Secretion of active-form Streptoverticillium mobaraense transglutaminase by Corynebacterium glutamicum: processing of the pro-transglutaminase by a cosecreted subtilisin-Like protease from Streptomyces albogriseolus.
The transglutaminase secreted by Streptoverticillium mobaraense is a useful enzyme in the food industry. A fragment of transglutaminase was secreted by Corynebacterium glutamicum when it was coupled on a plasmid to the promoter and signal peptide of a cell surface protein from C. glutamicum. We analyzed the signal peptide and the pro-domain of the transglutaminase gene and found that the signal peptide consists of 31 amino acid residues and the pro-domain consists of 45 residues. When the pro-domain of the transglutaminase was used, the pro-transglutaminase was secreted efficiently by C. glutamicum but had no enzymatic activity. However, when the plasmid carrying the S. mobaraense transglutaminase also encoded SAM-P45, a subtilisin-like serine protease derived from Streptomyces albogriseolus, the peptide bond to the C side of 41-Ser of the pro-transglutaminase was hydrolyzed, and the pro-transglutaminase was converted to an active form. Our findings suggest that C. glutamicum has potential as a host for industrial-scale protein production.
Kikuchi
Yoshimi
Y
Institute of Life Sciences, Ajinomoto Co., Inc., Kawasaki 210-8681, Japan. yoshimi_kikuchi@ajinomoto.com
Date
Masayo
M
Yokoyama
Kei-ichi
K
Umezawa
Yukiko
Y
Matsui
Hiroshi
H
D000595:Amino Acid Sequence; D001483:Base Sequence; D001709:Biotechnology / Q000379:methods; D003352:Corynebacterium / Q000201:enzymology* / Q000235:genetics*; D008969:Molecular Sequence Data; D010957:Plasmids; D021382:Protein Sorting Signals; D017422:Sequence Analysis, DNA; D013306:Streptomycetaceae / Q000201:enzymology* / Q000235:genetics; D011503:Transglutaminases / Q000235:genetics / Q000378:metabolism*
D021382:Protein Sorting Signals; D011503:Transglutaminases