Mot So Ung Dung Enzyme

8/9/2019 Mot So Ung Dung Enzyme

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Enzymes in Biotechnology


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3/29

Th tr-ng th gii cho mt s sn

phm ca cc phn ng enzyme

Sir ng cao fructose: 1 t $Aspartame: 850 triu $

Acrylamide: 300 triu $


4/29

Corn Starch Slurry (30-35% DS, pH 6.0-6.5, Ca2+ 50 ppm)

Sn xut sir ng cao fructose t tinh bt

LiquefactionThermostable E-AmylaseGelatinization (105C, 5 min)Dextrinization (95C, 2h)

Liquefied Starch DE 10-15

SaccharificationGlucoamylase(60C, pH 4.0-4.5, 24-72 h)

Glucose Syrups DE 95-96

IsomerizationGlucose isomerase(pH 7.5-8.0, 55-60C, 5 mM Mg2+)

High Fructose Corn Syrups (42% fructose)


5/29

Sn xut Glucose t tinh bt_______________________________________________________________

Liquefaction Saccharification DEGlucose

_______________________________________________________________

Acid Acid 92 85

Acid Glucoamylase 95 91

Acid/-amylase Glucoamylase 96 92

-Amylase/High pressure Glucoamylase 97 93

cooking/ -amylase

-Amylase (thermostable) Glucoamylase 97 94

-Amylase (thermostable) Glucoamylase 97-98.5 95-97.5

_______________________________________________________________


6/29

Chuyn i Glucose thnh Fructose

OHglucose

isomeraseOH

OH

HO

HO

O O

HO OH

OH

OH

HO


7/29

Qu trnh Deacylation Penicillinho hc & enzyme thnh 6-APA

CH3R C N

H

ON

O

S S

ON

CH3

COOH

Penicillin V or G

NH2 CH3CH3

COOH

(6-APA)

Penicillin acylase

Alkaline[Enzymatic]

CH3R C NH

ON

O

S

CH3

COOSiMe3

[Chemical]

[R=Ph or PhO]

PyridineMe3SiCl

PCl5ROHH2O


8/29

Cc enzyme:Penicillin G acylase (PGA)- Escherichia coli, Bacillus

megaterium,

Streptomyces lavendulae

Penicillin V acylases (PVA)- Beijerinckia indica var. Penicillium,

Fusarium sp., Pseudomonas

acidovorans

Cc enzyme c nh:

i sng: 500-2880 gi


9/29


10/29

S bin i Penicillin thnh 6-APAbng enzyme v Penicillin bn tng hp

CH3R C N

H

ON

O

S S

ON

CH3

COOH

Penicillin V or G

NH2 CH3CH3

COOH

(6-APA)

Penicillin acylase[Acylation] Acidic

Semisynthetic Penicillins

Penicillin acylase

Alkaline[Deacylation]


11/29

Tng r l ir- t l r

i r i l r

itril r t nd idase reacti ns


12/29


13/29

Ph-ng php ho hc: Nhm amino ca aspartic acid cn bo v ngn

cn phn ng ca n vi phn t khc ca aspartic acid

cho sn phm ph khng mong mun.

Chnh xc enantiomer n ca mi cht phn ng

phi -c s dng -c ho hc khng gian

stereochemistry mong mun ca aspartame (beta-

aspartame c v ng)


14/29

Ph-ng php enzyme:

Thermolysin thc y phn ng ch alpha-functionality

iu kin trung bnh, pH 6-8, 40 oC

Cbz, benzyloxycarbonyl


15/29

S sn t c t csinhhcca spartame

HO2C

CO2H

H2O

thermol sin

D, -phenylalanineMethyl ester

Cbz-aspartame

PhCH2OCNHO

+Ph

CO2MeH2N

HO2C

CNHPhCH2OCNH

O

CO2Me

Ph

O

N-Cbz-asparticacid

Cbz, benzyloxycarbonyl


16/29

L-Carnitine

Cht c ch thyroid

Cht gim bo (Slimming agent)

Thc phm b sung cho vn ng vin inkinh

Ch mt enantiomer ca thnh phn -c s

dng Hai con -ng x c tc sinh hc sn c snxut L-carnithine.

