cong nghe len men hien dai

4/20/2011

1

Cng ngh ln men

CDGD: Bi Hng QunBin son: Nguyn Minh Hin

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 iCng 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)..

4/20/2011

2

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 SV1. IM QU TRNH (30%)1.1. KIM TRA 15 PHT (15%)1.2. BO CO SEMINAR (15%): chn 1 trong 3 phng n sau1.2.1. 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 chiu 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).

2. IM THI KT THC MN HC (70%)Thi vit t lun

4/20/2011

3

NI DUNG CHI TIT HC PHN 2Phn 1. M uPhn 2. K thut ln men Phn 3: Cng ngh ln men ng dng

3.1. Ln men ethanol v cc ng dng3.2. Cng ngh sn xut acid hu c thc phm3.3. Cng ngh sn xut acid amin3.4. Cng ngh sn xut sinh khi vi sinh vt3.5. Cng ngh sn xut polysaccharide tVSV3.6. Cng ngh enzyme3.7. Cng ngh sx cc sp ln men truyn thng

www.gbd.edu.vn

PHN 1. M U

1.1. CNG NGH LN MEN 1.1.1. Khi nim v ln men (fermentation)1.1.2. Khi nim v CNLM (fermentation technology)1.2. PHN LOI CC SN PHM LN MEN

www.gbd.edu.vn

4/20/2011

4

(From latin fervere)

1.1.1. Khi nim v ln men (fermentation)

www.gbd.edu.vn

Quan im ca nh ha sinh hc: ln men l qu trnh sn sinhnng lng. Cc hp cht hu c hot ng vi vai tr va lcht cho, va l cht nhn in t. Ln men l qu trnh ymkh, nng lng c sn xut khng cn c oxy hoc cc chtnhn in t v c khc.Quan im ca Pasteur: 1857, ng cng b qu trnh ln menkhng phi cng trnh ca s cht nh nhng nh ha hcngh m l cng trnh ca s sng. ng a ra khi nim tnhk kh v i kh ca VSV v s ln men l h qu ca cucsng khng c khng kh

Theo ngha m rng: ln men l qu trnh nui cy VSV (c oxyhoc khng c oxy) thu nhn sinh khi, cc sn phm trao icht, thc hin s chuyn ha c cht.

1.1.1. Khi nim v ln men (tt)

4/20/2011

5

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).

Louis Pasteur (27.12.1822 28.9.1895) the father of the Microbiology


www.gbd.edu.vn

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


4/20/2011

6

The fermentation technology is the combinedapplication of the knowledge of processengineering, biochemistry and microbiology fordesigning or evaluation a fermentation process.

1.1.2. Khi nim v CNLM (fermentation technology)

Sn phm Sn lng (tn/nm)Acid foods (citric, lactic) 100,000Alcohol 1,000,000Amino acids 10 100,000Antibiotics 10 30,000Enzymes 0.1 3,000Pharmaceutical proteins 0.001 - 3

SCIENTIFIC KNOWLEDGESCIENTIFIC KNOWLEDGEBiochemistry

Microbiology

Molecular Biology

Process engineering

InformaticsStatistic

Immunology

Physiology

TOOLS / TECHNICAL ADVANCESTOOLS / TECHNICAL ADVANCES

Biosensors

Bioinformatics

Bioprocess

Protein engineeringExperimental design

Process analysis

ENVIRONMENTENVIRONMENTBioremediation

Environmental monitoringPollution control

Useful Useful ApplicationsApplications

FARMINGFARMINGYield

Animal healthFeed stocks

PHARMACYPHARMACYDiagnostics

VaccinesTherapeutics

FOODFOOD

4/20/2011

7

What I should do to do fermentation?Media preparation

Fermenter preparation

Sterilization

Adjusting parameters

Inoculation

Cultivation

Taking samples

Dev

elopm

ent t

he pr

odu

cer

stra

in

Harvest

Medium design

Physiology

Kinetics

Agitation/Aeration

Scale-up

conservation

Optimization

Fermenter typesOperation modes

CC SP TRAO I CHTSP TC BC 1: acid amin,vitamin, acid citric

SP TC BC 2: enzyme VSV,khng sinh

SP ln men: ru, acid lactic(ln men k kh)

C cht T bo (biomass)

1.2. PHN LOI CC SN PHM LN MEN T VSV

SINH KHI VSV(biomass): protein n bo(SPC), men bnh m, gingkhi ng (starter)

C cht Sn phm + T bo(SP TC)

4/20/2011

8

Phn loi sn phm ln men theo quan im kinh tBio-products

High value Lowproduct volume(MostlyMostly dependsdepends ononthethe substratesubstrate price)price)

Medium value Highproduct volume (TheTheprocessprocess isis relativelyrelativelyexpensiveexpensive andand somesomecomplexity)complexity)

Low value Highproduct volume(MostMost ofof thethe productionproductioncorrespondcorrespond totopurificationpurification processprocess)

Antibiotics Vitamins Enzymes Vaccines Steroids Hormones Other pharmaceutics

Amino acids Organic acids Biopolymers Baker yeast Microbial polysaccharides

Ethanol Biomass Methane Acetone Butane Fructose syrups Feeding products

Cost distributionHigh volume and Low value product Low volume and High value product

High investment cost (ph u t cao) High investment costHigh raw material cost (ph nguyn liu cao)

Cost are distributed in all production stages

High conversion efficiency (hiu qu chuyn i thp)

High recovery of the product in the fermentation stage

Low recovery cost (Ph quay vng thp) High purification costLow margin profit (li nhun thp) High margin profitOnly few regulatory problems (phi theo quy nh php lut)

Too much regulatory and legyslation restrictions, and high plant hygiene

Required not much R&D expenditures (khng tn nhiu tin cho RD Required too much R&D CostLow qualification of the labor force is acceptable (Ngi lao ng c th ko cn trnh chuyn mn cao)

Very high qualification of the labor force is nedeed

4/20/2011

9

PHN 2. K THUT LN MEN2.1. VSV TRONG CNG NGH LN MEN TP2.2. NG HC QU TRNH LN MEN 2.3. PHNG PHP & THIT B LN MEN2.4. CI TiN QU TRNH LN MEN

www.gbd.edu.vn

2.1. VSV TRONG CNG NGH LN MEN TP2.1.1. CC YU CU V GING VSV2.1.2. K THUT TO GING

2.1.3. K THUT NHN GING V SX GING2.1.4. K THUT KIM TRA GING VSV

2.1.5. K THUT NNG CAO CHT LNG GING2.1.6. K THUT BO QUN GING

4/20/2011

10

VSV TRONG CNG NGH LN MEN TPTP ln men vi s tham giaca VSV trong t nhin (TPln men truyn thng)

TP ln men vi s tham giaca VSV thun khit (TPln men cng nghip)

Sx th cng, quy m nh,nng sut khng cao Khng kim sot cqu trnh, cht lng chan nh v cha ng u. Mang bn sc m thc,kinh nghim, vn ha cami dn tc

Sx quy m ln, nng sut cao Ch ng cy 1 lng VSVvo nguyn liuKim sot c qu trnh lnmen, cht lng n nh &ng u

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 CNPhi 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

2.1.1. Yu cu ging VSV trong CNLM

4/20/2011

11

2.1.2. K thut to ging

Phn lp trong t nhin

PL trong k sx cng nghip

To ging VSV mi (p dng k thut di truyn)

PP Nguyn tc u im NhcPL trongt nhin

c c cht, c VSVphn gii c cht

ngun VSV phongph a dng

Tn thi gian,hot lc VSVcn thp

PL trongsx CN

Tnh thch nghi cao Hiu qu cao, VSV quen vi iukin sx cng nghip

/

ToVSV =KTDT

Tip hp, ti t hp To c gingVSV c c tnhmong mun

Yu cu trnh cao, thitb hin i

Cc trung tm lu tr ging trn th gii v Vit NamABBOTT: Abbott Lab, North Chicago, III.60064, USAATCC: America Type Culture Collector, 12301, ParklawDrive Rockvill Md20852, USAHIR: Food and Fermentation Divisio, Hokkatdo ProfecturalIndustrial Research Institute Saporo, JapanFERM: Fermentation Research Institute, Agency ofIndustrial Science and Technology Ministry of IndustrialTrade and Industry, Chiba, JapanVTCC, IMBT, National University, HaNoi, [email protected]; www.biotechvnu.edu.vn (Ngn hng gingquc gia, HN)

Bo tng ging: www.bacteriummuseum.org

4/20/2011

12

2.1.3. K thut nhn ging

Nhn ging trong PTN Nhn ging trong quy m sx ln

PPnhn ging khi ng truyn thng v hin i

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 VSVpH: 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

4/20/2011

13

Quan st i th Quan st vi thKim tra hot lc ging VSV (thoi ha)

Ging VSV b tp nhim, thoi ha phiphn lp li hoc thay ging khc

2.1. 4. K thut kim tra cht lng ca ging VSV

Kim tra thun ca gingthng xuyn (b nhim t ng gc)Kh trng MT dinh dng; vi ccMT c bo t cn kh trng trit hn. V d dit bo t Bac. subtilis1800C/60 - 90 (B nhim trong qtnhn ging)

Ging VSV b thoiha: do tc ngca mi trng bntrong v tc ngca nhng snphm TC dochnh VSV tit ra

2.1.5.1. Hun luyn thch nghi ging vi sinh vt Mi VSV u c kh nng thch nghi rt cao vi mi trng.

Nhng tc ng mi trng c lp i lp li nhiu ln to nntnh thch nghi bn vng.

Khi to c tnh thch nghi ca VSV phi lun duy tr tc ng mc to ra tnh thch nghi. c im ny khng bn.

Yu cu: ngi thc hin phi c tnh kin tr.

2.1.5. K thut nng cao cht lng ging

Khun lc nm men trn Hansen Agar c hm lng ng 26, 28% (phng php hun luyn ging)

4/20/2011

14

2.1.5.2. t bin VSVt bin bng tc nhn vt l ( tia U.V):Tia U.V c s dng rng ri trong cng ngh chn gingVSV. Tia U.V c kh nng to ra cc t bin c cht lng caohn chng loi ban u hng trm ln.

