title: Immobilization, Regiospecificity Characterization and Application of Aspergillus oryzae Lipase in the Enzymatic Synthesis of the Structured Lipid 1,3-Dioleoyl-2-Palmitoylglycerol.
abstract: The enzymatic synthesis of 1,3-dioleoyl-2-palmitoylglycerol (OPO), one of the main components of human milk fats, has been hindered by the relatively high cost of sn-1,3-specific lipases and the deficiency in biocatalyst stability. The sn-1,3-specific lipase from Aspergillus oryzae (AOL) is highly and efficiently immobilized with the polystyrene-based hydrophobic resin D3520, with a significant 49.54-fold increase in specific lipase activity compared with the AOL powder in catalyzing the synthesis of OPO through the acidolysis between palm stearin and oleic acid (OA). The optimal immobilization conditions were investigated, including time course, initial protein concentration and solution pH. The sn-1,3 specificity of lipases under different immobilization conditions was evaluated and identified as positively associated with the lipase activity, and the pH of the immobilization solution influenced the regiospecificity and synthetic activity of these lipases. Immobilized AOL D3520, as the biocatalyst, was used for the enzymatic synthesis of the structured lipid OPO through the acidolysis between palm stearin and OA. The following conditions were optimized for the synthesis of structured lipid OPO: 65 °C temperature; 1:8 substrate molar ratio between palm stearin and OA; 8% (w/w) enzyme load; 3.5% water content of the immobilized lipase; and 1 h reaction time. Under these conditions, highly efficient C52 production (45.65%) was achieved, with a tripalmitin content of 2.75% and a sn-2 palmitic acid (PA) proportion of 55.08% in the system.
authors.affiliation: College of Biosystems Engineering and Food Science, Zhejiang University, Hangzhou, China; Zhejiang Key Laboratory for Agro-Food Processing, Zhejiang University, Hangzhou, China.
authors.forename: Haiying
authors.initials: H
authors.lastname: Cai
authors.affiliation: College of Biosystems Engineering and Food Science, Zhejiang University, Hangzhou, China; Zhejiang Key Laboratory for Agro-Food Processing, Zhejiang University, Hangzhou, China.
authors.forename: Yang
authors.initials: Y
authors.lastname: Li
authors.affiliation: College of Biosystems Engineering and Food Science, Zhejiang University, Hangzhou, China; Zhejiang Key Laboratory for Agro-Food Processing, Zhejiang University, Hangzhou, China.
authors.forename: Minjie
authors.initials: M
authors.lastname: Zhao
authors.affiliation: Hangzhou Kangyuan Food Technology Co., Ltd, Hangzhou, China.
authors.forename: Guanwen
authors.initials: G
authors.lastname: Fu
authors.affiliation: College of Biosystems Engineering and Food Science, Zhejiang University, Hangzhou, China; Zhejiang Key Laboratory for Agro-Food Processing, Zhejiang University, Hangzhou, China.
authors.forename: Jia
authors.initials: J
authors.lastname: Lai
authors.affiliation: College of Biosystems Engineering and Food Science, Zhejiang University, Hangzhou, China; Zhejiang Key Laboratory for Agro-Food Processing, Zhejiang University, Hangzhou, China; Fuli Institute of Food Science, Zhejiang University, Hangzhou, China.
authors.forename: Fengqin
authors.initials: F
authors.lastname: Feng
mesh_terms: D001236:Aspergillus oryzae / Q000201:enzymology*; D002849:Chromatography, Gas; D004800:Enzymes, Immobilized / Q000737:chemistry / Q000378:metabolism*; D006801:Humans; D008049:Lipase / Q000737:chemistry* / Q000378:metabolism*; D013237:Stereoisomerism; D014280:Triglycerides / Q000737:chemistry* / Q000378:metabolism*
chemical_list: D004800:Enzymes, Immobilized; D014280:Triglycerides; D008049:Lipase; C095376:1,3-dioleoyl-2-palmitoylglycerol