Isolation and characterization of a β-galactosidase from a new Amazon forest strain of Aspergillus niger as a potential accessory enzyme for biomass conversion

dc.contributor.authorTonelotto, Mariana
dc.contributor.authorPirota, Rosangela Donizete Perpetua Buzon
dc.contributor.authorDelabona, Priscila Da Silva
dc.contributor.authorBarros, Georgia De Oliveira Figueiredo
dc.contributor.authorGolubev, Alexander M.
dc.contributor.authorPolikarpov, Igor
dc.contributor.authorFarinas, Cristiane Sanchez
dc.contributor.institutionUniversidade Federal de São Carlos (UFSCar)
dc.contributor.institutionEmpresa Brasileira de Pesquisa Agropecuária (EMBRAPA)
dc.contributor.institutionPNPI RAS Gatchina
dc.contributor.institutionUniversidade de São Paulo (USP)
dc.date.accessioned2022-04-28T18:59:18Z
dc.date.available2022-04-28T18:59:18Z
dc.date.issued2014-01-01
dc.description.abstractThe selection of enzyme-producing fungi is useful to obtain enzymes required to hydrolyze lignocellulosic material and thereby contribute to biomass conversion into fuels and chemicals. Besides cellulases, the presence of accessory enzymes in enzyme cocktails is necessary to enhance hydrolysis efficiency. This study evaluates the production, purification, and biochemical-kinetic characterization of β-galactosidase produced by a new strain of Aspergillus niger (P47C3) isolated from the Amazon Forest. The A. niger (P47C3) was cultured under SmF conditions and β-galactosidase was purified in a three-step purification, using an ultrafiltration membrane, ion exchange (TSK-SP), and gel filtration (Sephacryl S-200). The calculated molecular weight of the purified enzyme was 125 kDa. Optimum pH (4.0) and temperature (55°C) of β-galactosidase activity were determined. The values of the kinetic parameters obtained from p-nitrophenyl-β-D- galactopyranoside (PNPG) hydrolysis were 2.2 mM and 0.285 mM/min for Km and Vmax, respectively. The inhibition of PNPG hydrolysis by β-galactosidase in the presence of the inhibitor galactose gave a Ki value of 5.01 mM. As a precursor to elucidating the tertiary structure using X-ray diffraction, the β-galactosidase was crystallized using 0.2 M Tris-HCl buffer, with 12% PEG 4000 as the precipitation agent; the largest crystals were formed at pH 8.6. These results provide the basis for further structural and functional studies of this accessory enzyme to evaluate its potential biotechnological applications. © 2014 Informa UK, Ltd.en
dc.description.affiliationPrograma de Pós-graduação em Biotecnologia Universidade Federal de São Carlos (UFSCar), Rod. Washington Luis, Km 235, 13565-905, São Carlos/SP
dc.description.affiliationEmbrapa Instrumentação, Rua XV de Novembro, 1452, 13560-970, São Carlos/SP
dc.description.affiliationPetersburg Nuclear Physics Institute PNPI RAS Gatchina, Leningrad District 188300
dc.description.affiliationUniversidade do Estado de São Paulo (USP), Av. Trabalhador São-Carlense 400 Arnold Schimidt, 13566-590, São Carlos/SP
dc.format.extent13-22
dc.identifierhttp://dx.doi.org/10.3109/10242422.2013.801018
dc.identifier.citationBiocatalysis and Biotransformation, v. 32, n. 1, p. 13-22, 2014.
dc.identifier.doi10.3109/10242422.2013.801018
dc.identifier.issn1029-2446
dc.identifier.issn1024-2422
dc.identifier.scopus2-s2.0-84893154287
dc.identifier.urihttp://hdl.handle.net/11449/220044
dc.language.isoeng
dc.relation.ispartofBiocatalysis and Biotransformation
dc.sourceScopus
dc.subjectAccessory enzymes
dc.subjectAmazon
dc.subjectAspergillus niger
dc.subjectBiomass
dc.subjectβ-galactosidase
dc.titleIsolation and characterization of a β-galactosidase from a new Amazon forest strain of Aspergillus niger as a potential accessory enzyme for biomass conversionen
dc.typeArtigo

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