Tia U.V =260nm thng gy ra nhng t bin im caonht. y l loi bc x khng ion ha v gy ra nhng bini base cha nit trong gen.

t bin bng tc nhn ha hc (khng khng sinh):Dng mi trng c khng sinh tm cc dng t bin bnvng (khng khng sinh). Trn mi trng ny, cc t bo mncm vi khng sinh s b git cht, ch cn cc t bo t bini khng l tn ti

S lng khun lc/ nng khng sinh trong MT nui cy

Nhm

10-7 (khng bsung khng sinh)

0,01mg/ml 0,1mg/ml 0,5mg/ml

1 34 8 2Khng

xut hin khun

lc

2 58 5 3

3 107 7 5

4 Nhim Nhim 5

TN tin hnh vi vk E.coli. Nui cy E.coli nng pha long 10 -1 trong cc mi trng b sung khng sinh

4/20/2011

15

2.1.5.3. Lai ging nm menNguyn tc:T 2 ging nm men ging nhau ta c th to ra c ging nm men mi c nhng c tnh ca c 2 loi ban u bng cch cho chng tip xc vi nhau trong iu kin th nghim.Nhc im:T l thnh cng khng cao.S pha trn c tnh di truyn ch trong mt lai nht nh.Cc ging VSV khc nhau th khng th tin hnh qu trnhlai ging c.u im: n gin, d thc hin

Cch thc hin lai ging nm men1. 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 ngy3. Hai ging nm men lai vi nhau nu trn M2 xut hin

khun lc4. Kim tra ging thu nhn c phi l lng bi th bng

cch: Cy khun lc trn mi trng M2 vo mi trngThch acetat (M3). 48h

5. Lm tiu bn, quan st di knh hin vi, nu thy nangcha 4 bo t (t bo lng bi th) chng t lai thnhcng.

6. Kim tra tnh trng cn quan tm.

4/20/2011

16

2.1.5.4 S dng k thut di truyn hin iK thut bin i gen to ra cc sinh vt c tnh trng ch theo mun

Sng bn gen

4/20/2011

17

Mc ch: m bo c tnh cht ca ging (duy tr gn nhnguyn vn c tnh ban u ca ging VSV trc lc ctgi) tiu chun cho qu trnh sn xut.

Nguyn tc: lm chm qu trnh h hp v trao i cht VSV,ng thi ngn cn s sinh sn ca chng.

Phng php:Bc 1: Tin bo qun (thun ha ging). Chn chng VSV iu kin v giai on ti u cho bo qun.

Bc 2: Chn phng php thch hp cho bo qun.

2.1.6. K thut bo qun ging VSV

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 nhNhc 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 khongNguyn tc: c ch qu trnh h hp VSV trong c VSV ym

kh v hiu kh, hn ch tip xc vi oxy, ngn hin tngmt nc ca mi trng v VSV.

Bin php: 1 lp du khong hoc parafin lngu im: n gin, hiu qu cao, thi gian BQ di (1 nm)Nhc im: C ln du

4/20/2011

18

2.1.6.3. Bo qun ging trong ct, t sy khNguyn tc: S dng t, ct nh nhng gi th mang. Khi m mi trng gim ti thiu, VSV khng pht trin na. (tv ct l mi trng ti thiu)Cch thc hin: X l t, ct (ry u, ngm trong HCl hocH2SO4 m c 8-12h. Ra di vi nc cho n pH trung tnh,sy t, ct > 1000C. BQ iu kin v trng.VSV c nui mi trng thch. ct, t v trng vong nghim, lc u, sau rt qua ng nghim khc. Hn knming ngu im: thch hp BQ cc ging VSV trong x l mitrng, 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

2.1.6.4 Bo qun ging trong cc ht ng ccNguyn tc: s dng ht ng cc nh gi th mangThng s dng BQ cc nm si v VSV trong thc phmMc ch: gi VSV trng thi tim sinh.Cch thc hin: Ht ng cc ri, hp chn, nui nm mc trc

tip (3 5 ngy), sy t0 < 500C t m W

4/20/2011

19

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 t3. 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

2.1.6.6 Bo qun ging trong gelantine

Cch thc hin:1. Chun b mi trng: Mi trng N.B b sung 10%

gelantin v 5% acid ascorbic. Kh trng 1210C/ 152. Chun b ging VSV: nui ging VSV

3. Trn VSV vi mi trng.

4. Dng ng nh git v trng to thnh tng gitgelantine nh. Sy kh trong t ht chn khng

4/20/2011

20

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 (glycerin15%, saccharose 10% + gelantin 10%... Gip VSVkhng 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 trongcc 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 ccng ging theo quy m cng nghip.

Cch thc hin: ging, nhn ging trong mi trng lng, kimtra ging (c im sinh ha, sinh l), nu t tiu chun,my ng kh 24h, hn np li.

Thi gian bo qun: 20 nmu im: cht lng ging khng i, bo qun nhit

thng, thi gian bo qun diNhc im: chi ph ln

4/20/2011

21

2.2. NG HC QU TRNH LN MEN (the kinetics of the fermentation processes)2.2.1. PHNG TRNH MONOD2.2.2. CC YU T NH HNG N HIU SUT LM

Mc ch nghin cu ng hcThit k 1 qu trnh ln men minh gi hiu qu ca qt ln men ang thc hinCi tin qu trnh (cht lng, nng sut, gim chi ph)Nng cp hoc gim quy m sx

cells

biomass

CO2H2O

Products

nutrients

How to describe a fermentation process?Conversion process of nutrients to products of the microbial metabolism

CHmOl + aNH3 + bO2 cCHpOnNq + dCHrOsNt + eCO2 + fH2O (1)

(source of N& O2) (biomass)Metabolic by products or

Interest product

Substrate

Rest of the nutrients

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

4/20/2011

22

Carbon & Energy source

+ O2Nitrogen source

+

Other required nutrients

+ Cells Products+ + CO2 Heat+ H2O+

Monitor the consumption of the carbon source: Chemical analysis Chromatography

Determine O2 consumption:Paramagnetic O2 analyzer Mass spectrometerDissolved O2

Monitor the consumption of the nitrogen source:Chemical analysisNH3 electrodepH-statNitrate electrode

Monitor the consumption of essential nutrients:Chemical analysisIon selective electrodes

Measure cell components:ProteinDNA and RNACarbohydrates & LipidsEnzymes

Monitor product formation:Chemical analysesMass spectrometerpH and viscosityChromatography (gas and HPLC)Enzyme electrode

Monitor CO2 production:IR analysisMass spectrometer

Energy balanceHeat evolution

Direct estimation of cell mass: Cell dry or wet weight Optical density

Direct estimation of cell number: Microscope count Viable plate count Light scatteringD

irect

met

hods

Indi

rect

met

hods

Methods for monitoring fermentation kinetics

nh hng ca loi c cht ti Loi c cht max (h-1)Glucose 0,28Maltose 0,22Maltotriose 0,18

dtdX

X1

=

X: VSV (tb/ml)

S: c cht (g/L)

P: sn phm (g/L)

t: thi gian

ng cong sinh trngI. Adaptation (thch nghi)II. Exponential (bng n)III. Stationary (n nh)IV. Accelerated death (suy tn)Pha thch nghi = 0

Pha bng n = max Pha n nh = 0, do dX / dt = 0

: tc to sinh khi ring (Specific growth rate )

4/20/2011

23

SKS

s += max

Is K

SSK

S2max

++

=

2.2.1.Phng trnh Monod : tc to sinh khi ring (s giatng 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

.

KS: hng s Monod,l nng c chtkhi tc to thnhsinh khi = max/2.KS c x bngthc nghim.KS = f(chng VSVv c cht)

VSV C cht Ks (mg/L)Saccharomyces sp. Glucose 25E. coli Glucose 4

Lactose 20Aspergillus sp. Glucose 5Candida sp. Glycerol 4,5

Oxygen 0,042 0,45Pseudomonas sp. Methanol 0,7

Methane 0,4Hansenula sp. Methanol 120

Ribose 3

Phng trnh Monod: mi lin h gia nng c cht vtc 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]

2.2.1.Phng trnh Monod (tt)

4/20/2011

24

2.2.2.1. nh hng ca ging VSV2.2.2.2. nh hng ca nng c cht2.2.2.3. 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 li2.2.2.4. nh hng ca pHenzyme VSV ch hot ng tt pH nht nh.pH ti u ca mi VSV l khc nhau2.2.2.5. nh hng ca mt s yu t khc: cht c ch

2.2.2. Cc yu t nh hng n hiu sut ln men

Cells

IN OUTInternal

External T pH Medium Agit. Areac. Modo Oper.

Cell. Conc. Prod. Conc. Metabolites

2.2.2. Cc yu t nh hng n hiu sut ln men (tt)

4/20/2011

25

2.3 PHNG PHP V THIT B LN MEN2.3.1. Chun b mi trng ln men

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

www.gbd.edu.vn

2.3.1. Chun b mi trngNguyn 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)

www.gbd.edu.vn

4/20/2011

26

Cc bc chun b mi trng

2.3.2. Tit trng trong cng nghip ln men

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

4/20/2011

27

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 lngLn men b mt: LM trong cc khay vi mi trng lnghay mi trng c c cht rn hay xpTheo 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 menLn men hiu kh: thng kh bng h thng trc khuy csc kh (sx biomass)Ln men k kh (sn xut ru, bia)

2.3.3. PHNG PHP LN MEN

Ln men tnh c coi l 1 h khp kn do khngc tc ng ca cc yu t bn ngoi (tr kh)

Fermentation

Air

Air

Inoculation

Culture Media Preparation

Sterilization

Ln men tnh (Ln men theo m- Batch culture)

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 culturesNumber of generations or duplicationsDuplication timeYields: Product/Substrate (YP/S); Biomass / substrate (YX/S)Productivity: Volumetric productivity (P); Specific productivity (qP)

4/20/2011

28

Pha Lag: di t vi pht ti vi gi.Tlag = f (k mi trng: pH, To, MTnui cy v VSV).Tlag khng to ra sp, nh hng titng nng sut ca qt LM

LnX

time T lagNng sut = Lng SP mong mun (g/L)

Tng thi gian ln men (h)

Adaptation PhaseExponential PhaseStationary Phase

Clean & decontaminate Load

Sterilize CultureUnload

1. Correct ratio Inoculum volume(1/5 - 1/10) Fermentor volume

3. The inoculum should be inexponential phase at themoment of inoculation

=

Inoculum culture medium

Fermentor culture medium

2.

How to minimize the Lag phase ?

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

Mass Balance in batch culture

4/20/2011

29

Ln men theo m- Batch cultureu im Nhc im

n gin, hn ch c sly nhim.Thch hp bt unghin cu 1 sn phm.Cho php chn cc thng sv nghin cu c stng tc ca chng.X c MT dinh dng, ,tc ti u to thnh sp

Nng sut thp. Nng c chtcao c ch VSV.Khng kim sot c tc sinh trng ca VSV.Tnh ng u ca SP khng caoso vi cc PP khc.Chi ph ln (h thng tit trngbng hi nc, nc v sinh thitb, in, din tch, sc lao ng

Khc phc nhc im ca Batch culture bng Fed- Batch Culture v Continuous Culture

What is it?Starts with a batch culture which, after a specific time, is fed with freshmedium. The addition of fresh medium can be synchronized with thedepletion of the growth-limiting substrate from the starting culture medium.

Ln men tnh c b sung c cht (Fed-batch culture)

Air

1

2

Air

Feed sterilization

Harvesting

Fermentation

Fermentor inoculation

Preparation of the starting culture

medium

Fermentor sterilization

Feed additionPreparation of

the feed culture medium

4/20/2011

30

Ln men tnh c b sung c cht - Fed-batch cultureu im (Advantages) Nhc im (Disadvantages)

Thi gian ca cc k sinh trng ti u c ko di do cc cht c ch b pha long v cc cht dinh dng chnh c chuyn ha nh nhau mi thi im LMTng nng sut ca qu trnhdo gim 1 s cng on (rathit b, tit trng, loading vunloading).Sn phm c tnh ng nhtcao

Trang thit b t tin, yu cu k thut cao hn pp LM tnh Tch ly cc cht c ch s sinh trng to thnh trong qu trnh TC Accumulation of growth-inhibiting metabolitesNu s t bo/th tch mi trng qu ln th c kh nng Possibility of induction of contact inhibition if the number of cells per culture volume is too large

What is it?

