Clone and expression of β-1, 3-1, 4-glucanase in Pichia pastoris

β-1, 3-1, 4-glucanase, which can hydrolyzeβ-1, 3-1, 4-glucan by cutting off theβ-1, 4-glycosidic linkages and release shorter reducing sugar, has been applied in the brewing and animal feed additive industry.It can effectively improve digestibility of barley-based diets and reduce enteritis.In this report, the gene ofβ-1, 3-1, 4-glucanase, which was optimized according to Pichia pastoris condon usage, was amplified by polymerase chain reaction and ligated into the expression vector pPICZαA.The recombinant vector was lineared by Sac I and transformed into Pichia pastoris X-33 by electroporation.Then the optimal positive transformant, named X-33/pPICZαA-bgl, was screened by colony PCR and shake flask fermentation.The enzyme activity of recombinantβ-1, 3-1, 4-glucanase couldreach 308.5U/mL after 96h methanol induction.Results of SDS-PAGE showed that about 33ku protein of theβ-1, 3-1, 4-glucanase was expressed.Acidity and temperature optimal for this recombinant enzyme was pH5.0 and 50℃, respectively.