In vitro kinetic study of the squalestatin tetraketide synthase dehydratase reveals the stereochemical course of a fungal highly reducing polyketide synthase

Emma Liddle; Alan Scott; Li Chen Han; David Ivison; Thomas J. Simpson; Christine L. Willis; Russell J. Cox

doi:10.1039/c6cc10172k

In vitro kinetic study of the squalestatin tetraketide synthase dehydratase reveals the stereochemical course of a fungal highly reducing polyketide synthase

Emma Liddle
, Alan Scott
, Li Chen Han
, David Ivison
, Thomas J. Simpson
, Christine L. Willis
, Russell J. Cox^*

^*Corresponding author for this work

Institute of Organic Chemistry

University of Bristol

Research output: Contribution to journal › Article › Research › peer review

Abstract

Six potential diketide substrates for the squalestatin tetraketide synthase (SQTKS) dehydratase (DH) domain were synthesised as N-acetyl cysteamine thiolesters (SNAC) and tested in kinetic assays as substrates with an isolated DH domain. 3R-3-hydroxybutyryl SNAC 3R-16 was turned over by the enzyme, but its enantiomer was not. Of the four 2-methyl substrates only 2R,3R-2-methyl-3-hydroxybutyryl SNAC 2R,3R-8 was a substrate. Combined with stereochemical information from the isolated SQTKS enoyl reductase (ER) domain, our results provide a near complete stereochemical description of the first cycle of beta-modification reactions of a fungal highly reducing polyketide synthase (HR-PKS). The results emphasise the close relationship between fungal HR-PKS and vertebrate fatty acid synthases (vFAS).

Original language	English
Pages (from-to)	1727-1730
Number of pages	4
Journal	Chemical communications
Volume	53
Issue number	10
DOIs	https://doi.org/10.1039/c6cc10172k
Publication status	Published - 1 Jan 2017

ASJC Scopus subject areas

Catalysis
Electronic, Optical and Magnetic Materials
Ceramics and Composites
General Chemistry
Surfaces, Coatings and Films
Metals and Alloys
Materials Chemistry

Access to Document

10.1039/c6cc10172k

Cite this

@article{0a7116a1469f49a09f00efc27e3e0984,

title = "In vitro kinetic study of the squalestatin tetraketide synthase dehydratase reveals the stereochemical course of a fungal highly reducing polyketide synthase",

abstract = "Six potential diketide substrates for the squalestatin tetraketide synthase (SQTKS) dehydratase (DH) domain were synthesised as N-acetyl cysteamine thiolesters (SNAC) and tested in kinetic assays as substrates with an isolated DH domain. 3R-3-hydroxybutyryl SNAC 3R-16 was turned over by the enzyme, but its enantiomer was not. Of the four 2-methyl substrates only 2R,3R-2-methyl-3-hydroxybutyryl SNAC 2R,3R-8 was a substrate. Combined with stereochemical information from the isolated SQTKS enoyl reductase (ER) domain, our results provide a near complete stereochemical description of the first cycle of beta-modification reactions of a fungal highly reducing polyketide synthase (HR-PKS). The results emphasise the close relationship between fungal HR-PKS and vertebrate fatty acid synthases (vFAS).",

author = "Emma Liddle and Alan Scott and Han, \{Li Chen\} and David Ivison and Simpson, \{Thomas J.\} and Willis, \{Christine L.\} and Cox, \{Russell J.\}",

note = "Funding information: We thank EPSRC (EP/F066104/1) for LCMS equipment, BBSRC (BB/I003355/1) for funding AS and BrisSynBio Synthetic Biology Research Centre, (BB/L01386X/1) for funding L-CH. E. L. and D. I. were funded by the School of Chemistry, University of Bristol. We thank Dr J. E. Nettleship at the Oxford Protein Production Facility (OPPF) for assistance with protein production. Hao Yao and Oliver Piech (LUH) are thanked for technical assistance.",

year = "2017",

month = jan,

day = "1",

doi = "10.1039/c6cc10172k",

language = "English",

volume = "53",

pages = "1727--1730",

journal = "Chemical communications",

issn = "1359-7345",

publisher = "Chemical Society",

number = "10",

}

TY - JOUR

T1 - In vitro kinetic study of the squalestatin tetraketide synthase dehydratase reveals the stereochemical course of a fungal highly reducing polyketide synthase

AU - Liddle, Emma

AU - Scott, Alan

AU - Han, Li Chen

AU - Ivison, David

AU - Simpson, Thomas J.

AU - Willis, Christine L.

AU - Cox, Russell J.

N1 - Funding information: We thank EPSRC (EP/F066104/1) for LCMS equipment, BBSRC (BB/I003355/1) for funding AS and BrisSynBio Synthetic Biology Research Centre, (BB/L01386X/1) for funding L-CH. E. L. and D. I. were funded by the School of Chemistry, University of Bristol. We thank Dr J. E. Nettleship at the Oxford Protein Production Facility (OPPF) for assistance with protein production. Hao Yao and Oliver Piech (LUH) are thanked for technical assistance.

PY - 2017/1/1

Y1 - 2017/1/1

N2 - Six potential diketide substrates for the squalestatin tetraketide synthase (SQTKS) dehydratase (DH) domain were synthesised as N-acetyl cysteamine thiolesters (SNAC) and tested in kinetic assays as substrates with an isolated DH domain. 3R-3-hydroxybutyryl SNAC 3R-16 was turned over by the enzyme, but its enantiomer was not. Of the four 2-methyl substrates only 2R,3R-2-methyl-3-hydroxybutyryl SNAC 2R,3R-8 was a substrate. Combined with stereochemical information from the isolated SQTKS enoyl reductase (ER) domain, our results provide a near complete stereochemical description of the first cycle of beta-modification reactions of a fungal highly reducing polyketide synthase (HR-PKS). The results emphasise the close relationship between fungal HR-PKS and vertebrate fatty acid synthases (vFAS).

AB - Six potential diketide substrates for the squalestatin tetraketide synthase (SQTKS) dehydratase (DH) domain were synthesised as N-acetyl cysteamine thiolesters (SNAC) and tested in kinetic assays as substrates with an isolated DH domain. 3R-3-hydroxybutyryl SNAC 3R-16 was turned over by the enzyme, but its enantiomer was not. Of the four 2-methyl substrates only 2R,3R-2-methyl-3-hydroxybutyryl SNAC 2R,3R-8 was a substrate. Combined with stereochemical information from the isolated SQTKS enoyl reductase (ER) domain, our results provide a near complete stereochemical description of the first cycle of beta-modification reactions of a fungal highly reducing polyketide synthase (HR-PKS). The results emphasise the close relationship between fungal HR-PKS and vertebrate fatty acid synthases (vFAS).

UR - https://www.scopus.com/pages/publications/85011103494

U2 - 10.1039/c6cc10172k

DO - 10.1039/c6cc10172k

M3 - Article

C2 - 28106181

AN - SCOPUS:85011103494

SN - 1359-7345

VL - 53

SP - 1727

EP - 1730

JO - Chemical communications

JF - Chemical communications

IS - 10

ER -

In vitro kinetic study of the squalestatin tetraketide synthase dehydratase reveals the stereochemical course of a fungal highly reducing polyketide synthase

Abstract

ASJC Scopus subject areas

Access to Document

Other files and links

Cite this