Biosynthesis of the tunicamycin antibiotics proceeds via unique exo-glycal intermediates

Filip J. Wyszynski; Seung Seo Lee; Tomoaki Yabe; Hua Wang; Juan Pablo Gomez-Escribano; Mervyn J. Bibb; Soo Jae Lee; Gideon J. Davies; Benjamin G. Davis

doi:10.1038/NCHEM.1351

Biosynthesis of the tunicamycin antibiotics proceeds via unique exo-glycal intermediates

Filip J. Wyszynski^*, Seung Seo Lee, Tomoaki Yabe, Hua Wang, Juan Pablo Gomez-Escribano, Mervyn J. Bibb, Soo Jae Lee, Gideon J. Davies, Benjamin G. Davis

^*Corresponding author for this work

Chemistry

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

Abstract

The tunicamycins are archetypal nucleoside antibiotics targeting bacterial peptidoglycan biosynthesis and eukaryotic protein N-glycosylation. Understanding the biosynthesis of their unusual carbon framework may lead to variants with improved selectivity. Here, we demonstrate in vitro recapitulation of key sugar-manipulating enzymes from this pathway. TunA is found to exhibit unusual regioselectivity in the reduction of a key alpha,beta-unsaturated ketone. The product of this reaction is shown to be the preferred substrate for TunF-an epimerase that converts the glucose derivative to a galactose. In Streptomyces strains in which another gene (tunB) is deleted, the biosynthesis is shown to stall at this exo-glycal product. These investigations confirm the combined TunA/F activity and delineate the ordering of events in the metabolic pathway. This is the first time these surprising exo-glycal intermediates have been seen in biology. They suggest that construction of the aminodialdose core of tunicamycin exploits their enol ether motif in a mode of C-C bond formation not previously observed in nature, to create an 11-carbon chain.

Original language	English
Pages (from-to)	539-546
Number of pages	8
Journal	Nature Chemistry
Volume	4
Issue number	7
DOIs	https://doi.org/10.1038/NCHEM.1351
Publication status	Published - Jul 2012

Keywords

MANIPULATION
STREPTOMYCES
SUBSTRATE-SPECIFICITY
DERIVATIVES
HETEROLOGOUS EXPRESSION
GENE-CLUSTER
IDENTIFICATION
GLCNAC C4 EPIMERASE
DTDP-GLUCOSE 4,6-DEHYDRATASE
MASS-SPECTROMETRY

Access to Document

10.1038/NCHEM.1351

Cite this

@article{5e2dc65247964d0caf2eb40222e309a9,

title = "Biosynthesis of the tunicamycin antibiotics proceeds via unique exo-glycal intermediates",

abstract = "The tunicamycins are archetypal nucleoside antibiotics targeting bacterial peptidoglycan biosynthesis and eukaryotic protein N-glycosylation. Understanding the biosynthesis of their unusual carbon framework may lead to variants with improved selectivity. Here, we demonstrate in vitro recapitulation of key sugar-manipulating enzymes from this pathway. TunA is found to exhibit unusual regioselectivity in the reduction of a key alpha,beta-unsaturated ketone. The product of this reaction is shown to be the preferred substrate for TunF-an epimerase that converts the glucose derivative to a galactose. In Streptomyces strains in which another gene (tunB) is deleted, the biosynthesis is shown to stall at this exo-glycal product. These investigations confirm the combined TunA/F activity and delineate the ordering of events in the metabolic pathway. This is the first time these surprising exo-glycal intermediates have been seen in biology. They suggest that construction of the aminodialdose core of tunicamycin exploits their enol ether motif in a mode of C-C bond formation not previously observed in nature, to create an 11-carbon chain.",

keywords = "MANIPULATION, STREPTOMYCES, SUBSTRATE-SPECIFICITY, DERIVATIVES, HETEROLOGOUS EXPRESSION, GENE-CLUSTER, IDENTIFICATION, GLCNAC C4 EPIMERASE, DTDP-GLUCOSE 4,6-DEHYDRATASE, MASS-SPECTROMETRY",

