完整後設資料紀錄
DC 欄位語言
dc.contributor.authorChiu, Hsien-Taien_US
dc.contributor.authorLin, Yu-Chinen_US
dc.contributor.authorLee, Meng-Naen_US
dc.contributor.authorChen, Yi-Linen_US
dc.contributor.authorWang, Mei-Sinen_US
dc.contributor.authorLai, Chia-Chunen_US
dc.date.accessioned2014-12-08T15:10:13Z-
dc.date.available2014-12-08T15:10:13Z-
dc.date.issued2009en_US
dc.identifier.issn1742-206Xen_US
dc.identifier.urihttp://hdl.handle.net/11536/7800-
dc.identifier.urihttp://dx.doi.org/10.1039/b912395ben_US
dc.description.abstractThe indolocarbazole family of natural products has attracted great attention because of their unique structural features and potential therapeutic applications. Structurally distinct in the family, K-252a is characterized by an unusual dihydrostreptose moiety cross-bridged to K-252c aglycone with two C-N linkages. K-252a has served as a valuble lead for treatments of various cancers and neurodegenerative disorders. Recent cloning of the nok gene cluster for biosyntheses of K-252a and its analogs from Nocardiopsis sp. K-252 (NRRL15532) has revealed the nokABCD genes indispensible for K-252c biosynthesis and the key gene (nokL) coding for N-glycosylation. Herein, we report the first, successful demonstration of in vitro sugar transferase activity of indolocarbazole N-glycosyltransferase (NokL) by use of soluble protein expressed from Escherichia coli. Notably, NokL was found to exhibit peculiar mode of substrate promiscuity. Moreover, NokA and NokB reactions were biochemically characterized thoroughly by natual and alternative (e. g. fluoro-) substrates and by ammonium hydroxide (NH(4)OH). Interestingly, the in vitro expression of NokA revealed high substrate stereoselectivity, giving several indole-3-pyruvic acid-derived compounds, including indol-3-carboxaldehyde (ICA) and indole-3-acetic acid. The use of NH(4)OH successfully dissected the in vitro NokA/NokB coupled reaction, revealing mechanistic insight into the enzymes and their cross-talking relationship. Also, a simple, useful method to synthesize K-252d, ICA and chromopyrrolic acid (the NokB product) was developed by the E. coli expression systems of NokL, NokA and NokA/NokB, respectively. Together with NokA and NokB, NokL may serve as a useful tool for combinatorial engineering of K-252a and its analogs for improved therapeutic values.en_US
dc.language.isoen_USen_US
dc.titleBiochemical characterization and substrate specificity of the gene cluster for biosyntheses of K-252a and its analogs by in vitro heterologous expression system of Escherichia colien_US
dc.typeArticleen_US
dc.identifier.doi10.1039/b912395ben_US
dc.identifier.journalMOLECULAR BIOSYSTEMSen_US
dc.citation.volume5en_US
dc.citation.issue10en_US
dc.citation.spage1192en_US
dc.citation.epage1203en_US
dc.contributor.department生物科技學系zh_TW
dc.contributor.departmentDepartment of Biological Science and Technologyen_US
dc.identifier.wosnumberWOS:000269835400012-
dc.citation.woscount3-
顯示於類別:期刊論文


文件中的檔案:

  1. 000269835400012.pdf

若為 zip 檔案,請下載檔案解壓縮後,用瀏覽器開啟資料夾中的 index.html 瀏覽全文。