Purification and characterization of four key enzymes from a feather-degrading Bacillus subtilis from the gut of tarantula Chilobrachys guangxiensis |
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| Institution: | 1. Proteomic Laboratory, Ezequiel Dias Foundation, FUNED, 80 Conde Pereira Carneiro St., Belo Horizonte, MG 30510-010, Brazil;2. Departamento de Ciências Fisiológicas, Centro de Ciências da Saúde, Universidade Federal do Espírito Santo, 1468 Marechal Campos Av., Vitória, ES 29043-900, Brazil;3. Computational Mass Spectrometry Group, Carlos Chagas Institute, Fiocruz, 3775 Professor Algacyr Munhoz Mader St., Curitiba, PR 81350-010, Brazil;4. Department of Pathology, University of Michigan, Ann Arbor, MI 48109, USA;5. Departamento de Bioquímica, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, 6627 Antônio Carlos Av., Belo Horizonte, MG 31270-901, Brazil;6. Department of Chemical Physiology and Molecular and Cellular Neurobiology, The Scripps Research Institute, 10550 N. Torrey Pines Rd., La Jolla, CA 92037, USA |
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| Abstract: | A feather-degrading bacterium was isolated from the gut of the tarantula Chilobrachys guangxiensis, and was classified as Bacillus subtilis (named Bacillus subtilis CH-1) according to both the phenotypic characteristics and 16S rRNA profile. The improved culture conditions for feather-degrading were 10.0 g l−1 mannitol, 10.0 g l−1 tryptone, 0.1 g l−1 MgCl2, 0.4 g l−1 KH2PO4, 0.3 g l−1 K2HPO4, 0.5 g l−1 NaCl, and 2.0 g l−1 intact feather, with pH 8.5 and 37 °C. In the optimized medium, the intact black feather was completely degraded by Bacillus subtilis CH-1 in 24 h. Furthermore, four kinds of enzymes which include extracellular protease Vpr, peptidase T, γ-glutamyl transpeptidase and glyoxalmethylglyoxal reductase were identified as having principal roles. Simultaneously, the relationship between the disulfide bond reducing activity (DRT) and the keratinase activity (KT) in B. subtilis CH-1 fermentation system was discussed. This is the first report for a feather-degrading enteric bacterium from tarantula. The identification of the enzymes shines a light on further understanding the molecular mechanism of feather-degrading by microbes. |
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| Keywords: | Keratinase Enteric bacteria Enzyme system |
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