Investigation of the prevalence and catalytic activity of rubredoxin-fused alkane monooxygenases (AlkBs).


Journal

Journal of inorganic biochemistry
ISSN: 1873-3344
Titre abrégé: J Inorg Biochem
Pays: United States
ID NLM: 7905788

Informations de publication

Date de publication:
06 2021
Historique:
received: 23 12 2020
revised: 28 01 2021
accepted: 21 02 2021
pubmed: 23 3 2021
medline: 18 1 2022
entrez: 22 3 2021
Statut: ppublish

Résumé

Interest in understanding the environmental distribution of the alkane monooxygenase (AlkB) enzyme led to the identification of over 100 distinct alkane monooxygenase (AlkB) enzymes containing a covalently bound, or fused, rubredoxin. The rubredoxin-fused AlkB from Dietzia cinnamea was cloned as a full-length protein and as a truncated protein with the rubredoxin domain deleted. A point mutation (V91W) was introduced into the full-length protein, with the goal of assessing how steric bulk in the putative substrate channel might affect selectivity. Based on activity studies with alkane and alkene substrates, the rubredoxin-fused AlkB oxidizes a similar range of alkane substrates relative to its rubredoxin domain-deletion counterpart. Oxidation of terminal alkenes generated both an epoxide and a terminal aldehyde. The products of V91W-mutant-catalyzed oxidation of alkenes had a higher aldehyde-to-epoxide ratio than the products formed in the presence of the wild type protein. These results are consistent with this mutation causing a structural change impacting substrate positioning.

Identifiants

pubmed: 33752122
pii: S0162-0134(21)00056-8
doi: 10.1016/j.jinorgbio.2021.111409
pmc: PMC8557626
mid: NIHMS1687019
pii:
doi:

Substances chimiques

Alkanes 0
Alkenes 0
Bacterial Proteins 0
Rubredoxins 0
Mixed Function Oxygenases EC 1.-

Types de publication

Journal Article Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov't

Langues

eng

Sous-ensembles de citation

IM

Pagination

111409

Subventions

Organisme : NIGMS NIH HHS
ID : R01 GM130989
Pays : United States

Informations de copyright

Copyright © 2021 Elsevier Inc. All rights reserved.

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Auteurs

Shoshana C Williams (SC)

Department of Chemistry, Barnard College, 3009 Broadway, New York, NY 10027, USA.

Allison P Forsberg (AP)

Department of Chemistry, Barnard College, 3009 Broadway, New York, NY 10027, USA.

Juliet Lee (J)

Department of Chemistry, Barnard College, 3009 Broadway, New York, NY 10027, USA.

Christina L Vizcarra (CL)

Department of Chemistry, Barnard College, 3009 Broadway, New York, NY 10027, USA.

Allison J Lopatkin (AJ)

Department of Biology, Barnard College, 3009 Broadway, New York, NY 10027, USA; Data Science Institute Columbia University, New York, NY 10027 USA.

Rachel N Austin (RN)

Department of Chemistry, Barnard College, 3009 Broadway, New York, NY 10027, USA. Electronic address: raustin@barnard.edu.

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Classifications MeSH