Starts with a batch culture to which, after a specific period, fresh culture mediumis added, and cells plus exhaust medium are removed at a rate that keeps cultureconditions constant. When the interval between successive additions andextractions decreases, the culture is named continuous and its volume remainsconstant.

Ln men lin tc - Continuous culture

Fermentation

Preparation of starting culture medium

Feed preparation

Harvesting

Fermentor inoculation

Fermentor sterilization 1

Feed addition2

Feed sterilization

Fresh culture medium

Air

Spent culture medium, cells, Air

4/20/2011

31

Ln men lin tc - Continuous cultureu im Nhc im

Dch ln men lun c b sungnn cc cht c ch b pha longTng nng sut ca qu trnh doYX/S, YP/S khng i (= constant)iu khin v kim sot c tc c cht chnh vo thit b nnduy tr YP/S ti uTng nng sut ca qu trnh dogim 1 s cng on (ra thit b,tit trng, loading v unloading).Sn phm c tnh ng nht cao

Nguy c vy nhim cao Kh nng xut hin t bin cao Chi ph thit b t tinSo vi cc pp khc, pp ny cha c nhiu kinh nghim p dng quy m cng nghip

PHN LOI THIT B LN MENTheo nguyn l hot ng: fermenter hot ng lintc, gin on, bn lin tc

Theo cu trc thit b: fermenter c cnh khuy(Shaking Tank), khng c cnh khuyTheo 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

2.3.4. Thit b ln men

4/20/2011

32

Mechanical agitation

Variety of de impelents

Baffles **

Working volume = (60-70) % of V tankHeight Diameter ratio H/D =1; H/D > 1

Heat transfer **

Fermentor: Shaking tank

Anchor

Propeller Rushton Turbine

Palette

Rake anchor

Coiled helix

Some types of impellers

Baffles

Airlift

CYTOLIFT

V= 580 ml

H= 38 cm

D= 10 cm

4/20/2011

33

Heat transfer in bioreactors

a) Continuous jacketb) External coiled jacketc) Internal coiled jacketd) Coiled bafflese) With external heat interchanger

4/20/2011

34

Fermentor ln men bia ti Jangjing Beer. Chp bi BHQ 2007

www.gbd.edu.vn

600 BC XIX century XX century Spontaneous, natural processes

Submerged Anaerobic & Superficial Aerobic cultures

Submerged Aerobic Culture

CHmOl + aNH3 + bO2 ycCHpOnNq + zCHrOsNt + dCO2 + cH2O

The oxygen demand of the cell and the low solubility of oxygen inwater, required a continuous air supply into the fermentor and acontinuous transfer of oxygen from the gas phase into the liquidphase

How is the oxygen transfer into the fermentor?

Cn x nhu cu oxy

4/20/2011

35

Oxygen Mass Balance:

O2 in - O2 out = O2 metabolized + O2 cumulativeO2 in = yO2 inQAirAirO2 out = yO2 outQEAEA O2 metabolized = qO2XV

O2 cumulative = d(CO2V)/dt = VdCO2/dt Assuming: QAirAir = QAEAE

yO2QAirAir = V(qO2X + dCO2/dt)

OTR Oxygen Transfer Rate

Flow meter

pO2

Condenser

Bioreactor0.2 -Filter

yO2in, Qair, Air

yO2out, QEA , EA

O2 probe

Pmanometer

Motor, P

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

4/20/2011

36

1. Diffusion of the O2 from the bulk gasto the gas liquid interface

2. Transport across the gas liquidinterface

3. Diffusion of the O2 through arelatively stagnant liquid regionadjacent to the gas bubble

4. Transport of O2 through the well-mixed liquid to a relatively unmixedliquid region surrounding the cells.

5. Diffusion throught the stagnant region surrounding the cells.

6. Transport from the liquid to the pellet cell aggregate

7. Diffusive transport of oxygen into the pellet

8. Transport across the cell envelope

9. Transport from the cell envelope to the intracellular reaction site. e.gmitochondria

5, 6 and 7 are relevant only for processes in which pellets or cell aggregates appear

General mechanism of O2 transfer

How we can improve the YP/X? Strain development approach- Fermentation process optimization

2.4. CI TIN QU TRNH LN MEN

4/20/2011

37

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 VSV3.6. Cng ngh enzyme 3.7. Cng ngh sx cc sp ln men truyn thng

www.gbd.edu.vn

3.1. LN MEN ETHANOL V CC NG DNG3.1.1. Cng ngh sn xut ethanol

3.1.2. Cng ngh sn xut ru cao

3.1.3. Cng ngh sn xut bia

3.1.4. Cng ngh sn xut ru vang

www.gbd.edu.vn

4/20/2011

38

3.1.1.1.GII THIU V ETHANOL

3.1.1.2. NGUYN LIU SN XUT ETHANOL3.1.1.3. PP SN XUT ETHANOL

3.1.1.4. SN XUT ETHANOL BNG PP LN MEN3.1.1.5. CHNG CT & TINH CH ETHANOL SAU LN MEN3.1.1.6. SN PHM CA QU TRNH LM ETHANOL 3.1.1.7. ETHANOL GREENFUEL

3.1.1. Cng ngh sn xut ethanol

3.1.1.1. GII THIU V ETHANOL

Cc hnh nh kho c v vic lm ru t khi nnnng nghip xut hin (ngi Xume, 4000 B.C) Tm thy ru trong lng m ca ngi Ai CpCTHH: 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

4/20/2011

39

nh hng ca ETHANOL vi sc khe nginh hng ca ETHANOL vi sc khe ngi

Ung t khng gy nh hng n tim nhng ung nhiu c thlm t lit h thng thn kinh v gy nh hng xu n h thngtun hon mu.Lng ru trong mu: 0.5 - 2 c th gy say

4 c th dn n cht ngi.Hp th nhanh vo thnh d dy (29%) (xy ra nhanh nht khid dy rng) phn cn li c hp thu rut non.Ethanol c hp thu s c oxy ho 90-98 %. Ngi trngthnh c th oxy ho khong 10 ml/h, ph thuc vo nng cntrong mu. Ung nhiu s gy s hp thu nhanh hn tc oxyho, dn n gy c.Ethanol khng c oxy ho c th ri c th theo phi v thn,n cng c th c tm thy trong m hi, nc mt, mt, ncming lng nh.

nh hng ca ETHANOL vi sc khe nginh hng ca ETHANOL vi sc khe ngi

Methanol (ru g) l mt cht c cht ngi, gy cho nhngngi nghin ru tnh trng m hay cht.

Methanol dn dn thm vo rut non khi nhm methyl caaspartame gp enzyme chymotripsin.

Mt nh gi ca EPA v trng thi ca methanol cho rngmethanol c xem nh mt c cht c tch lu v t l bitit rt thp.

Trong c th, methanol c oxy ho thnh formaldehyt vacid formic; c hai cht chuyn ha ny u l cht c thnkinh cht ngi. Liu lng tiu th cho php khong 7,8mg / ngy

4/20/2011

40

nh hng ca FURFUROL vi sc khe nginh hng ca FURFUROL vi sc khe ngiL cht c vi h thn kinh, gy nhc u, bun nn.

Ung ru c hm lng furfurol cao qu mc cho phpgy tch t trong dch ca phi (ph phi).Fufurol c th gy d ng da, nu s d ng pht trin cao,c th gy nga v ni mn da. Tnh trng trn nu tiptc c th gy mt v gic, t li, au u, mt v rngmnh. Tnh trng ko di c th nh hng ti gan.

Tip xc lu di vi furfurol c nguy c gy ung th vnguy c i vi sinh sn.

nh hng ca ETHANOL vi VSVnh hng ca ETHANOL vi VSV

Ethanol xm nhp vo mng t bo VSV, gyng t mt s protein, nh hng n qu trnhTC ca t bo. VSV b cht hoc b c ch

Sn lng ethanol trn TG

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 trinBraxin (BNDES)

www.gbd.edu.vn

4/20/2011

41

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 X1975-1978, 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 lnnh, tng cng sut 40-80 triu lt/ nm (cha k lng ru dnnu t do).1997, sn lng ru quc doanh t 17 triu lt/ 1997. Nu kc 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 rucn t tinh bt chim 30 - 40%.

9/2007, Cty Petrosetco k tha thun hp tc thnh lplin doanh xy dng nh my sx Ethanol vi Tp onItochu Nht Bn. Nh my d nh t ti KCN HipPhc - TP.HCM vi cng sut 100 triu lt Ethanol/ nmt sn lt.

Vi 1,2 triu tn sn lt xut khu hng nm, Vit Nam cth sn xut c t nht 400 triu lt Ethanol/ nm.

Ethanol s dng pha vo xng (p ng 50% nhu cuca th trng xng Vit Nam)

TNH HNH SN XUT ETHANOL VN

4/20/2011

42

Sx ung c chaethanol (vodka, liqueur,vang)Sn xut acid acetic

Lnh vc phi thc phm

VAI TR CA ETHANOLLnh vc thc phm

Dc hcY hc: cht st trngNng lngCN ha hc

3.1.1.2. NGUYN LIU SN XUT ETHANOL

-

NGUYN LIU sx Ethanol theo quy m CNNguyn 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 maWhey (lactoserum)

4/20/2011

43

C2H4 + H2O

PP LN MEN

C2H5OH

Xc tc

PP HA HC

C6H12O6

VSV

2C2H5OH + 2CO2

3.1.1.3. PP SN XUT ETHANOL

Tinh bt/Cellulose

1.1.1.4. SN XUT ETHANOL BNG PP LN MEN

4/20/2011

44

Ch tiu Thnh phn ho hc ca mt s nguyn liu cha tinh bt trong sx ethanol (%)

Khoai ty Ng Sn Go t Tm Carbohydrate 12 - 21 68,4 74,74 69,2 42,0Protein 1,2 3,2 8,3 0,205 7,3 5,3X th 0,5 1,5 4,1 1,11 0,5 22,5Lipid 0,1 0,3 5,1 0,41 1,2 2,0Nc 72 -80 12,5 13,12 11 11,5

3.1.1.4.1. LM ethanol t nguyn liu cha tinh bt

PP sx Nguyn liu

VSV c im ca pp Phm vi p dng

LM truyn thng

Nguyn liu cha tinh bt

Bnh men n gin, hiu sut thp

H gia nh

Amylose VK lactic + mc+ men

i hi k thut cao khi cy VSV vo bnh ln men

Quy m sx CN

Mycomalt Ch phm mc + men

Tn din tch, cng sc

Quy m sx va & CN

LM CN Amylase + men

Yu cu thit b hin i, hiu sut cao

Quy m sx CN

LM ethanol t nguyn liu cha tinh bt bng pp ln men

4/20/2011

45

1,5 2,5%

Quy trnh sx ru theo pp ln men truyn thng

Hnh nh trong sx ru th cng ti VN

4/20/2011

46

Quy trnh sx ru theo pp mycomalt

Ch phm VSV (mc)

Men S. c

Phng php mycomalt: g ng ha v ru ha din ra 2 thit b.

