Enhanced extracellular production of recombinant proteins in Escherichia coli by co-expression with Bacillus cereus phospholipase C. Issue 1 (December 2017)

• Record Type: Journal Article
• Authors: Su, Lingqia; Jiang, Qi; Yu, Lingang; Wu, Jing
• Is Part Of: Microbial cell factories. Volume 16:Issue 1(2017)
• Page Start: 1
• Page End: 11
• Publication Date: 2017-12
• Subjects: Bacillus cereus phospholipase C -- Co-expression -- Extracellular protein production -- Membrane permeability -- Foam formation; Microbial biotechnology -- Periodicals; Recombinant proteins -- Synthesis -- Periodicals; 660.62
• DOI: 10.1186/s12934-017-0639-3
• Languages: English
• ISSNs: 1475-2859

Abstract

Abstract Background Our laboratory has reported a strategy for improving the extracellular production of recombinant proteins through co-expression withThermobifida fusca cutinase, which increases membrane permeability via its phospholipid hydrolysis activity. However, the foam generated by the lysophospholipid product makes the fermentation process difficult to control in a fermentor. Phospholipase C (PLC) catalyzes the hydrolysis of phospholipids to produce sn1, 2-diacylglycerides and organic phosphate, which do not induce foam formation. Therefore, co-expression withBacillus cereus PLC was investigated as a method to improve the extracellular production of recombinant proteins. Results WhenB. cereus PLC was expressed inEscherichia coli without its signal peptide, 95.3% of the total PLC activity was detected in the culture supernatant. PLC expression enhanced membrane permeability without obvious cell lysis. Then, six test enzymes, three secretory and three cytosolic, were co-expressed withB. cereus PLC. The enhancement of extracellular production correlated strongly with the molecular mass of the test enzyme. Extracellular production ofStreptomyces sp. FA1 xylanase (43 kDa), which had the lowest molecular mass among the secretory enzymes, was 4.0-fold that of its individual expression control. Extracellular production of glutamate decarboxylase (51 kDa), which had the lowest molecular mass among the cytosolic enzymes, reached 26.7 U/mL; 88.3% of the total activity produced. This strategy was effectively scaled up using a 3-L fermentor. No obvious foam was generated during this fermentation process. Conclusions This is the first study to detail the enhanced extracellular production of recombinant proteins through co-expression with PLC. This new strategy, which is especially appropriate for lower molecular mass proteins, allows large-scale protein production in an easily controlled fermentation process.