uu.seUppsala University Publications
Change search
CiteExportLink to record
Permanent link

Direct link
Cite
Citation style
  • apa
  • ieee
  • modern-language-association
  • vancouver
  • Other style
More styles
Language
  • de-DE
  • en-GB
  • en-US
  • fi-FI
  • nn-NO
  • nn-NB
  • sv-SE
  • Other locale
More languages
Output format
  • html
  • text
  • asciidoc
  • rtf
Shuffling Active Site Substate Populations Affects Catalytic Activity: The Case of Glucose Oxidase
Forschungszentrum Julich, Inst Complex Syst Struct Biochem, D-52425 Julich, Germany..
Rhein Westfal TH Aachen, Inst Mol Biotechnol, Worringerweg 1, D-52074 Aachen, Germany.;Aquila Biolabs GmbH, Arnold Sommerfeld Ring 2, D-52499 Baesweiler, Germany..
Uppsala University, Disciplinary Domain of Science and Technology, Biology, Department of Cell and Molecular Biology, Structure and Molecular Biology.ORCID iD: 0000-0002-3190-1173
Rhein Westfal TH Aachen, Inst Mol Biotechnol, Worringerweg 1, D-52074 Aachen, Germany..
Show others and affiliations
2017 (English)In: ACS Catalysis, ISSN 2155-5435, E-ISSN 2155-5435, Vol. 7, no 9, 6188-6197 p.Article in journal (Refereed) Published
Abstract [en]

Glucose oxidase has wide applications in the pharmaceutical, chemical, and food industries. Many recent studies have enhanced key properties of this enzyme using directed evolution, yet without being able to reveal why these mutations are actually beneficial. This work presents a synergistic combination of experimental and computational methods, indicating how mutations, even when distant from the active site, positively affect glucose oxidase catalysis. We have determined the crystal structures of glucose oxidase mutants containing molecular oxygen in the active site. The catalytically important His516 residue has been previously shown to be flexible in the wild-type enzyme. The molecular dynamics simulations performed in this work allow us to quantify this floppiness, revealing that His516 exists in two states: catalytic and noncatalytic. The relative populations of these two substates are almost identical in the wild-type enzyme, with His516 readily shuffling between them. In the glucose oxidase mutants, on the other hand, the mutations enrich the catalytic His516 conformation and reduce the flexibility of this residue, leading to an enhancement in their catalytic efficiency. This study stresses the benefit of active site preorganization with respect to enzyme conversion rates by reducing molecular reorientation needs. We further suggest that the computational approach based on Hamiltonian replica exchange molecular dynamics, used in this study, may be a general approach to screening in silico for improved enzyme variants involving flexible catalytic residues.

Place, publisher, year, edition, pages
AMER CHEMICAL SOC , 2017. Vol. 7, no 9, 6188-6197 p.
Keyword [en]
molecular dynamics, Hamiltonian replica exchange, X-ray, enzyme floppiness, active-site preorganization, side-chain dynamics, anticorrelated motions
National Category
Biochemistry and Molecular Biology
Identifiers
URN: urn:nbn:se:uu:diva-335408DOI: 10.1021/acscatal.7b01575ISI: 000410005700072OAI: oai:DiVA.org:uu-335408DiVA: diva2:1163122
Funder
Swedish Research Council, 2015-04928EU, European Research Council, 306474
Available from: 2017-12-06 Created: 2017-12-06 Last updated: 2017-12-06Bibliographically approved

Open Access in DiVA

No full text

Other links

Publisher's full text

Authority records BETA

Kamerlin, Shina C. Lynn

Search in DiVA

By author/editor
Kamerlin, Shina C. Lynn
By organisation
Structure and Molecular Biology
In the same journal
ACS Catalysis
Biochemistry and Molecular Biology

Search outside of DiVA

GoogleGoogle Scholar

doi
urn-nbn

Altmetric score

doi
urn-nbn
Total: 19 hits
CiteExportLink to record
Permanent link

Direct link
Cite
Citation style
  • apa
  • ieee
  • modern-language-association
  • vancouver
  • Other style
More styles
Language
  • de-DE
  • en-GB
  • en-US
  • fi-FI
  • nn-NO
  • nn-NB
  • sv-SE
  • Other locale
More languages
Output format
  • html
  • text
  • asciidoc
  • rtf