Biosynthesis of the pyrrolidine protein synthesis inhibitor anisomycin involves novel gene ensemble and cryptic biosynthetic steps

Xiaoqing Zheng, Qiuxiang Cheng, Fen Yao, Xiaozheng Wang, Lingxin Kong, Bo Cao, Min Xu, Shuangjun Lin, Zixin Deng, Yit Heng Chooi, Delin You

Research output: Contribution to journalArticle

6 Citations (Scopus)

Abstract

The protein synthesis inhibitor anisomycin features a unique benzylpyrrolidine system and exhibits diverse biological and pharmacologic activities. Its biosynthetic origin has remained obscure for more than 60 y, however. Here we report the identification of the biosynthetic gene cluster (BGC) of anisomycin in Streptomyces hygrospinosus var. beijingensis by a bioactivity-guided high-throughput screening method. Using a combination of bioinformatic analysis, reverse genetics, chemical analysis, and in vitro biochemical assays, we have identified a core four-gene ensemble responsible for the synthesis of the pyrrolidine system in anisomycin: aniQ, encoding a aminotransferase that catalyzes an initial deamination and a later reamination steps; aniP, encoding a transketolase implicated to bring together an glycolysis intermediate with 4-hydroxyphenylpyruvic acid to form the anisomycin molecular backbone; aniO, encoding a glycosyltransferase that catalyzes a cryptic glycosylation crucial for downstream enzyme processing; and aniN, encoding a bifunctional dehydrogenase that mediates multistep pyrrolidine formation. The results reveal a BGC for pyrrolidine alkaloid biosynthesis that is distinct from known bacterial alkaloid pathways, and provide the signature sequences that will facilitate the discovery of BGCs encoding novel pyrrolidine alkaloids in bacterial genomes. The biosynthetic insights from this study further set the foundation for biosynthetic engineering of pyrrolidine antibiotics.

Original languageEnglish
Pages (from-to)4135-4140
Number of pages6
JournalProceedings of the National Academy of Sciences of the United States of America
Volume114
Issue number16
DOIs
Publication statusPublished - 18 Apr 2017

Fingerprint

Anisomycin
Protein Synthesis Inhibitors
Alkaloids
Genes
Multigene Family
Transketolase
High-Throughput Screening Assays
Bacterial Genomes
Reverse Genetics
Glycosyltransferases
Deamination
Streptomyces
Glycolysis
Transaminases
Computational Biology
Glycosylation
Oxidoreductases
pyrrolidine
Anti-Bacterial Agents
Enzymes

Cite this

Zheng, Xiaoqing ; Cheng, Qiuxiang ; Yao, Fen ; Wang, Xiaozheng ; Kong, Lingxin ; Cao, Bo ; Xu, Min ; Lin, Shuangjun ; Deng, Zixin ; Chooi, Yit Heng ; You, Delin. / Biosynthesis of the pyrrolidine protein synthesis inhibitor anisomycin involves novel gene ensemble and cryptic biosynthetic steps. In: Proceedings of the National Academy of Sciences of the United States of America. 2017 ; Vol. 114, No. 16. pp. 4135-4140.
@article{70a9e0e4c729446685bd76af026f50d0,
title = "Biosynthesis of the pyrrolidine protein synthesis inhibitor anisomycin involves novel gene ensemble and cryptic biosynthetic steps",
abstract = "The protein synthesis inhibitor anisomycin features a unique benzylpyrrolidine system and exhibits diverse biological and pharmacologic activities. Its biosynthetic origin has remained obscure for more than 60 y, however. Here we report the identification of the biosynthetic gene cluster (BGC) of anisomycin in Streptomyces hygrospinosus var. beijingensis by a bioactivity-guided high-throughput screening method. Using a combination of bioinformatic analysis, reverse genetics, chemical analysis, and in vitro biochemical assays, we have identified a core four-gene ensemble responsible for the synthesis of the pyrrolidine system in anisomycin: aniQ, encoding a aminotransferase that catalyzes an initial deamination and a later reamination steps; aniP, encoding a transketolase implicated to bring together an glycolysis intermediate with 4-hydroxyphenylpyruvic acid to form the anisomycin molecular backbone; aniO, encoding a glycosyltransferase that catalyzes a cryptic glycosylation crucial for downstream enzyme processing; and aniN, encoding a bifunctional dehydrogenase that mediates multistep pyrrolidine formation. The results reveal a BGC for pyrrolidine alkaloid biosynthesis that is distinct from known bacterial alkaloid pathways, and provide the signature sequences that will facilitate the discovery of BGCs encoding novel pyrrolidine alkaloids in bacterial genomes. The biosynthetic insights from this study further set the foundation for biosynthetic engineering of pyrrolidine antibiotics.",
keywords = "Anisomycin biosynthetic pathway, Cryptic glycosylation, Protein synthesis inhibitor, Pyrrolidine antibiotics, Streptomyces",
author = "Xiaoqing Zheng and Qiuxiang Cheng and Fen Yao and Xiaozheng Wang and Lingxin Kong and Bo Cao and Min Xu and Shuangjun Lin and Zixin Deng and Chooi, {Yit Heng} and Delin You",
year = "2017",
month = "4",
day = "18",
doi = "10.1073/pnas.1701361114",
language = "English",
volume = "114",
pages = "4135--4140",
journal = "National Academy of Sciences, Proceedings",
issn = "0027-8424",
publisher = "NATL ACAD SCIENCES",
number = "16",