author = "Wyszynski, {Filip J.} and Lee, {Seung Seo} and Tomoaki Yabe and Hua Wang and Gomez-Escribano, {Juan Pablo} and Bibb, {Mervyn J.} and Lee, {Soo Jae} and Davies, {Gideon J.} and Davis, {Benjamin G.}",

year = "2012",

month = jul,

doi = "10.1038/NCHEM.1351",

language = "English",

volume = "4",

pages = "539--546",

journal = "Nature Chemistry",

issn = "1755-4330",

publisher = "Nature Publishing Group",

number = "7",

}

TY - JOUR

T1 - Biosynthesis of the tunicamycin antibiotics proceeds via unique exo-glycal intermediates

AU - Wyszynski, Filip J.

AU - Lee, Seung Seo

AU - Yabe, Tomoaki

AU - Wang, Hua

AU - Gomez-Escribano, Juan Pablo

AU - Bibb, Mervyn J.

AU - Lee, Soo Jae

AU - Davies, Gideon J.

AU - Davis, Benjamin G.

PY - 2012/7

Y1 - 2012/7

N2 - The tunicamycins are archetypal nucleoside antibiotics targeting bacterial peptidoglycan biosynthesis and eukaryotic protein N-glycosylation. Understanding the biosynthesis of their unusual carbon framework may lead to variants with improved selectivity. Here, we demonstrate in vitro recapitulation of key sugar-manipulating enzymes from this pathway. TunA is found to exhibit unusual regioselectivity in the reduction of a key alpha,beta-unsaturated ketone. The product of this reaction is shown to be the preferred substrate for TunF-an epimerase that converts the glucose derivative to a galactose. In Streptomyces strains in which another gene (tunB) is deleted, the biosynthesis is shown to stall at this exo-glycal product. These investigations confirm the combined TunA/F activity and delineate the ordering of events in the metabolic pathway. This is the first time these surprising exo-glycal intermediates have been seen in biology. They suggest that construction of the aminodialdose core of tunicamycin exploits their enol ether motif in a mode of C-C bond formation not previously observed in nature, to create an 11-carbon chain.

AB - The tunicamycins are archetypal nucleoside antibiotics targeting bacterial peptidoglycan biosynthesis and eukaryotic protein N-glycosylation. Understanding the biosynthesis of their unusual carbon framework may lead to variants with improved selectivity. Here, we demonstrate in vitro recapitulation of key sugar-manipulating enzymes from this pathway. TunA is found to exhibit unusual regioselectivity in the reduction of a key alpha,beta-unsaturated ketone. The product of this reaction is shown to be the preferred substrate for TunF-an epimerase that converts the glucose derivative to a galactose. In Streptomyces strains in which another gene (tunB) is deleted, the biosynthesis is shown to stall at this exo-glycal product. These investigations confirm the combined TunA/F activity and delineate the ordering of events in the metabolic pathway. This is the first time these surprising exo-glycal intermediates have been seen in biology. They suggest that construction of the aminodialdose core of tunicamycin exploits their enol ether motif in a mode of C-C bond formation not previously observed in nature, to create an 11-carbon chain.

KW - MANIPULATION

KW - STREPTOMYCES

KW - SUBSTRATE-SPECIFICITY

KW - DERIVATIVES

KW - HETEROLOGOUS EXPRESSION

KW - GENE-CLUSTER

KW - IDENTIFICATION

KW - GLCNAC C4 EPIMERASE

KW - DTDP-GLUCOSE 4,6-DEHYDRATASE

KW - MASS-SPECTROMETRY

UR - http://www.scopus.com/inward/record.url?scp=84862881045&partnerID=8YFLogxK

U2 - 10.1038/NCHEM.1351

DO - 10.1038/NCHEM.1351

M3 - Article

SN - 1755-4330

VL - 4

SP - 539

EP - 546

JO - Nature Chemistry

JF - Nature Chemistry

IS - 7

ER -

Biosynthesis of the tunicamycin antibiotics proceeds via unique exo-glycal intermediates

Abstract

Keywords

Access to Document

Other files and links

Cite this