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(Referência obtida automaticamente do Web of Science, por meio da informação sobre o financiamento pela FAPESP e o número do processo correspondente, incluída na publicação pelos autores.)

Characterization of a beta-galactosidase from Bacillus subtilis with transgalactosylation activity

Texto completo
Autor(es):
Carneiro, Lara A. B. C. [1] ; Yu, Li [2] ; Dupree, Paul [2] ; Ward, Richard J. [3]
Número total de Autores: 4
Afiliação do(s) autor(es):
[1] Univ Sao Paulo, FMRP, Dept Bioquim & Imunol, BR-14049900 Ribeirao Preto, SP - Brazil
[2] Univ Cambridge, Dept Biochem, Cambridge CB2 1QW - England
[3] Univ Sao Paulo, FFCLRP, Dept Quim, Ave Bandeirantes 3900, BR-14040901 Ribeirao Preto, SP - Brazil
Número total de Afiliações: 3
Tipo de documento: Artigo Científico
Fonte: International Journal of Biological Macromolecules; v. 120, n. A, p. 279-287, DEC 2018.
Citações Web of Science: 2
Resumo

Microbial beta-galactosidases (EC 3.1.2.23) have applications in the production of galacto-oligosaccharides, which are established prebiotic food ingredients. The p-galactosidase from Bacillus subtilis (YesZ) was expressed as a heterologous protein in Escherichia coil, and presented an optimum activity at pH 6.5 and 40 degrees C. The catalytic constants K-m and V-max of the enzyme were 8.26 mM and 1.42 mu mol . min(-1) . mg-(1) against pNP-beta-D-galactopyranoside, respectively. Structural characterization revealed that YesZ is a homotrimer in solution, and homology modeling suggested that the YesZ conserves a Cys cluster zinc binding site. Flame photometry experiments confirmed the presence of bound zinc in the recombinant enzyme, and YesZ activity was inhibited by 1 mM zinc, copper and silver ions. Transgalactosylation activity of YesZ was observed with the synthetic substrate p-NP-beta-Gal in the presence of a o-xylose acceptor, producing a beta-D-galactopyranosyl-(1 -> 4)-D-xylopyranose disaccharide. Analysis of this disaccharide by MALDI-ToF-MS/MS suggested a beta-1,4 glycosidic linkage between a non-reducing galactose residue and the xylose. The p-galactosidase YesZ from B. subtilis is a candidate for enzymatic synthesis showing favorable thermostability (with residual activity of 50% after incubation at 30 degrees C for 25 h) and transgalactosylation activity. (C) 2018 Published by Elsevier B.V. (AU)

Processo FAPESP: 12/24147-8 - Desenho racional de enzimas multifuncionais para despolimerização do material lignocelulósico
Beneficiário:Lara Aparecida Buffoni de Campos Carneiro
Linha de fomento: Bolsas no Brasil - Doutorado
Processo FAPESP: 10/18850-2 - Identificação, caracterização e engenharia de enzimas que degradam a parede celular das plantas
Beneficiário:Richard John Ward
Linha de fomento: Auxílio à Pesquisa - Temático