Enhanced xyloglucan-specific endo-β-1,4-glucanase efficiency in an engineered CBM44-XegA chimera
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| 024 | 7 | 0 | |a 10.1007/s00253-014-6324-0 |2 doi |
| 035 | |a (NATIONALLICENCE)springer-10.1007/s00253-014-6324-0 | ||
| 245 | 0 | 0 | |a Enhanced xyloglucan-specific endo-β-1,4-glucanase efficiency in an engineered CBM44-XegA chimera |h [Elektronische Daten] |c [Gilvan Furtado, Camila Santos, Rosa Cordeiro, Lucas Ribeiro, Luiz de Moraes, André Damásio, Maria Polizeli, Marcos Lourenzoni, Mário Murakami, Richard Ward] |
| 520 | 3 | |a Xyloglucan-specific endo-β-1,4-glucanases (Xegs, EC 3.2.1.151) exhibit high catalytic specificity for β-1,4 linkages of xyloglucan, a branched hemicellulosic polysaccharide abundant in dicot primary cell walls and present in many monocot species. In nature, GH12 Xegs are not associated with carbohydrate-binding modules (CBMs), and here, we have investigated the effect of the fusion of the xyloglucan-specific CBM44 on the structure and function of a GH12 Xeg from Aspergillus niveus (XegA). This fusion presented enhanced catalytic properties and conferred superior thermal stability on the XegA. An increased k cat (chimera, 177.03s−1; XegA, 144.31s−1) and reduced KM (chimera, 1.30mgmL−1; XegA, 1.50mgmL−1) resulted in a 1.3-fold increase in catalytic efficiency of the chimera over the parental XegA. Although both parental and chimeric enzymes presented catalytic optima at pH 5.5 and 60°C, the thermostabilitiy of the chimera at 60°C was greater than the parental XegA. Moreover, the crystallographic structure of XegA together with small-angle X-ray scattering (SAXS) and molecular dynamics simulations revealed that the spatial arrangement of the domains in the chimeric enzyme resulted in the formation of an extended binding cleft that may explain the improved kinetic properties of the CBM44-XegA chimera. | |
| 540 | |a Springer-Verlag Berlin Heidelberg, 2015 | ||
| 690 | 7 | |a GH12 glycosylhydrolase |2 nationallicence | |
| 690 | 7 | |a X-ray crystallography |2 nationallicence | |
| 690 | 7 | |a Small-angle X-ray scattering |2 nationallicence | |
| 700 | 1 | |a Furtado |D Gilvan |u Departamento de Bioquímica e Imunologia, FMRP-USP, Universidade de São Paulo, Ribeirão Preto, SP, Brazil |4 aut | |
| 700 | 1 | |a Santos |D Camila |u Laboratório Nacional de Biociências, Centro Nacional de Pesquisa em Energia e Materiais, Giuseppe Maximo Scolfaro, 10000, 13083-970, Campinas, SP, Brazil |4 aut | |
| 700 | 1 | |a Cordeiro |D Rosa |u Laboratório Nacional de Biociências, Centro Nacional de Pesquisa em Energia e Materiais, Giuseppe Maximo Scolfaro, 10000, 13083-970, Campinas, SP, Brazil |4 aut | |
| 700 | 1 | |a Ribeiro |D Lucas |u Departamento de Bioquímica e Imunologia, FMRP-USP, Universidade de São Paulo, Ribeirão Preto, SP, Brazil |4 aut | |
| 700 | 1 | |a de Moraes |D Luiz |u Computational Modelling Group: Simulation and Evolution of Biomolecules, Fundação Oswaldo Cruz—Fiocruz-CE, Fortaleza, CE, Brazil |4 aut | |
| 700 | 1 | |a Damásio |D André |u Laboratório Nacional de Ciência e Tecnologia do Bioetanol (CTBE), Centro Nacional de Pesquisas em Energia e Materiais, Campinas, SP, Brazil |4 aut | |
| 700 | 1 | |a Polizeli |D Maria |u Departamento de Biologia, FFCLRP-USP, Universidade de São Paulo, Ribeirão Preto, SP, Brazil |4 aut | |
