Industrial Biotransformations

Lipase
Pseudomonas cepacia
N
Boc
1
OH
1 = (R,S)-N-(tert-butoxycarbonyl)-3-hydroxymethylpiperidine
2 = succinic anhydride
3 = (R)-N-(tert-butoxycarbonyl)-3-hydroxymethylpiperidine
4 = (S)-hemisuccinate
Fig. 3.1.1.3 – 1
1) Reaction conditions
[1]: 0.46 M, 99 g · L –1 [215.29 g · mol –1 ]
[2]: 0.39 M, 39 g · L –1 [100.07 g · mol –1 ]
T: 20 °C
medium: biphasic: water–toluene
reaction type: hydrolysis
catalyst: immobilized enzyme (Amano lipase PS)
enzyme: triacylglycerol lipase
strain: Pseudomonas cepacia
CAS (enzyme): [9001-62-1]
2) Remarks
O
O
O
E
EC 3.1.1.3
Bristol-Myers Squibb
● The lipase of Pseudomonas cepacia from various commercial sources, especially the lipase PS
from Amano, was found to be the best enzyme for the stereospecific hydrolysis of the (R,S)
esters.
● Enzymatic resolution of the (R,S) esters by lipase P from Pseudomonas fluorescens was reported
for the large-scale production (Roche) as a process with low substrate concentration and chromatographic
separation. Such processes will have limited practical use in large-scale industrial
application.
● The process for enzymatic resolution followed by easy separation involved lipase PS/PP
enzyme-catalyzed esterification of (R,S)-alcohol with succinic anhydride followed by separation
of the unreacted R-alcohol from the S-hemisuccinate by extraction with base (5 %
NaHCO 3). Subsequent hydrolysis of the S-hemisuccinate with NaOH provided the desired Salcohol.
● By using toluene as solvent, the S-alcohol was isolated in 23 % yield (maximum theoretical
yield is 50 %) and having ee >95 % The E-value was found to be 65–70.
● The reaction rate was also increased by increasing the amount of succinic anhydride.
● The reaction can be controlled by choice of solvent, amount of succinic anhydride and amount
of enzyme.
318
+
N
Boc
2 3 4
OH
+
N
Boc
O
O
COOH