title: Expression of the carboxypeptidase T gene from Thermoactinomyces vulgaris in stable protoplast type L-forms of Proteus mirabilis.
abstract: The structural gene of the carboxypeptidase T (cpt) was successfully expressed in cell wall-less L-form cells of Proteus mirabilis. The DNA sequence encoding the PhoA leader peptide was fused with a truncated cpt gene encoding the mature enzyme. The modified gene in a pUC-based kanamycin resistance vector under the control of the lac promoter was transformed into L-form cells of P. mirabilis. They were able to produce the recombinant CpT both as a secretory and as a cell-bound insoluble form. The co-secretory processing of the PhoA leader peptide was quite efficient. The yield of the secreted CpT was not less than 20 mg l-1 and should be improvable.
authors.affiliation: Laboratory of Protein Chemistry, Institute of Microbial Genetics (GNII Genetika), Moscow, Russia.
authors.forename: A M
authors.initials: AM
authors.lastname: Bushueva
authors.forename: A B
authors.initials: AB
authors.lastname: Shevelev
authors.forename: J
authors.initials: J
authors.lastname: Gumpert
authors.forename: G G
authors.initials: GG
authors.lastname: Chestukhina
authors.forename: A V
authors.initials: AV
authors.lastname: Serkina
authors.forename: C
authors.initials: C
authors.lastname: Hoischen
authors.forename: M V
authors.initials: MV
authors.lastname: Matz
authors.forename: M V
authors.initials: MV
authors.lastname: Kuryatova
authors.forename: V M
authors.initials: VM
authors.lastname: Stepanov
mesh_terms: D000595:Amino Acid Sequence; D001426:Bacterial Proteins; D001483:Base Sequence; D002268:Carboxypeptidases / Q000235:genetics*; D007740:L Forms / Q000235:genetics*; D008848:Micromonosporaceae / Q000235:genetics*; D008969:Molecular Sequence Data; D011513:Proteus mirabilis / Q000235:genetics*; D011994:Recombinant Proteins / Q000378:metabolism
chemical_list: D001426:Bacterial Proteins; D011994:Recombinant Proteins; D002268:Carboxypeptidases; C076614:CPT protein, Thermoactinomyces vulgaris