chapter 5 Iterative Type I Polyketide Synthases for Enediyne Core Biosynthesis

Geoffrey P. Horsman; Steven G. Van Lanen; Ben Shen

doi:10.1016/S0076-6879(09)04605-9

chapter 5 Iterative Type I Polyketide Synthases for Enediyne Core Biosynthesis

Geoffrey P. Horsman, Steven G. Van Lanen, Ben Shen

Research output: Chapter in Book/Report/Conference proceeding › Chapter › peer-review

19 Scopus citations

Abstract

Enediyne natural products are extremely potent antitumor antibiotics with a remarkable core structure consisting of two acetylenic groups conjugated to a double bond within either a 9- or 10-membered ring. Biosynthesis of this fascinating scaffold is catalyzed in part by an unusual iterative type I polyketide synthase, PKSE, that is shared among all enediyne biosynthetic pathways whose gene clusters have been sequenced to date. The PKSE is unusual in two main respects: (1) it contains an acyl carrier protein (ACP) domain with no sequence homology to any known proteins, and (2) it is self-phosphopantetheinylated by an integrated phosphopantetheinyl transferase (PPTase) domain. The unusual domain architecture and biochemistry of the PKSE hold promise both for the rapid identification of new enediyne natural products and for obtaining fundamental catalytic insights into enediyne biosynthesis. This chapter describes methods for rapid PCR-based classification of conserved enediyne biosynthetic genes, heterologous production of 9-membered PKSE proteins and isolation of the resulting polyene product, and in vitro characterization of the PKSE ACP domain.

Original language	English
Title of host publication	Complex Enzymes in Microbial Natural Product Biosynthesis, Part B
Subtitle of host publication	Polyketides, Aminocoumarins and Carbohydrates
Editors	David Hopwood
Pages	97-112
Number of pages	16
Edition	B
DOIs	https://doi.org/10.1016/S0076-6879(09)04605-9
State	Published - 2009

Publication series

Name	Methods in Enzymology
Number	B
Volume	459
ISSN (Print)	0076-6879

Bibliographical note

Funding Information:
This work was supported in part by National Institute of Health (NIH) grants CA78747 and CA113297. G.P.H. is the recipient of an Natural Sciences and Engineering Research Council (Canada) (NSERC) postdoctoral fellowship, and S.V.L. is the recipient of an NIH postdoctoral fellowship (CA1059845).

ASJC Scopus subject areas

Biochemistry
Molecular Biology

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10.1016/S0076-6879(09)04605-9

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Horsman, G. P., Van Lanen, S. G., & Shen, B. (2009). chapter 5 Iterative Type I Polyketide Synthases for Enediyne Core Biosynthesis. In D. Hopwood (Ed.), Complex Enzymes in Microbial Natural Product Biosynthesis, Part B: Polyketides, Aminocoumarins and Carbohydrates (B ed., pp. 97-112). (Methods in Enzymology; Vol. 459, No. B). https://doi.org/10.1016/S0076-6879(09)04605-9

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abstract = "Enediyne natural products are extremely potent antitumor antibiotics with a remarkable core structure consisting of two acetylenic groups conjugated to a double bond within either a 9- or 10-membered ring. Biosynthesis of this fascinating scaffold is catalyzed in part by an unusual iterative type I polyketide synthase, PKSE, that is shared among all enediyne biosynthetic pathways whose gene clusters have been sequenced to date. The PKSE is unusual in two main respects: (1) it contains an acyl carrier protein (ACP) domain with no sequence homology to any known proteins, and (2) it is self-phosphopantetheinylated by an integrated phosphopantetheinyl transferase (PPTase) domain. The unusual domain architecture and biochemistry of the PKSE hold promise both for the rapid identification of new enediyne natural products and for obtaining fundamental catalytic insights into enediyne biosynthesis. This chapter describes methods for rapid PCR-based classification of conserved enediyne biosynthetic genes, heterologous production of 9-membered PKSE proteins and isolation of the resulting polyene product, and in vitro characterization of the PKSE ACP domain.",

author = "Horsman, {Geoffrey P.} and {Van Lanen}, {Steven G.} and Ben Shen",

note = "Funding Information: This work was supported in part by National Institute of Health (NIH) grants CA78747 and CA113297. G.P.H. is the recipient of an Natural Sciences and Engineering Research Council (Canada) (NSERC) postdoctoral fellowship, and S.V.L. is the recipient of an NIH postdoctoral fellowship (CA1059845).",

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chapter 5 Iterative Type I Polyketide Synthases for Enediyne Core Biosynthesis. / Horsman, Geoffrey P.; Van Lanen, Steven G.; Shen, Ben.
Complex Enzymes in Microbial Natural Product Biosynthesis, Part B: Polyketides, Aminocoumarins and Carbohydrates. ed. / David Hopwood. B. ed. 2009. p. 97-112 (Methods in Enzymology; Vol. 459, No. B).

Research output: Chapter in Book/Report/Conference proceeding › Chapter › peer-review

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chapter 5 Iterative Type I Polyketide Synthases for Enediyne Core Biosynthesis

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