Engineering of a microbial cell factory for the extracellular production of catalytically active phospholipase a2 of streptomyces violaceoruber

Hyun Jae Lee, Ara Cho, Yeji Hwang, Jin Byung Park, Sun Ki Kim

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7 Scopus citations

Abstract

Phospholipase A2 (PLA2) from Streptomyces violaceoruber is a lipolytic enzyme used in a wide range of industrial applications including production of lysolecithins and enzymatic degumming of edible oils. We have therefore investigated expression and secretion of PLA2 in two workhorse microbes, Pichia pastoris and Escherichia coli. The PLA2 was produced to an activity of 0.517 ± 0.012 U/ml in the culture broth of the recombinant P. pastoris. On the other hand, recombinant E. coli BL21 star (DE3), overexpressing the authentic PLA2 (P-PLA2), showed activity of 17.0 ± 1.3 U/ml in the intracellular fraction and 21.7 ± 0.7 U/ml in the culture broth. The extracellular PLA2 activity obtained with the recombinant E. coli system was 3.2-fold higher than the corresponding value reached in a previous study, which employed recombinant E. coli BL21 (DE3) overexpressing codon-optimized PLA2. Finally, we observed that the extracellular PLA2 from the recombinant E. coli P-PLA2 culture was able to hydrolyze 31.1 g/l of crude soybean lecithin, an industrial substrate, to a conversion yield of approximately 95%. The newly developed E. coli-based PLA2 expression system led to extracellular production of PLA2 to a productivity of 678 U/l·h, corresponding to 157-fold higher than that obtained with the P. pastoris-based system.

Original languageEnglish
Pages (from-to)1244-1251
Number of pages8
JournalJournal of Microbiology and Biotechnology
Volume30
Issue number8
DOIs
StatePublished - Aug 2020

Bibliographical note

Funding Information:
This research was financially supported by the National Research Foundation of Korea (NRF) Grant (2019R1C1C1003521) funded by the Korean Ministry of Science, ICT and Future Planning, and also by the Chung-Ang University Graduate Research Scholarship in 2019. Ara Cho, Yeji Hwang, and Jin-Byung Park were supported by the Marine Biomaterials Research Center grant from the Marine Biotechnology Program [No. D11013214H480000100] funded by the Ministry of Oceans and Fisheries, Korea.

Publisher Copyright:
© 2020 by The Korean Society for Microbiology and Biotechnology

Keywords

  • Escherichia coli
  • Extracellular production
  • Phospholipase A
  • Pichia pastoris

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