文献类型：期刊文献

英文题名：Comparison of the expression of phospholipase D from Streptomyces halstedii in different hosts and its over-expression in Streptomyces lividans

作者：Tao, Xinyi[1];Zhao, Ming[1];Zhang, Ying[1];Liu, Min[1];Liu, Qinghai[1];Wang, Wei[1];Wang, Feng-Qing[1];Wei, Dongzhi[1]

机构：[1]East China Univ Sci & Technol, Newworld Inst Biotechnol, State Key Lab Bioreactor Engn, 130 Meilong Rd, Shanghai 200237, Peoples R China

年份：2019

卷号：366

期号：5

外文期刊名：FEMS MICROBIOLOGY LETTERS

收录：;WOS:【SCI-EXPANDED(收录号:WOS:000469763400011)】；

基金：This research was financially supported by National Natural Science Foundation of China (81603056).

语种：英文

外文关键词：phospholipase D; Streptomyces lividans; Pichia pastoris; cytotoxic; dual-promoter

摘要：Phospholipase D (PLD) proteins from Streptomyces species are useful biocatalysts for synthesizing phospholipid derivatives relevant for the pharmaceutical and food industry from low-cost phosphatidylcholine. The overexpression of PLD in a recombinant strain is necessary to achieve large-scale PLD production. In this study, we investigated the feasibility of expressing PLD from Streptomyces halstedii in different hosts. The enzymatic activity of PLD reached 69.12 U/mL in the homologous Streptomyces lividans host, which was around 50-fold higher than that in the original host. Meanwhile, in Escherichia coli and Pichia pastoris, PLD expression was poor and showed obvious toxicity to cells, which may have been one of the reasons for low levels of PLD observed in heterologous hosts. An induced (Ptip)/constitutive (PermE*) dual-promoter expression system in S. lividans was constructed, which could achieve constitutive expression with PLD enzymatic activity of 13.41 U/mL under non-induced conditions and yield the highest PLD enzymatic activity of 68.33 U/mL with 2 mu g/mL thiostreptone. The concentration of the expensive inducer was significantly reduced to only 10% of that used in the original expression system without affecting the protein expression level, which provided a good foundation for subsequent industrial applications to reduce production costs.