Abstract
ABSTRACTl-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.
Highlights
L-alpha-glycerylphosphorylcholine (GPC; sn-glycero-3phosphocholine), a common choline compound found in the brain, is a derivative of phosphatidylcholine (PC; 1,2-diacyl-sn-glycero-3-phosphocholine) that is synthesised via the deacylation of sn-1 and sn-2
This study describes a candidate phospholipase for potential applications involving the modification of phospholipids and vegetable oil degumming
Water degumming only removes hydratable phospholipids, and acid and super degumming only remove a portion of the non-hydratable phospholipids (NHPs), resulting in large amounts of NHPs remaining in vegetable oils [4]
Summary
L-alpha-glycerylphosphorylcholine (GPC; sn-glycero-3phosphocholine), a common choline compound found in the brain, is a derivative of phosphatidylcholine (PC; 1,2-diacyl-sn-glycero-3-phosphocholine) that is synthesised via the deacylation of sn-1 and sn-2. A process through which phospholipase hydrolyses the acyl bonds of phospholipids and transforms NHPs into their hydratable forms, has been applied to the vegetable-oil refining process to enhance product yields and reduce wastewater generation and operating costs [5]. PLB catalysed the production of GPC through the deacylation of PC as the substrate, whereas phosphoglycerate, which is more hydrophilic than lysophospholipids, is obtained from NHPs in vegetable oils and is efficiently removed via centrifugation. The plb gene (hereafter referred to as plb) from S. cerevisiae was cloned and expressed in Pichia pastoris GS115, and recombinant PLB (rPLB) was purified, characterised and tested for the production of GPC and peanut oil degumming. Our work will be beneficial for the large-scale production of rPLB and its further application in phospholipid modification and vegetable oil degumming
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