Abstract
The cyclohexenone ring A of tetracyclines exhibits unique structural features not observed among other aromatic polyketides. These substitutions include the C2 primary amide, C4 dimethylamine, and the C12a tertiary alcohol. Here we report the identification and reconstitution of the minimum set of enzymes required for the biosynthesis of anhydrotetracycline (ATC, 5), the first intermediate in the tetracycline biosynthetic pathway that contains the fully functionalized ring A. Using a combination of in vivo and in vitro approaches, we confirmed OxyL, OxyQ, and OxyT to be the only enzymes required to convert 6-methylpretetramid 1 into 5. OxyL is a NADPH-dependent dioxygenase that introduces two oxygen atoms into 1 to yield the unstable intermediate 4-keto-ATC 2. The aminotransferase OxyQ catalyzes the reductive amination of C4-keto of 2, yielding 4-amino-ATC 3. Furthermore, the N, N-dimethyltransferase OxyT catalyzes the formation of 5 from 3 in a (S)-adenosylmethionine (SAM)-dependent manner. Finally, a "non-natural" anhydrotetracycline derivative was generated, demonstrating that our heterologous host/vector pair can be a useful platform toward the engineered biosynthesis of tetracycline analogues.
References
Feb 1, 1989·Journal of Bacteriology·C BinnieM J Butler
Jun 15, 1964·Biochemical and Biophysical Research Communications·P A MillerN Bohonos
Jan 1, 1969·Journal of the American Chemical Society·J R McCormick, E R Jensen
Apr 8, 1994·Gene·E S KimD H Sherman
Apr 1, 2000·Applied and Environmental Microbiology·T NakanoR Katsumata
Aug 15, 2000·Angewandte Chemie·C w ChangH w Liu
Sep 29, 2000·Organic Letters·E R RafananB Shen
Jul 18, 2002·Biochemistry·Huawei ChenHung-Wen Liu
Apr 20, 1965·Journal of the American Chemical Society·J R MCCORMICKN O SJOLANDER
Sep 9, 2005·The Journal of Biological Chemistry·Natasa Peric-ConchaIain S Hunter
Apr 7, 2006·Applied and Environmental Microbiology·Wenjun ZhangYi Tang
Jul 13, 2007·Journal of the American Chemical Society·Wenjun ZhangYi Tang
Jul 17, 2007·The Journal of Biological Chemistry·Wenjun ZhangYi Tang
Citations
May 18, 2012·Journal of the American Chemical Society·Yit-Heng ChooiYi Tang
Nov 17, 2009·Journal of the American Chemical Society·Lauren B PickensYi Tang
Apr 2, 2010·Natural Product Reports·Hui ZhouYi Tang
Jun 5, 2010·The Journal of Biological Chemistry·Lauren B Pickens, Yi Tang
Aug 31, 2011·PloS One·Katherine S Ryan
Aug 28, 2012·Journal of Bioscience and Bioengineering·Bingji MaJixun Zhan
Dec 30, 2015·ACS Chemical Biology·Yiguang ZhuChangsheng Zhang
Nov 15, 2008·Metabolic Engineering·Lauren B Pickens, Yi Tang
Mar 1, 2012·IUBMB Life·Dayu YuJixun Zhan
Jun 23, 2015·Chemistry & Biology·Kevin J ForsbergGautam Dantas
Jun 27, 2013·Metabolic Engineering·Tao ZhuDelin You
May 20, 2016·ACS Chemical Biology·Lei SunJixun Zhan
Feb 1, 2017·Biotechnology Journal·Simon J MoorePaul S Freemont
Oct 2, 2012·Angewandte Chemie·Peng WangYi Tang
Apr 29, 2016·Microbiology and Molecular Biology Reviews : MMBR·Chris GreeningColin J Jackson
Feb 9, 2010·Chembiochem : a European Journal of Chemical Biology·Madan K KharelJürgen Rohr
Apr 1, 2010·Natural Product Reports·Carlos OlanoJosé A Salas
May 26, 2020·Microbial Cell Factories·Jia WangQipeng Yuan
May 28, 2009·Chembiochem : a European Journal of Chemical Biology·Peng WangYi Tang
Jun 7, 2020·The Journal of Biological Chemistry·Yamini MathurAmrita B Hazra
Dec 4, 2019·Angewandte Chemie·Vincent C FäsekeChristof Sparr
Dec 29, 2020·Microbiological Research·Xing XiaoJunmin Zhang
Feb 13, 2019·ACS Chemical Biology·Tadeja LukežičRolf Müller
Apr 30, 2013·Journal of the American Chemical Society·Peng WangYi Tang
May 10, 2013·ACS Synthetic Biology·Bijan Zakeri, Timothy K Lu
Dec 24, 2019·ACS Omega·Lixia PanDengfeng Yang