Sn xut ethanol bng pp ln men (tt)

Thy phn tinh btbng malt, ch phmnm mc

Quy trnh sx ru t ng theo pp mycomalt

4/20/2011

47

Quy trnh sx ru theo pp amylose

VSV (VK, mc, men)ng ha, ru ha

Phng php amylose: g ngha v ru ho xy ra gn nhng thi, trong cng thit b

Sn xut ethanol bng pp ln men (tt)

Thy phn tinhbt bng enzyme

VSV

Men Saccharomyces cerevisiae

VK Zymononas mobilis

VK E.coli ( chuyn gen)

Quy trnh sx ethanol theo quy m CN

4/20/2011

48

Nghin trc

Nghin bt mn Nghin cao p

THIT B NGHIN NGUYN LIU (tinh bt)My nghin ba kiu git nc

4/20/2011

49

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 vkch thc ca ht tinh bt)

Phng phpGin on Bn lin tucLin tc

Ln men ethanol t TB theo quy m CNNU NGUYN LIU H HA - GELATINISATION

LN MEN ETHANOL T TB - THY PHN TB = enzyme

Chuyn tbt thnhng, cung cp ccht cho qt ln menQuyt nh phnln hiu sut thuhi ru.

DCH HA

NG HA

4/20/2011

50

LN MEN ETHANOL T TB THNG S K THUT CA MI TRNG LN MEN

Dch ln men Yu cu Nit trong dch ln men (nit hu c, v c)

12oP 140 -150 mg/L 16oP 200 mg/L18oP > 280 mg/L

Dch ln men t sn 300 mg/LDch LM t r ng 150 200 mg/L

Nng cht kh: 16 18%Hm lng ng kh: 90 -100 g/LKhong (Nit, Mg, P)Chnh pH: pHop 4,5 - 5

THNG S K THUT TI CTY RU BNH TYTrnh t cng vic To (oC) pht

Xung bt: 1080 L nc + 300kg go (xay 1mm) + 445g CaCl2 + 283mL Termamyl

32 10

Nng nhit 32 83 40Gi nhit 83 50Nng nhit 83 95 15Gi nhit, cho vo 120mL H2SO4 95 30Bm v h nhit dch nu bng corbin sang thng ng ha, m bo To thng ng ha l 58oC

95 58 30

Cho 32g San super, o trn, gi nhit 56 58 60dch khng lm mt mu iod, h nhit 56 30 40Bm sang thng ln men, + 50g ure

4/20/2011

51

THNG S K THUT TI CTY RU BNH TYTrnh t cng vic To (oC) pht

Ln men: t l men 8 10%

Kim tra mu sau 4 ngy ln men: Bal, chuaKt thc ln men: Bal 10%v/vLc bng my ly tmChng ct (600mm Hg) 68 - 72Lc lnh ru thnh phm 5oC

Ch tiu Mt r sx cng nghipc VN

Nng lng th (MJ/kg VCK) 9,47 9,87VCK (%) 75,9 76,7ng kh (%) 34,6 39Saccharose (g/100g VCK) 23,3 21,2Glucose (g/100g VCK) 5,1 8,7Fructose (g/100g VCK) 6,2 8,47Nit (g/100g VCK) 0,46 0,29Protein th (%) 2,87 1,80Khong (g/100g VCK) 11,7 6,2Ca (g/100g VCK) 0,48 0,44Mg (g/100g VCK) 0,32 0,18K (g/100g VCK) 1,94 0,92P (mg/100g VCK) 47 30Na (mg/kg VCK) 661 75S (g/100g VCK) 0,3 0,33Cu (g/kg VCK) 1,58 3,01Fe (mg/kg VCK) 97 417Mn (mg/kg VCK) 52 52Zn (mg/kg VCK) 3,8 12,6THNH PHN HA HC CA MT R

3.1.1.4.2. LM ethanol t nguyn liu cha ng

(mt r)

4/20/2011

52

Quy trnh sx ethanol t mt r

LN MEN ETHANOL T MT R

4/20/2011

53

LN MEN ETHANOL T MT R

3.1.1.4.3. LM ethanol t nguyn liu cha CELLULOSE

4/20/2011

54

CELLULOSE

4/20/2011

55

LN MEN ETHANOL T CELLULOSE - THY PHN

Tc nhn thy phn

Acid

Ch phm nm mc

Ch phm enzyne cellulase (thy phn lin kt 1,4-beta-D-glycosidic trong cellulose, lichenin v beta-D-glucans)Kt hp acid v enzyme

Thy phn cellulose bng enzyme

4/20/2011

56

Thy phn cellulose v hemicellulose bng mc Trichoderma reseii

S quy trnh LM ethanol t nguyn liu cha CELLULOSE

4/20/2011

57

3.1.1.5. CHNG CT (DISTILLATION) & TINH CH (RETIFICATION) ETHANOL SAU LN MEN

Qu trnh thu ethanol v tp cht d bay hi tdch 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

CHNG CT ETHANOL LIN TC (continuous rectifier )

B: ct tinh luynC, D: b phn ngng tR: b phn lm lnh spE: my xc nh v cn

U: thit b thu hi nhit

4/20/2011

58

Thit b chng ct ti nh my ru Bc Kinh TQ

Dy chuyn ng chai ti nh my ru Bc Kinh TQ

3.1.1.6. CC SN PHM CA QU TRNH LM ETHANOL

Sn phm ph

Men

CO2Hm ru

Sn phm: C2H5OHAldehydes: by oxidation of alcoholAcids: by oxidation of aldehydesFusel Oil: by conversion of freeamino acids in water to higheralcoholsEasters: by esterification of alcoholsand acidsVolatile sulphur compounds: bycombination of sulphate and sulphurwith amino acids

4/20/2011

59

CH TIU K THUT CA ETHANOLCC SN PHM CA QU TRNH LN MEN ETHANOL

Nng ru 15oC > 96,2Acid hu c (tnh theo mg acetic/l ) < 15Furfurol 0Metanol 0Aldehyde tng s (mg/L) < 4Ru cao phn t (mg/L) < 4Este (mg/L) < 30Hm lng kim loi ng ( Cu mg/l ) < 30Xyanhydric ( HCN mg/l cn 100o )

(vi cn sx t khoai m

4/20/2011

60

CC SN PHM PH CA QU TRNH LN MEN ETHANOL HM RU

Thnh phn Dch hm t nguyn liuKhoai ty Khoai m

Cht kh (%) 6 3,3Protein th (% tng cht kh) 27 14,2Lipid 2,7 1,8Cellulose 8,1 2,8Cht chit phi nit 49,9 74,2Tro 12,6 6,7

HNG S DNG HM RUThc n cho gia scNguyn liu lm phn vi sinhNguyn liu sx vitamin bng VSV

CC SN PHM PH CA QU TRNH LN MEN ETHANOL CO2

Quy trnh x l CO2

Oxi ha

Ra nc

Tch nc

Lm ngui Tch du

X l bng than hot tnh, silicagel

Nn ha lngKMnO4/ K2Cr2O7

Sn phm

B sung vobia lon, ncngt c gas,ChampagneNui to

4/20/2011

61

SX ETHANOL BNG PP LN MEN HIU SUT THU HI ETHANOL

3.1.1.7. ETHANOL - GREENFUEL

Nhin liu truyn thng ang cn kit

Nhin liu sinh hc

Mt r, bp, ph phm nng nghip,

cellulose

4/20/2011

62

Nguyn liu Nhin liu sinh hc/ tng lng xng du

EU Bp, c ci ng, ma, sn

Nm 2010: 5,7%

M Bp Nm 2012: 24 t ltPhp C ci ng Nm 2010: 7%n Cy Jatropha,

PongamiaNm 2012: 5,%

Trung Quc Cy la min Tng 12 triu tn nhin liu sinh hc/nm

Philippines Ma, Cy la minThi Lan Ma, sn Nm 2010: 3 triu ltBrasil Ma

CHNH SCH PHT TRIN GREENFUEL

Production of bioethanol from sugar cane or sugar beet

This is the simplest of all the processes for producing bioethanol byfermentation. Sugar cane has an energy production per hectare that issubstantially higher than the other feedstocks considered here, but theprocess conversion efficiency is only about 0.35-0.40 GJ bioethanol perGJ feedstock. The sugar cane residue or bagasse can be burned togenerate electricity, producing about 0.08 GJ electricity per GJ feedstock.

4/20/2011

63

Production of bioethanol from corn using wet or dry milling

Wet milling, several types of residues are produced; dry millingproduces only one type of animal feed product. The starches werebroken down into C6 sugars. The current conversion efficiency of bothprocess routes is about 0.55 GJ of ethanol per GJ wheat (USDA 2002).The processes are well established but there is some limited scope forefficiency improvements.

The process is similar and very slow. The wheat is first crushed ormilled. In its passive form, malting is a process by which undercontrolled conditions of temperature and humidity, enzymes present inthe wheat break down starches into C6 sugars. These sugars arewashed out of the wheat with water, whilst the leftover residue can besold for animal feed. Fermentation at 32-35oC; pH 5.2. Ethanol isproduced at 10-15% concentration and the solution is distilled toproduce ethanol at higher concentrations.

Production of Bioethanol from wheat from malting and fermentation

4/20/2011

64

Production of bioethanol from wood or straw by acid hydrolysis and fermentation

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

4/20/2011

65

SC water: supercritical water

4/20/2011

66

9 FUEL ICONSUnleaded PetrolSuper UnleadedHigh OctaneDieselBio-EthanolBio-DieselCNGLPGLeaded Four Star.

3.1.2. Cng ngh sn xut ru cao

3.1.2.1. SX RU RUM T MA

www.gbd.edu.vn

4/20/2011

67

Husks

All the discarded husks ofthe sugar cane. The first stepin making rum is to crush thecane to get out all the juice-you can see they make a lotof rum here at River's.

SX RU RUM T MA

After all the sugar canejuice is squeezed out,the next step in theprocess of making rumis to boil it down ingiant kettles (ex: copperkettles) - like the onesyou see here.

SX RU RUM T MA

4/20/2011

68

After the sugar cane juice is boileddown in the coppers it has toferment for a couple of days.

SX RU RUM T MA

CondenserThe final step is the condenser, where the alcohol getsseperated out thru coils and a cooling process. Whatcomes out is the final product- pure white rum.

SX RU RUM T MT R

4/20/2011

69

SX RU RUM T MA

Whisky: The difference between rum and whiskymanufacture begins with the starting point. Whisky startswith barley that must be malted before it is convertedfrom a starch to a sugar that can be fermented. Afterfermentation, it is also distilled usually using a pot still orsingle column.

Cognac: The sugar that is fermented to make cognac,come from grapes. As is the French style, only grapesgrown in a particular area in France can be used. First thegrapes are fermented into wine and this is distilled inspecial stills to yield cognac.Brandy is made the same way but because the grapesarent grown in the Cognac region, it can not be calledcognac. All spirits are aged in a special barrel prior toblending and bottling.