| 700 | 1 | |a Lourenzoni |D Marcos |u Computational Modelling Group: Simulation and Evolution of Biomolecules, Fundação Oswaldo Cruz—Fiocruz-CE, Fortaleza, CE, Brazil |4 aut | |
| 700 | 1 | |a Murakami |D Mário |u Laboratório Nacional de Biociências, Centro Nacional de Pesquisa em Energia e Materiais, Giuseppe Maximo Scolfaro, 10000, 13083-970, Campinas, SP, Brazil |4 aut | |
| 700 | 1 | |a Ward |D Richard |u Laboratório Nacional de Ciência e Tecnologia do Bioetanol (CTBE), Centro Nacional de Pesquisas em Energia e Materiais, Campinas, SP, Brazil |4 aut | |
| 773 | 0 | |t Applied Microbiology and Biotechnology |d Springer Berlin Heidelberg |g 99/12(2015-06-01), 5095-5107 |x 0175-7598 |q 99:12<5095 |1 2015 |2 99 |o 253 | |
| 856 | 4 | 0 | |u https://doi.org/10.1007/s00253-014-6324-0 |q text/html |z Onlinezugriff via DOI |
| 898 | |a BK010053 |b XK010053 |c XK010000 | ||
| 900 | 7 | |a Metadata rights reserved |b Springer special CC-BY-NC licence |2 nationallicence | |
| 908 | |D 1 |a research-article |2 jats | ||
| 949 | |B NATIONALLICENCE |F NATIONALLICENCE |b NL-springer | ||
| 950 | |B NATIONALLICENCE |P 856 |E 40 |u https://doi.org/10.1007/s00253-014-6324-0 |q text/html |z Onlinezugriff via DOI | ||
| 950 | |B NATIONALLICENCE |P 700 |E 1- |a Furtado |D Gilvan |u Departamento de Bioquímica e Imunologia, FMRP-USP, Universidade de São Paulo, Ribeirão Preto, SP, Brazil |4 aut | ||
| 950 | |B NATIONALLICENCE |P 700 |E 1- |a Santos |D Camila |u Laboratório Nacional de Biociências, Centro Nacional de Pesquisa em Energia e Materiais, Giuseppe Maximo Scolfaro, 10000, 13083-970, Campinas, SP, Brazil |4 aut | ||
| 950 | |B NATIONALLICENCE |P 700 |E 1- |a Cordeiro |D Rosa |u Laboratório Nacional de Biociências, Centro Nacional de Pesquisa em Energia e Materiais, Giuseppe Maximo Scolfaro, 10000, 13083-970, Campinas, SP, Brazil |4 aut | ||
| 950 | |B NATIONALLICENCE |P 700 |E 1- |a Ribeiro |D Lucas |u Departamento de Bioquímica e Imunologia, FMRP-USP, Universidade de São Paulo, Ribeirão Preto, SP, Brazil |4 aut | ||
| 950 | |B NATIONALLICENCE |P 700 |E 1- |a de Moraes |D Luiz |u Computational Modelling Group: Simulation and Evolution of Biomolecules, Fundação Oswaldo Cruz—Fiocruz-CE, Fortaleza, CE, Brazil |4 aut | ||
| 950 | |B NATIONALLICENCE |P 700 |E 1- |a Damásio |D André |u Laboratório Nacional de Ciência e Tecnologia do Bioetanol (CTBE), Centro Nacional de Pesquisas em Energia e Materiais, Campinas, SP, Brazil |4 aut | ||
| 950 | |B NATIONALLICENCE |P 700 |E 1- |a Polizeli |D Maria |u Departamento de Biologia, FFCLRP-USP, Universidade de São Paulo, Ribeirão Preto, SP, Brazil |4 aut | ||
| 950 | |B NATIONALLICENCE |P 700 |E 1- |a Lourenzoni |D Marcos |u Computational Modelling Group: Simulation and Evolution of Biomolecules, Fundação Oswaldo Cruz—Fiocruz-CE, Fortaleza, CE, Brazil |4 aut | ||
| 950 | |B NATIONALLICENCE |P 700 |E 1- |a Murakami |D Mário |u Laboratório Nacional de Biociências, Centro Nacional de Pesquisa em Energia e Materiais, Giuseppe Maximo Scolfaro, 10000, 13083-970, Campinas, SP, Brazil |4 aut | ||
| 950 | |B NATIONALLICENCE |P 700 |E 1- |a Ward |D Richard |u Laboratório Nacional de Ciência e Tecnologia do Bioetanol (CTBE), Centro Nacional de Pesquisas em Energia e Materiais, Campinas, SP, Brazil |4 aut | ||
| 950 | |B NATIONALLICENCE |P 773 |E 0- |t Applied Microbiology and Biotechnology |d Springer Berlin Heidelberg |g 99/12(2015-06-01), 5095-5107 |x 0175-7598 |q 99:12<5095 |1 2015 |2 99 |o 253 | ||