期刊
APPLIED BIOCHEMISTRY AND BIOTECHNOLOGY
卷 162, 期 6, 页码 1574-1584出版社
SPRINGER
DOI: 10.1007/s12010-010-8939-7
关键词
(S)-naproxen; Bacillus subtilis carboxylesterase; Enantioselective hydrolysis; Naproxen methyl ester
资金
- National Natural Science Foundation of China [20773038, 20902023]
- Ministry of Science and Technology, People's Republic of China [2009CB724706, 2009ZX09501-016]
- China National Special Fund for State Key Laboratory of Bioreactor Engineering [2060204]
- Shanghai Leading Academic Discipline Project [B505]
An (S)-enantioselective esterase from Bacillus subtilis ECU0554, named BsENP01, has been cloned and over-expressed in a heterologous host Escherichia coli BL21. BsE-NP01 was shown to be a carboxylesterase with a molecular mass of about 32 kDa, and temperature and pH optima at 50 degrees C and 8.5, respectively. It could catalyze the selective hydrolysis of the (S)-enantiomer of racemic naproxen methyl ester, giving optically pure (S)-naproxen with 98% enantiomeric excess. A mechanic-grinding approach to substrate dispersion was also reported, which was considered to be an alternative to take the place of deleterious surfactants such as Tween-80, with improved performance of the hydrolysis reaction. Batch production of (S)-naproxen was repeatedly carried out in a solid-water biphasic system at 2-L scale, achieving an average total yield of about 85% after ten runs with complete recycling of (R)-substrate.
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