Study on expression of thermostable β-galactosidase in Kluyveromyces lactis and Pichia pastoris expression system
Thermostable β- galactosidase gene bgaB from Geobacillus stearothermophilus were cloned into pKLAC1 and pPIC9k downstreams of the α- mating factor domain, resulting in construction of eukaryotic expression vectors of Kluyveromyces lactis and Pichia pastoris expression systems.Introduction of the linearized expression cassettes in K.lactis and P.pastoris cells was achieved by electrotransformation, and expression cassettes were inserted into the host genomes at the homology locus.Recombinant enzymes were purified by Ni2+ -NTA affinity chromatography,identified by western blot and the enzyme property was characterized.The results suggested that the recombinant thermostable β-galactosidases were capable of expression in yeast expression system and the stability was preserved well, but recombinants can not be secreted effectively.
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