Production of native-type Streptoverticillium mobaraense transglutaminase in Corynebacterium glutamicum.
We previously observed secretion of active-form transglutaminase in Corynebacterium glutamicum by coexpressing the subtilisin-like protease SAM-P45 from Streptomyces albogriseolus to process the prodomain. However, the N-terminal amino acid sequence of the transglutaminase differed from that of the native Streptoverticillium mobaraense enzyme. In the present work we have used site-directed mutagenesis to generate an optimal SAM-P45 cleavage site in the C-terminal region of the prodomain. As a result, native-type transglutaminase was secreted.
Date
Masayo
M
Institute of Life Sciences, Ajinomoto Co., Inc., 1-1 Suzuki-cho, Kawasaki-ku 210-8681, Japan.
Yokoyama
Kei-ichi
K
Umezawa
Yukiko
Y
Matsui
Hiroshi
H
Kikuchi
Yoshimi
Y
D000595:Amino Acid Sequence; D001483:Base Sequence; D003352:Corynebacterium / Q000201:enzymology* / Q000235:genetics*; D004269:DNA, Bacterial / Q000235:genetics; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D017434:Protein Structure, Tertiary; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics; D017384:Sequence Deletion; D013302:Streptomyces / Q000201:enzymology* / Q000235:genetics*; D011503:Transglutaminases / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics*
D004269:DNA, Bacterial; D011994:Recombinant Proteins; D011503:Transglutaminases