3.6.1. Cc khi nim c bn (tt) Enz ngoi bo (exoenzyme): chim a s, vsv tng hp enz ri y ra ngoi t bo cht Enz ni bo (endoenzyme): vsv tng hp enz ri gi li trong t bo cht Enz bn th (constitutive enzyme): vsv tng hp enz, m khng ph thuc vo iu kin mi trng v iu kin nui cy Enz cm ng (inductive enzyme): thng l enz ni bo. Cm ng ph thuc vo thnh phn thc n trong mi trng. Khi mi trng c cht cm ng, vsv ch sinh tng hp nhng g cn thit, vi hm lng cho n s dng
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3.6.1. Cc khi nim c bn (tt) N V HOT CA ENZYME IU: 1 n v hot IU l lng enzyme cn thit : xc tc chuyn ha 1 mol c cht hoc xc tc to thnh 1 mol sn phm trong 1 pht cc iu kin xc nh (pH, nhit , nng , c cht) Mt n v hot ca Termamyl 120L ( amylase) l lng enzyme cn thu phn 5,26 g tinh bt trong mt gi (cht nn l tinh bt ho tan, Canxi 0,0043M trong 720 37oC ti pH 5,6) Lng enz xc tc cng thp th hot tnh enz cng cao
3.6.2. Ngun thu nhn ch phm enzyme VSV: cc VSV ny c xp loi l VSV an ton GRAS (generally recognized as safe) Thc vt ng vt
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3.6.3. Sn xut enz t VSV 3.6.3.1. Cc VSV s dng trong sn xut enzyme 3.6.3.2. Quy trnh sx enz t VSV 3.6.3.2.1. Thu nhn sinh khi th 3.6.3.2.2. Ph v t bo 3.6.3.2.3. C c v bc u tinh sch enz 3.6.3.2.4. Tinh sch enz 3.6.3.2.5. B sung cht bo v, n nh enz 3.6.3.3. Cc yu t nh hng ti sx enz t VSV 3.6.3.4. nh gi cht lng ch phm enz
Chng loi: Hot tnh Gii php ci thin hot tnh Thi gian thu nhn Gi thnh Kh nng pht trin cng ngh
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In leather processing; ion food processing; in manufacture of liquid glue;in degumminga of silk; in clarification of beer protein haze and as an adjunct to soap for cleaning in laundries; in tenderizing meat; in recovery of silver from spent film (photography).
Protease c s dng nhiu nht. Protease dng trong nc tng, nc mm phi chu c acid, mui. Protease dng trong bt git phi chu c kim
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Ln men
Loi b vch, mnh v tb (Ly tm/lc) C c (Siu lc/ta) B sung cht bo v, cht n nh
Sy phun Enz lng Enz bt
3.6.3.2. Quy trnh sx enz t VSV (tt) LN MEN B MT (VN) (Solid state fermentation) u: u t t
PP LN MEN
LN MEN B SU (TG) (Submerged fermentation) u: Lng c cht st thp Nhc: din tch ln Canh trng thu nhn c T ng ha, c gii ha l t hp ca nhiu enzyme Khng cn lao ng th cng p dng ph bin Thu nhn 1 enzyme vt tr i so vi cc enzyme khc Hot tnh enzyme cao (tnh theo hm lng c cht trong mi trng hoc theo g cht kh ca canh trng) Fermenter 5L
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C c
CANH TRNG LNG Enz ngoi bo: lc, ly tm, thu phn lng, dch enz th l canh trng lng Enz ni bo: ly canh trng , ly tm, lc thu sinh khi, ph v mng t bo trch ly enzyme. Ly tm, lc, dch lng l enzyme th
CANH TRNG RN (thu nhn enz ni bo, p dng cho nm mc v x khun) Ly canh trng em nghin ph v thnh t bo (nu canh trng ti xp th khng cn nghin), trch ly bng nc mui 0,9%, lc, ly phn lng.
