Industrial Biotransformations

Subtilisin
Bacillus licheniformis
O
N
1) Reaction conditions
[1]: 0.198 M, 80.68 g · L –1 [407.45 g · mol –1 ]
pH: 7.5
T: 40 °C
medium: two-phase: aqueous/organic
reaction type: ester bond hydrolysis
catalyst: solubilized enzyme
enzyme: subtilisin Carlsberg (hydrolase)
strain: Bacillus licheniformis
CAS (enzyme): [9014-01–1]
2) Remarks
O
O O
S
(R/S)-1
COOEt
i) + NaOH; - EtOH
1 = 2-benzyl-3-[[1-methyl-1-((morpholin-4-yl)carbonyl)ethyl]sulfonyl]propionic
acid-ethyl ester
2 = 2-benzyl-3-[[1-methyl-1-((morpholin-4-yl)carbonyl)ethyl]sulfonyl]propionic
acid
Fig. 3.4.21.62 – 1
● The EC number was formerly 3.4.4.16 and 3.4.21.14.
EC 3.4.21.62
● On a 17 kg scale (R,S)-1 was reacted as an 8 % aqueous suspension using solid Optimase ® M
440 (6.5 % with respect to (R,S)-1, pH 7.5, 40 °C, 69 h) to yield 43 % of (S)-2.
● Alternatively, (R,S)-1 is reacted as a 10 % solution in an aqueous buffer / tert-butyl methyl ether
mixture using Protease ® L 660 (10 % with respect to (R,S)-1, pH 7.5, 40°C) to give 46 % of
(S)-2.
● Optimase ® M 440 is a granular subtilisin preparation used in detergent formulations.
● Protease ® L 660 is a stabilized, water-miscible, liquid food-grade subtilisin preparation used in
detergent formulations.
360
E
O
O
N
N
O
O O
S
O
O O
S
(S)-1
+
(R)-2
COOEt
COOH
Hoffmann La-Roche AG