4/20/2011

70

3.1.3.1. GII THIU V BIA

3.1.3.2. NGUYN LIU SN XUT BIA3.1.3.3. VSV TRONG QU TRNH LN MEN BIA3.1.3.4. CNG NGH LN MEN BIA SN PHM

3.1.3. Cng ngh sn xut bia

www.gbd.edu.vn

3.1.3.1. GII THIU V BIA - LCH S PHT TRIN7000 B.C, Ngi Babylon (a ra vn t lut php quy nhv vic bn ru bia) pha ch c 20 loi bia.3000 B.C, ngi Ai Cp c i pht minh ra ng ht ungbia cn ln cn v la m500 400 B.C, bia l sn phm dnh cho ngi lao ng768: 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 imch, hoa houblon v nc1857: Pasteur nghin cu qu trnh ln men ethanol, thanhtrng bia 63oC/ 20 301883: Hansen v ging men thun khit

4/20/2011

71

GII THIU V BIA Sn lng biaSn lng bia trn th gii: >120 t lt/nm

Lng bia tiu th:

CHLB c 170L/ngi/nm

VN: 5 10 L/ngi/nm

Babylonia's table (6000 BC): Describe the beer preparation

Lut sx bia

Nguyn liu Ph gia Cng ngh Sn phm

c: 100%malt i mch

Php: s dng30% th liuVN: 2030%go

B sung caramen

B sung acid

Men ging khng c t bo cht

GII THIU V BIA

4/20/2011

72

GII THIU V BIA - CC HNG LN SX BIA TRN TH GIIAnherue busch (M)Heineken (H Lan)Miller (M)Kirin (Nht)Fosters (c)Danone (Php)Brahma (Brasile)Guiness (Anh) (bia en)SAB (Nam Phi) Pilsen (Tip)Carlsberg (an Mch) (men chm)

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

PHN LOI BIA THEO NH SX

4/20/2011

73

PHN LOI BIA THEO NH SX

PHN LOI BIA THEO MU SC VNG EN XANHwww.gbd.edu.vn

4/20/2011

74

GII THIU V BIA

Bia l sn phm ca qu trnh ln men ethanol

t dch nha, khng qua chng ct

Weizen

Bia en Munich

3.1.3.2. NGUYN LIU SN XUT BIA

Nguyn liu chnh: malt ai mch

Th liu: nguyn liu giu glucid

Hoa Houblon

Nc

4/20/2011

75

Nguyn liu chnh sn xut cc loi bia.

i mch ny mm v sy n m bt buc

Ht i mch

Ht malt

Ny mm

Hot hov tch luhenzyme(amilase,protase)

H enzyme trong ht malt s lmnhim v ct cc hp cht caophn t trong ni nh ca htthnh cc cht c trng lngphn t thp hn (dextrin, acidamin, pepton, ng n) hotan vo nc tr thnh chtchit ca dch ng.

NGUYN LIU SN XUT BIA - MALT I MCH

Ch tiu cht lng malt Mu sc Vng rm

Mi v Thm

chit sut > 785

Hot lc enz > 285 oWK

m < 4,5pH nc malt 5,8 6,3

Tg ng ha 10 15

NGUYN LIU SN XUT BIA - MALT I MCHCh tiu cht lng malt (tt)

Mu nc malt 2,8 3,5 oEBC

Pr tng s 9 13 %

Pr ha tan Min 4,5 %

Ch s Kolback 42 48%

nht M 1,6 cp

xp Min 84%

C ht >2,5mm >92%

C ht

4/20/2011

76

NGUYN LIU SN XUT BIA - TH LIU

TH LIUCHA NG:saccharose, syrusthu phn tinh bt(maltose,glucose)

TH LIU

TH LIU CHATINH BT: ht imch, go, tm(VN), bp tch phi(chu u), khoaim, khoai ty, tinhbt

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 chtca sn phm (tng bnca keo)To ra chng loi bia ccc mc cht lngkhc nhau


NHC IM nh hng cho qu trnh nu(do th liu c t hoc ko cenzyme)nh hng ti qu trnh lnmen (Protein ha tan trong dchnha gim, khng cn i thnhphn dinh dng)

www.gbd.edu.vn

4/20/2011

77

TH LIU S DNG TI VN - GOcu trc ht tinh bt cht, nhit h ha cao, kh nghaBia Heineken: 100% malt i mchBia Tiger: 15% goBia Si Gn: 20% go

TH LIU S DNG TI CHU U BPTch phi loi cht bo

Cht bo ph hy h bt, nh hng n cht lngcm quan


NGUYN LIU SN XUT BIA - HOA HOUBLONNguyn liu khng th thay thtrong cng ngh sx biaTo v ng du, hng thmc trng Tng kh nng to v gi bt Tng bn keo v tnh nnh sinh hc ca sn phm.Trong sx bia ch s dng hoa cicha th phn (c ht lupulin) dohoa c v hoa ci th phn chm lng cht mi v cht to vthp (khng c ht lupulin)

4/20/2011

78

Houblon to hng

3,85 5,14% acid

Houblon to v ng

8,62 9,31% acid

HOUBLON

NGUYN LIU SN XUT BIA - HOA HOUBLON

Ch tiu Hoa vin Hoa cao

Mi v Thm c trng, khng mi l

Thm c trng, khng mi l

Alpha acid 8 0,3 30 2

m 200 cht nh terpene, ester, ketone, hp cht cha lu hunh)

0,5 2

Polyphenol 2 -5Protein 12 18Lipide 2 - 4Cellulose 40 50Pectin 1 2Tro 1 - 2

4/20/2011

79

nh gi cht lng hoa houplonCh s Loi 1 Loi 2 Loi 3Mu hoa Vng vng ng Vng lc Vng xanh

Mu ht lupulin

Vng ng nh Vng, vng sm Vng sm

Mi Thm d chu, c trng

Thm, khng c mi tp cht

Hi nng

Cnh hoa To u chc, khng rch

Cnh hoa c th b rch (1,5cm), c chm c ph

Rch nhiu, chm c ph nhiu

Tp cht

4/20/2011

80

Hoa dng ht, vin: W 4 -6%, bt hoa kh (c th thm 20% bentonote SiO2ri to thnh vin

Hoa cao trch ly; dngst c hm lngpolyphenol thp, lmgim bn keo

Hoa kh (W 4 -6%)

Nghin

Ry (2-8mm)

Bao gi (chn khng,

CO2, N2)

Bo qun

Bt hoa kh

Quyt nh n cht lng biaChim 90 95% trong bia

YU CU V NC TRONG SN XUT BIAYU CU CMQUAN: trong, khng cmi v l

YU CU VI SINH:khng c E.coli vStreptococci/100 mlnc.

YU CU HO LpH 7.0 cng 2-4mg/L kim

4/20/2011

81

MEN NI(S. cerevisiae)

MEN CHM(S. carlsbergensis)

Nhit ln men 10 - 25 0 - 10Kh nng ln men Mnh, xy ra trn b

mt mi trngMnh, xy ra trong lng mt

Kh nng s dng ng 3C rafinose

33% 100%

Kh nng lng khi ln men kt thc

Cc tb kt chm,chui to thnh lpdy ni ln b mtmt, bia trong chm

Cc tb kt chm,chui lng xungy thng ln menrt nhanh, bia ttrong nhanh hn

3.1.3.3. VSV TRONG LN MEN BIA

Ta ch ta p ch a t

M alt a i ma ch /The lieu

Ng h ie n

Na u d ch n h a

Lo c

u n so i v i h o a h o u blo n

Ta ch b a h o u b lo n

La m la n h , b a o h o a O 2

Le n me n ch n h

Le n me n p h u

X ly la m tro n g b ia

Ba o h o a CO 2 , ta n g tr

Ch ie t ch ai / lon

Th an h tru n g

N c

Houblon

O 2 v o tru n g

CO 2

Chai / lon

Na m me n Nh a n g io n g

Sa n p h a m b iachai / lon

Ta p ch a t

Ba malt

Ba h o u b lo n

Sin h k h o in a m me n

Ca n

Ca n

3.1.3.4. CNG NGH SX BIA

4/20/2011

82

CNG NGH SN XUT BIA

3.1.3.4.1. Sx bia NGHINMc 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 mCu trc ht tinh btThit bPhng php nghin

Nghin th liu: go

Nguyn tc: nghin go cng mn cng tt

4/20/2011

83

Sn phm nghin malt: v tru, ht tm ln, tm nh, bt.Mc nghin MALTNghin 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 malt (% khi lng)Ch tiu Nghin trc (lc

bng ni lc)Nghin ba (lc bng thit b lc khung bn)

Tru 15 18 9 12Ht tm ln 12 15 12 15Ht tm nh 30 35 30 35Bt 25 - 35 40 - 45

Trch ly cc cht chit c Mthp (ng, acid amin, vitamine,khong) t malt i mch v th liu vo ncThy phn mt s cht c Mln thnh cc cht c Mnh (ccht cho nm men)

3.1.3.4.2. Sx bia NU DCH NHA

Cc phn ng xy ra trong qu trnh nu dch nhaThy 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 ngnThy phn mt s cht hu c c cha acid phosphoric: tothnh acid phosphoric

4/20/2011

84

THNG S K THUTT 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

3.1.3.4.2. Sx bia NU DCH NHA (tt)

CNG NGH SX BIA Nh my Bia LIDA500kg nguyn liu/m thu c 3100 3300L dch lnmen. 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 15tng t 40oC n 72oC, 72oC/20. Trong 10 tng n83oC, 83oC/5. Trong 5, h xung 72oC, 72oC/20.Trong 20 tng ln 100oC, 100oC/15. Trong 5, h xung65 67oC

Ni malt: 370 kg malt + 1480 1850 lit nc + 300gCaCl2. Dng H3PO4 chnh pH 5,3 -5,4. Khi nu goc 60 th nu malt. 40oC/20. Trong 15, Tng n50oC, 50oC/20. Trong 10 tng n 65 67oC .

4/20/2011

85

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 nugo v malt l 210. Sau lc dch v ra b (40 50). Dch lc c un si trong 120. Trong thi gianun si, cho hoa houblon 3 ln. Thi gian cho hoa: ln 1sau 10 un, ln 2 sau 30, ln 3 sau 75. B sung 250mLcaramen chnh mu dch ng. + 2g ZnCl2 bsung nguyn t vi lng cho men.

Sau 120 un, chuyn qua lng 25 tch cn, cc chtkhng ha tan nh xc hoa, cht kt ta

Lm lnh nhanh.

