Cloning, expression and characterisation of phospholipase B from Saccharomyces cerevisiae and its application in the synthesis of l -alpha-glycerylphosphorylcholine and peanut oil degumming

l-alpha-glycerylphosphorylcholine (GPC) has been shown to enhance cognitive performance. Meanwhile, vegetable oils must be refined to remove the impurities for them to be edible. Phospholipase B (PLB), having the ability of hydrolyzing both the sn-1 and sn-2 acyl ester bonds of phospholipids, can pr...

Full description

Saved in:
Bibliographic Details
Published inBiotechnology, biotechnological equipment Vol. 32; no. 4; pp. 968 - 973
Main Authors Liu, Yihan, Li, Mingjie, Huang, Lin, Gui, Shuang, Jia, Leibo, Zheng, Dong, Fu, Yu, Zhang, Yutong, Rui, Jinqiu, Lu, Fuping
Format Journal Article
LanguageEnglish
Published Sofia Taylor & Francis Ltd 04.07.2018
Taylor & Francis Group
Subjects
characterisation
Cloning
Cognitive ability
Degumming
Enzymatic activity
Fermentation
Fungi
Impurities
l-alpha-glycerylphosphorylcholine
Optimization
Peanut oil
Peanuts
pH effects
Phospholipase
Phospholipase B
Phospholipids
Phosphorus
Pichia pastoris
Saccharomyces cerevisiae
Substrates
Vegetable oils
Vegetables
Yeast
Online AccessGet full text

Cover

More Information
Summary:l-alpha-glycerylphosphorylcholine (GPC) has been shown to enhance cognitive performance. Meanwhile, vegetable oils must be refined to remove the impurities for them to be edible. Phospholipase B (PLB), having the ability of hydrolyzing both the sn-1 and sn-2 acyl ester bonds of phospholipids, can produce GPC using PC as substrate and transform the non-hydratable phospholipids into their hydratable forms. The Saccharomyces cerevisiae plb gene, which encodes PLB, was cloned and expressed in Pichia pastoris GS115 to produce recombinant PLB (rPLB). Fermentation optimisation yielded rPLB activity levels as high as 1723 U/mL. rPLB demonstrated maximum enzymatic activity at 40 °C and pH 5.5 and was stable at temperatures between 30 and 40 °C and pH values between 5.0 and 6.0. rPLB synthesised GPC with a conversion rate of 17% (w/w) and exhibited high degumming activity towards peanut oil, decreasing the phosphorus content from 91.8 to 3.7 mg/kg within 3 h. This study describes a candidate phospholipase for potential applications involving the modification of phospholipids and vegetable oil degumming.
ISSN:1310-2818
1314-3530
DOI:10.1080/13102818.2018.1455536