Optimization and characterization of the depolymerase gene from Fusarium perfringens Expression in Pichia pastoris

Abstract

poly (butylene succinate) is a biodegradable plastic that can replace traditional plastics, which is helpful to solve the problem of white pollution, the key to the rapid and effective degradation of microorganisms or depolymerization Enzymes.The recombinant Pichia pastoris expression system was constructed to realize (FSC)EFFICIENT EXPRESSION.Codon Optimization Based on Pichia pastoris preferenceFSCGene codons of Pichia PastorisX33Achieving its recombinant expression in.The fermentation conditions of the strain were optimized by single factor experiments..The results show that the optimizedFSCIn gene sequence157One base changed,G + CContent59.6%Drop48.3%, Sequence homology is77.34%;Constructed recombinant expression vectorPpiczAlpha- FSCTransferred to Pichia PastorisX33, Combined with resistance plate screening,SDS-PAGEAndWestern BoltAnd the enzyme activity of shake flask fermentation was determined.1.Recombinant strain with high enzyme yieldL1;Further determine its fermentation conditions:Medium InitiationPH 6.0, Shaker speed220 r/minMethanol supplement1%, Inoculation amount8%Training time72 h, Incubation temperature30Under the optimized conditions, the enzyme activity of fermentation broth was up110 U/ml.