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Discovery of a Novel Alginate Lyase from Nitratiruptor sp SB155-2 Thriving at Deep-sea Hydrothermal Vents and Identification of the Residues Responsible for Its Heat Stability

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タイトル: Discovery of a Novel Alginate Lyase from Nitratiruptor sp SB155-2 Thriving at Deep-sea Hydrothermal Vents and Identification of the Residues Responsible for Its Heat Stability
著者: Inoue, Akira 著作を一覧する
Anraku, Moe 著作を一覧する
Nakagawa, Satoshi 著作を一覧する
Ojima, Takao 著作を一覧する
発行日: 2016年 7月23日
出版者: American Society for Biochemistry and Molecular Biology (ASBMB)
誌名: Journal of Biological Chemistry (JBC)
巻: 291
号: 30
開始ページ: 15551
終了ページ: 15563
出版社 DOI: 10.1074/jbc.M115.713230
抄録: Extremophiles are expected to represent a source of enzymes having unique functional properties. The hypothetical protein NIS_0185, termed NitAly in this study, was identified as an alginate lyase-homolog protein in the genomic database of epsilon-Proteobacteria Nitratiruptor sp.SB155-2, which was isolated from deep-sea hydrothermal vents at a water depth of 1,000 m. Among the characterized alginate lyases in the polysaccharide lyase family 7 (PL-7), the amino acid sequence of NitAly showed the highest identity (39%) with that of red alga Pyropia yezoensis alginate lyase PyAly. Recombinant NitAly (rNitAly) was successfully expressed in Escherichia coli. Purified rNitAly degraded alginate in an endolytic manner. Among alginate block types, polyM was preferable to polyG and polyMG as a substrate, and its end degradation products were mainly tri-, tetra-, and penta-saccharides. The optimum temperature and pH values were 70 degrees C and around 6, respectively. Ahigh concentration of NaCl (0.8-1.4 M) was required for maximum activity. In addition, a 50% loss of activity was observed after incubation at 67 degrees C for 30 min. Heat stability was decreased in the presence of 5 mM DTT, and Cys-80 and Cys-232 were identified as the residues responsible for heat stability but not lyase activity. Introducing two cysteines into PyAly based on homology modeling using Pseudomonas aeruginosa alginate lyase PA1167 as the template enhanced its heat stability. Thus, NitAly is a functional alginate lyase, with its unique optimum conditions adapted to its environment. These insights into the heat stability of NitAly could be applied to improve that of other PL-7 alginate lyases.
Rights: "This research was originally published in Journal of Biological Chemistry (JBC). Inoue, Akira, Anraku, Moe, Nakagawa, Satoshi, Ojima, Takao. Discovery of a Novel Alginate Lyase from Nitratiruptor sp SB155-2 Thriving at Deep-sea Hydrothermal Vents and Identification of the Residues Responsible for Its Heat Stability. Journal of Biological Chemistry (JBC). 2016; 291:15551pp-15563pp. © the American Society for Biochemistry and Molecular Biology."
資料タイプ: article
URI: http://hdl.handle.net/2115/66755
出現コレクション:雑誌発表論文等 (Peer-reviewed Journal Articles, etc)

提供者: 井上 晶

 

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