title: Cloning, expression and characterization of a lipase gene from the Candida antarctica ZJB09193 and its application in biosynthesis of vitamin A esters.
abstract: Lipase is one of the most important industrial enzymes, which has been widely used in the preparation of food additives, cosmetics and pharmaceuticals industries. In order to obtain a large amount of lipase, the lipase gene from Candida antarctica ZJB09193 was cloned, and expressed in Pichia pastoris with the vector pPICZαA. Under the optimal conditions, the yield of recombinant lipase in the culture broth reached 3.0 g/L. After purification, the properties of recombinant lipase were studied: the optimum pH and temperature were pH 8.0 and 52°C, Ca(2+) activated the activity of lipase, and the apparent K(m) and V(max) values for p-nitrophenyl acetate were 0.34 mM and 7.36 μmol min(-1) mg(-1), respectively. Furthermore, the recombinant lipase was immobilized on pretreated textile for biosynthesis of vitamin A esters. In a system of n-hexane, 0.3 g immobilized recombinant lipase was used in the presence of 0.06 g vitamin A acetate and 0.55 mmol fatty acid (nine different fatty acids were tested). The yield of all vitamin A esters exceeded 78% in 7h at 30°C except using lactic acid and hexanoic acid as substrates. After optimization, the yield of vitamin A palmitate reached 87%. This study has the potential to be developed into industrial application.
authors.affiliation: Institute of Bioengineering, Zhejiang University of Technology, Hangzhou, Zhejiang 310014, People's Republic of China.
authors.forename: Zhi-Qiang
authors.initials: ZQ
authors.lastname: Liu
authors.forename: Xiao-Bo
authors.initials: XB
authors.lastname: Zheng
authors.forename: Su-Ping
authors.initials: SP
authors.lastname: Zhang
authors.forename: Yu-Guo
authors.initials: YG
authors.lastname: Zheng
mesh_terms: D002118:Calcium / Q000378:metabolism; D002175:Candida / Q000201:enzymology* / Q000235:genetics; D003001:Cloning, Molecular; D020536:Enzyme Activators / Q000378:metabolism; D004795:Enzyme Stability; D004800:Enzymes, Immobilized / Q000378:metabolism; D004951:Esterification; D004952:Esters / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008049:Lipase / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D010843:Pichia / Q000235:genetics; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D013696:Temperature; D014801:Vitamin A / Q000031:analogs & derivatives* / Q000378:metabolism*
chemical_list: D020536:Enzyme Activators; D004800:Enzymes, Immobilized; D004952:Esters; D011994:Recombinant Proteins; D014801:Vitamin A; D008049:Lipase; D002118:Calcium