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Title: Structural insights into dehydratase substrate selection for the borrelidin and fluvirucin polyketide synthases

Abstract

Abstract Engineered polyketide synthases (PKSs) are promising synthetic biology platforms for the production of chemicals with diverse applications. The dehydratase (DH) domain within modular type I PKSs generates an α,β-unsaturated bond in nascent polyketide intermediates through a dehydration reaction. Several crystal structures of DH domains have been solved, providing important structural insights into substrate selection and dehydration. Here, we present two DH domain structures from two chemically diverse PKSs. The first DH domain, isolated from the third module in the borrelidin PKS, is specific towards a trans-cyclopentane-carboxylate-containing polyketide substrate. The second DH domain, isolated from the first module in the fluvirucin B1 PKS, accepts an amide-containing polyketide intermediate. Sequence-structure analysis of these domains, in addition to previously published DH structures, display many significant similarities and key differences pertaining to substrate selection. The two major differences between BorA DH M3, FluA DH M1 and other DH domains are found in regions of unmodeled residues or residues containing high B-factors. These two regions are located between α3–β11 and β7–α2. From the catalytic Asp located in α3 to a conserved Pro in β11, the residues between them form part of the bottom of the substrate-binding cavity responsible for binding to acyl-ACP intermediates.

Authors:
; ; ; ; ; ; ; ; ; ; ; ;
Publication Date:
Research Org.:
Lawrence Berkeley National Laboratory (LBNL), Berkeley, CA (United States)
Sponsoring Org.:
USDOE Office of Science (SC), Biological and Environmental Research (BER); USDOE Office of Energy Efficiency and Renewable Energy (EERE), Sustainable Transportation Office. Bioenergy Technologies Office (BETO)
OSTI Identifier:
1619394
Alternate Identifier(s):
OSTI ID: 1526606
Grant/Contract Number:  
AC02-05CH11231
Resource Type:
Published Article
Journal Name:
Journal of Industrial Microbiology and Biotechnology
Additional Journal Information:
Journal Name: Journal of Industrial Microbiology and Biotechnology Journal Volume: 46 Journal Issue: 8; Journal ID: ISSN 1367-5435
Publisher:
Oxford University Press
Country of Publication:
Germany
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; 42 ENGINEERING

Citation Formats

Barajas, Jesus F., McAndrew, Ryan P., Thompson, Mitchell G., Backman, Tyler W. H., Pang, Bo, de Rond, Tristan, Pereira, Jose H., Benites, Veronica T., Martín, Héctor García, Baidoo, Edward E. K., Hillson, Nathan J., Adams, Paul D., and Keasling, Jay D. Structural insights into dehydratase substrate selection for the borrelidin and fluvirucin polyketide synthases. Germany: N. p., 2019. Web. doi:10.1007/s10295-019-02189-z.
Barajas, Jesus F., McAndrew, Ryan P., Thompson, Mitchell G., Backman, Tyler W. H., Pang, Bo, de Rond, Tristan, Pereira, Jose H., Benites, Veronica T., Martín, Héctor García, Baidoo, Edward E. K., Hillson, Nathan J., Adams, Paul D., & Keasling, Jay D. Structural insights into dehydratase substrate selection for the borrelidin and fluvirucin polyketide synthases. Germany. https://doi.org/10.1007/s10295-019-02189-z
Barajas, Jesus F., McAndrew, Ryan P., Thompson, Mitchell G., Backman, Tyler W. H., Pang, Bo, de Rond, Tristan, Pereira, Jose H., Benites, Veronica T., Martín, Héctor García, Baidoo, Edward E. K., Hillson, Nathan J., Adams, Paul D., and Keasling, Jay D. Thu . "Structural insights into dehydratase substrate selection for the borrelidin and fluvirucin polyketide synthases". Germany. https://doi.org/10.1007/s10295-019-02189-z.
@article{osti_1619394,
title = {Structural insights into dehydratase substrate selection for the borrelidin and fluvirucin polyketide synthases},
author = {Barajas, Jesus F. and McAndrew, Ryan P. and Thompson, Mitchell G. and Backman, Tyler W. H. and Pang, Bo and de Rond, Tristan and Pereira, Jose H. and Benites, Veronica T. and Martín, Héctor García and Baidoo, Edward E. K. and Hillson, Nathan J. and Adams, Paul D. and Keasling, Jay D.},
abstractNote = {Abstract Engineered polyketide synthases (PKSs) are promising synthetic biology platforms for the production of chemicals with diverse applications. The dehydratase (DH) domain within modular type I PKSs generates an α,β-unsaturated bond in nascent polyketide intermediates through a dehydration reaction. Several crystal structures of DH domains have been solved, providing important structural insights into substrate selection and dehydration. Here, we present two DH domain structures from two chemically diverse PKSs. The first DH domain, isolated from the third module in the borrelidin PKS, is specific towards a trans-cyclopentane-carboxylate-containing polyketide substrate. The second DH domain, isolated from the first module in the fluvirucin B1 PKS, accepts an amide-containing polyketide intermediate. Sequence-structure analysis of these domains, in addition to previously published DH structures, display many significant similarities and key differences pertaining to substrate selection. The two major differences between BorA DH M3, FluA DH M1 and other DH domains are found in regions of unmodeled residues or residues containing high B-factors. These two regions are located between α3–β11 and β7–α2. From the catalytic Asp located in α3 to a conserved Pro in β11, the residues between them form part of the bottom of the substrate-binding cavity responsible for binding to acyl-ACP intermediates.},
doi = {10.1007/s10295-019-02189-z},
journal = {Journal of Industrial Microbiology and Biotechnology},
number = 8,
volume = 46,
place = {Germany},
year = {Thu Aug 01 00:00:00 EDT 2019},
month = {Thu Aug 01 00:00:00 EDT 2019}
}

Journal Article:
Free Publicly Available Full Text
Publisher's Version of Record
https://doi.org/10.1007/s10295-019-02189-z

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Cited by: 7 works
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