Purification, Characterization, and cDNA Cloning of a Prominent β-Glucosidase from the Gut of the Xylophagous Cockroach Panesthia angustipennis spadica

• Arakawa Gaku

  Insect-mimetics Research Unit, National Institute of Agrobiological Sciences

• Kamino Kei

  Biological Resource Center, National Institute of Technology and Evaluation

• Tokuda Gaku

  Tropical Biosphere Research Center, University of the Ryukyus

• Watanabe Hirofumi

  Insect-mimetics Research Unit, National Institute of Agrobiological Sciences Molecular Biomimetics Research Unit, Institute of Agrobiological Sciences, National Agriculture and Food Research Organization

書誌事項

タイトル別名

• Purification, Characterization, and cDNA Cloning of a Prominent β-Glucosidase from the Gut of the Xylophagous Cockroach <i>Panesthia angustipennis spadica</i>

抄録

Abstract: In this study, a β-glucosidase (PaBG1b) with high specific activity was purified from gut extracts of the wood-feeding cockroach Panesthia angustipennis spadica using Superdex 75 gel filtration chromatography and High-Trap phenyl hydrophobic chromatography. The protein was purified 14-fold to a single band identified by sodium dodecyl sulfate polyacrylamide gel electrophoresis, with an apparent molecular mass of 56.7 kDa. The specific activity of the purified enzyme was 708 μmol/min/mg protein using cellobiose as substrate. To the best of our knowledge, this is the highest specific activity reported among β-glucosidases to date. The purified PaBG1b showed optimal activity at pH 5.0 and retained more than 65 % of the activity between pH 4.0 and 6.5. The activity was stable up to 50 °C for 30 min. Kinetic studies on cellobiose revealed that the K_m was 5.3 mM, and the V_max was 1,020 μmol/min/mg. The internal amino acid sequence of PaBG1b was analyzed, and two continuous sequences (a total of 39 amino acids) of the C-terminal region were elucidated. Based on these amino acid sequences, a full-length cDNA (1,552 bp) encoding 502 amino acids was isolated. The encoded protein showed high similarity to β-glucosidases from glycoside hydrolase family 1. Thus, the current study demonstrated the potential of PaBG1b for application in enzymatic biomass-conversion as a donor gene for heterologous recombination of cellulase-producing agents (fungi or bacteria) or an additive enzyme for cellulase products based on the high-performance of PaBG1b as a digestive enzyme in cockroaches.

収録刊行物

• Ｊｏｕｒｎａｌ　ｏｆ　Ａｐｐｌｉｅｄ　Ｇｌｙｃｏｓｃｉｅｎｃｅ

  Ｊｏｕｒｎａｌ　ｏｆ　Ａｐｐｌｉｅｄ　Ｇｌｙｃｏｓｃｉｅｎｃｅ 63 (3), 51-59, 2016

  日本応用糖質科学会

キーワード

• glycoside hydrolase family 1
• cellobiase
• wood-feeding cockroach
• β-glucosidase

詳細情報

• CRID: 1390001206294034816
• NII論文ID: 40020932813
• NII論文ID: 130005261412
• NII書誌ID: AA11809133
• DOI: 10.5458/jag.jag.jag-2016_006
• ISSN: 18807291
• ISSN: 13447882
• NDL書誌ID: 027599054
• Web Site: https://ndlsearch.ndl.go.jp/books/R000000004-I027599054
• Web Site: https://www.jstage.jst.go.jp/article/jag/63/3/63_jag.JAG-2016_006/_pdf
• 本文言語コード: en
• データソース種別:
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  • NDL
  • Crossref
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  • KAKEN
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