TY - JOUR
T1 - Diene incorporation by a dehydratase domain variant in modular polyketide synthases
AU - Hobson, Christian
AU - Jenner, Matthew
AU - Jian, Xinyun
AU - Griffiths, Daniel
AU - Roberts, Douglas M.
AU - Rey-Carrizo, Matias
AU - Challis, Gregory L.
N1 - Funding Information:
This research was supported by the Biotechnology and Biological Sciences Research Council (grant nos. BB/L021692/1, BB/K002341/1 and BB/M017982/1 to G.L.C.). M.J. and D.G. were supported by a Biotechnology and Biological Sciences Research Council Discovery Fellowship (grant no. BB/R012121/1) and the EPSRC MOAC Doctoral Training Centre (grant no. EP/F500378/1), respectively. M.R.-C. was supported by a mobility grant from the Government of Andorra (grant no. AM025-AND-2015). C.H. was supported by a PhD studentship from the University of Warwick. G.L.C. was the recipient of a Wolfson Research Merit Award from the Royal Society (grant no. WM130033).
Publisher Copyright:
© 2022, The Author(s), under exclusive licence to Springer Nature America, Inc.
PY - 2022/12
Y1 - 2022/12
N2 - Modular polyketide synthases (PKSs) are biosynthetic assembly lines that construct structurally diverse natural products with wide-ranging applications in medicine and agriculture. Various mechanisms contribute to structural diversification during PKS-mediated chain assembly, including dehydratase (DH) domain-mediated elimination of water from R and S-configured 3-hydroxy-thioesters to introduce E- and Z-configured carbon–carbon double bonds, respectively. Here we report the discovery of a DH domain variant that catalyzes the sequential elimination of two molecules of water from a (3R, 5S)-3,5-dihydroxy thioester during polyketide chain assembly, introducing a conjugated E,Z-diene into various modular PKS products. We show that the reaction proceeds via a (2E, 5S)-2-enoyl-5-hydroxy-thioester intermediate and involves an additional universally conserved histidine residue that is absent from the active site of most conventional DH domains. These findings expand the diverse range of chemistries mediated by DH-like domains in modular PKSs, highlighting the catalytic versatility of the double hotdog fold. [Figure not available: see fulltext.]
AB - Modular polyketide synthases (PKSs) are biosynthetic assembly lines that construct structurally diverse natural products with wide-ranging applications in medicine and agriculture. Various mechanisms contribute to structural diversification during PKS-mediated chain assembly, including dehydratase (DH) domain-mediated elimination of water from R and S-configured 3-hydroxy-thioesters to introduce E- and Z-configured carbon–carbon double bonds, respectively. Here we report the discovery of a DH domain variant that catalyzes the sequential elimination of two molecules of water from a (3R, 5S)-3,5-dihydroxy thioester during polyketide chain assembly, introducing a conjugated E,Z-diene into various modular PKS products. We show that the reaction proceeds via a (2E, 5S)-2-enoyl-5-hydroxy-thioester intermediate and involves an additional universally conserved histidine residue that is absent from the active site of most conventional DH domains. These findings expand the diverse range of chemistries mediated by DH-like domains in modular PKSs, highlighting the catalytic versatility of the double hotdog fold. [Figure not available: see fulltext.]
UR - http://www.scopus.com/inward/record.url?scp=85138027973&partnerID=8YFLogxK
U2 - 10.1038/s41589-022-01127-y
DO - 10.1038/s41589-022-01127-y
M3 - Article
C2 - 36109649
AN - SCOPUS:85138027973
SN - 1552-4450
VL - 18
SP - 1410
EP - 1416
JO - Nature Chemical Biology
JF - Nature Chemical Biology
IS - 12
ER -