Food and Fermentation Industries >

2022 , Vol. 48 >Issue 3: 30 - 37

DOI: https://doi.org/10.13995/j.cnki.11-1802/ts.028010

Heterologous expression of surfactant-like peptides in Escherichia coli

  • MENG Shujuan ,
  • LI Huinan ,
  • PANG Zhengjun ,
  • MU Jing ,
  • WANG Fenghuan
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  • (School of Light Industry,Beijing Technology and Business University,Beijing 100048,China)

Received date: 2021-05-18

  Revised date: 2021-06-22

  Online published: 2022-03-04

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Abstract

In this study, we investigated the expression of a novel surfactant-like peptide (A6K) in Escherichia coli and explored its purification process. Firstly, the synthetic DNA fragment (A6K)15 encoding the new surfactant-like peptide A6K was amplified and inserted into four different prokaryotic expression plasmids, pET-28a (+), pET-28a-CLCⅠ, pET-28a-SUMO and pT7-GST-His. All the constructed vectors were verified by restriction enzyme digestion and sequencing. Then, the indicating vector pET28a-CLCⅠ-(A6K)15 which contain a fluorescent protein label was transformed into E. coli strain BL21(DE3), BL21(DE3) plysS and Rosetta (DE3) to find the best expression host. The IPTG concentration, induction temperature and induction time were optimized to obtain the best expression conditions. Finally, the expression product was purified by Ni-NTA chelation chromatography, and the purified (A6K)15 was detected by SDS-PAGE and Western blot using the 6×His tag in (A6K)15. The product concentration was determined by the BCA method for economic benefit analysis. The results indicated that four different recombinant expression vectors containing the target gene (A6K)15 were constructed. The optimal vector pET-28a-SUMO-(A6K)15 and the optimal host BL21(DE3) were screened out. The best expression condition for the surfactant-like peptide (A6K)15 was IPTG 1.0 mmol/mL, induction temperature 30 ℃, and induction time 12 h. SDS-PAGE and Western blot analysis proved the molecule of Polypeptide (A6K)15. In this study, the novel surfactant-like peptide (A6K)15 expression vector was successfully constructed and explored to obtain the optimal expression conditions for its large-scale preparation and purification. This research provides potential ideas for the production of new surface-active peptides by biological methods.

Key words： surfactant-like peptides; recombinant vector; fusion expression; purification

Cite this article

MENG Shujuan , LI Huinan , PANG Zhengjun , MU Jing , WANG Fenghuan . Heterologous expression of surfactant-like peptides in Escherichia coli[J]. Food and Fermentation Industries, 2022 , 48(3) : 30 -37 . DOI: 10.13995/j.cnki.11-1802/ts.028010

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