title: Overproduction of soluble recombinant transglutaminase from Streptomyces netropsis in Escherichia coli.
abstract: A novel microbial transglutaminase (TGase) from the cultural filtrate of Streptomyces netropsis BCRC 12429 (Sn) was purified. The specific activity of the purified TGase was 18.2 U/mg protein with an estimated molecular mass of 38 kDa by sodium dodecyl sulfate polyacrylamide gel electrophoresis analysis. The TGase gene of S. netropsis was cloned and an open reading frame of 1,242 bp encoding a protein of 413 amino acids was identified. The Sn TGase was synthesized as a precursor protein with a preproregion of 82 amino acid residues. The deduced amino acid sequence of the mature S. netropsis TGase shares 78.9-89.6% identities with TGases from Streptomyces spp. A high level of soluble Sn TGase with its N-terminal propeptide fused with thioredoxin was expressed in E. coli. A simple and efficient process was applied to convert the purified recombinant protein into an active enzyme and showed activity equivalent to the authentic mature TGase.
authors.affiliation: Institute of Molecular Biology, National Chung Hsing University, Taichung, 40227, Taiwan, Republic of China.
authors.forename: Yu-Jen
authors.initials: YJ
authors.lastname: Yu
authors.forename: Shih-Cheng
authors.initials: SC
authors.lastname: Wu
authors.forename: Hung-Hsiang
authors.initials: HH
authors.lastname: Chan
authors.forename: Yu-Cheng
authors.initials: YC
authors.lastname: Chen
authors.forename: Zong-Yu
authors.initials: ZY
authors.lastname: Chen
authors.forename: Ming-Te
authors.initials: MT
authors.lastname: Yang
mesh_terms: D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D001483:Base Sequence; D003001:Cloning, Molecular; D004795:Enzyme Stability; D004926:Escherichia coli / Q000235:genetics* / Q000378:metabolism; D015870:Gene Expression; D008969:Molecular Sequence Data; D008970:Molecular Weight; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D013302:Streptomyces / Q000201:enzymology*; D011503:Transglutaminases / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*
chemical_list: D001426:Bacterial Proteins; D011994:Recombinant Proteins; D011503:Transglutaminases