Chuyn dch vo tank cy men trc

Mc chTch b b maltRa b malt

Phng phpRa b malt bng nc nngToop = 78 80oC

Cc yu t nh hng n qu trnh lcMc nghin maltNhit 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)

3.1.3.4.3. Sx bia Lc dch nha

4/20/2011

86

So snh hiu qu s dng thit b lcCh tiu Ni lc Lc khung bnMng lc B malt Vi cotonChiu cao mng lc 30 60cmTc lc 0,5 4,5m/s 1 2m/sThi gian lc 60 110 20 50; P 0,3 barLng nc ra Tn nhiu ncThi gian ra b 60 120,

ra ti a 3 ln90 140. P 0,4 1,2 bar

Thi gian tho b 30 40 pht Tho b t ngTn tht cht chit Thp Cao hn m b Cao hn ThpMc c gii ha v t ng ha

Tt Tn nhn cng

3.1.3.4.3. Sx bia Lc dch nha (tt)

MC CHChit 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 xun cht lng sn phm

3.1.3.4.4. Sx bia un si dch nha vi hoa Houplon

4/20/2011

87

LNG HOA HOUPLON B SUNG (Tnh theo acid) 7 20 g acid/hL

THI GIAN UN SI: 60 120PHNG 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

3.1.3.4.4. Sx bia un si dch nha vi hoa Houplon (tt)

Nguyn l: Bm dch nha vo thitb hnh tr ng, dung dch chuynng theo ng xon c. Dng lcly tm lng (lng xoy tm)

3.1.3.4.5. Sx bia Tch b hoa Houplon

4/20/2011

88

Thnh phn dch nhapH 5,2 5,4Nng cht kh 10 13oPtGlucose 5 15 g/LFructose 0,5 2 g/LSaccharose 2 6g/LMaltose 45 65g/LTrisaccharide (maltotriose, raffinose) /Dextrin /Beta glucan v gum 200 800mg/LNit tng 500 1.100mg/LNit amin t do (FAN) 100 250mg/LCc cht khc (acid nucleic, vitamine, khong, polyphenol)

/

MC CHH nhit dch nha, chun b cho ln men

Cung cp oxy cn thit cho sinh trng ca men

YU CULm 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

3.1.3.4.6. Sx bia Lm ngui v np oxy vo dch nha

4/20/2011

89

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. Cyqua 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 dch800L, 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

MC CH LN MEN: Chuyn dch nha thnh bia

Ln men chnh

Ln men ph

Chuyn ha ng thnh ethanol, CO2 v cc sn phm ph, sp trung gianTc 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 haCO2 cho bia ti, n nh hngv, cht lng, trong cho snphm, tng gi tr cm quan

3.1.3.4.7. Sx bia NHN GING MEN & LN MEN (tt)

4/20/2011

90

CC YU T NH HNG N LN MEN CHNHMi trng ln men (dch nha): pH, nng cht kh, thnh phn ha hc

Nm men

iu kin ln men: To,

THNG S K THUT0,5L 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

3.1.3.4.7. Sx bia NHN GING MEN & LN MEN (tt)Ln men chnh Primary fermentation

THNG S K THUT LM PH4 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

3.1.3.4.7. Sx bia NHN GING MEN & LN MEN (tt)Ln men Ln men ph Secondary fermentation

4/20/2011

91

Hnh nh thit b ln men bia

MC CHTch trit cc phn t rn lng, khuych tn trong biaTng trong, Lm n nh thnh phn c hcTng bn keo v bn sinh hc cho sn phm

YU CUNhit lm trong: 1 2oC

THIT B LCThit b lc khung bn: r, d vn hnh, bi v VSV d xm nhpThit b lc ly tm, lc da, lc nn.

3.1.3.4.8. Sx bia Lm trong bia

4/20/2011

92

Cc cht lm trongCho 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

3.1.3.4.8. Sx bia Lm trong bia (tt)

Lc khung bn

MC CHChun ha hm lng CO2 cho bia chai v bia lon

YU CUNhit : 0 1oC

P = 0,15 -0,2 MPa

Hm lng CO2 trong bia non: 4-4,5g/l. Nu khng t tiu chun: b sung thm CO2

3.1.3.4.9. Sx bia Bo ha CO2

4/20/2011

93

MC CHKo di thi gian BQ sn phmD vn chuyn v s dng

YU CUNhit : 0 1oCP = 0,15 -0,2 MPa

Quy trnh cng ngh phn xng chit

3.1.3.4.10. Sx bia Chit v thanh trng bia

Ch tiu cm quan Yu cuMu sc Vng rm sng trong Trong sut bt Bt trng, mn lu tanMi Thm c trngV ng du, m , c hu, c trng

Ch tiu vi sinh Yu cuVi khun k kh 0VKHK

4/20/2011

94

Ch tiu ha l Yu cuCht ha tan ban u (% khi lng) 10,2 0,2ng st (%) 1,8 2,2 cn 4,3 0,2pH 4,0 4,2 chua (mL NaOH/100mL bia) 1,6 0,2 mu (EBC) 7,0 7,2CO2 (g/L) 4,0 4,5 bn ca bt (giy) 150 250Oxi < 3ppm ng 20oBUDiacetyl < 0,1ppm

Sa hnh nh my bia ti Bc Kinh - TQ

4/20/2011

95

3.1.4. Cng ngh sn xut ru vang

3.2. Cng ngh sn xut acid hu c thc phm3.2.1. Sx acid lactic3.2.2. Sx acid acetic3.2.3. Sx acid citric

www.gbd.edu.vn

4/20/2011

96

3.2.1. Sx acid lactic

3.2.1.1. GII THIU V ACID LACTIC

3.2.1.2. VAI TR CA ACID LACTIC3.2.1.3. C CHT SN XUT ACID LACTIC

3.2.1.4. VSV LN MEN ACID LACTIC3.2.1.5. CNG NGH SN XUT ACID LACTIC3.2.1.6. THU NHN V TINH SCH SN PHM

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 khoahc Php) sx acid lactic bng pp ln men

3.2.1.1. GII THIU V ACID LACTIC

C3H6O3 CH3-CHOH-COOHM= 98,08; khng mu, mi nh, ht m cao, Tosi = 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 pdng vo cc lnh vc khc nhau

4/20/2011

97

(dng L c trong c th ngi)

T l D:L = 50:50 (Hn hpraxemic (k hiu l DL-acidlactic)Raxemic c dng lng, tantrong nc, cn, khng tantrong CHCl3, To si = 122oC,nhit nng chy 16,8oC

Raxemic ch c c khi tinhnh tng hp hu c

D-acid lactic L-acid lactic

LACTIC TRONG LNH VC THC PHM

LACTIC TRONG LNH VC PHI THC PHM

3.2.1.2. VAI TR CA ACID LACTIC

Sn lng acid lactic c tnh khong50 ngn tn/nm, khong 2/3 sn lngc sn xut bng pp ln men

4/20/2011

98

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, thchhp lm chai rt nng, khay s dng trong l vi ba, ccloi si v thit b in t chu nhit

3.2.1.2. VAI TR CA ACID LACTIC (tt)

4/20/2011

99

3.2.1.3. C CHT SN XUT ACID LACTIC

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 RPha long mt r (1mt r : 3 nc)

Than hot tnh

Dch trong

Mi trng ln men lactic

B

Mt r x l muH2O 5% H2SO4 (w/w)

Mt r x l h keo

H2O Khong

Na2CO3/NaOH

3.2.1.3. C CHT SN XUT ACID LACTIC (tt)

4/20/2011

100

X L NGUYN LIU CHA TINH BT

Nit


Thy phn

Mi trng ln men lactic

Nguyn liu cha cc ng n gin

CaCO3

3.2.1.3. C CHT SN XUT ACID LACTIC (tt)

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 lBacterium 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 lacticng hnh

4/20/2011

101

Cc VSV chnh s dng trong cng nghip sx acid lactic hin nay:Lactobacillus casei.Streptococcus thermophilus.Lactobacillus acidophilus.Lactobacillus bulgaricus.Lactobacillus delbrueckiiChng c th s dng tinh bt: L. amylophilus, L. amylovorus

3.2.1.4. VSV LN MEN ACID LACTIC (tt)

C ch chuyn ha glucose ca VK lactic ng hnh

4/20/2011

102

VK Top pHop c im sinh ha SPLb. casei 35-

39oC5 7 +: Glu, Fructose, Lactose,

GalactoseLM chm: Maltose, Saccha., -: Manitol, Dextrin

D acid lactic

Lb. bulgaricus

45-48oC

5,4 +: Glu, Lactose, Galactose,-: Maltose, Manitol, Dextrin

L acid lactic

Lb. acidophilus

45-50oC

5,5 6,5

+: glucose, fructose, galactose, maltose

D, L acid lactic

Lb. delbrueckii

35 -45oC

4,6 -5,4

+: Glu, fructose, galactose, maltose-: lactose

L acid lactic

Strep. thermophilus

43 -46oC

5,8 6,0

+: Glu, fructose, galactose, maltose

L acid lactic


Strep. thermophilus

Lb. acidophilus


Lactobacillus bulgaricusLactobacillus bulgaricus

Lb. delbrueckiidelbrueckii (2(2 7 7 0,40,40,80,8m)m)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

4/20/2011

103

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, NaMn2+, Mg2+, Fe2+ c tc ng tch cc ln s pht trin

v sinh ra acid lactic ca VK Lactobacillus


Nguyn liu

X l

Mi trng ln men

Ln men lacticVK lactic 3-5%v/v

Mui lactac

Thu nhn lactic

Acid lactic

3.2.1.5. CNG NGH SN XUT ACID LACTIC

4/20/2011

104

Commercial lactic acid is producednaturally by fermentation ofcarbohydrates such as glucose,sucrose, or lactose.

Sn xut L-Lactic Acid (lactide) bng PP ln men trc tip khoai ty Hokkaido

4/20/2011

105

Sago Industry: PLA (Poly Lactic Acid) production incorporated with Sago

3.2.1.5. CNG NGH SN XUT ACID LACTIC (tt)

4/20/2011

106

Nh my B&G, sx lactic ln nhtChu TQ, cung cp cho thtrng thc n gia sc v sx PLA(Poly-Lactic Acid)

GALACTIC, sx L(+) lactic theo ppLM ln th 2 trn TG, xut khu >90% sn phm cho > 50 nc.

