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Hokkaido University Collection of Scholarly and Academic Papers >
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Two Novel Glycoside Hydrolases Responsible for the Catabolism of Cyclobis-(1→6)-α-nigerosyl
| Title: | Two Novel Glycoside Hydrolases Responsible for the Catabolism of Cyclobis-(1→6)-α-nigerosyl |
| Authors: | Tagami, Takayoshi Browse this author | | Miyano, Eri Browse this author | | Sadahiro, Juri Browse this author | | Okuyama, Masayuki Browse this author →KAKEN DB | | Iwasaki, Tomohito Browse this author | | Kimura, Atsuo Browse this author →KAKEN DB |
| Keywords: | actinobacteria | | bacterial metabolism | | carbohydrate metabolism | | enzyme catalysis | | enzyme kinetics | | gene expression | | glycoside hydrolase | | glycosyltransferase | | cyclic oligosaccharide | | starch |
| Issue Date: | 5-Aug-2016 |
| Publisher: | American Society for Biochemistry and Molecular Biology (ASBMB) |
| Journal Title: | Journal of Biological Chemistry (JBC) |
| Volume: | 291 |
| Issue: | 32 |
| Start Page: | 16438 |
| End Page: | 16447 |
| Publisher DOI: | 10.1074/jbc.M116.727305 |
| PMID: | 27302067 |
| Abstract: | The actinobacterium Kribbella flavida NBRC 14399(T) produces cyclobis-(1 -> 6)-alpha-nigerosyl (CNN), a cyclic glucotetraose with alternate alpha-(1 -> 6)- and alpha-(1 -> 3)-glucosidic linkages, from starch in the culture medium. We identified gene clusters associated with the production and intracellular catabolism of CNN in the K. flavida genome. One cluster encodes 6-alpha-glucosyl-transferase and 3-alpha-isomaltosyltransferase, which are known to coproduce CNN from starch. The other cluster contains four genes annotated as a transcriptional regulator, sugar transporter, glycoside hydrolase family (GH) 31 protein (Kfla1895), and GH15 protein (Kfla1896). Kfla1895 hydrolyzed the alpha-(1 -> 3)-glucosidic linkages of CNN and produced isomaltose via a possible linear tetrasaccharide. The initial rate of hydrolysis of CNN (11.6 s(-1)) was much higher than that of panose (0.242 s(-1)), and hydrolysis of isomaltotriose and nigerose was extremely low. Because Kfla1895 has a strong preference for the alpha-(1 -> 3)-isomaltosyl moiety and effectively hydrolyzes the alpha-(1 -> 3)-glucosidic linkage, it should be termed 1,3-alpha-isomaltosidase. Kfla1896 effectively hydrolyzed isomaltose with liberation of beta-glucose, but displayed low or no activity toward CNN and the general GH15 enzyme substrates such as maltose, soluble starch, or dextran. The k(cat)/K-m for isomaltose (4.81 +/- 0.18 s(-1) mM(-1)) was 6.9- and 19-fold higher than those for panose and isomaltotriose, respectively. These results indicate that Kfla1896 is a new GH15 enzyme with high substrate specificity for isomaltose, suggesting the enzyme should be designated an isomaltose glucohydrolase. This is the first report to identify a starch-utilization pathway that proceeds via CNN. |
| Rights: | This research was originally published in Journal of Biological Chemistry (JBC). Tagami, Takayoshi; Miyano, Eri; Sadahiro, Juri; Okuyama, Masayuki; Iwasaki, Tomohito; Kimura, Atsuo. Two Novel Glycoside Hydrolases Responsible for the Catabolism of Cyclobis-(1→6)-α-nigerosyl. Journal of Biological Chemistry (JBC). 2016; Vol:pp.16438-16447.©the American Society for Biochemistry and Molecular Biology. |
| Type: | article |
| URI: | http://hdl.handle.net/2115/66973 |
| Appears in Collections: | 農学院・農学研究院 (Graduate School of Agriculture / Faculty of Agriculture) > 雑誌発表論文等 (Peer-reviewed Journal Articles, etc)
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Submitter: 木村 淳夫
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