Biosynthesis of Cytosporones in Leotiomycetous Filamentous Fungi

Li Li; Weimao Zhong; Hang Liu; Patricia Espinosa-Artiles; Ya Ming Xu; Chen Wang; Jose Manuel Verdugo Robles; Tiago Antunes Paz; Marielle Cascaes Inácio; Fusheng Chen; Yuquan Xu; A. A.Leslie Gunatilaka; István Molnár

doi:10.1021/jacs.3c14066

Biosynthesis of Cytosporones in Leotiomycetous Filamentous Fungi

Li Li, Weimao Zhong, Hang Liu, Patricia Espinosa-Artiles, Ya Ming Xu, Chen Wang, Jose Manuel Verdugo Robles, Tiago Antunes Paz, Marielle Cascaes Inácio, Fusheng Chen, Yuquan Xu^*, A. A.Leslie Gunatilaka^*, István Molnár^*

^*Corresponding author for this work

BA53 Industrial biotechnology and food

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

5 Citations (Scopus)

Abstract

Polyketides with the isochroman-3-one pharmacophore are rare among fungal natural products as their biosynthesis requires an unorthodox S-type aromatic ring cyclization. Genome mining uncovered a conserved gene cluster in select leotiomycetous fungi that encodes the biosynthesis of cytosporones, including isochroman-3-one congeners. Combinatorial biosynthesis in total biosynthetic and biocatalytic formats in Saccharomyces cerevisiae and in vitro reconstitution of key reactions with purified enzymes revealed how cytosporone structural and bioactivity diversity is generated. The S-type acyl dihydroxyphenylacetic acid (ADA) core of cytosporones is assembled by a collaborating polyketide synthase pair. Thioesterase domain-catalyzed transesterification releases ADA esters, some of which are known Nur77 modulators. Alternatively, hydrolytic release allows C6 hydroxylation by a flavin-dependent monooxygenase, yielding a trihydroxybenzene moiety. Reduction of the C9 carbonyl by a short chain dehydrogenase/reductase initiates isochroman-3-one formation, affording cytosporones with cytotoxic and antimicrobial activity. Enoyl di- or trihydroxyphenylacetic acids are generated as shunt products, while isocroman-3,4-diones are formed by autoxidation. The cytosporone pathway offers novel polyketide biosynthetic enzymes for combinatorial synthetic biology to advance the production of “unnatural” natural products for drug discovery.

Original language	English
Pages (from-to)	6189-6198
Number of pages	10
Journal	Journal of the American Chemical Society
Volume	146
Issue number	9
DOIs	https://doi.org/10.1021/jacs.3c14066
Publication status	Published - 6 Mar 2024
MoE publication type	A1 Journal article-refereed

Funding

This research was supported by the National Natural Science Foundation of China (No. 31501453 to L.L.); the Science and Technology Department of Hubei Province, China (No. 2019AHB067 to L.L.); the Center for Scientific Computing (NCC/GridUNESP) of the São Paulo State University (to T.A.P.); the National Natural Science Foundation of China (32070053 and 22277137 to Y.X.), the Joint Genomics Institute of the U.S. Department of Energy (WIP ID 1349 to I.M.); the U.S. National Institutes of Health (NIGMS 5R01GM114418 to I.M.); and the VTT Technical Research Center of Finland (to I.M.).

Keywords

Fungi/genetics
Saccharomyces cerevisiae/metabolism
Polyketide Synthases/metabolism
Polyketides/chemistry
Biological Products/metabolism

Access to Document

10.1021/jacs.3c14066

Cite this

@article{1040583a4f484edb92c720d813fe0e21,

title = "Biosynthesis of Cytosporones in Leotiomycetous Filamentous Fungi",

abstract = "Polyketides with the isochroman-3-one pharmacophore are rare among fungal natural products as their biosynthesis requires an unorthodox S-type aromatic ring cyclization. Genome mining uncovered a conserved gene cluster in select leotiomycetous fungi that encodes the biosynthesis of cytosporones, including isochroman-3-one congeners. Combinatorial biosynthesis in total biosynthetic and biocatalytic formats in Saccharomyces cerevisiae and in vitro reconstitution of key reactions with purified enzymes revealed how cytosporone structural and bioactivity diversity is generated. The S-type acyl dihydroxyphenylacetic acid (ADA) core of cytosporones is assembled by a collaborating polyketide synthase pair. Thioesterase domain-catalyzed transesterification releases ADA esters, some of which are known Nur77 modulators. Alternatively, hydrolytic release allows C6 hydroxylation by a flavin-dependent monooxygenase, yielding a trihydroxybenzene moiety. Reduction of the C9 carbonyl by a short chain dehydrogenase/reductase initiates isochroman-3-one formation, affording cytosporones with cytotoxic and antimicrobial activity. Enoyl di- or trihydroxyphenylacetic acids are generated as shunt products, while isocroman-3,4-diones are formed by autoxidation. The cytosporone pathway offers novel polyketide biosynthetic enzymes for combinatorial synthetic biology to advance the production of “unnatural” natural products for drug discovery.",