Saccharomyces cerevisiae


17/29

Tngh p L-Carnitine

O

Cl

reductaseO

Cl OC8H17

HO HO

OC8H17K-chloroacetoacetic

acidoctyl ester(R)-K-chloro-F-hydroxybutanoic

acidoctyl ester

HO HO

Me3N OHL-carnitine

hydroxylaseO

Me3N OHMe3N

HO H O

OHL-carnitineK-butyrobetaine


18/29

Tngh pNaproxene

CH3O CH3O

CO2H

*

CH3O CH3O

CO2H

*(D/L)

CH3O

CO2H

*

CH3O

O

C CH2CH3

CH3O

CO2H

*

biocatalysts

multistep resolution

1) Tartaricacid2) r23) Hydrolysis


19/29

TnghpCalcium ntagonist Drug

DiltiazemOMe

O

MeO2C

esterase

racemate

O

HO2C

OMe

(R,R)

(S, S)

DiltiazemNH O

O

O

OMe

S


20/29

Tngh p L-malic Acid L-AsparticAcid

T Fumaric Acid

HO

HO2C

CO2H

H2O

fumarase HO2C

CO2H

H

fumaric acid L-malic acid

HO2C

CO2H

HO2C

HCO2HHO

fumaric acid L-asparticacid

NH3

aspartase


21/29

Tngh pCatechol t GlucoseT- ng thchmi trng

benzene

a

cumene phenol

catechol

protocatechuicacid

3-dehydroshi imicacid

D-glucose

acetone

b

c

hydroquinone

d dd

HO

HO

OH

HOOH

OH

OH

O

CO2HOH

OH

OH

OH

HO

O

CO2H

(a) propylene, solidH3PO4 catalyst, 200-260C, 400-600psi.(b) O2, 80-130C thenSO2, 60-100C.

(c) 70% H2O2, DTA, Fe2+ orCo2, 70-80C.

(d) E. liAB2834/pKD136/pKD9.069A, 37C.Draths andFrost, 1995


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Xylanase-tr gi p x l lm trng

____________________________________________________

Sequence after Pulp Total active chlorineEnzyme treatment consumption

decrease (%)

____________________________________________________

(CD)EDED Softwood kraft 21(CD)EoDED Pine kraft 18.4

(CD)EpDEpD Birch kraft 18

(CD)EopDEpD Pine kraft 12

DEopDED Softwood kraft 15___________________________________________________

C, elemental chlorine (Cl2), D, chlorine dioxide (ClO3), E,alkaline extraction (NaOH), Eo/Ep, oxygen/hydrogen peroxide

reinforced alkaline extraction


24/29

Mannitol

Ph gia thc phm

Lm gim kh nng kt tinh ca -ng v-c s dng ko di tui th ca thc

phm.

S dng trong ko cao su

To cng thc d-c phm ca cc vin ngm

v ht bt. Ngn nga hp th m t khng kh, thhin tuet vi cc tnh cht nn c hc, khng

t-ng tc vi cc thnh phn hot ng v v


25/29


26/29

Qu trnh hydro ho D-Fructose

H2, catalyst

D-Fructose D-Sorbitol D-Mannitol

H2C OH

C O

HO CH

HC OH

HC OH

H2C OH

H2C OH

HC OH

HO CH

HC OH

HC OH

H2C OH

H2C OH

HO CH

HO CH

HC OH

HC OH

H2C OH

+


27/29

Chu trnh chuyn i Heterofermentative

ca Fructose thnh Mannitol

Fructose

NADPH + H+

Lactate

ADP

NADP+

Acetate

CO2

Glyceraldehyde - 3-P

Glucose 6-P

Pyruvate

NAD+

ATP

2 ADP

2 ATP

Acetyl - P

NADPH + H+

NADP+6 - Phosphogluconate

Ribulose 5-P

Xylulose 5-P

NAD+

NADH + H+

NADH + H+

Fructose 6-P

2 Fructose

2 Mannitol

ADP

ATP


28/29

S chuyn ho Fructose thnhMannitol

bng enzyme

Mannitol 2-Dehydrogenase

D-fructose Mannitol

CH2OH

O

HO H

H OH

H OH

CH2OH

CH2OH

HO H

HO H

H OH

H OH

CH2OH

NAD(P)H NAD(P)


29/29

S chuyn i Fructose thnh

Mannitol bng enzyme vi s ti tocofactor lin tc

Mannitol Dehydrogenase

Glucose + H20

NADH NAD+

D-Fructose

Gluconic acid

Mannitol

Glucose Dehydrogenase

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Mot So Ung Dung Enzyme