Characterization of yrpC Gene Product of Bacillus subtilis IFO 3336 as Glutamate Racemase Isozyme

• 資料種別: 記事
• タイトル: Characterization of yrpC Gene Product of Bacillus subtilis IFO 3336 as Glutamate Racemase Isozyme
• 著者・編者: Makoto Ashiuchi; Kenji Soda; Haruo Misono
• 著者標目: Makoto Ashiuchi; Kenji Soda; Haruo Misono
• タイトル（掲載誌）: Bioscience, Biotechnology, and Biochemistry
• 巻号年月日等（掲載誌）: 63(5) (通号 716) 1999.05
• 掲載巻: 63
• 掲載号: 5
• 掲載通号: 716
• 掲載ページ: 792～798
• 掲載年月日（W3CDTF）: 1999-05
• ISSN（掲載誌）: 0916-8451
• ISSN-L（掲載誌）: 0916-8451
• 出版事項（掲載誌）: Tokyo : Japan Society for Bioscience
• 出版地（国名コード）: JP
• 本文の言語コード: eng
• NDLC: ZR7
• 対象利用者: 一般
• 所蔵機関: 国立国会図書館
• 請求記号: Z53-G223
• 連携機関・データベース: 国立国会図書館 : 国立国会図書館雑誌記事索引; https://ndlsearch.ndl.go.jp
• 書誌ID（NDLBibID）: 4743726; http://id.ndl.go.jp/bib/4743726
• 整理区分コード: 632

• 要約等: Glr, the glutamate racemase of <i>Bacillus subtilis</i> (formerly <i>Bacillus natto</i>) IFO 3336 encoded by the <i>glr</i> gene, and YrpC, a protein encoded by the <i>yrpC</i> gene, which is located at a different locus from that of the <i>glr</i> gene in the <i>B. subtilis</i> genome, share a high sequence similarity. The <i>yrpC</i> gene complemented the <small>D</small>-glutamate auxotrophy of <i>Escherichia coli</i> WM335 cells defective in the glutamate racemase gene. Glutamate racemase activity was found in the extracts of <i>E. coli</i> WM335 clone cells harboring a plasmid, pYRPC1, carrying its gene. Thus, the <i>yrpC</i> gene encodes an isozyme of glutamate racemase of <i>B. subtilis</i> IFO 3336. YrpC is mostly found in an inactive inclusion body in <i>E. coli</i> JM109/pYRPC1 cells. YrpC was solubilized readily, but glutamate racemase activity was only slightly restored. We purified YrpC from the extracts of <i>E. coli</i> JM109/pYRPC2 cells using a Glutathione S-transferase Gene Fusion System to characterize it. YrpC is a monomeric protein and contains no cofactors, like Glr. Enzymological properties of YrpC, such as the substrate specificity and optimum pH, are also similar to those of Glr. The thermostability of YrpC, however, is considerably lower than that of Glr. In addition, YrpC showed higher affinity and lower catalytic efficiency for <small>L</small>-glutamate than Glr. This is the first example showing the occurrence and properties of a glutamate racemase isozyme.<br>
• DOI: 10.1271/bbb.63.792
• オンライン閲覧公開範囲: インターネット公開
• 連携機関・データベース: 科学技術振興機構 : J-STAGE; http://www.jstage.jst.go.jp

• 要約等: Glr, the glutamate racemase of <i>Bacillus subtilis</i> (formerly <i>Bacillus natto</i>) IFO 3336 encoded by the <i>glr</i> gene, and YrpC, a protein encoded by the <i>yrpC</i> gene, which is located at a different locus from that of the <i>glr</i> gene in the <i>B. subtilis</i> genome, share a high sequence similarity. The <i>yrpC</i> gene complemented the <small>D</small>-glutamate auxotrophy of <i>Escherichia coli</i> WM335 cells defective in the glutamate racemase gene. Glutamate racemase activity was found in the extracts of <i>E. coli</i> WM335 clone cells harboring a plasmid, pYRPC1, carrying its gene. Thus, the <i>yrpC</i> gene encodes an isozyme of glutamate racemase of <i>B. subtilis</i> IFO 3336. YrpC is mostly found in an inactive inclusion body in <i>E. coli</i> JM109/pYRPC1 cells. YrpC was solubilized readily, but glutamate racemase activity was only slightly restored. We purified YrpC from the extracts of <i>E. coli</i> JM109/pYRPC2 cells using a Glutathione S-transferase Gene Fusion System to characterize it. YrpC is a monomeric protein and contains no cofactors, like Glr. Enzymological properties of YrpC, such as the substrate specificity and optimum pH, are also similar to those of Glr. The thermostability of YrpC, however, is considerably lower than that of Glr. In addition, YrpC showed higher affinity and lower catalytic efficiency for <small>L</small>-glutamate than Glr. This is the first example showing the occurrence and properties of a glutamate racemase isozyme.<br>
• DOI: 10.1271/bbb.63.792; https://doi.org/10.1271/bbb.63.792
• 関連情報（URI）: http://id.ndl.go.jp/bib/4743726; https://ndlsearch.ndl.go.jp/books/R000000004-I4743726; http://www.tandfonline.com/doi/pdf/10.1271/bbb.63.792
• 参照: Amino Acid Racemases: Functions and Mechanisms; https://cir.nii.ac.jp/crid/1360002219160499584
• 連携機関・データベース: 国立情報学研究所 : CiNii Research; https://cir.nii.ac.jp/
• 提供元機関・データベース: Japan Link Center; https://japanlinkcenter.org/top; 雑誌記事索引データベース; https://ndlsearch.ndl.go.jp; Crossref; https://www.crossref.org; PubMed; https://pubmed.ncbi.nlm.nih.gov; CiNii Articles
• 書誌ID（NDLBibID）: 4743726; http://id.ndl.go.jp/bib/4743726
• NII論文ID: 110002679566