Using the pimeloyl-CoA synthetase adenylation fold to synthesize fatty acid thioesters

Menglu Wang; Lucile Moynié; Peter J. Harrison; Van Kelly; Andrew Piper; James H. Naismith; Dominic J. Campopiano

doi:10.1038/nchembio.2361

Using the pimeloyl-CoA synthetase adenylation fold to synthesize fatty acid thioesters

Menglu Wang, Lucile Moynié, Peter J. Harrison, Van Kelly, Andrew Piper, James H. Naismith, Dominic J. Campopiano

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Abstract

Biotin is an essential vitamin in plants and mammals, functioning as the carbon dioxide carrier within central lipid metabolism. Bacterial pimeloyl-CoA synthetase (BioW) acts as a highly specific substrate-selection gate, ensuring the integrity of the carbon chain in biotin synthesis. BioW catalyzes the condensation of pimelic acid (C7 dicarboxylic acid) with CoASH in an ATP-dependent manner to form pimeloyl-CoA, the first dedicated biotin building block. Multiple structures of Bacillus subtilis BioW together capture all three substrates, as well as the intermediate pimeloyl-adenylate and product pyrophosphate (PP_i), indicating that the enzyme uses an internal ruler to select the correct dicarboxylic acid substrate. Both the catalytic mechanism and the surprising stability of the adenylate intermediate were rationalized through site-directed mutagenesis. Building on this understanding, BioW was engineered to synthesize high-value heptanoyl (C7) and octanoyl (C8) monocarboxylic acid-CoA and C8 dicarboxylic-CoA products, highlighting the enzyme's synthetic potential.

Original language	English
Pages (from-to)	660-667
Number of pages	10
Journal	Nature Chemical Biology
Volume	13
Issue number	6
Early online date	17 Apr 2017
DOIs	https://doi.org/10.1038/nchembio.2361
Publication status	Published - Jun 2017

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10.1038/nchembio.2361

Wang_2017_NCB_Pimeloyl-CoA_AAM
© 2017 the Authors. This work has been made available online in accordance with the publisher’s policies. This is the author created, accepted version manuscript following peer review and may differ slightly from the final published version. The final published version of this work is available at https://doi.org/10.1038/nchembio.2361
Accepted author manuscript, 3.43 MB

The Human SPT Complex: The Human Serine Palmitoyltransferase (SPT) Complex; Specificity, Structure, Regulation and Inhibition
Naismith, J. (PI)
BBSRC
31/12/14 → 29/01/18
Project: Standard
Exploring the mechanism: Exploring the mechanism and scope of the enzymatic formation of five membered ring
Naismith, J. (PI), Botting, C. H. (CoI), Koehnke, J. A. J. G. (CoI) & Schwarz-Linek, U. (CoI)
BBSRC
1/10/13 → 30/09/17
Project: Standard
Transport and Polymerisation INV Award: Transport and polymerisation of bacterial polysaccharides
Naismith, J. (PI)
The Wellcome Trust
1/07/13 → 30/06/20
Project: Standard

Cite this

@article{7861ea2838f3448393c875e27ba239a9,

title = "Using the pimeloyl-CoA synthetase adenylation fold to synthesize fatty acid thioesters",

abstract = "Biotin is an essential vitamin in plants and mammals, functioning as the carbon dioxide carrier within central lipid metabolism. Bacterial pimeloyl-CoA synthetase (BioW) acts as a highly specific substrate-selection gate, ensuring the integrity of the carbon chain in biotin synthesis. BioW catalyzes the condensation of pimelic acid (C7 dicarboxylic acid) with CoASH in an ATP-dependent manner to form pimeloyl-CoA, the first dedicated biotin building block. Multiple structures of Bacillus subtilis BioW together capture all three substrates, as well as the intermediate pimeloyl-adenylate and product pyrophosphate (PPi), indicating that the enzyme uses an internal ruler to select the correct dicarboxylic acid substrate. Both the catalytic mechanism and the surprising stability of the adenylate intermediate were rationalized through site-directed mutagenesis. Building on this understanding, BioW was engineered to synthesize high-value heptanoyl (C7) and octanoyl (C8) monocarboxylic acid-CoA and C8 dicarboxylic-CoA products, highlighting the enzyme's synthetic potential.",

author = "Menglu Wang and Lucile Moyni{\'e} and Harrison, {Peter J.} and Van Kelly and Andrew Piper and Naismith, {James H.} and Campopiano, {Dominic J.}",