keywords = "Fungi/genetics, Saccharomyces cerevisiae/metabolism, Polyketide Synthases/metabolism, Polyketides/chemistry, Biological Products/metabolism",

author = "Li Li and Weimao Zhong and Hang Liu and Patricia Espinosa-Artiles and Xu, \{Ya Ming\} and Chen Wang and \{Verdugo Robles\}, \{Jose Manuel\} and Paz, \{Tiago Antunes\} and \{Cascaes In{\'a}cio\}, Marielle and Fusheng Chen and Yuquan Xu and Gunatilaka, \{A. A.Leslie\} and Istv{\'a}n Moln{\'a}r",

note = "Publisher Copyright: {\textcopyright} 2024 American Chemical Society.",

year = "2024",

month = mar,

day = "6",

doi = "10.1021/jacs.3c14066",

language = "English",

volume = "146",

pages = "6189--6198",

journal = "Journal of the American Chemical Society",

issn = "0002-7863",

publisher = "American Chemical Society ACS",

number = "9",

}

TY - JOUR

T1 - Biosynthesis of Cytosporones in Leotiomycetous Filamentous Fungi

AU - Li, Li

AU - Zhong, Weimao

AU - Liu, Hang

AU - Espinosa-Artiles, Patricia

AU - Xu, Ya Ming

AU - Wang, Chen

AU - Verdugo Robles, Jose Manuel

AU - Paz, Tiago Antunes

AU - Cascaes Inácio, Marielle

AU - Chen, Fusheng

AU - Xu, Yuquan

AU - Gunatilaka, A. A.Leslie

AU - Molnár, István

PY - 2024/3/6

Y1 - 2024/3/6

N2 - Polyketides with the isochroman-3-one pharmacophore are rare among fungal natural products as their biosynthesis requires an unorthodox S-type aromatic ring cyclization. Genome mining uncovered a conserved gene cluster in select leotiomycetous fungi that encodes the biosynthesis of cytosporones, including isochroman-3-one congeners. Combinatorial biosynthesis in total biosynthetic and biocatalytic formats in Saccharomyces cerevisiae and in vitro reconstitution of key reactions with purified enzymes revealed how cytosporone structural and bioactivity diversity is generated. The S-type acyl dihydroxyphenylacetic acid (ADA) core of cytosporones is assembled by a collaborating polyketide synthase pair. Thioesterase domain-catalyzed transesterification releases ADA esters, some of which are known Nur77 modulators. Alternatively, hydrolytic release allows C6 hydroxylation by a flavin-dependent monooxygenase, yielding a trihydroxybenzene moiety. Reduction of the C9 carbonyl by a short chain dehydrogenase/reductase initiates isochroman-3-one formation, affording cytosporones with cytotoxic and antimicrobial activity. Enoyl di- or trihydroxyphenylacetic acids are generated as shunt products, while isocroman-3,4-diones are formed by autoxidation. The cytosporone pathway offers novel polyketide biosynthetic enzymes for combinatorial synthetic biology to advance the production of “unnatural” natural products for drug discovery.

AB - Polyketides with the isochroman-3-one pharmacophore are rare among fungal natural products as their biosynthesis requires an unorthodox S-type aromatic ring cyclization. Genome mining uncovered a conserved gene cluster in select leotiomycetous fungi that encodes the biosynthesis of cytosporones, including isochroman-3-one congeners. Combinatorial biosynthesis in total biosynthetic and biocatalytic formats in Saccharomyces cerevisiae and in vitro reconstitution of key reactions with purified enzymes revealed how cytosporone structural and bioactivity diversity is generated. The S-type acyl dihydroxyphenylacetic acid (ADA) core of cytosporones is assembled by a collaborating polyketide synthase pair. Thioesterase domain-catalyzed transesterification releases ADA esters, some of which are known Nur77 modulators. Alternatively, hydrolytic release allows C6 hydroxylation by a flavin-dependent monooxygenase, yielding a trihydroxybenzene moiety. Reduction of the C9 carbonyl by a short chain dehydrogenase/reductase initiates isochroman-3-one formation, affording cytosporones with cytotoxic and antimicrobial activity. Enoyl di- or trihydroxyphenylacetic acids are generated as shunt products, while isocroman-3,4-diones are formed by autoxidation. The cytosporone pathway offers novel polyketide biosynthetic enzymes for combinatorial synthetic biology to advance the production of “unnatural” natural products for drug discovery.

KW - Fungi/genetics

KW - Saccharomyces cerevisiae/metabolism

KW - Polyketide Synthases/metabolism

KW - Polyketides/chemistry

KW - Biological Products/metabolism

UR - https://www.scopus.com/pages/publications/85186193482

U2 - 10.1021/jacs.3c14066

DO - 10.1021/jacs.3c14066

M3 - Article

C2 - 38386630

AN - SCOPUS:85186193482

SN - 0002-7863

VL - 146

SP - 6189

EP - 6198

JO - Journal of the American Chemical Society

JF - Journal of the American Chemical Society

IS - 9

ER -

Biosynthesis of Cytosporones in Leotiomycetous Filamentous Fungi

Abstract

Funding

Keywords

Access to Document

Other files and links

Fingerprint

Cite this