Cloning, nucleotide sequencing, and expression of the beta-galactosidase-encoding gene (lacA) from Aspergillus oryzae.
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.
Ito
Yoshiyuki
Y
Laboratory of Lactic Acid Bacteria Research, Food Functionality Research Institute, Meiji Dairies Corporation, Odawara 250-0862, Japan. YOSHIYUKI_ITOU@MEIJI-MILK.COM
Sasaki
Takashi
T
Kitamoto
Katsuhiko
K
Kumagai
Chieko
C
Takahashi
Kohjiro
K
Gomi
Katsuya
K
Tamura
Gakuzo
G
D000595:Amino Acid Sequence; D001236:Aspergillus oryzae / Q000235:genetics*; D001483:Base Sequence; D003001:Cloning, Molecular; D005091:Exons; D007438:Introns; D008969:Molecular Sequence Data; D017385:Sequence Homology; D001616:beta-Galactosidase / Q000737:chemistry / Q000235:genetics*
D001616:beta-Galactosidase