RECOMBINANT PLASMID PDSBCM, TRANSFORMED MICROORGANISMS THEREWITH, AND METHOD FOR PRODUCING ALKALINE PROTEASE VAPK

摘要

PURPOSE: A recombinant plasmid pDSBCm, transformed microorganisms therewith, and a method for producing alkaline protease VapK are provided, thereby increasing the productivity of alkaline protease VapK. CONSTITUTION: The recombinant plasmid pDSBCm comprising a vapk gene dimer encoding alkaline protease VapK is constructed by the steps of: inserting a par gene into a recombinant plasmid pSBCm comprising the vapk gene to construct a plasmid pSP1, wherein the plasmid stability is improved; constructing a plasmid pSPR1 comprising the vapk gene in the reverse direction; digesting the plasmid pSPR1 with EcoRI, treating it with an endonuclease Bal31, and ligating it with a ligase to construct a plasmid pSPR-47, wherein the gene size is decreased; and digesting the plasmid pSPR-47 with HindIII and inserting the vapk gene into the plasmid pSPR-47 to produce the recombinant plasmid pDSBCm. The host strain Vibrio metschnikovii is transformed by the plasmid pDSBCm.