title: Comparative analysis of three hyperthermophilic GH1 and GH3 family members with industrial potential.
abstract: Beta-glucosidases (BGLs) are enzymes of great potential for several industrial processes, since they catalyze the cleavage of glucosidic bonds in cellobiose and other short cellooligosaccharides. However, features such as good stability to temperature, pH, ions and chemicals are required characteristics for industrial applications. This work aimed to provide a comparative biochemical analysis of three thermostable BGLs from Pyrococcus furiosus and Thermotoga petrophila. The genes PfBgl1 (GH1 from P. furiosus), TpBgl1 (GH1 from T. petrophila) and TpBgl3 (GH3 from T. petrophila) were cloned and proteins were expressed in Escherichia coli. The purified enzymes are hyperthermophilic, showing highest activity at temperatures above 80°C at acidic (TpBgl3 and PfBgl1) and neutral (TpBgl1) pHs. The BGLs showed greatest stability to temperature mainly at pH 6.0. Activities using a set of different substrates suggested that TpBgl3 (GH3) is more specific than GH1 family members. In addition, the influence of six monosaccharides on BGL catalysis was assayed. While PfBgl1 and TpBgl3 seemed to be weakly inhibited by monosaccharides, TpBgl1 was activated, with xylose showing the strongest activation. Under the conditions tested, TpBgl1 showed the highest inhibition constant (Ki=1100.00mM) when compared with several BGLs previously characterized. The BGLs studied have potential for industrial use, specifically the enzymes belonging to the GH1 family, due to its broad substrate specificity and weak inhibition by glucose and other saccharides.
authors.affiliation: Laboratório Nacional de Ciência e Tecnologia do Bioetanol (CTBE), Centro Nacional de Pesquisa em Energia e Materiais (CNPEM), Campinas, SP, Brazil.
authors.forename: Junio
authors.initials: J
authors.lastname: Cota
authors.affiliation: Laboratório Nacional de Ciência e Tecnologia do Bioetanol (CTBE), Centro Nacional de Pesquisa em Energia e Materiais (CNPEM), Campinas, SP, Brazil.
authors.forename: Thamy L R
authors.initials: TL
authors.lastname: Corrêa
authors.affiliation: Laboratório Nacional de Ciência e Tecnologia do Bioetanol (CTBE), Centro Nacional de Pesquisa em Energia e Materiais (CNPEM), Campinas, SP, Brazil.
authors.forename: André R L
authors.initials: AR
authors.lastname: Damásio
authors.affiliation: Laboratório Nacional de Ciência e Tecnologia do Bioetanol (CTBE), Centro Nacional de Pesquisa em Energia e Materiais (CNPEM), Campinas, SP, Brazil.
authors.forename: José A
authors.initials: JA
authors.lastname: Diogo
authors.affiliation: Laboratório Nacional de Ciência e Tecnologia do Bioetanol (CTBE), Centro Nacional de Pesquisa em Energia e Materiais (CNPEM), Campinas, SP, Brazil.
authors.forename: Zaira B
authors.initials: ZB
authors.lastname: Hoffmam
authors.affiliation: Centro de Ciências Naturais e Humanas, Universidade Federal do ABC, Santo André, SP, Brazil.
authors.forename: Wanius
authors.initials: W
authors.lastname: Garcia
authors.affiliation: Laboratório Nacional de Ciência e Tecnologia do Bioetanol (CTBE), Centro Nacional de Pesquisa em Energia e Materiais (CNPEM), Campinas, SP, Brazil; Departamento de Física, Instituto de Biociências, Letras e Ciências Exatas - IBILCE, UNESP - Univ Estadual Paulista, São José do Rio Preto, SP, Brazil.
authors.forename: Leandro C
authors.initials: LC
authors.lastname: Oliveira
authors.affiliation: Department of Microbiology and Molecular Genetics, Oklahoma State University - OSU, Stillwater, OK, USA.
authors.forename: Rolf A
authors.initials: RA
authors.lastname: Prade
authors.affiliation: Laboratório Nacional de Ciência e Tecnologia do Bioetanol (CTBE), Centro Nacional de Pesquisa em Energia e Materiais (CNPEM), Campinas, SP, Brazil. Electronic address: fabio.squina@bioetanol.org.br.
authors.forename: Fabio M
authors.initials: FM
authors.lastname: Squina
mesh_terms: D000704:Analysis of Variance; D055162:Biocatalysis / Q000187:drug effects; D004187:Disaccharides / Q000378:metabolism; D004492:Edetic Acid / Q000494:pharmacology; D006863:Hydrogen-Ion Concentration; D006868:Hydrolysis / Q000187:drug effects; D007221:Industry; D007700:Kinetics; D011134:Polysaccharides / Q000378:metabolism; D020027:Pyrococcus furiosus / Q000201:enzymology; D013379:Substrate Specificity / Q000187:drug effects; D013696:Temperature; D001617:beta-Glucosidase / Q000378:metabolism*
chemical_list: D004187:Disaccharides; D011134:Polysaccharides; D004492:Edetic Acid; D001617:beta-Glucosidase