In Vitro Biosynthetic Studies of Bottromycin Expand the Enzymatic Capabilities of the YcaO Superfamily

Christopher J. Schwalen, Graham A. Hudson, Simone Kosol, Nilkamal Mahanta, Gregory L. Challis, Douglas A. Mitchell

Research output: Contribution to journalArticleResearchpeer-review

Abstract

The bottromycins belong to the ribosomally synthesized and posttranslationally modified peptide (RiPP) family of natural products. Bottromycins exhibit unique structural features, including a hallmark macrolactamidine ring and thiazole heterocycle for which divergent members of the YcaO superfamily have been biosynthetically implicated. Here we report the in vitro reconstitution of two YcaO proteins, BmbD and BmbE, responsible for the ATP-dependent cyclodehydration reactions that yield thiazoline- and macrolactamidine-functionalized products, respectively. We also establish the substrate tolerance for BmbD and BmbE and systematically dissect the role of the follower peptide, which we show serves a purpose similar to canonical leader peptides in directing the biosynthetic enzymes to the substrate. Lastly, we leverage the expanded capabilities of YcaO proteins to conduct an extensive bioinformatic survey to classify known YcaO chemistry. This analysis predicts new functions remain to be uncovered within the superfamily.

Original languageEnglish
Pages (from-to)18154-18157
Number of pages4
JournalJournal of the American Chemical Society
Volume139
Issue number50
DOIs
Publication statusPublished - 20 Dec 2017

Cite this

Schwalen, Christopher J. ; Hudson, Graham A. ; Kosol, Simone ; Mahanta, Nilkamal ; Challis, Gregory L. ; Mitchell, Douglas A. / In Vitro Biosynthetic Studies of Bottromycin Expand the Enzymatic Capabilities of the YcaO Superfamily. In: Journal of the American Chemical Society. 2017 ; Vol. 139, No. 50. pp. 18154-18157.
@article{2f9ee41b8f894c34b46998c9044343d1,
title = "In Vitro Biosynthetic Studies of Bottromycin Expand the Enzymatic Capabilities of the YcaO Superfamily",
abstract = "The bottromycins belong to the ribosomally synthesized and posttranslationally modified peptide (RiPP) family of natural products. Bottromycins exhibit unique structural features, including a hallmark macrolactamidine ring and thiazole heterocycle for which divergent members of the YcaO superfamily have been biosynthetically implicated. Here we report the in vitro reconstitution of two YcaO proteins, BmbD and BmbE, responsible for the ATP-dependent cyclodehydration reactions that yield thiazoline- and macrolactamidine-functionalized products, respectively. We also establish the substrate tolerance for BmbD and BmbE and systematically dissect the role of the follower peptide, which we show serves a purpose similar to canonical leader peptides in directing the biosynthetic enzymes to the substrate. Lastly, we leverage the expanded capabilities of YcaO proteins to conduct an extensive bioinformatic survey to classify known YcaO chemistry. This analysis predicts new functions remain to be uncovered within the superfamily.",
author = "Schwalen, {Christopher J.} and Hudson, {Graham A.} and Simone Kosol and Nilkamal Mahanta and Challis, {Gregory L.} and Mitchell, {Douglas A.}",
year = "2017",
month = "12",
day = "20",
doi = "10.1021/jacs.7b09899",
language = "English",
volume = "139",
pages = "18154--18157",
journal = "Journal of the American Chemical Society",
issn = "0002-7863",
publisher = "ACS Publications",
number = "50",

}

In Vitro Biosynthetic Studies of Bottromycin Expand the Enzymatic Capabilities of the YcaO Superfamily. / Schwalen, Christopher J.; Hudson, Graham A.; Kosol, Simone; Mahanta, Nilkamal; Challis, Gregory L.; Mitchell, Douglas A.

In: Journal of the American Chemical Society, Vol. 139, No. 50, 20.12.2017, p. 18154-18157.

Research output: Contribution to journalArticleResearchpeer-review

TY - JOUR

T1 - In Vitro Biosynthetic Studies of Bottromycin Expand the Enzymatic Capabilities of the YcaO Superfamily

AU - Schwalen, Christopher J.

AU - Hudson, Graham A.

AU - Kosol, Simone

AU - Mahanta, Nilkamal

AU - Challis, Gregory L.

AU - Mitchell, Douglas A.

PY - 2017/12/20

Y1 - 2017/12/20

N2 - The bottromycins belong to the ribosomally synthesized and posttranslationally modified peptide (RiPP) family of natural products. Bottromycins exhibit unique structural features, including a hallmark macrolactamidine ring and thiazole heterocycle for which divergent members of the YcaO superfamily have been biosynthetically implicated. Here we report the in vitro reconstitution of two YcaO proteins, BmbD and BmbE, responsible for the ATP-dependent cyclodehydration reactions that yield thiazoline- and macrolactamidine-functionalized products, respectively. We also establish the substrate tolerance for BmbD and BmbE and systematically dissect the role of the follower peptide, which we show serves a purpose similar to canonical leader peptides in directing the biosynthetic enzymes to the substrate. Lastly, we leverage the expanded capabilities of YcaO proteins to conduct an extensive bioinformatic survey to classify known YcaO chemistry. This analysis predicts new functions remain to be uncovered within the superfamily.

AB - The bottromycins belong to the ribosomally synthesized and posttranslationally modified peptide (RiPP) family of natural products. Bottromycins exhibit unique structural features, including a hallmark macrolactamidine ring and thiazole heterocycle for which divergent members of the YcaO superfamily have been biosynthetically implicated. Here we report the in vitro reconstitution of two YcaO proteins, BmbD and BmbE, responsible for the ATP-dependent cyclodehydration reactions that yield thiazoline- and macrolactamidine-functionalized products, respectively. We also establish the substrate tolerance for BmbD and BmbE and systematically dissect the role of the follower peptide, which we show serves a purpose similar to canonical leader peptides in directing the biosynthetic enzymes to the substrate. Lastly, we leverage the expanded capabilities of YcaO proteins to conduct an extensive bioinformatic survey to classify known YcaO chemistry. This analysis predicts new functions remain to be uncovered within the superfamily.

UR - http://www.scopus.com/inward/record.url?scp=85038608835&partnerID=8YFLogxK

U2 - 10.1021/jacs.7b09899

DO - 10.1021/jacs.7b09899

M3 - Article

VL - 139

SP - 18154

EP - 18157

JO - Journal of the American Chemical Society

JF - Journal of the American Chemical Society

SN - 0002-7863

IS - 50

ER -