Expanding the promiscuity of a natural-product glycosyltransferase by directed evolution

Gavin J. Williams; Changsheng Zhang; Jon S. Thorson

doi:10.1038/nchembio.2007.28

Expanding the promiscuity of a natural-product glycosyltransferase by directed evolution

Gavin J. Williams, Changsheng Zhang, Jon S. Thorson

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

231 Scopus citations

Abstract

Natural products, many of which are decorated with essential sugar residues, continue to serve as a key platform for drug development. Adding or changing sugars attached to such natural products can improve the parent compound's pharmacological properties, specificity at multiple levels, and/or even the molecular mechanism of action. Though some natural-product glycosyltransferases (GTs) are sufficiently promiscuous for use in altering these glycosylation patterns, the stringent specificity of others remains a limiting factor in natural-product diversification and highlights a need for general GT engineering and evolution platforms. Herein we report the use of a simple high-throughput screen based on a fluorescent surrogate acceptor substrate to expand the promiscuity of a natural-product GT via directed evolution. Cumulatively, this study presents variant GTs for the glycorandomization of a range of therapeutically important acceptors, including aminocoumarins, flavonoids and macrolides, and a potential template for engineering other natural-product GTs.

Original language	English
Pages (from-to)	657-662
Number of pages	6
Journal	Nature Chemical Biology
Volume	3
Issue number	10
DOIs	https://doi.org/10.1038/nchembio.2007.28
State	Published - Oct 20 2007

Bibliographical note

Funding Information:
We are grateful to the School of Pharmacy Analytical Instrumentation Center for analytical support, H.-W. Liu (University of Texas-Austin) for plasmid pET28/ OleD and S. Singh for helpful discussions. This work was supported in part by the US National Institutes of Health grants AI52218 and U19 CA113297. J.S.T. is a University of Wisconsin H.I. Romnes Fellow.

Funding

We are grateful to the School of Pharmacy Analytical Instrumentation Center for analytical support, H.-W. Liu (University of Texas-Austin) for plasmid pET28/ OleD and S. Singh for helpful discussions. This work was supported in part by the US National Institutes of Health grants AI52218 and U19 CA113297. J.S.T. is a University of Wisconsin H.I. Romnes Fellow.

Funders	Funder number
National Institutes of Health (NIH)	U19 CA113297
National Institute of Allergy and Infectious Diseases	R01AI052218

ASJC Scopus subject areas

Molecular Biology
Cell Biology

Access to Document

10.1038/nchembio.2007.28

Cite this

@article{be98b7dd5448427f8354e4638427d733,

title = "Expanding the promiscuity of a natural-product glycosyltransferase by directed evolution",

abstract = "Natural products, many of which are decorated with essential sugar residues, continue to serve as a key platform for drug development. Adding or changing sugars attached to such natural products can improve the parent compound's pharmacological properties, specificity at multiple levels, and/or even the molecular mechanism of action. Though some natural-product glycosyltransferases (GTs) are sufficiently promiscuous for use in altering these glycosylation patterns, the stringent specificity of others remains a limiting factor in natural-product diversification and highlights a need for general GT engineering and evolution platforms. Herein we report the use of a simple high-throughput screen based on a fluorescent surrogate acceptor substrate to expand the promiscuity of a natural-product GT via directed evolution. Cumulatively, this study presents variant GTs for the glycorandomization of a range of therapeutically important acceptors, including aminocoumarins, flavonoids and macrolides, and a potential template for engineering other natural-product GTs.",

author = "Williams, {Gavin J.} and Changsheng Zhang and Thorson, {Jon S.}",

note = "Funding Information: We are grateful to the School of Pharmacy Analytical Instrumentation Center for analytical support, H.-W. Liu (University of Texas-Austin) for plasmid pET28/ OleD and S. Singh for helpful discussions. This work was supported in part by the US National Institutes of Health grants AI52218 and U19 CA113297. J.S.T. is a University of Wisconsin H.I. Romnes Fellow.",

year = "2007",

month = oct,

day = "20",

doi = "10.1038/nchembio.2007.28",

language = "English",

volume = "3",

pages = "657--662",

number = "10",

}

TY - JOUR

T1 - Expanding the promiscuity of a natural-product glycosyltransferase by directed evolution

AU - Williams, Gavin J.

AU - Zhang, Changsheng

AU - Thorson, Jon S.

N1 - Funding Information: We are grateful to the School of Pharmacy Analytical Instrumentation Center for analytical support, H.-W. Liu (University of Texas-Austin) for plasmid pET28/ OleD and S. Singh for helpful discussions. This work was supported in part by the US National Institutes of Health grants AI52218 and U19 CA113297. J.S.T. is a University of Wisconsin H.I. Romnes Fellow.

PY - 2007/10/20

Y1 - 2007/10/20

N2 - Natural products, many of which are decorated with essential sugar residues, continue to serve as a key platform for drug development. Adding or changing sugars attached to such natural products can improve the parent compound's pharmacological properties, specificity at multiple levels, and/or even the molecular mechanism of action. Though some natural-product glycosyltransferases (GTs) are sufficiently promiscuous for use in altering these glycosylation patterns, the stringent specificity of others remains a limiting factor in natural-product diversification and highlights a need for general GT engineering and evolution platforms. Herein we report the use of a simple high-throughput screen based on a fluorescent surrogate acceptor substrate to expand the promiscuity of a natural-product GT via directed evolution. Cumulatively, this study presents variant GTs for the glycorandomization of a range of therapeutically important acceptors, including aminocoumarins, flavonoids and macrolides, and a potential template for engineering other natural-product GTs.

AB - Natural products, many of which are decorated with essential sugar residues, continue to serve as a key platform for drug development. Adding or changing sugars attached to such natural products can improve the parent compound's pharmacological properties, specificity at multiple levels, and/or even the molecular mechanism of action. Though some natural-product glycosyltransferases (GTs) are sufficiently promiscuous for use in altering these glycosylation patterns, the stringent specificity of others remains a limiting factor in natural-product diversification and highlights a need for general GT engineering and evolution platforms. Herein we report the use of a simple high-throughput screen based on a fluorescent surrogate acceptor substrate to expand the promiscuity of a natural-product GT via directed evolution. Cumulatively, this study presents variant GTs for the glycorandomization of a range of therapeutically important acceptors, including aminocoumarins, flavonoids and macrolides, and a potential template for engineering other natural-product GTs.

UR - http://www.scopus.com/inward/record.url?scp=34548685673&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=34548685673&partnerID=8YFLogxK

U2 - 10.1038/nchembio.2007.28

DO - 10.1038/nchembio.2007.28

M3 - Article

C2 - 17828251

AN - SCOPUS:34548685673

SN - 1552-4450

VL - 3

SP - 657

EP - 662

JO - Nature Chemical Biology

JF - Nature Chemical Biology

IS - 10

ER -

Expanding the promiscuity of a natural-product glycosyltransferase by directed evolution

Abstract

Bibliographical note

Funding

ASJC Scopus subject areas

Access to Document

Other files and links

Fingerprint

Cite this