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Molecular cloning and characterization of 58 kDa chitinase gene fromSerratia marcescens KCTC 2172

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Abstract

A chitinase gene (pCHi58) encoding a 58 kDa chitinase was isolated from theSerratia marcescens KCTC 2172 cosmid library. The chitinase gene consisted of a 1686 bp open reading frame that encoded 562 amino acids.Escherichia coil harboring the pChi58 gene secreted a 58 kDa chitinase into the culture supernatant. The 58 kDa chitinase was purified using a chitin affinity column and mono-S column. A nucleotide andN-terminal amino acid sequence analysis showed that the 58 kDa chitinase had a leader peptide consisting of 23 amino acids which was cleaved prior to the 24th alanine. The 58 KDa chitinase exhibited a 98% similarity to that ofS. marcescens QMB 1466 in its nuclotide sequence. The chitinolytic patterns of the 58 kDa chitinase released N,N′-diacetyl chitobiose (NAG2) as the major hydrolysis end-product with a trace amount ofN-acetylglucosamine. When a 4-methylumbellyferyl-N-acetylglucosamin monomer, dimmer, and tetramer were used as substrates, the 58 kDa chitinase did not digest the 4-Mu-NAG monomer (analogue of NAG2), thereby indicating that the 58 kDa chitinase was likely an endochitinase. The optimum reaction temperature and pH of the enzyme were 50°C and 5.0, respectively.

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Authors and Affiliations

  1. Department of Microbiological Engineering, Chinju National University, 660-758, Chinju, Korea

    Sang Wan Gal & S. W. Lee

  2. Department of Food and Nutrition, Sila University, 617-736, Pusan, Korea

    Y. J. Choi

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  1. Sang Wan Gal
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  2. S. W. Lee
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  3. Y. J. Choi
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Correspondence to Sang Wan Gal.

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Gal, S.W., Lee, S.W. & Choi, Y.J. Molecular cloning and characterization of 58 kDa chitinase gene fromSerratia marcescens KCTC 2172. Biotechnol. Bioprocess Eng. 7, 38–42 (2002). https://doi.org/10.1007/BF02935878

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  • DOI: https://doi.org/10.1007/BF02935878

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