White Biotechnology: Production of Chiral Compounds through Enzymatic Processes, Slides of Biochemistry

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Module 10:

White Biotechnology

Lecture38:

White Biotechnology

(Contd.)

D

-Hydantoinase

NH

N

H

O

NH

N

H

O

NH

2

N

H

O

E

D Hydantoinase

O

O

H

NH

N

H

O

NH

2

N

H

CO

H 2

O

H

O

H

O

H

pH = 8.0, medium: aqueousreaction type: carboxylic acid amide hydrolysiscatalyst: immobilized whole cellsstrain:

Bacillus brevis

developer: Kanegafuchi; ee = 99%; yield = 99%

*** The unreacted L-hydantoins are readily racemized under the reaction condition* Quantitaive conversion is achieved because of the in situ racemization**

O

O

D-Hydantoinase/carbamoylase

NH

N H

O

NH

N H

O

O

NH

2

N H

O

CO

2 H

N H

2

CO

H 2

E

E

O H

O

O H

O H

O H

subs conc: 40g/L; temp: 38

oC

subs conc: 40g/L; temp: 38 CpH = 8.0, medium: aqueousreaction type: carboxylic acid amide hydrolysiscatalyst: whole cellsstrain:

Pseudomonas sp.

developer: Dr. Vig Medicaments; ee = 99%; yield =80%

The strain contains both enzymes (hydantoinase and carbamoylase)

HN

O

O

β

-lactamase

N

N H

O

CO

2

NH

3

E

(+)

(-)

subs conc: 200g/Lreaction type: carboxylic acid amide hydrolysiscatalyst: immobilized enzymestrain:

Aureobacterium

sp

strain:

Aureobacterium

sp.

developer: Celltech Group plc.

Lactamase/Racemase

N H

O

NH

2

N H

O

NH

2

NH

E

N H

2

N H

2

N

H

2

NH

2 CO

2 H

E

E

Subs conc: 100g/L; pH = 8.0-9.0; temp: 40

o C

reaction type: Carboxylic acid amide hydrolysis/racemizationreaction type: Carboxylic acid amide hydrolysis/racemizationcatalyst: suspended whole cellsenzyme: E1 = L-lysisne-1,6-lactame hydrolase

E2 = caprolactam racemase

strain:

Cryptococcus laurentii/ Achromobacter obae

i^

99%

developer: Toray Industries Inc. ee = 99%

*** The lactamase and racemase are active at same pH.* The products is used as nutrient and food suppliment**

The products is used as nutrient and food suppliment

Nit il

CN

O

E

MeNH

Nit

rilase

CN

CN

CO

2

-NH

4

N

E

2-methylglutaronitrile

4-cyanaopentanoicacid ammonium salt

MeNH

2

Pd/C, H

2

Xolvone

subs conc: 130 g/L; pH = 7.0; temp: 30

o

C

reaction type: nitrile hydrolysiscatalyst: immobilized whole cells

y

strain: E coli transformant which expresses

Acidovorax facilis

developer: Du Pont de nemours & Co. yield = 98.7%

Xolvone is a precision cleaning solvent. It is not flammable, completely miscible with water,has a good toxicological profile and is readily biodregadable.

Dehalogenase

Cl

CO

H 2

Cl

CO

H 2

OH

CO

2 H

E

+H

O 2

HCl

-HCl

lactic acid

subs conc: 11 g/L; pH = 7.4; temp: 30

o C

reaction type: Enantioselective C-halide hydrolysiscatalyst: suspended whole cells

2 h l

id d h l

enzyme: 2-haloacid dehalogenasestrain: Pseudomonas putidadeveloper: Astra Zeneca; yield = 42%; ee = 99%

The manin application of (S)-2-chloropropionic acid is in the synthesis of Cl

Cl

O

CO

H 2

Cl

Me

O

CO H

The

manin application of (S) 2 chloropropionic acid is in the synthesis of

following enantiopure phenoxypropionic acid herbicides.

Cl

O

C

O

H 2

N

CF

3

O

O

CO

2 H

O

O

O

CO

H 2

dichloprop

mecoprop

3

2

N

O

O

CO

2

fluazifop

fenoxaprop

Dehalogenase/ application

Cl

CO

H 2

Br

CO

2 H

OH

CO H

2

CO

2 H

CO

H 2

(S)-2-chloropropionic acid

(S)-2-bromopropionic acid

(S)-lactic acid

Br

CO

2 H

Br

OH

CO

2 H

Br

(S) 2 3 dibromopropionic acid

(S) 3 b

2 h d

i^

i^

id

Br

OH

(S)

-2,3-dibromopropionic acid

(S)-3-bromo-2-hydroxy-propionic acid

CO

2 H

CO

2 H

(S)-2-bromo-butyric acid

(S)-2-hydroxy-butyric acid

The above chiral products are synthesized by the deghalogenase strategy