}

Biosynthesis of the pyrrolidine protein synthesis inhibitor anisomycin involves novel gene ensemble and cryptic biosynthetic steps. / Zheng, Xiaoqing; Cheng, Qiuxiang; Yao, Fen; Wang, Xiaozheng; Kong, Lingxin; Cao, Bo; Xu, Min; Lin, Shuangjun; Deng, Zixin; Chooi, Yit Heng; You, Delin.

In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 114, No. 16, 18.04.2017, p. 4135-4140.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Biosynthesis of the pyrrolidine protein synthesis inhibitor anisomycin involves novel gene ensemble and cryptic biosynthetic steps

AU - Zheng, Xiaoqing

AU - Cheng, Qiuxiang

AU - Yao, Fen

AU - Wang, Xiaozheng

AU - Kong, Lingxin

AU - Cao, Bo

AU - Xu, Min

AU - Lin, Shuangjun

AU - Deng, Zixin

AU - Chooi, Yit Heng

AU - You, Delin

PY - 2017/4/18

Y1 - 2017/4/18

N2 - The protein synthesis inhibitor anisomycin features a unique benzylpyrrolidine system and exhibits diverse biological and pharmacologic activities. Its biosynthetic origin has remained obscure for more than 60 y, however. Here we report the identification of the biosynthetic gene cluster (BGC) of anisomycin in Streptomyces hygrospinosus var. beijingensis by a bioactivity-guided high-throughput screening method. Using a combination of bioinformatic analysis, reverse genetics, chemical analysis, and in vitro biochemical assays, we have identified a core four-gene ensemble responsible for the synthesis of the pyrrolidine system in anisomycin: aniQ, encoding a aminotransferase that catalyzes an initial deamination and a later reamination steps; aniP, encoding a transketolase implicated to bring together an glycolysis intermediate with 4-hydroxyphenylpyruvic acid to form the anisomycin molecular backbone; aniO, encoding a glycosyltransferase that catalyzes a cryptic glycosylation crucial for downstream enzyme processing; and aniN, encoding a bifunctional dehydrogenase that mediates multistep pyrrolidine formation. The results reveal a BGC for pyrrolidine alkaloid biosynthesis that is distinct from known bacterial alkaloid pathways, and provide the signature sequences that will facilitate the discovery of BGCs encoding novel pyrrolidine alkaloids in bacterial genomes. The biosynthetic insights from this study further set the foundation for biosynthetic engineering of pyrrolidine antibiotics.

AB - The protein synthesis inhibitor anisomycin features a unique benzylpyrrolidine system and exhibits diverse biological and pharmacologic activities. Its biosynthetic origin has remained obscure for more than 60 y, however. Here we report the identification of the biosynthetic gene cluster (BGC) of anisomycin in Streptomyces hygrospinosus var. beijingensis by a bioactivity-guided high-throughput screening method. Using a combination of bioinformatic analysis, reverse genetics, chemical analysis, and in vitro biochemical assays, we have identified a core four-gene ensemble responsible for the synthesis of the pyrrolidine system in anisomycin: aniQ, encoding a aminotransferase that catalyzes an initial deamination and a later reamination steps; aniP, encoding a transketolase implicated to bring together an glycolysis intermediate with 4-hydroxyphenylpyruvic acid to form the anisomycin molecular backbone; aniO, encoding a glycosyltransferase that catalyzes a cryptic glycosylation crucial for downstream enzyme processing; and aniN, encoding a bifunctional dehydrogenase that mediates multistep pyrrolidine formation. The results reveal a BGC for pyrrolidine alkaloid biosynthesis that is distinct from known bacterial alkaloid pathways, and provide the signature sequences that will facilitate the discovery of BGCs encoding novel pyrrolidine alkaloids in bacterial genomes. The biosynthetic insights from this study further set the foundation for biosynthetic engineering of pyrrolidine antibiotics.

KW - Anisomycin biosynthetic pathway

KW - Cryptic glycosylation

KW - Protein synthesis inhibitor

KW - Pyrrolidine antibiotics

KW - Streptomyces

UR - http://www.scopus.com/inward/record.url?scp=85017608597&partnerID=8YFLogxK

U2 - 10.1073/pnas.1701361114

DO - 10.1073/pnas.1701361114

M3 - Article

VL - 114

SP - 4135

EP - 4140

JO - National Academy of Sciences, Proceedings

JF - National Academy of Sciences, Proceedings

SN - 0027-8424

IS - 16

ER -