In vitro synthesis of phospholipids with yeast phospholipase B, a phospholipid deacylating enzyme

• Main Authors: Yasuo Watanabe, Itsuki Kobayashi, Takanori Ohnaka, Seiya Watanabe
• Format: Article
• Language: English
• Published: Elsevier 2018-06-01
• Series: Biotechnology Reports
• Subjects: Phospholipid deacylating enzyme; Saccharomyces cerevisiae phospholipase B; Enzyme-mediated phospholipid synthesis; Transacylation; Esterification
• Online Access: http://www.sciencedirect.com/science/article/pii/S2215017X17303107

The gene encoding the Saccharomyces cerevisiae phospholipid deacylation enzyme, phospholipase B (ScPLB1), was successfully expressed in E. coli. The enzyme (Scplb1p) was engineered to have a histidine-tag at the C-terminal end and was purified by metal (Ni) affinity chromatography. Enzymatic properties, optimal pH, and substrate specificity were similar to those reported previously. For example, deacylation activity was observed in acidic pH in the absence of Ca2+ and was additive in neutral pH in the presence of Ca2+, and the enzyme had the same substrate priority as reported previously, with the exception of PE, suggesting that yeast phospholipase B could be produced in its native structure in bacterial cells. Scplb1p retained transacylation activity in aqueous medium, and esterified lysophosphatidylcholine with free fatty acid to form phosphatidylcholine in a non-aqueous, glycerin medium. We propose that phospholipase B could serve as an additional tool for in vitro enzyme-mediated phospholipid synthesis.