Preparation Of Lipases In Pichia And Screening Of Combi-Lipase System

In the lipase enzyme-catalyzed reactions, in the oil-water interface reactions such as the promotion of ester hydrolysis, while it may be enzymatic synthesis in the organic phase and transesterification. In this way, it provided a possibility to use cooking oil or industrial waste oils and low-carbon alcohol as raw materials for synthesis of biodiesel. The immobilized lipase enzyme can enhance the stability of lipase and recovery efficiency, in order to achieve the effective use of lipases. The aim of this project is to product the lipases and to select the lipases. The lipase is selected by the new combi-lipase immobilization method for the multi-layer, orientation and controllable proportional mode. And as a basis of this theory.In this study, Pichia pastoris was used as an expression vector to prepare the lipases from Candida sp 99-125 , Candida antarctica lipase and Rhizopus oryzae lipase, the amount of which is 18. By this experiment The p-nitrophenyl laurate and p-nitrophenyl octanoate were used to determine lipase-to-medium lipids The hydrolysis ability of p-nitrophenylbutyrate was determined by lipase on short chain lipase.Screening the lipase with the advantage of hydrolysis. The esterification activity of lipase was determined by oleic acid, palmitic acid, lauric acid and octanoic acid respectively. The highest hydrolysis ability lipase and the highest esterification activity lipase were as the combi-lipase system.Of the composite effect. And through the design of the ball surface with the support of the stent protein to match the anchoring domain to achieve the effect of immobilization. The results of the final experiment showed that the lipase enzyme of Rhizopus oryzae had good hydrolytic activity to long chain, medium chain and short chain lipids. The Candida antarctica lipse CALB has a good esterification activity for long and medium chain fatty acids, and the combination of the two forms of complex enzymes also in the medium and short chain fatty acid esterification reaction has a composite enhancement effect.