Improvement of 2'-Hydroxybiphenyl-2-sulfinate Desulfinase, an Enzyme Involved in the Dibenzothiophene Desulfurization Pathway, from Rhodococcus erythropolis KA2-5-1 by Site-Directed Mutagenesis
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概要
- 論文の詳細を見る
In the microbial dibenzothiophene desulfurization pathway, 2′-hydroxybiphenyl-2-sulfinate is converted to 2-hydroxybiphenyl and sulfinate by desulfinase (DszB) at the last step, and this reaction is rate-limiting for the whole pathway. The catalytic activity and thermostability of DszB were enhanced by the two amino acid substitutions. Based on information on the 3-D structure of DszB and a comparison of amino acid sequences between DszB and reported thermophilic and thermostable homologs (TdsB and BdsB), two amino acid residues, Tyr63 and Gln65, were selected as targets to mutate and improve DszB. These two residues were replaced by several amino acids, and the promising mutant enzymes were purified and their properties were examined. Among the wild-type and mutant enzymes, Y63F had higher catalytic activity but similar thermostability, and Q65H showed higher thermostability but less catalytic activity and affinity for the substrate. To compensate for these drawbacks, the double mutant enzyme Y63F-Q65H was purified and its properties were investigated. This mutant enzyme showed higher thermostability without loss of catalytic activity or affinity for the substrate. These superior properties of the mutant enzyme have also been confirmed with resting cells harboring the mutant gene.
- 社団法人 日本農芸化学会の論文
- 2007-11-23
著者
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Izumi Yoshikazu
Dep. Of Biotechnology Tottori Univ.
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Tanokura Masaru
Department Of Applied Biological Chemistry Graduate School Of Agricultural And Life Sciences The Uni
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Lee Woo
Department Of Biological Scienceand Basic Science Research Institute Sungkyunkwan University
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Ohshiro Takashi
Department Of Biotechnology Faculty Of Engineering Tottori University
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Izumi Yoshikazu
Department Of Agricultural Chemistry Kyoto University
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Tanokura Masaru
Department Of Applied Biological Chemistry Graduate School Of Agricultural And Life Sciences Univers
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OHKITA Ryo
Department of Biotechnology, Tottori University
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TAKIKAWA Takeshi
Department of Biotechnology, Tottori University
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MANABE Masanori
Department of Biotechnology, Tottori University
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Ohkita Ryo
Department Of Biotechnology Tottori University
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Manabe Masanori
Department Of Biotechnology Tottori University
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Takikawa Takeshi
Department Of Biotechnology Tottori University
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Lee Woo
Department Of Applied Biological Chemistry Graduate School Of Agricultural And Life Sciences Univers
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