title: Chemical synthesis of the gene for microbial transglutaminase from Streptoverticillium and its expression in Escherichia coli.
abstract: The gene coding for microbial transglutaminase (TGase) from Streptoverticillium, which consists of 331 amino acids, was chemically synthesized. The codons have been substituted for those mainly favored in yeast. Our strategy involved the construction of the TGase gene in five sections (54 oligomers) that contained unique restriction enzyme sites at both ends, which could readily be ligated to form the full-length product. The chemically synthesized gene was inserted downstream from the ompA signal peptide of the E. coli expression vector, pIN-III-ompA, which carries lpp and lac promotors. The resultant plasmid directed the expression of TGase, with the activity being secreted mainly into the periplasmic space of E. coli. The induced gene product was identical with native TGase in size and in immunological properties, though the enzyme activity was low.
authors.affiliation: Food Research & Development Laboratories, Ajinomoto Co., Inc., Kawasaki, Japan.
authors.forename: S
authors.initials: S
authors.lastname: Takehana
authors.forename: K
authors.initials: K
authors.lastname: Washizu
authors.forename: K
authors.initials: K
authors.lastname: Ando
authors.forename: S
authors.initials: S
authors.lastname: Koikeda
authors.forename: K
authors.initials: K
authors.lastname: Takeuchi
authors.forename: H
authors.initials: H
authors.lastname: Matsui
authors.forename: M
authors.initials: M
authors.lastname: Motoki
authors.forename: H
authors.initials: H
authors.lastname: Takagi
mesh_terms: D000595:Amino Acid Sequence; D001483:Base Sequence; D003001:Cloning, Molecular; D004269:DNA, Bacterial; D004926:Escherichia coli / Q000235:genetics*; D005798:Genes, Bacterial; D005813:Genes, Synthetic; D008969:Molecular Sequence Data; D010957:Plasmids; D013306:Streptomycetaceae / Q000201:enzymology* / Q000235:genetics; D011503:Transglutaminases / Q000235:genetics*
chemical_list: D004269:DNA, Bacterial; D011503:Transglutaminases