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Purification and characterization of Vibrio parahaemolyticus extracellular chitinase and chitin oligosaccharide deacetylase involved in the production of heterodisaccharide from chitin

  • Biotechnologically Relevant Enzymes and Proteins
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Abstract

A chitin-degrading bacterial strain, KN1699, isolated from Yatsu dry beach (Narashino, Chiba Prefecture, Japan), was identified as Vibrio parahaemolyticus. Treatment of powdered chitin with crude enzyme solution prepared from the supernatant of KN1699 cultures yielded a disaccharide, β-d-N-acetylglucosaminyl-(1,4)-d-glucosamine (GlcNAc-GlcN), as the primary chitin degradation product. The extracellular enzymes involved in the production of this heterodisaccharide, chitinase (Pa-Chi; molecular mass, 92 kDa) and chitin oligosaccharide deacetylase (Pa-COD; molecular mass, 46 kDa), were isolated from the crude enzyme solution, and their hydrolysis specificities were elucidated. These studies confirmed that (1) Pa-Chi hydrolyzes chitin to produce (GlcNAc)2 and (2) Pa-COD hydrolyzes the acetamide group of reducing end GlcNAc residue of (GlcNAc)2. These findings indicate that GlcNAc-GlcN is produced from chitin by the cooperative hydrolytic reactions of both Pa-Chi and Pa-COD.

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Acknowledgment

This work was supported by a grant from the 21st Century Center of Excellence (COE) Program of the Ministry of Education, Science, Sports, and Culture (Japan) to promote advanced scientific research.

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Authors and Affiliations

  1. Department of Biological Chemistry, College of Bioresource Sciences, Nihon University, 1866 Kameino, Fujisawa, Kanagawa, 252-8510, Japan

    K. Kadokura, A. Rokutani, M. Yamamoto, T. Ikegami, T. Oku & T. Nishio

  2. Department of Marine Sciences and Resource, College of Bioresource Sciences, Nihon University, 1866 Kameino, Fujisawa, Kanagawa, 252-8510, Japan

    H. Sugita & S. Itoi

  3. Division of Organic Chemistry, National Institute of Health Sciences (NIHS), 1-8-1 Kamiyoga, Setagaya-ku, Tokyo, 158-8501, Japan

    W. Hakamata

Authors
  1. K. Kadokura
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  2. A. Rokutani
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  3. M. Yamamoto
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  4. T. Ikegami
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  5. H. Sugita
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  8. T. Oku
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  9. T. Nishio
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Correspondence to T. Nishio.

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Kadokura, K., Rokutani, A., Yamamoto, M. et al. Purification and characterization of Vibrio parahaemolyticus extracellular chitinase and chitin oligosaccharide deacetylase involved in the production of heterodisaccharide from chitin. Appl Microbiol Biotechnol 75, 357–365 (2007). https://doi.org/10.1007/s00253-006-0831-6

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  • DOI: https://doi.org/10.1007/s00253-006-0831-6

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