Role of Tyrosine 114 of L-Methionine γ-lyase from Pseudomonas putida

• INOUE Hiroyuki

  Department of Bioresources Chemistry, Faculty of Agriculture, Okayama University

• INAGAKI Kenji

  Department of Bioresources Chemistry, Faculty of Agriculture, Okayama University

• ADACHI Naoki

  Department of Bioresources Chemistry, Faculty of Agriculture, Okayama University

• TAMURA Takashi

  Department of Bioresources Chemistry, Faculty of Agriculture, Okayama University

• ESAKI Nobuyoshi

  Institute for Chemical Research, Kyoto University

• SODA Kenji

  Department of Biotechnology, Faculty of Engineering, Kansai University

• TANAKA Hidehiko

  Department of Bioresources Chemistry, Faculty of Agriculture, Okayama University

書誌事項

タイトル別名

• Role of Tyrosine 114 of L-Methionine .GAMMA.-lyase from Pseudomonas putida.
• Role of Tyrosine 114 of L Methionine ガンマ lyase from Pseudomonas putida
• Role of Tyrosine 114 of<scp>L</scp>-Methionine γ-lyase from<i>Pseudomonas putida</i>

抄録

  L-Methionine γ-lyase from Pseudomonas putida has a conserved tyrosine residue (Tyr114) in the active site as in all known sequences of γ-family pyridoxal 5′-phosphate dependent enzymes. A mutant form of L-methionine γ-lyase in which Tyr114 was replaced by phenylalanine (Y114F) resulted in 910-fold decrease in k_cat for α,γ-elimination of L-methionine, while the K_m remained the same as the wild type enzyme. The Y114F mutant had the reduced k_cat by only 28- and 16-fold for substrates with an electron-withdrawing group at the γ-position, namely O-acetyl-L-homoserine and L-methionine sulfone, respectively, and also the similar reduction of k_cat for α,β-elimination and deamination substrates. The hydrogen exchange reactions of substrate and the spectral changes of the substrate-enzyme complex catalyzed by the mutant enzyme suggested that γ-elimination process for L-methionine is the rate-limiting determination step in α,γ-elimination overall reaction of the Y114F mutant. These results indicate that Tyr114 of L-methionine γ-lyase is important in γ-elimination of the substrate.<br>

収録刊行物

• Ｂｉｏｓｃｉｅｎｃｅ，　Ｂｉｏｔｅｃｈｎｏｌｏｇｙ，　ａｎｄ　Ｂｉｏｃｈｅｍｉｓｔｒｙ

  Ｂｉｏｓｃｉｅｎｃｅ，　Ｂｉｏｔｅｃｈｎｏｌｏｇｙ，　ａｎｄ　Ｂｉｏｃｈｅｍｉｓｔｒｙ 64 (11), 2336-2343, 2000

  公益社団法人 日本農芸化学会

キーワード

• general acid catalyst
• <small>L</small>-methionine γ-lyase
• reaction mechanism
• site-directed mutagenesis

詳細情報

• CRID: 1390001206473671168
• NII論文ID: 110002679823
• NII書誌ID: AA10824164
• DOI: 10.1271/bbb.64.2336
• COI: 1:CAS:528:DC%2BD3cXos1Srt7c%3D
• ISSN: 13476947
• ISSN: 09168451
• NDL書誌ID: 5590395
• PubMed: 11193400
• Web Site: https://ndlsearch.ndl.go.jp/books/R000000004-I5590395
• Web Site: https://www.tandfonline.com/doi/pdf/10.1271/bbb.64.2336
• 本文言語コード: en
• データソース種別:
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  • PubMed
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• 抄録ライセンスフラグ: 使用不可