Cloning, nucleotide sequencing, and expression of the beta-galactosidase-encoding gene (lacA) from Aspergillus oryzae

摘要

lacA coding for beta-galactosidase (beta-gal) was cloned from the genomic DNA of Aspergillus oryzae RIB40. There were 9 exons in lacA and the coding region of 3,015 bp encoded a protein of 1,005 aa with a deduced molecular mass of 109,898. A.oryzae lacA was highly homologous to fungal beta-gals, with the highest aa identity of 70.7% to A.niger lacA, and also showed significant identity to acid beta-gals belonging to family 35 glycosyl hydrolases. Approximately 10 copies of lacA under control of A.oryzae glaA promoter were integrated into the chromosome of A.oryzae M-2-3. The recombinant strain expressed more than 700-fold of the beta-gal activity as compared to the wild type strain under induction by maltose.