A novel high-yield process of phospholipase D-mediated phosphatidylserine production with cyclopentyl methyl ether. (March 2018)

• Record Type: Journal Article
• Authors: Qin, Wen; Wu, Chengjin; Song, Wei; Chen, Xiulai; Liu, Jia; Luo, Qiuling; Liu, Liming
• Is Part Of: Process biochemistry. Volume 66(2018)
• Page Start: 146
• Page End: 149
• Publication Date: 2018-03
• Subjects: PC phosphatidylcholine -- PS phosphatidylserine -- PA phosphatidic acid -- ScPLD Streptomyces chromofuscus phospholipase D -- CPME cyclopentyl methyl ether -- 2-MeTHF 2-methyltetrahydrofuran -- E. coli Escherichia coli -- PG phosphatidylglycerol -- PE phosphatidylethanolamine -- PLD phospholipase D -- SDS-PAGE sodium dodecyl sulfate polyacrylamide gel electrophoresis -- HPLC high performance liquid chromatography -- NCBI National Center of Biotechnology Information; Non-aqueous biocatalysis -- Phosphatidylserine -- Phospholipase D -- Cyclopentyl methyl ether; Biochemical engineering -- Periodicals; Biotechnology -- Periodicals; Biochemistry -- periodicals; Biotechnology -- periodicals; Chemical Engineering -- periodicals; Génie biochimique -- Périodiques; Biotechnologie -- Périodiques; Biochemical engineering; Biotechnology; Periodicals; 660.63
• DOI: 10.1016/j.procbio.2017.10.017
• Languages: English
• ISSNs: 1359-5113

Abstract

Graphical abstract: Highlights: Cyclopentyl methyl ether is an eco-friendly and green solvent that suitable for phosphatidylserine (PS) production. This novel process produces PS with high-efficiency and industrialized potential successfully. The phospholipids are easily purified from cyclopentyl methyl ether by evaporation. The biocatalyst is available and productive. Abstract: In this study, an eco-friendly and high-efficient method for the bioconversion of phosphatidylserine (PS) from phosphatidylcholine (PC) and l -serine was developed. After the strain screening, the Streptomyces chromofuscus phospholipase D ( Sc PLD) was chosen and expressed in Escherichia coli BL21 with pET28a (+). After optimization, the maximal specific productivity and Sc PLD hydrolysis activity reached 9049.6 U/g and 10.6 U/mL, respectively, while induction with 2 g/L lactose, at 25 °C for 12 h. The maximal conversions of PC to PS was 93.4% with a final PS concentration of 106.2 g/L (53.1 g/L/h) under theoptimal conditions: 0.153 M PC in CPME, 2.3 M l-ser in sodium acetate buffer, 2.0 g lyophilized cells/mol PC, 100 mM Ca 2+, 40 °C, and 2 h of incubation. After scale-up to 1 L, the conversions and yield maintained at 85.4% and 97.2 g/L. Therefore, the enzymatic process showed great potential for industrialization of PS.