University of Oulu

Lisette Van Tassel, Antti Moilanen, Lloyd W. Ruddock, Efficient production of wild-type lipase B from Candida antarctica in the cytoplasm of Escherichia coli, Protein Expression and Purification, Volume 165, 2020, 105498, ISSN 1046-5928,

Efficient production of wild-type lipase B from Candida antarctica in the cytoplasm of Escherichia coli

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Author: Van Tassel, Lisette1; Moilanen, Antti1; Ruddock, Lloyd W.1
Organizations: 1Faculty of Biochemistry and Molecular Medicine and Biocenter Oulu, University of Oulu, Aapistie 7, 90220, Oulu, Finland
Format: article
Version: accepted version
Access: open
Online Access: PDF Full Text (PDF, 1.2 MB)
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Language: English
Published: Elsevier, 2020
Publish Date: 2020-09-12


Candida antarctica lipase B (CalB) is a very efficient catalyst and is used in a wide range of industries from food flavour to pharmaceutical, and biodiesel manufacturing. It has a high degree of enantioselective and regioselective substrate specificity and is stable over a wide range of biophysical conditions including pH, temperature and solvent conditions. High-level expression of biologically active wild-type CalB has been problematic, partly due to folding events. Consequently, focus has been on modified CalB, which has allowed orders of magnitude increase in yields of protein. However, these modifications alter the quaternary structure of the protein. Here we produce soluble wild-type CalB in high yields in the cytoplasm of E.coli using a catalyzed system for cytoplasmic disulfide bond formation both in shake flasks and in fermentation in chemically defined media. The CalB produced had the expected stereospecific activity and had a higher activity than CalB from a commercial source.

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Series: Protein expression & purification
ISSN: 1046-5928
ISSN-E: 1096-0279
ISSN-L: 1046-5928
Volume: 165
Article number: 105498
DOI: 10.1016/j.pep.2019.105498
Type of Publication: A1 Journal article – refereed
Field of Science: 1182 Biochemistry, cell and molecular biology
Funding: This work was supported by the Academy of Finland [grant number 266457] and Biocenter Oulu.
Copyright information: © 2019. This manuscript version is made available under the CC-BY-NC-ND 4.0 license