THNG S K THUTMi trng ln men

10 15oBxpH = f (pHop ca VK s dng ln men)

iu kin ln menTo = f (Top ca VK s dng ln men)Fermenter c cnh khuy v thi khDuy tr pHop trong sut qu trnh ln men

VSVS dng chng VK lactic ng hnhT l ging 3-5% (v/v), mt t bo 106 tb/ml

3.2.1.5. CNG NGH SN XUT ACID LACTIC (tt)

4/20/2011

107

3.2.1.6. THU NHN V TINH SCH LACTIC

Cng ngh hin iCng ngh hin iPP thm tch in &trao i ionPP thm tch in &trao i ionPP tch phaPP tch pha

Cng ngh truyn thngCng ngh truyn thng

Dch sau ln men (lactate canci)Dch sau ln men (lactate canci)

un dch n 80un dch n 80--9090ooCCCaCOCaCO33

Chnh pHChnh pHdchdch 6.56.53 5 h

Lc khung bnLc khung bn

Thit b to kt ta CaThit b to kt ta Ca2+2+lactate (ln 1)lactate (ln 1)Dch lcDch lc

Dch lcDch lc

C c dch lcC c dch lc

sinh khisinh khi

7070--8080ooCCKt taKt ta

Thit b to kt ta CaThit b to kt ta Ca2+2+lactate (ln 2)lactate (ln 2)

Lactic Lactic

Tinh sch lactic theo cng ngh truyn thngTinh sch lactic theo cng ngh truyn thng

Chnh pHChnh pHdchdch 6.56.5NaNa22COCO33

Thm tch inThm tch in

NaNa++ lactate (lng)lactate (lng) sinh khisinh khiTb C c chn khngTb C c chn khng

Thm tch in trch lyThm tch in trch ly

Dung dch acid lacticDung dch acid lactic

Ct trao i ionCt trao i ion

LcLc Dch trongDch trong

C c n 40 C c n 40 70% W70% Wban uban uNaNa22COCO33

Chnh pHChnh pHdchdch 6.56.5

Thit b tch phaThit b tch pha

PhaPha huhu cc (cht(cht hphp thth lactic)lactic) ++PhaPha haha tantan ((NaNa22COCO33 ++ HH22COCO33))

ChngChng ctct chnchn khngkhng

80 240oC; 2 100mmHg

(c trialkyamin & (c trialkyamin & COCO22 (P= 75pSi)(P= 75pSi)

Dch sau ln men (lactate canci)Dch sau ln men (lactate canci)

Lactic tinh khit (99%)Lactic tinh khit (99%) PP tch phaPP thm tch in

& trao i ion

3 5 h

4/20/2011

108

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%

SN PHM ACID LACTIC

3.2.2. Sx acid acetic

3.2.2.1. GII THIU V ACID ACETIC

3.2.2.2. VAI TR CA ACID ACETIC3.2.2.3. C CHT SN XUT ACID ACETIC

3.2.2.4. VSV LN MEN ACID ACETIC3.2.2.5. CNG NGH SN XUT ACID ACETIC3.2.2.6. THU NHN V TINH SCH SN PHM

www.gbd.edu.vn

4/20/2011

109

3.2.2.1. GII THIU V ACID ACETIC (ETHANOIC ACID)

C2H4O2 CH3-COOHL carboxylic acid n gin nht v cu toM= 60,05 To to tinh th = 16,7oC n mn nhiu kim loi, gy bng daGim n (acetic 4 8%)

ACETIC TRONG LNH VC THC PHM

ACETIC TRONG LNH VC PHI THC PHM

3.2.2.2. VAI TR CA ACID ACETIC

Nhu cu v acetic acid khong 6.5 triutn/nm (Mt/a). Khong 4 Mt/a csn xut t cng nghip ha du hoct cc ngun sinh hc.

Tnh ti 2006, VN nhp khu acidacetic 100%

4/20/2011

110

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 lmgim phi c sx bng pp ln men.Mi v v ca gim ph thuc vo nguynliu v cch sx.

3.2.2.2. VAI TR CA ACID ACETIC (tt)

TRONG LNH VC PHI THC PHMCN ha cht: s dng nhiu trong thuc nSx polyethylene terephthalate (s dng trong nc giikht ng chai)Sx cellulose acetate: s dng trong phim chp hnhSx polyvinyl acetate: dng trong keo dn gS dng lm dung mi pha ha chtSx vinyl acetate monomer dng trong snCng nghip ch bin cao suTng hp cht do t siTrong gia nh: acetic long lm sch cn bn.

3.2.2.2. VAI TR CA ACID ACETIC (tt)

4/20/2011

111

3.2.2.3. C CHT SN XUT ACID ACETIC


Cellulose (s dng Clostridium lentocellum SG6 nghin cu ca H Osmania, n ) Nguyn liu cha ng

Cn, ru vang

3.2.2.4. VSV LN MEN ACID ACETIC

VK hiu khSau 12h, sinh khi tng 17 triu lnChuyn ha ru thnh acid acetic

4/20/2011

112

3.2.2.5. CNG NGH SN XUT ACID ACETIC

PP HA HC (3)Methanol carbonizationButane oxidationAcetaldehyde oxidation

PP SINH HC

PP HA HCCho ti 1960s, acetic acid c sx ch yu bng pp oxi hacc ngun hydrocarbon.

1960s, PP sx acetic quy m cng nghip (methanolcarbonization).1970s, Monsanto ci tin pp ny, gim chi ph ph sx.

Hin nay, s dng pp methanol carbonization, kt hpmethanol v carbon monoxide to thnh acetic acid. Lngacetic sx t pp ny chim 80% tng sn lng

3.2.2.5. CNG NGH SN XUT ACID ACETIC (tt)

4/20/2011

113

CATIVA acetic acid plants in Hull, UK (left), and Chongqing, China (right)

At the heart of this success is BP's proprietary andtrademarked CATIVA process for acetic acidmanufacture, which, with a market share of morethan 25 per cent of the 10 million tones of aceticacid produced globally each year, can justifiablyclaim to be leader of the pack.

4/20/2011

114

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 va ra cng ngh ln men nhanh.


3.5.2.5.1. Phng php ln men chm (PP Orleans)3.5.2.5.2. Phng php ln men nhanh

3.5.2.5.3. Phng php ln men chm3.5.2.5.4. Phng php kt hp


4/20/2011

115

S sn xut gim go bng pp ln men acetic


Mi trng dinh dngGim: B sung vo mi trng ban u acid homi trng, ngn chn s pht trin vi sinh vt c hi, bsung 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 bsung 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 sinhtrng nh: glucoza, cao nm men, pepton...

Gim n Cc yu t nh hng

4/20/2011

116

nh hng ca oxyCn lng khng kh ln cho qutrnh 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 cn1mol O2.Qua thc t nhiu tc gi cho thylng oxy cn thit phi ln gp ilng oxy ho theo l thuyt.


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 vikhun v lm gim hiu sut qu trnh ln men do s bayhi acid acetic v ru etylic.


nh hng ca tc nhn sinh hc(Ln gim, b gim, rui gim

4/20/2011

117

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 Comdidamycoderma, Acetobacter xylium, Acetobacter aceti, lngim (2), b gim, rui gim (3) gy ra.Khc phc (1,3): v sinh thit b ln men sch, thanhtrng Pasteur dch nhit 60 -70oC ri cho ln men li,cho thm vo K2S2O5 vi liu lng 5 15gam/100 lt gimkt 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 lnn 40 500C ri lc loi b ln gim.

Gim n Cc h hng thng gp

3.5.2.5.1. PP LN MEN CHM (PHP - PP Orleans)Xut x: t 1670 vng trng nhoni ting ti Php (dch vang nho trong nhng thng g h np dndn b c, c mng trng ph trnb mt cha VK Acetobacterorleaneuse v tr nn chua).Hin nay ch cn mt vi ncchm pht trin cn s dngphng php ny, trong c VN

4/20/2011

118

Thng ln men acetic theo pp Orleans (150 L)

Nguyn liu: dch nhoLn men trong thng gsi 250 300 litV dch ln men: 1/2 - 2/3 VthngMi trng ln men: 12 14oBxVSV: Ace. orleaneuseThi gian ln men: 10 20ngy 5 tunHm lng acid acetic tc: 5 -6%

3.5.2.5.1. PP LN MEN CHM (PHP - PP Orleans) (tt)

u im

D thc hin

Thit b khng phc tp

Sn phm thm

Nhc imThi gian ln men diHm lng acid acetic thp Nng sut thp, kh c gii ha, hin i haTn din tchSau 20 ngy, acetic b oxi ha (ha hc v sinh hc)

Khc phc s oxi ha aceticTrong dd acetic phi cn 5% cnThanh trng PasteurB sung 0,2% NaCl tiu dit ln dm

3.5.2.5.1. PP LN MEN CHM (PHP - PP Orleans) (tt)

4/20/2011

119

Xut x: Shiizenbachi tm ra nm 1923.Nguyn tc: to ra b mt tip xc ln hn gia VKAcetobacter vi khng kh oxy ha ru bng cch s dngvt 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, thtch 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 lcht d bay hi), thit b phc tp, kh iu chnh thit bthng kh.

3.5.2.5.2. PP LN MEN NHANH (C)

B1.Thanh trng cht mang

B2. Gn VSV vo cht mang

B3. Cho dch ln men tip xc vi VSV c nh

Nhc im

Thi kh lm oxi ha ru v acetic

u imThi gian ln men nhanhHm lng acid acetic cao Thit b n gint tn din tch

3.5.2.5.2. PP LN MEN NHANH (C) (tt)

4/20/2011

120

Thit b ln men gim (Vinegar Generator)

Thng ln men g, hnhtr, H=2,5 6m; =1,2 3m; H= 2 C h thng thi kh tdi ln trnMT ln men: Cn, 0,1 0,5% ng, P, N, K VSV: Ace. xylinum gnvo cht mang (mt ca,li bp) Tgian LM: 8 - 10 ngy

Hm lng acid acetict c: 8 -10%

3.5.2.5.2. PP LN MEN NHANH (C) (tt)

H thng ln men acetic cgi l acetator.Cho dung dch ln men vothit b v thi kh rt mnh.Khi trong dung dch ln mens to ra th huyn ph v dungdch ln men. Hai th ny luntrn ln vo nhau v nh vyqu trnh oxy ho xy ra rtmnh lit. u im: hiu sut chuynho c khi t c 98 99%.

Thit b ln men chm

3.5.2.5.3. PP LN MEN CHM (Submerged Process)

4/20/2011

121

Cu to h thng ln men: phn trn l lp m (lp chtmang cha VSV), lp gia l 1 thng cha dung dch saukhi ln men phn trn chy xung, di cng l hthng thi kh mnh, kh s c thi qua phn dung dchny ri chuyn ngc ln phn trn (ging thit b lnmen chm). u im: hiu sut ln men rt cao, nng acidacetic 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

3.5.2.5.3. PP LN MEN KT HP

Bnh ln men hi lu, c sc kh ti PTN Vi Sinh H NL. VK Acetobacter aceti c c nh trncht mang l b ma. C cht l dung dch nc xoi

4/20/2011

122

NNG CAO NNG ACETIC TRONG PP LN MENPP CHNG CT BNG NHITPP CHNG CT BNG MUI

S dung mui c kh nng phn ly mnh (CaCl2 hocCH3COONa). Hiu sut thu c s caoPP CHNG CT TRCH LYChn 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 ctPP CHNG CT CHN KHNG

BLND-101 / V-101

Blending PFLN-101 / PF-101

Plate & Frame FiltrationINX-101 / C-101

Ion Exchange

CMPS-101 / G-101

CF Compressor

HSRL-101 / ST-101

Heat Sterilization

FRMN-101 / F-101

Fermentation

AFLN-101 / AF-101

Air Filtration

AFLN-102 / AF-102

Air Filtration

STRG-101 / V-103

Storage Tank

C-Source

Water 1

S-103

S-104S-105

S-106

S-107 S-108

S-109

S-112

AirS-115

S-117

S-119

S-120

S-121

Citric Acid Production - Recovery Based on Precipitation

BLND-103 / V-104

Blending

Nutrients

HSRL-102 / ST-102

Heat Sterilization

S-102

S-110

S-111

Water 2 RVFL-101 / RVF-101Rotary Vacuum Filtration

RXN-101 / R-101

Product Precipitation

RVFL-102 / RVF-102

Rotary Vacuum FiltrationRXN-102 / R-102

Gypsum Formation

RVFL-103 / RVF-103

Rotary Vacuum Filtration

RVFL-104 / RVF-104

Rotary Vacuum Filtration

RDRNG-101 / RDR-101

Rotary Drying

S-101

S-113 Biomass

S-118

Lime

S-123

S-124

S-125

S-126

S-127

Sulfuric AcidS-129

S-130

S-131GypsumS-137

S-138

S-139

S-140

S-141

Product

CCRN-101 / R-103

Cont. Crystallization

S-122

S-128S-133

S-134

3.2.3. Sx

acid citric

4/20/2011

123

Elements $/kg $/ao %Raw material 0,40 3 949 000 17,79Depreciation and 1,09 10 866 000 48,96MaintenanceSalaries 0,27 2 668 000 12,02Operation supply 0,00 19 000 0,09Lab./QA 0,04 400 000 1,80Waste treatment 0,13 1 290 000 5,81Aux. facilities 0,30 3 003 000 13,53Total 2,22 22 195 000 100,00

