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Expression of Bacillus amyloliquefaciens transglutaminase in recombinant E. coli under the control of a bicistronic plasmid system in DO-stat fed-batch bioreactor cultivations.
Duarte, Lovaine Silva; Matte, Carla Roberta; Dall Cortivo, Paulo Roberto; Nunes, José Eduardo Sacconi; Barsé, Laisa Quadros; 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, Porto Alegre, RS, ZC 91501-970, Brazil.
  • Matte CR; Biotechnology, Bioprocess, and Biocatalysis Group, Food Science and Technology Institute, Federal University of Rio Grande Do Sul, Av. Bento Gonçalves 9500, PO Box 15090, Porto Alegre, RS, ZC 91501-970, Brazil.
  • Dall Cortivo PR; Biotechnology, Bioprocess, and Biocatalysis Group, Food Science and Technology Institute, Federal University of Rio Grande Do Sul, Av. Bento Gonçalves 9500, PO Box 15090, Porto Alegre, RS, ZC 91501-970, Brazil.
  • Nunes JES; 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, Porto Alegre, ZC 90650-001, 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, Porto Alegre, ZC 90650-001, 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, Porto Alegre, ZC 90650-001, 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, Porto Alegre, RS, ZC 91501-970, Brazil. mazayub@ufrgs.br.
Braz J Microbiol ; 52(3): 1225-1233, 2021 Sep.
Article en En | MEDLINE | ID: mdl-34008152
We studied the expression of Bacillus amyloliquefaciens transglutaminase cloned in Escherichia coli BL21(DE3)pLysS harboring the plasmid pBAD/3C/bTGase, a bicistronic expression system, in bioreactor cultivation. Batch and fed-batch controlled as DO-stat strategies were employed for the production of the recombinant enzyme. In 30 h-batch cultivations using Terrific broth (TB), 6 g/L of biomass and 3.12 U/mgprotein of transglutaminase activity were obtained. DO-stat fed-batch cultivations under the control of oxygen concentration (DO-stat) using TB as medium but fed with glucose allowed the increment in biomass formation (17.5 g/L) and enzyme activity (6.43 U/mgprotein). DO-stat fed-batch using mineral medium (M9) and fed with glucose under the same conditions produced even higher enzymatic activity (9.14 U/mgprotein). The pH effect was investigated, and the best enzymatic activity could be observed at pH 8. In all cultivations, the bicistronic system remained stable, with 100% of plasmid-bearing cells. These results show that E. coli bearing bicistronic plasmid constructs to express recombinant TGase could be cultivated in bioreactors under DO-stat fed-batch using mineral medium and it is a promising strategy in future optimizations to produce this important enzyme.
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Transglutaminasas / Escherichia coli Idioma: En Revista: Braz J Microbiol Año: 2021 Tipo del documento: Article País de afiliación: Brasil Pais de publicación: Brasil
Texto completo
Añadir a Mi BVS
Imprimir
XML
PubMed Links
Buscar en Google

Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Transglutaminasas / Escherichia coli Idioma: En Revista: Braz J Microbiol Año: 2021 Tipo del documento: Article País de afiliación: Brasil Pais de publicación: Brasil