Functional and translational analyses of a beta-glucosidase gene (glycosyl hydrolase family 1) isolated from the gut of the lower termite Reticulitermes flavipes |
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| Authors: | Michael E. Scharf Elena S. Kovaleva Sanjay Jadhao James H. Campbell George W. Buchman Drion G. Boucias |
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| Affiliation: | 1. Entomology and Nematology Department, University of Florida, PO Box 110620, Gainesville, FL 32611-0620, USA;2. Chesapeake-PERL Inc., Savage, MD 20763, USA;1. Section for Sustainable Biotechnology, Aalborg University Copenhagen, AC Meyers Vaenge 15, DK-2450 Copenhagen SV, Denmark;2. Bioproducts, Sciences & Engineering Laboratory, Washington State University TriCities, 2710 Crimson Way, Richland, WA 99354, USA;1. Department of Chemistry, Faculty of Philosophy, Sciences and Languages of Ribeirão Preto, University of São Paulo, Ribeirão Preto, SP 14040-901, Brazil;2. Department of Biochemistry, Faculty of Medicine of Ribeirão Preto, University of São Paulo, Ribeirão Preto, SP 14049-900, Brazil;3. Carlos Chagas Institute, Fiocruz, Curitiba, PR 81350-010, Brazil;4. Department of Biology, Faculty of Philosophy, Sciences and Languages of Ribeirão Preto, University of São Paulo, Ribeirão Preto, SP 14040-901, Brazil;5. Brazilian Biosciences National Laboratory, National Center for Research in Energy and Materials, Campinas, SP 13083-100, Brazil;1. Department of Biotechnology, The University of Tokyo, 1-1-1 Yayoi, Bunkyo-ku, Tokyo 113-8657, Japan;2. National Institute of Agrobiological Sciences, 1-2 Owashi, Tsukuba, Ibaraki 305-8634, Japan;3. Tropical Biosphere Research Center, COMB, University of the Ryukyus, Nishihara, Okinawa 903-0213, Japan;4. Institute of Agrobiological Sciences, National Agriculture and Food Research Organization (reorganized from 2 in April, 2016), Japan;1. College of Plant Sciences, Jilin University, Changchun 130062, China;2. Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, Tianjin 300308, China;1. Laboratório Nacional de Ciência e Tecnologia do Bioetanol (CTBE), Centro Nacional de Pesquisa em Energia e Materiais (CNPEM), Campinas, SP, Brazil;2. Centro de Ciências Naturais e Humanas, Universidade Federal do ABC, Santo André, SP, Brazil;3. Departamento de Física, Instituto de Biociências, Letras e Ciências Exatas – IBILCE, UNESP - Univ Estadual Paulista, São José do Rio Preto, SP, Brazil;4. Department of Microbiology and Molecular Genetics, Oklahoma State University – OSU, Stillwater, OK, USA |
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| Abstract: | This research focused on digestive beta-glucosidases from glycosyl hydrolase family (GHF) 1 from the gut of the lower termite Reticulitermes flavipes. In preceding studies on R. flavipes, we characterized beta-glucosidase activity across the gut and its inhibition by carbohydrate-based inhibitors, and subsequently we identified two partial beta-glucosidase cDNA sequences from a host gut cDNA library. Here, we report on the full-length cDNA sequence for one of the R. flavipes beta-glucosidases (RfBGluc-1), the expression of its mRNA in the salivary gland and foregut, the production of recombinant protein using a baculovirus–insect expression system, optimal recombinant substrate specificity profiles and parameters, and significant inhibition by the established beta-glucosidase inhibitor cellobioimidazole. We also report the partial cDNA sequence for a second gut beta-glucosidase (RfBGluc-2), and show that like RfBGluc-1 its mRNA is localized mainly in the salivary gland. Other results for RfBGluc-1 showing activity against laminaribose, a component of microbial cell walls, suggest that RfBGluc-1 may serve dual functions in cellulose digestion and immunity. These findings provide important information that will enable the testing of hypotheses related to collaborative host–symbiont lignocellulose digestion, and that contributes to the development of next-generation termiticides and novel biocatalyst cocktails for use in biomass-to-bioethanol applications. |
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