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A novel thermostable GH5 ß-xylosidase from Thermogemmatispora sp. T81.
Tomazini, Atilio; Higasi, Paula; Manzine, Livia R; Stott, Matthew; Sparling, Richard; Levin, David B; Polikarpov, Igor.
Afiliação
  • Tomazini A; Sao Carlos Institute of Physics, University of Sao Paulo, São Carlos, São Paulo, Brazil.
  • Higasi P; Sao Carlos Institute of Physics, University of Sao Paulo, São Carlos, São Paulo, Brazil.
  • Manzine LR; Sao Carlos Institute of Physics, University of Sao Paulo, São Carlos, São Paulo, Brazil.
  • Stott M; School of Biological Sciences, University of Canterbury, New Zealand.
  • Sparling R; Department of Microbiology, University of Manitoba, Winnipeg, Manitoba, Canada.
  • Levin DB; Department of Biosystems Engineering, University of Manitoba, Winnipeg, Manitoba, Canada.
  • Polikarpov I; Sao Carlos Institute of Physics, University of Sao Paulo, São Carlos, São Paulo, Brazil. Electronic address: ipolikarpov@ifsc.usp.br.
N Biotechnol ; 53: 57-64, 2019 Nov 25.
Article em En | MEDLINE | ID: mdl-31299302
A glycoside hydrolase family 5 (GH5) subfamily 22 gene, designated T81Xyl5_22A, was identified in the genome of the aerobic thermophilic bacterium, Thermogemmatispora sp. T81 (locus A4R35_07040). The gene was cloned and heterologously expressed in Escherichia coli and the gene product characterized biochemically. The recombinant enzyme had an optimal catalytic activity at pH5.0 and 65 °C, and was active against beechwood xylan and rye arabinoxylan. It yielded only xylose molecules as products of beechwood xylan hydrolysis, indicating that it is a GH5 family ß-d-xylosidase. Using 4-nitrophenyl ß-d-xylopyranoside (pNPX) as a substrate, the KM, Vmax, kcat and kcat/KM kinetic parameters were determined as 0.25 ±â€¯0.03 mM, 889.47 ±â€¯28.54 U/mg, 39.20 s-1 and 156.8 mM-1 s-1, respectively. Small-angle X-ray scattering (SAXS) data enabled reconstruction of the enzyme's low-resolution molecular envelope and revealed that it formed dimers in solution. As far as we are aware, this is the first description of a thermostable bacterial GH5 family ß-d-xylosidase.
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Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Temperatura / Xilosidases / Chloroflexi Idioma: En Revista: N Biotechnol Assunto da revista: BIOLOGIA MOLECULAR / ENGENHARIA BIOMEDICA Ano de publicação: 2019 Tipo de documento: Article País de afiliação: Brasil País de publicação: Holanda
Texto completo
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Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Temperatura / Xilosidases / Chloroflexi Idioma: En Revista: N Biotechnol Assunto da revista: BIOLOGIA MOLECULAR / ENGENHARIA BIOMEDICA Ano de publicação: 2019 Tipo de documento: Article País de afiliação: Brasil País de publicação: Holanda