Purification, Characterization and Gene Analysis of N-Acetylglucosaminidase from Enterobacter sp. G-1
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概要
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Enterobacter sp. G-1 is a bacterium isolated previously as a chitinase-producing bacterium. We found this bacterium also produced N-acetylglucosaminidase and characterized that in this study. Extracellular N-acetylglucosaminidase of 92.0 kDa was purified near homogeneity by 8.57-fold from Enterobacter sp. G-1. The optimum temperature and the optimum pH of the purified N-acetylglucosaminidase was 45℃ and 6.0,respectively. The N-terminal amino acid sequence of 23 residues of N-acetylglucosaminidase was identified. Based on the N-terminal sequence, we amplified pieces of the DNA fragments by PCR. Using these PCR products as probes, we screened the genomic library and successfully isolated the entire N-acetylglucosaminidase gene (designated nag1) from Enterobacter sp. G-1. The nucleotide sequence of the nag1 gene was found to consist of 2,655 bp encoding a protein of 885 amino acid residues. Comparison of the deduced amino acid sequence from the nag1 gene found 97.3% identity with chitobiase from Serratia marcescens, 54.4% identity with N, N'-diacetylchitobiase from Vibrio harveyi, and 42.7% identity with N-acetylglucosaminidase (ExoI) from Vibrio furnissii. Enzymatic activity assay of N-acetylglucosaminidase indicated stronger activity toward PNP-GlcNAc than PNP-(GlcNAc)_2 or PNP-(GlcNAc)_3.
- 社団法人日本農芸化学会の論文
- 1999-07-23
著者
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Takahashi N
Plant Functions Laboratory Riken
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NAKAGAWA Tsuyoshi
Research Institute of Molecular Genetics, Shimane University
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KAWAMUKAI Makoto
Department of Applied Bioscience and Biotechnology, Faculty of Life and Environmental Science
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MATSUDA Hideyuki
Department of Applied Bioscience and Biotechnology, Faculty of Life and Environmental Science
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Tanaka Katsunori
Department of Applied Bioscience and Biotechnology, Faculty of Life and Environmental Science, Shima
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Tanaka K
Tottori Univ. Tottori
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Nakano Takeshi
Riken Advanced Sci. Inst.
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MATSUO Yasuhiro
Ariake National College of Technology
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Takahashi Nobutaka
Frontier Research Program The Institute Of Physical And Chemical Research (rlken)
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Park Jae
Department Of Applied Bioscience And Biotechnology Faculty Of Life And Environmental Science
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MATSUO Yasuhiro
Department of Applied Bioscience and Biotechnology, Faculty of Life and Environmental Science
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KURITA Masako
Department of Applied Bioscience and Biotechnology, Faculty of Life and Environmental Science
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PARK JaeKweon
Department of Applied Bioscience and Biotechnology, Faculty of Life and Environmental Science
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Nakagawa Tetsuto
Department Of Bioscience And Biotechnology Division Of Bioresource And Bioenvironmental Sciences Gra
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Nakagawa Tsuyoshi
Dep. Of Molecular And Functional Genomics Center For Integrated Res. In Sci. Shimane Univ.
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Tanaka Kiyoshi
Coll. Environ. Health Azabu Univ.
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NAKANO Takeshi
The Institute of Physical and Chemical Research (RIKEN)
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Kurita Masako
Department Of Applied Bioscience And Biotechnology Faculty Of Life And Environmental Science
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Kawamukai Makoto
Department Of Applied Bioscience And Biotechnology Faculty Of Life And Environmental Science Shimane
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Nakagawa T
Department Of Bioscience And Biotechnology Division Of Bioresource And Bioenvironmental Sciences Gra
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Matsuo Yasuhiro
Department Of Applied Bioscience And Biotechnology Faculty Of Life And Environmental Science Shimane
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Morita K
Department Of Applied Bioscience And Biotechnology Faculty Of Life And Environmental Science Shimane
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Kawamukai M
Department Of Applied Bioscience And Biotechnology Faculty Of Life And Environmental Science Shimane
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Takahashi Nobuyuki
Hokkaido University Of Education
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Matsuda H
Department Of Applied Bioscience And Biotechnology Faculty Of Life And Environmental Science Shimane
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Matsuda Hideyuki
Department Of Applied Bioscience And Biotechnology Faculty Of Life And Environmental Science Shimane
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Kawamukai Makoto
Department Of Life Science And Biotechnology Faculty Of Life And Environmental Science Shimane Unive
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Kikuchi Takao
Coll. Environ. Health Azabu Univ.
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Nakagawa Tsuyoshi
Department Of Molecular And Functional Genomics Center For Integrated Research In Science Shimane Un
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Kawamukai Makoto
Department Of Life Science And Biotechnology Faculty Of Life And Environmental Science Shimane Unive
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Park Jae
Department of Advanced Materials Science and Engineering, Sungkyunkwan University, Suwon, Kyunggi-do 440-746, South Korea
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Tanaka Kyoko
Graduate School of Life and Environmental Sciences, Kyoto Prefectural University
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