Identifying the minimal enzymes required for anhydrotetracycline biosynthesis

Journal of the American Chemical Society
Wenjun ZhangJixun Zhan

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

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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 28, 2012·Journal of Bioscience and Bioengineering·Bingji MaJixun Zhan
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
Feb 1, 2017·Biotechnology Journal·Simon J MoorePaul S Freemont
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
May 28, 2009·Chembiochem : a European Journal of Chemical Biology·Peng WangYi Tang
Dec 4, 2019·Angewandte Chemie·Vincent C FäsekeChristof Sparr
Apr 30, 2013·Journal of the American Chemical Society·Peng WangYi Tang
May 10, 2013·ACS Synthetic Biology·Bijan Zakeri, Timothy K Lu

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