Identification of genes encoding squalestatin S1 biosynthesis and: In vitro production of new squalestatin analogues

B. Bonsch; V. Belt; C. Bartel; N. Duensing; M. Koziol; C. M. Lazarus; A. M. Bailey; T. J. Simpson; R. J. Cox

doi:10.1039/c6cc02130a

Identification of genes encoding squalestatin S1 biosynthesis and: In vitro production of new squalestatin analogues

B. Bonsch
, V. Belt
, C. Bartel
, N. Duensing
, M. Koziol
, C. M. Lazarus
, A. M. Bailey
, T. J. Simpson
, R. J. Cox^*

^*Corresponding author for this work

Institute of Organic Chemistry

University of Bristol

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

Abstract

A gene cluster responsible for the biosynthesis of squalestatin S1 (SQS1, 1) was identified by full genome sequencing of two SQS1-producing ascomycetes: Phoma sp. C2932 and unidentified fungus MF5453. A transformation protocol was established and a subsequent knockout of one PKS gene from the cluster led to loss of SQS1 production and enhanced concentration of an SQS1 precursor. An acyltransferase gene from the cluster was expressed in E. coli and the expressed protein MfM4 shown to be responsible for loading acyl groups from CoA onto the squalestatin core as the final step of biosynthesis. MfM4 appears to have a broad substrate selectivity for its acyl CoA substrate, allowing the in vitro synthesis of novel squalestatins.

Original language	English
Pages (from-to)	6777-6780
Number of pages	4
Journal	Chemical communications
Volume	52
Issue number	41
DOIs	https://doi.org/10.1039/c6cc02130a
Publication status	Published - 1 Jan 2016

ASJC Scopus subject areas

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

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10.1039/c6cc02130a

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@article{672e1dd516324f6db3300245e6dbc87a,

title = "Identification of genes encoding squalestatin S1 biosynthesis and: In vitro production of new squalestatin analogues",

abstract = "A gene cluster responsible for the biosynthesis of squalestatin S1 (SQS1, 1) was identified by full genome sequencing of two SQS1-producing ascomycetes: Phoma sp. C2932 and unidentified fungus MF5453. A transformation protocol was established and a subsequent knockout of one PKS gene from the cluster led to loss of SQS1 production and enhanced concentration of an SQS1 precursor. An acyltransferase gene from the cluster was expressed in E. coli and the expressed protein MfM4 shown to be responsible for loading acyl groups from CoA onto the squalestatin core as the final step of biosynthesis. MfM4 appears to have a broad substrate selectivity for its acyl CoA substrate, allowing the in vitro synthesis of novel squalestatins.",

author = "B. Bonsch and V. Belt and C. Bartel and N. Duensing and M. Koziol and Lazarus, \{C. M.\} and Bailey, \{A. M.\} and Simpson, \{T. J.\} and Cox, \{R. J.\}",

note = "Funding information: We thank EPSRC (EP/F066104/1), Leibniz Universitat Hannover and DFG (INST 187/621) for funding LCMS instruments. CB Thanks the MINAS programme of Lower Saxony for funding. We thank the University of Bristol Genomics facility for Illumina genome sequencing.",

year = "2016",

month = jan,

day = "1",

doi = "10.1039/c6cc02130a",

language = "English",

volume = "52",

pages = "6777--6780",

journal = "Chemical communications",

issn = "1359-7345",

publisher = "Chemical Society",

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TY - JOUR

T1 - Identification of genes encoding squalestatin S1 biosynthesis and

T2 - In vitro production of new squalestatin analogues

AU - Bonsch, B.

AU - Belt, V.

AU - Bartel, C.

AU - Duensing, N.

AU - Koziol, M.

AU - Lazarus, C. M.

AU - Bailey, A. M.

AU - Simpson, T. J.

AU - Cox, R. J.

N1 - Funding information: We thank EPSRC (EP/F066104/1), Leibniz Universitat Hannover and DFG (INST 187/621) for funding LCMS instruments. CB Thanks the MINAS programme of Lower Saxony for funding. We thank the University of Bristol Genomics facility for Illumina genome sequencing.

PY - 2016/1/1

Y1 - 2016/1/1

N2 - A gene cluster responsible for the biosynthesis of squalestatin S1 (SQS1, 1) was identified by full genome sequencing of two SQS1-producing ascomycetes: Phoma sp. C2932 and unidentified fungus MF5453. A transformation protocol was established and a subsequent knockout of one PKS gene from the cluster led to loss of SQS1 production and enhanced concentration of an SQS1 precursor. An acyltransferase gene from the cluster was expressed in E. coli and the expressed protein MfM4 shown to be responsible for loading acyl groups from CoA onto the squalestatin core as the final step of biosynthesis. MfM4 appears to have a broad substrate selectivity for its acyl CoA substrate, allowing the in vitro synthesis of novel squalestatins.

AB - A gene cluster responsible for the biosynthesis of squalestatin S1 (SQS1, 1) was identified by full genome sequencing of two SQS1-producing ascomycetes: Phoma sp. C2932 and unidentified fungus MF5453. A transformation protocol was established and a subsequent knockout of one PKS gene from the cluster led to loss of SQS1 production and enhanced concentration of an SQS1 precursor. An acyltransferase gene from the cluster was expressed in E. coli and the expressed protein MfM4 shown to be responsible for loading acyl groups from CoA onto the squalestatin core as the final step of biosynthesis. MfM4 appears to have a broad substrate selectivity for its acyl CoA substrate, allowing the in vitro synthesis of novel squalestatins.

UR - https://www.scopus.com/pages/publications/84970005923

U2 - 10.1039/c6cc02130a

DO - 10.1039/c6cc02130a

M3 - Article

C2 - 27056201

AN - SCOPUS:84970005923

SN - 1359-7345

VL - 52

SP - 6777

EP - 6780

JO - Chemical communications

JF - Chemical communications

IS - 41

ER -

Identification of genes encoding squalestatin S1 biosynthesis and: In vitro production of new squalestatin analogues

Abstract

ASJC Scopus subject areas

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