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Cloning and expression of the Bacillus amyloliquefaciens transglutaminase gene in E. coli using a bicistronic vector construction.
Duarte, Lovaine Silva; Barsé, Laísa Quadros; Dalberto, Pedro Ferrari; da Silva, William Tadeu Santos; Rodrigues, Rafael Costa; Machado, Pablo; Basso, Luiz Augusto; Bizarro, Cristiano Valim; Ayub, Marco Antônio Záchia.
Afiliación
  • Duarte LS; Biotechnology, Bioprocess, and Biocatalysis Group, Food Science and Technology Institute, Federal University of Rio Grande do Sul, Av. Bento Gonçalves 9500, PO Box 15090, ZC 91501-970, Porto Alegre, RS, Brazil.
  • Barsé LQ; Centro de Pesquisas em Biologia Molecular e Funcional (CPBMF), Pontifícia Universidade Católica do Rio Grande do Sul (PUCRS), 92A TECNOPUC Building, 4592 Av. Bento Gonçalves, ZC 90650-001, Porto Alegre, Brazil.
  • Dalberto PF; Centro de Pesquisas em Biologia Molecular e Funcional (CPBMF), Pontifícia Universidade Católica do Rio Grande do Sul (PUCRS), 92A TECNOPUC Building, 4592 Av. Bento Gonçalves, ZC 90650-001, Porto Alegre, Brazil.
  • da Silva WTS; Biotechnology, Bioprocess, and Biocatalysis Group, Food Science and Technology Institute, Federal University of Rio Grande do Sul, Av. Bento Gonçalves 9500, PO Box 15090, ZC 91501-970, Porto Alegre, RS, Brazil.
  • Rodrigues RC; Biotechnology, Bioprocess, and Biocatalysis Group, Food Science and Technology Institute, Federal University of Rio Grande do Sul, Av. Bento Gonçalves 9500, PO Box 15090, ZC 91501-970, Porto Alegre, RS, Brazil.
  • Machado P; Centro de Pesquisas em Biologia Molecular e Funcional (CPBMF), Pontifícia Universidade Católica do Rio Grande do Sul (PUCRS), 92A TECNOPUC Building, 4592 Av. Bento Gonçalves, ZC 90650-001, Porto Alegre, Brazil.
  • Basso LA; Centro de Pesquisas em Biologia Molecular e Funcional (CPBMF), Pontifícia Universidade Católica do Rio Grande do Sul (PUCRS), 92A TECNOPUC Building, 4592 Av. Bento Gonçalves, ZC 90650-001, Porto Alegre, Brazil.
  • Bizarro CV; Centro de Pesquisas em Biologia Molecular e Funcional (CPBMF), Pontifícia Universidade Católica do Rio Grande do Sul (PUCRS), 92A TECNOPUC Building, 4592 Av. Bento Gonçalves, ZC 90650-001, Porto Alegre, Brazil.
  • Ayub MAZ; Biotechnology, Bioprocess, and Biocatalysis Group, Food Science and Technology Institute, Federal University of Rio Grande do Sul, Av. Bento Gonçalves 9500, PO Box 15090, ZC 91501-970, Porto Alegre, RS, Brazil. Electronic address: mazayub@ufrgs.br.
Enzyme Microb Technol ; 134: 109468, 2020 Mar.
Article en En | MEDLINE | ID: mdl-32044021
Transglutaminases (TGases) are a class of transferases widely used in the food and biotechnology industries. In this work, we describe the production of recombinant Bacillus amyloliquefaciens TGase in Escherichia coli, obtaining the protein in its soluble and active form. In order to reduce TGase activity inside host cells and consequently its toxicity, we constructed a bicistronic plasmid containing the B. amyloliquefaciens TGase gene fused to the inhibitory Streptomyces caniferus prodomain. To make the enzyme active and avoid the need of prodomain removal in vitro, we also cloned the 3C protease gene into the same plasmid. After a fast single-step purification protocol, we obtained a partially purified recombinant TGase with 37 mU/mg protein activity, that crosslinked bovine serum albumin (BSA). This is the first report on the expression of B. amyloliquefaciens TGase in E. coli in its mature and active form.
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Plásmidos / Transglutaminasas / Clonación Molecular / Bacillus amyloliquefaciens / Vectores Genéticos Idioma: En Revista: Enzyme Microb Technol Año: 2020 Tipo del documento: Article País de afiliación: Brasil Pais de publicación: Estados Unidos
Texto completo
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Plásmidos / Transglutaminasas / Clonación Molecular / Bacillus amyloliquefaciens / Vectores Genéticos Idioma: En Revista: Enzyme Microb Technol Año: 2020 Tipo del documento: Article País de afiliación: Brasil Pais de publicación: Estados Unidos