Journal of Microbiology and Biotechnology

  • 제11권1호
  • /
  • Pages.118-123
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  • 2001
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  • 1017-7825(pISSN)
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  • 1738-8872(eISSN)
한국미생물·생명공학회 (The Korean Society for Microbiology and Biotechnology)
권호 표지

Effect of His192 Mutation on the Activity of Alginate Lyase A1-III from Sphingomonas Species A1

  • Yoon, Hye-Jin (Department of Chemistry, College of Natural Science, Seoul National University) ;
  • Choi, Yong-Jin (Graduate School of Biotechnology, Korea University) ;
  • Osamu Miyake (Research Institute for Food Science, Kyoto University) ;
  • Wataru Hashimoto (Research Institute for Food Science, Kyoto University) ;
  • Kousaku Murata (Research Institute for Food Science, Kyoto University) ;
  • Bunzo Mikami (Research Institute for Food Science, Kyoto University)
  • 발행 : 2001.02.01
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초록

The alginate lyase A1-III gene of Sphingomonas species A1 is composed of 1,077 nucleotides, encoding a protein (359 amino acids) with a molecular mass of 40,322 Da. Recombinant A1-III expressed in Escherichia coli exhibited the same full enzymatic activity as native A1-III. In order to identify the critical residue for activity, a site-directed mutation was introduced into the A1-III gene (H192A, His192->Ala). Recombinant A1-III (H192A) exhibited a significant decrease in enzyme activity (one-thirty thousandth of that of A1-III), without any conformational change, as detected by the CD spectra in the far UV region. Also, the chemical modification of wild-type A1-III with methyl 4-nitro benzene sulfonate resulted in a 40% decrease from the initial activity, whereas the same modification of A1-III (H192A) produced no change in the activity. The role of His192 on the catalytic process was also explored based on a model of A1-III docked with mannuronic acid into the active site.

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