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3.6.3.2.2. Ph v t bo PP PH V THNH T BO VSV VT L: nghin bng bi thy tinh 0.2 0.3mm, nn p lc HA HC: trn tb vi cc cht oxi ha kh mnh nh benzen, hexan, to thnh l thng trn mng tb Nhc im: cc dung mi hu c, cht ty ra c th lm v hot, bin tnh enz SINH HC: t phn, enz HA SINH KT HP
3.6.3.2.3. Loi b vch, mnh v tb 1.Tch acid nucleic v lipid: S dng nuclease tch nucleic. Tch lipid bng ct lc nhi vi hoc bng thy tinh c xp nh 2. Ly tm: 30.000 v/p trong 45 u: thch hp tch enz khi tb, mnh v tb Nhc: chi ph cao 3. Tch dch 2 lp 4. Lc Lc su: depth filter: gi tb, mnh v tb trn b mt mng v su bn trong mng Lc mng: gi tb, mnh v tb trn b mt mng. 0.2 - 10m Lc mng r, thao tc n gin nhng p nn, bt l lc, thng mng lc. C th dng mng nhiu lp, l lc gim dn
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Th t xp mng trong thit b lc mng (kch thc l mng lc t ngoi vo trong 5, 1, 0.45m)
Phase dextran cha tb, mnh v PP tch dch 2 lp t s dng trong cng nghip
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3.6.3.2.4. C c v bc u tinh sch enzyme Dung mi hu c (ethanol, propanol, acetone) lm lch cn bng tng tc, gi m ha tan ca Pr. Tin hnh 4 10oC. Vdung mi/ Vdd enz = (2.5 3)/1 Mui (Na2SO4, (NH4)2SO4, NaCl) + nc lm mt lp v hydrate ca Pr. Cc Pr lin kt vi nhau, kt ta. Tch mui = pp thm tch Ta pI: ti pI, Pr c ha tan thp, hydrate ha ca Pr min lm tng tng tc gia cc Pr, dn n to ta
Ta
Polymer: c s dng rng ri. PEG MW 6000, 12000 t l 5 -15% w/v. Tin hnh 37oC. C ch ging dung mi hu c
3.6.3.2.4. C c v bc u tinh sch enz (tt) Thm tch: qt siu lc tch cc LMW (thnh phn m, peptid, mui, etylic, a.a) ra khi dd Pr. Tin hnh loi mui v lc trc khi thm tch. Lin tc b sung dung mi sch vo dd Pr ly nhng phn t c LMW ra khi dch nh qt thm thu. tch nhng cht tch in ra khi nhng cht khng tch in v ngc li
Thm tch
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3.6.3.2.4. C c v bc u tinh sch enz (tt) C dch enz = cht trao i ion C dch enz = siu lc
3.6.3.2.4. C c v bc u tinh sch enz (tt) C dch enz = cht trao i ion Nguyn l: iu chnh pH v lc ion to ta cha Pr gn kt vi cht trao i ion mang du ngc vi Pr. Ly tm tch ta, gii phng Pr khi cht trao i ion, ra cht trao i ion ti s dng Cht trao i ion (cht mang) Diethyaminoethyl (DEAE) Quaternary ammonium (Q) Quaternary aminonium (QAE) Carboxymethyl (CM) Methyl sulphonate (S) Sulphopropyl (SP) Kiu trao i ion Trao i anion Trao i anion Trao i anion Trao i cation Trao i cation Trao i cation
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3.6.3.2.4. C c v bc u tinh sch enz (tt) C dch enz = siu lc u: khng lm mt hot tnh Pr, hiu sut tch cao. Nhc: d b bt l lc Mng lc c 1 20nm tch Pr c MW t 1 300kDa. Vt liu mng siu lc: ch to t mng cellulose acetate hoc cellulose nitrate, polyvinyl, polycarbonate.
Sc k lc gel (gel filtration) Sc k trao i ion (ion exchange chromatography) Sc k i lc (affined chromatography) in di (electrophoresis)
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Nguyn tc hot ng ca cc sc k thng dng K thut sc k Bn cht Sc k trao i ion Khc bit v in tch trn mt Pr gi tr pH quy nh Sc k lc gel Khc bit v hnh dng v MW ca phn t Sc k i lc Da vo tng tc c hiu gia Pr v ligand tng thch Sc k tng tc Khc bit v tnh k nc trn b mt k nc Pr Sc k ng in Tch theo im ng in ca tng phn t Pr Sc k trn nn Tng tc tng hp gia Pr v cht hydroxyapatide mang Ca-phosphate
3.6.3.2.6. B sung cht bo v, cht n nh Cc cht bo v: propylene glycol, glycerol, sorbitol. Dch enz c c: thng b sung thm ammoniumsulfate, sodium sulfate, sodium chloride to ta hoc tinh th enz. gi enz dng huyn ph b sung silicon dioxide
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3.6.3.3. Cc yu t nh hng ti sx enz t VSV CC YU T NH HNG TI QU TRNH LN MEN PP B MT M CHIU CAO MI TRNG NHIT CUNG CP OXY pH THI GIAN THIT B
