Change search
ReferencesLink to record
Permanent link

Direct link
Furaldehyde substrate specificity and kinetics of Saccharomyces cerevisiae alcohol dehydrogenase 1 variants
Uppsala University, Disciplinary Domain of Science and Technology, Chemistry, Department of Chemistry - BMC, Biochemistry.
Show others and affiliations
2014 (English)In: Microbial Cell Factories, ISSN 1475-2859, Vol. 13, 112- p.Article in journal (Other academic) Published
Abstract [en]


A previously discovered mutant of Saccharomyces cerevisiae alcohol dehydrogenase 1 (Adh1p) was shown to enable a unique NADH-dependent reduction of 5-hydroxymethylfurfural (HMF), a well-known inhibitor of yeast fermentation. In the present study, site-directed mutagenesis of both native and mutated ADH1 genes was performed in order to identify the key amino acids involved in this substrate shift, resulting in Adh1p-variants with different substrate specificities.


In vitro activities of the Adh1p-variants using two furaldehydes, HMF and furfural, revealed that HMF reduction ability could be acquired after a single amino acid substitution (Y295C). The highest activity, however, was reached with the double mutation S110P Y295C. Kinetic characterization with both aldehydes and the in vivo primary substrate acetaldehyde also enabled to correlate the alterations in substrate affinity with the different amino acid substitutions.


We demonstrated the key role of Y295C mutation in HMF reduction by Adh1p. We generated and kinetically characterized a group of protein variants using two furaldehyde compounds of industrial relevance. Also, we showed that there is a threshold after which higher in vitro HMF reduction activities do not correlate any more with faster in vivo rates of HMF conversion, indicating other cell limitations in the conversion of HMF.

Place, publisher, year, edition, pages
2014. Vol. 13, 112- p.
National Category
Biochemistry and Molecular Biology
URN: urn:nbn:se:uu:diva-229869DOI: 10.1186/s12934-014-0112-5ISI: 000340803100001OAI: diva2:738158
Available from: 2014-08-15 Created: 2014-08-15 Last updated: 2014-09-22Bibliographically approved

Open Access in DiVA

fulltext(711 kB)122 downloads
File information
File name FULLTEXT01.pdfFile size 711 kBChecksum SHA-512
Type fulltextMimetype application/pdf

Other links

Publisher's full text

Search in DiVA

By author/editor
Janfalk Carlsson, Åsa
By organisation
In the same journal
Microbial Cell Factories
Biochemistry and Molecular Biology

Search outside of DiVA

GoogleGoogle Scholar
Total: 122 downloads
The number of downloads is the sum of all downloads of full texts. It may include eg previous versions that are now no longer available

Altmetric score

Total: 434 hits
ReferencesLink to record
Permanent link

Direct link