3.2.3. Sx acid citric chi ph sx

Costo de produccin

0,1%1,8%

5,8%

12,0%

17,8%13,5%

49,0%

Materias primas

Depreciacin

Salarios

Sum de operacin

Lab/Aseg. Calidad

Tratam. de residual

Fac. auxiliares

3.3. Cng ngh sn xut acid amin3.3.1. Sx Lysine

3.3.2. Sx Glutamic

4/20/2011

124

CC PP SX ACID AMINTHY PHN PROTEINTNG HP HA HCCHUYN HA SINH HCLN MEN

NG DNG CA ACID AMINThc phmHa hcY hcCc lnh vc khc

3.3. Cng ngh sn xut acid amin

4/20/2011

125

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 LYSINE3.3.1.4. LN MEN LYSINE3.3.1.5. THU NHN V TINH SCH 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 emgip tr n ngon ming, hp th dinh dngtt, pht trin chiu cao; cht ph gia thc n;thc phm chc nng, dch truyn amin, bsung vo thc n gia sc

3.3.1.1. GII THIU V LYSINE

4/20/2011

126

LYSINE (2,6-diaminohexanoic acid) ( amino acid)C6H14N2O2, M= 146.188, cu hnh L v D

CH2 CH2 CH2 CH2 CH COOH| | NH2 NH2

3.3.1.1. GII THIU V LYSINE (tt)

1985, Nht Bn sx lysine qui m ln bngCorynebacteriumglutamicumTng sn lng trn TG:1triu tn/nm

MT ln menGlucose: 50g (10 12%)tinh bt ng ( loi bo): 20 gNgun nit: ure, cc loi mui amon hoc ncamoniac: (NH4)2SO4: 25g; Urea: 1 gKhong: mui Photpho 0,008 0,02mg/l;KH2PO4: 2.5gMgSO4.7H2O: 0.75g (0,03 0,5%)biotin 7,5g/l; treonin 40mg/lduy tr pH = 7.0 bng NaOH 2MNhit : 28 300C.

3.3.1.2. C CHT SN XUT LYSINE

C cht: mt r, methanol, glucose.

4/20/2011

127

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 nngng ho glucose, fructose, maltose, saccharose.Khng c kh nng ng ho lactose, rafinose, pentose

3.3.1.4. VSV LN MEN LYSINE

VSV C cht Y (g/L)Cyronebacterium glutacium b-6

R ng 32oC/48h 100

C.glutacium H-8241 10%Sucrose 32oC/72h 48C.glutamicum 18%Glucose 27oC/70h 60C.lactofermentum AJ 12592

Glucose 31,5oC/48h 11,8

Brevibacterium lactofermentum AJ 12937

Glucose 31,5oC/58h 120,5

B.lactofermetum Glucose 32oC/72h 48,8Bacillus methanolicus

Methanol 32oC/48h

3.3.1.4. VSV LN MEN LYSINE (tt)

4/20/2011

128

Sinh khi & cc sp lin quan trong sx lysine bng C. glutamicumATCC 13287 theo thi gian khi nui cy theo m (mmol/mol)Sn phm 5.8 h 6.9 h 8.1 h 9.2 hBiomass 86.1 68.1 55.5 56.2Lysine 0.0 191.0 181.9 184.0Valine 0.0 0.0 8.6 15.2Alanine 0.0 0.0 3.1 17.4Glycine 8.3 0.0 1.7 3.9Glutamate 0.0 0.0 0.0 0.0

3.3.1.4. VSV LN MEN LYSINE (tt)

Corynebacterium glutamicum

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

4/20/2011

129

3.3.1.4. LN MEN LYSINE

Thit b ln men c cnhkhuy v thi kh lin tcQu trnh gm hai pha: 18 24 h u (Pha to thnh sinhkhi) + 40 60h sau (Phato thnh lysine)

Cng ngh sinh hc LM L-Lysine c ci thin khngngng v s tin b v kthut to ra s lngln lysine p ng nhu cungy nay

Lysine C. glutamicum

4/20/2011

130

Asp: AspactatAsp-P: beta Aspactyl phosphateASA: Aspactate beta SinialdehydeDAP: DihydropicolinasyntetaseAK: aspartokinase Lys: Lysine

Hom: HomogerinThr: ThreoninIle: IlexinMet: MethionineHSD: homoserinedehydrogenase

4/20/2011

131

4/20/2011

132

Pilot-Scale Batch Production of High Lysine Content Animal Feed Suppliment (20h/m)

Pilot-Scale Continuous Production of High Lysine Content Animal Feed Suppliment

4/20/2011

133

Variant pathways for the synthesis of m-DAP/lysine in plants and bacteria. m-DAP/lysine biosynthesis genes present in chlamydiae are boxed

McCoy A. J. et.al. PNAS 2006;103:17909-17914

2006 by National Academy of Sciences

4/20/2011

134

3.3.1.6. THU NHN V TINH SCH LYSINEly tm dch ln men, loi sinh khi v cht khng tanDch 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 amoniacun nng dch cha lysine loi amoniacH 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 thDch sau khi ly tm s ti kt tinh thu nhn trit lysine c trong dch ln menHa 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 GLUTAMIC3.3.2.4. LN MEN GLUTAMIC3.3.2.5. CC YU T NH HNG3.3.2.5. THU NHN V TINH SCH GLUTAMIC

4/20/2011

135

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

VAI TR CA ACID GLUTAMICTrong y hc v dc hc: nng thp, acid glutamic l cht an thn, thuc trnhc u, c tc dng b noTrong thc phm: mui ca acid glutamic (glutamate) lcht iu v l tngM chnh (bt ngt) c s dng ti hu ht cc nctrn TGSn lng khong 1,6 triu tn/nm.Cc dng sn phm: cht iu v 621, ht nm, bt ngtcnh to, bt ngt xay nhHnh thc s dng: thc phm ch bin tm p bt

ngt, b sung trc tip vo thc phm

3.3.2.1. GII THIU V GLUTAMIC (tt)

4/20/2011

136

1860, Ritthaussen, c, x thnhphn 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 cca.a

1889, Kikunae Ikeda (Nht) tch a.glutamic v iu ch natri glutamatet rong bin 21/4/1909, ng ng k patent s9440 ti Anh: sx cht to v 1920, ngi Trung Quc cng tmc cng ngh sx glutamate


M chnh t nhin Mg/100g M chnh t nhin Mg/100gTo 2240 Bp ci 100Formate 1206 Nm 67Ch xanh 668 u tng 66Mc 146 Khoai lang 60C chua 140 Tm 43S 132 Hn 41Ng 130 C rt 33Khoai ty 102 Sa m 22To 102 Sa b 2


4/20/2011

137

Bt ngt - cht iu v E621.Bt ngt c v Umami - l mt trong 5 v chnh mcon ngi vn cm nhn trong ba n hng ngycng vi chua, ngt, mn, ng."iu v: c php s dng trong thc phmgip iu ha, lm tng v ngon ca mn n, khngcung cp dinh dngCht iu v c php s dng: E621, E627(disodium inosinate) v E631 (disodium guanylate)627 v 631 ("siu bt ngt) c "ngt" cao hnnhiu so vi 621.


1970s, JECFA - y ban hn hp v Ph giathc phm ca T chc Y t th gii (WHO) vT chc Lng nng (FAO) nh gi tnh anton ca m chnh v a ra quy nh v lngs dng hng ngy: 0-120mg/kg th trng1987, Hi ngh ln th 31 ca JECFA nh gili an ton ca m chnh v a ra kt lunliu dng khng xc nh

n bt ngt c

hi khng?


Tr em n bt ngt c

hi khng?

Ro cn ngn Glutamate xm nhp vo no tr:Ro cn rutRo cn nhau thaiRo cn noTr em tiu ha m chnh ging ngi ln nnkhng tm thy mi nguy hi no i vi tr em

4/20/2011

138

Nguyn liu cha tinh bt: sn, ng, goNguyn 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

3.3.2.2. C CHT SN XUT GLUTAMIC

3.3.2.2. C CHT SN XUT GLUTAMIC (tt)

4/20/2011

139

MT 1

Mt r: 20%; Urea: 2%; K2HPO4: 0,05%; MgSO4.7H2O: 0,03%; CaCO3: 1%; pH: 6,8 7,8

MT2Saccharose: 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

3.3.2.2. C CHT SN XUT GLUTAMIC (tt)

Brevibacterium

Micrococcus

Arthrobacter

Brevibacterium flavumCorynebacterium 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

Corynebacterium glutamicum

3.3.2.3. VSV LN MEN GLUTAMIC

4/20/2011

140

3.3.2.4. PHNG PHP SN XUT GLUTAMIC

3.3.2.4.1. PP HA HC: (thy phn protein t bt m, 3.3.2.4.2. PP SINH HC: (ln men)

Sx glutamic v bt ngt bng pp ha hc

3.3.2.4.1. PP HA HC

4/20/2011

141

1957, Nht sx glutamictheo quy m cng nghipbng pp ln men

3.3.2.4.2. PP SINH HC: (ln men)PHNG PHP LN MEN

Ln men tnhLn 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 - 8Sc kh: 40 85mg O2/L.min. nu khng sc kh th sn phm to ra lactate.Thi gian: 40 60gSn phm: 100 150 g/L

Sx glutamic v bt ngt bng pp ln men t nguyn liu tinh bt

Men ging l Corynebacterium glutamicum

3.3.2.4.2. PP SINH HC (tt)

4/20/2011

142

Sx glutamic v bt ngt bng pp ln men t glucose

FBP: Fructose 1-6 BiphosphateGAP: Glyceraldehyde 3 phosphatePEP: phospho enolpyruvateAcCoA: acetyl coenzyme AIsoCit: IsoCitrateKG: alpha ketoglutanateGlu: glutamateSuc: SuccinateFum: FumarateOxa: oxaloacetate

4/20/2011

143

3.3.2.4.2. PP SINH HC (tt)

4/20/2011

144

3.3.2.4.2. PP SINH HC (tt)

3.3.2.4.2. PP SINH HC (tt)

4/20/2011

145

pHop = 6,8 8

iu chnh pH bngNH4+ (ure, ncNH3, kh NH3,NH4Cl)

Cung cp oxy: (hiu kh bt buc)Thiu oxy: sn phm ch yu lacid lacti

cong nghe len men hien dai

Documents