%A Yang,Peizhou
%A Wu,Yun
%A Jiang,Suwei
%A Zheng,Zhi
%A Hou,Zhigang
%A Mu,Dongdong
%A Xiao,Wei
%A Jiang,Shaotong
%A Yang,Yung-Hun
%D 2019
%J Frontiers in Bioengineering and Biotechnology
%C
%F
%G English
%K Serratia marcescens,Phospholipase A,Enzymatic degumming,Rapeseed oil,enzyme characterization,Lecitase Ultra
%Q
%R 10.3389/fbioe.2019.00272
%W
%L
%M
%P
%7
%8 2019-October-17
%9 Original Research
%#
%! Phospholipase degumming crude oil
%*
%<
%T Effective Expression of the Serratia marcescens Phospholipase A1 Gene in Escherichia coli BL21(DE3), Enzyme Characterization, and Crude Rapeseed Oil Degumming via a Free Enzyme Approach
%U https://www.frontiersin.org/articles/10.3389/fbioe.2019.00272
%V 7
%0 JOURNAL ARTICLE
%@ 2296-4185
%X Crude oil degumming by phospholipid removal is crucial to guarantee oil quality. Phospholipase degumming could produce green vegetable oil by reducing energy consumption and protecting the environment. To develop a novel phospholipase for oil degumming, we cloned the Serratia marcescens outer membrane phospholipase A gene (OM-PLA1) and expressed its 33 KDa protein in engineered Escherichia coli BL21(DE3). OM-PLA1 activity reached 18.9 U mL−1 with the induction of 0.6 mM isopropyl β-D-1-thiogalactopyranoside for 4 h. The optimum temperature and pH were 50°C and 7.5, respectively. Mg2+, Ca2+, Co2+, and Mn2+ at 0.1 mM L−1 significantly increased OM-PLA1 activity. The kinetic equations of OM-PLA1 and Lecitase Ultra were y = 13.7x+0.74 (Km = 18.53 mM, Vmax = 1.35 mM min−1) and y = 24.42x+0.58 (Km = 42.1 mM, Vmax = 1.72 mM min−1), respectively. The phosphorus content decreased from 22.6 to 9.3 mg kg−1 with the addition of 15 units of free recombinant OM-PLA1 into 150 g of crude rapeseed oil. OM-PLA1 has the close degumming efficiency with Lecitase Ultra. The S. marcescens outer membrane phospholipase gene (OM-PLA1) possessed higher substrate affinity and catalytic efficiency than Lecitase Ultra. This study provides an alternative approach to achieve crude vegetable oil degumming with enzymatic technology.