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An enzyme-trap approach allows isolation of intermediates in cobalamin biosynthesis

Deery, Evelyne, Schroeder, Susanne, Lawrence, Andrew D., Taylor, Samantha L, Seyedarabi, Arefeh, Waterman, Jitka, Wilson, Keith S, Brown, David G., Geeves, Michael A., Howard, Mark J., and others. (2012) An enzyme-trap approach allows isolation of intermediates in cobalamin biosynthesis. Nature Chemical Biology, 8 (11). pp. 933-940. ISSN 1552-4450. (doi:10.1038/nchembio.1086) (The full text of this publication is not currently available from this repository. You may be able to access a copy if URLs are provided) (KAR id:31428)

The full text of this publication is not currently available from this repository. You may be able to access a copy if URLs are provided.
Official URL:
http://dx.doi.org/10.1038/nchembio.1086

Abstract

The biosynthesis of many vitamins and coenzymes has often proven difficult to elucidate owing to a combination of low abundance and kinetic lability of the pathway intermediates. Through a serial reconstruction of the cobalamin (vitamin B 12) pathway in Escherichia coli and by His tagging the terminal enzyme in the reaction sequence, we have observed that many unstable intermediates can be isolated as tightly bound enzyme-product complexes. Together, these approaches have been used to extract intermediates between precorrin-4 and hydrogenobyrinic acid in their free acid form and permitted the delineation of the overall reaction catalyzed by CobL, including the formal elucidation of precorrin-7 as a metabolite. Furthermore, a substrate-carrier protein, CobE, that can also be used to stabilize some of the transient metabolic intermediates and enhance their onward transformation, has been identified. The tight association of pathway intermediates with enzymes provides evidence for a form of metabolite channeling.

Item Type: Article
DOI/Identification number: 10.1038/nchembio.1086
Uncontrolled keywords: article; biosynthesis; catalysis; chemical reaction; complex formation; enzyme activity; enzyme substrate; Escherichia coli; metabolite; nonhuman; priority journal; carrier protein; cobalamin; cobamamide; precorrin; unclassified drug; uroporphyrinogen
Subjects: Q Science
Divisions: Divisions > Division of Natural Sciences > Biosciences
Depositing User: Susan Davies
Date Deposited: 09 Oct 2012 10:50 UTC
Last Modified: 16 Nov 2021 10:09 UTC
Resource URI: https://kar.kent.ac.uk/id/eprint/31428 (The current URI for this page, for reference purposes)

University of Kent Author Information

Deery, Evelyne.

Creator's ORCID: https://orcid.org/0000-0002-8189-678X
CReDIT Contributor Roles:

Schroeder, Susanne.

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Lawrence, Andrew D..

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Brown, David G..

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Geeves, Michael A..

Creator's ORCID: https://orcid.org/0000-0002-9364-8898
CReDIT Contributor Roles:

Howard, Mark J..

Creator's ORCID:
CReDIT Contributor Roles:

Warren, Martin J..

Creator's ORCID: https://orcid.org/0000-0002-6028-6456
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