Industrial Biotransformations

Dehydrogenase
Geotrichum candidum
1 = 4-chloro-3-oxo-butanoic acid methyl ester
2 = 4-chloro-3-hydroxy-butanoic acid methyl ester
Fig. 1.1.X.X – 1
1) Reaction conditions
[1]: 0.066 M, 10 g · L –1 [150.56 g · mol –1 ]
pH: 6.8
T: 28 °C
medium: aqueous
reaction type: reduction of keto group
catalyst: suspended whole cells
enzyme: dehydrogenase
strain: Geotrichum candidum SC 5469
2) Remarks
Cl
● In initial experiments an enantiomeric excess of 96.9 % was reached. After heat-treatment (at
50 °C for 30 min) of the cells prior to use the ee increased to 99 %.
● The oxidoreductase was also isolated and purified 100-fold. It is NADP-dependant and the
regeneration of cofactor in case of the isolated enzyme is carried out with glucose dehydrogenase.
The isolated enzyme was immobilized on Eupergit C.
3) Flow scheme
Not published.
O
1
O
4) Process parameters
yield: 95 %
ee: 99 %
reactor type: batch
reactor volume: 750 L
capacity: multi kg
down stream processing: filtration and extraction
enzyme supplier: Bristol-Myers Squibb, USA
company: Bristol-Myers Squibb, USA
O
E
+NADPH,H +
Cl
OH
O
(S)-2
O
EC 1.1.X.X
Bristol-Myers Squibb
187