note = "This work was supported by grants from the Biotechnology and Biological Sciences Research Council (BBSRC, BB/M003493/1) (DJC & JHN) and Wellcome Trust (WT100209MA (J.H.N.). The native mass spectrometry data was acquired on an instrument funded by an Engineering and Physical Sciences Research Council (EPSRC) grant to the University of Edinburgh (EP/K039717/1). J.H.N. is a Royal Society Wolfson Merit Award Holder and 1000 talent scholar at Sichuan University. M.W. was funded by a University of Edinburgh PhD scholarship.",

year = "2017",

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doi = "10.1038/nchembio.2361",

language = "English",

volume = "13",

pages = "660--667",

journal = "Nature Chemical Biology",

issn = "1552-4450",

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AU - Wang, Menglu

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AU - Harrison, Peter J.

AU - Kelly, Van

AU - Piper, Andrew

AU - Naismith, James H.

AU - Campopiano, Dominic J.

N1 - This work was supported by grants from the Biotechnology and Biological Sciences Research Council (BBSRC, BB/M003493/1) (DJC & JHN) and Wellcome Trust (WT100209MA (J.H.N.). The native mass spectrometry data was acquired on an instrument funded by an Engineering and Physical Sciences Research Council (EPSRC) grant to the University of Edinburgh (EP/K039717/1). J.H.N. is a Royal Society Wolfson Merit Award Holder and 1000 talent scholar at Sichuan University. M.W. was funded by a University of Edinburgh PhD scholarship.

PY - 2017/6

Y1 - 2017/6

N2 - Biotin is an essential vitamin in plants and mammals, functioning as the carbon dioxide carrier within central lipid metabolism. Bacterial pimeloyl-CoA synthetase (BioW) acts as a highly specific substrate-selection gate, ensuring the integrity of the carbon chain in biotin synthesis. BioW catalyzes the condensation of pimelic acid (C7 dicarboxylic acid) with CoASH in an ATP-dependent manner to form pimeloyl-CoA, the first dedicated biotin building block. Multiple structures of Bacillus subtilis BioW together capture all three substrates, as well as the intermediate pimeloyl-adenylate and product pyrophosphate (PPi), indicating that the enzyme uses an internal ruler to select the correct dicarboxylic acid substrate. Both the catalytic mechanism and the surprising stability of the adenylate intermediate were rationalized through site-directed mutagenesis. Building on this understanding, BioW was engineered to synthesize high-value heptanoyl (C7) and octanoyl (C8) monocarboxylic acid-CoA and C8 dicarboxylic-CoA products, highlighting the enzyme's synthetic potential.

AB - Biotin is an essential vitamin in plants and mammals, functioning as the carbon dioxide carrier within central lipid metabolism. Bacterial pimeloyl-CoA synthetase (BioW) acts as a highly specific substrate-selection gate, ensuring the integrity of the carbon chain in biotin synthesis. BioW catalyzes the condensation of pimelic acid (C7 dicarboxylic acid) with CoASH in an ATP-dependent manner to form pimeloyl-CoA, the first dedicated biotin building block. Multiple structures of Bacillus subtilis BioW together capture all three substrates, as well as the intermediate pimeloyl-adenylate and product pyrophosphate (PPi), indicating that the enzyme uses an internal ruler to select the correct dicarboxylic acid substrate. Both the catalytic mechanism and the surprising stability of the adenylate intermediate were rationalized through site-directed mutagenesis. Building on this understanding, BioW was engineered to synthesize high-value heptanoyl (C7) and octanoyl (C8) monocarboxylic acid-CoA and C8 dicarboxylic-CoA products, highlighting the enzyme's synthetic potential.

U2 - 10.1038/nchembio.2361

DO - 10.1038/nchembio.2361

M3 - Article

SN - 1552-4450

VL - 13

SP - 660

EP - 667

JO - Nature Chemical Biology

JF - Nature Chemical Biology

IS - 6

ER -

Using the pimeloyl-CoA synthetase adenylation fold to synthesize fatty acid thioesters

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Projects

The Human SPT Complex: The Human Serine Palmitoyltransferase (SPT) Complex; Specificity, Structure, Regulation and Inhibition

Exploring the mechanism: Exploring the mechanism and scope of the enzymatic formation of five membered ring

Transport and Polymerisation INV Award: Transport and polymerisation of bacterial polysaccharides

Cite this