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3.6.4. C nh enzyme 3.6.4.1. Gii thiu chung v c nh enz 3.6.4.2. Cht mang gn enz 3.6.4.3. Phng php c nh enz 3.6.4.4. Cc dng bnh phn ng v enz c nh 3.6.4.5. Cc ng dng ca enz c nh 3.6.4.6. C nh enz v c nh t bo VSV
3.6.4.1. Gii thiu chung v c nh enz u im Khi nim: gn enz ln 1 cht mang tr ha hc, to enz khng tan trong nc Ti s dng nhiu ln (60 70 ln) m hot tnh enz gim khng ng k Khng ln enz trong sn phm C th gn ng thi nhiu enz ln cht mang thc hin chui phn ng. T c th xy dng m hnh h a enz trong t bo nghin cu C th c gii ha, t ng ha C th hi lu lp li phn ng nu sn phm cha t yu cu
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3.6.4.2. Cht mang gn enzyme Cht mang hu c Tng hp ha hc: bn vi tc ng MT v VSV Polymer: poly-acrylamid, poly-styren, poly-acetate Cht mang v c
bn vi tc ng MT, kh gn
Cc oxit kim loi T nhin: khng bn vi tc ng (Al, Mg, Ti), cao lanh, than hot MT v VSV tnh, si bng Dn xut cellulose (si bng, d m thy tinh bo, li ng), dn xut chitin, chitosan, collagen, tinh bt, alginate, oligosaccharide (dextran)
3.6.4.2. Cht mang gn enzyme (tt) Yu cu ca cht mang gn enzyme Gn cht vi enz, khng c tch ra khi enz trong khi phn ng D tm, gi r Khng lm thay i hot tnh ca enz Bn trong mi trng
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3.6.4.3. Phng php c nh enzyme PP vt l (lin kt vt l) s hp ph, to khun gel, bao bc mng bn thm PP ha hc (lin kt ng ha tr gia enz v cht mang)
3.6.4.3. Phng php c nh enz (tt) - PP vt l PP hp ph: trn cht hp ph vi enz. Enz hp ph trn b mt cht mang nh lin kt (ion, k nc, tnh in, Vander Walls) u: n gin, s dng nhiu cht mang khc nhau) Nhc: khng bn, nu thay i pH, To t ngt s gii hp ph (tch enz ra) PP vi ti (micro capsul): gn v nht enz trong mng bn th m. C th nht a h enz trong vi ti. ng dng: vi ti urease cha bnh thn, catalase cha co c. Gn vi ti enz vo cc c quan khng tng hp c enz, vi ti khng i theo mu.
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3.6.4.3. Phng php c nh enz (tt) - PP ha hc Gn cht mang qua cht hot ha trung gian (2 giai on). PP s dng ph bin. Thng s dng vi cht mang v c nh si thy tinh, si silicate G1: hot ha cht mang G2: Gn cht mang hot ha vi enz
Cht mang + Cht hot ha Cht mang Cht hot ha
+
3.6.4.4. Cc dng bnh phn ng v enzyme c nh Enzyme c nh l lng huyn ph Enzyme c nh trn mng mng, xp nhiu lp Enzyme gn trn si cellulose (xp cc si thng ng, b si) Enzyme c nh gn trn mng mng, kch thc ln, cun xon Lp enzyme c nh c chiu cao ng k, dy
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3.6.4.4. Cc dng bnh phn ng v enzyme c nh (tt) Thit b (Bioreactor) Thp hnh tr, h>> , h ln c thi gian phn ng gia c cht S vi enzyme E ng dn c cht nh, c van y tho sn phm, c th iu chnh V Cu to: Bnh c 2 lp v, gia c nc iu chnh nhit phn ng. ng hi lu ln nh thp.
3.6.4.5. ng dng ca enzyme c nh Sn xut fructose bng enzyme glucose isomerase c nh Sn xut L-acid amin Sn xut sa bo philactose Urease c nh dng vi ti cha bnh thn, phn gii urea S dng enzyme glucose oxydase nh lng nhanh glucose trong dch sinh hc Enzyme pectinase c nh
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Glucoseisomerase c tch t x khun Actinomyces Cht mang: silicrom (dn xut cellulose) Cht hot ha: glutaraldehyte
C NH T BO VSV Khng phi tch chit enz, khng tn cng sc Cc enz hot ng trong mi trng t bo (dng t nhin), thun li, ph hp hn enz c tch chit ra Tn dng c a h enz trong t bo C kh ha, t ng ha, sn xut lin tc Ph hot ng ca enz trong t bo thng kh rng Lng enz c th tng ln khi t bo sng
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3.6.4.6. C nh enz v c nh t bo VSV (tt) Enzyme c nh T bo VSV c nh u gn ln cht mang C th ti s dng Cc cht mang dng cho enz c nh u c th dng cho c nh t bo VSV Thit b c th tng t nhau Cht mang gn trc tip Cht mang khng gn trc tip vi enz hoc gn ln cht vi enz m gn vi mng t bo hot ha) VSV. Enz trng thi t do, c th dch chuyn trong t bo Khng cn cung cp Cn cung cp dinh dng dinh dng nui enz nui t bo
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