Abstract
An (S)-enantioselective esterase from Bacillus subtilis ECU0554, named BsE-NP01, 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 °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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Acknowledgment
This work was financially supported by the National Natural Science Foundation of China (Nos. 20773038 and 20902023), the Ministry of Science and Technology, People’s Republic of China (Nos. 2009CB724706 and 2009ZX09501-016), China National Special Fund for State Key Laboratory of Bioreactor Engineering (No. 2060204) and Shanghai Leading Academic Discipline Project, Project (No. B505).
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Liu, X., Xu, JH., Pan, J. et al. Efficient Production of (S)-Naproxen with (R)-Substrate Recycling Using an Overexpressed Carboxylesterase BsE-NP01. Appl Biochem Biotechnol 162, 1574–1584 (2010). https://doi.org/10.1007/s12010-010-8939-7
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DOI: https://doi.org/10.1007/s12010-010-8939-7