Change search
ReferencesLink to record
Permanent link

Direct link
NMR Spectroscopic Analysis Reveals Extensive Binding Interactions of Complex Xyloglucan Oligosaccharides with the Cellvibrio japonicus Glycoside Hydrolase Family 31 a-Xylosidase
KTH, School of Biotechnology (BIO), Glycoscience.
Show others and affiliations
2012 (English)In: Chemistry - A European Journal, ISSN 0947-6539, E-ISSN 1521-3765, Vol. 18, no 42, 13395-13404 p.Article in journal (Refereed) Published
Abstract [en]

The study of the interaction of glycoside hydrolases with their substrates is fundamental to diverse applications in medicine, food and feed production, and biomass-resource utilization. Recent molecular modeling of the a-xylosidase CjXyl31A from the soil saprophyte Cellvibrio japonicus, together with protein crystallography and enzyme-kinetic analysis, has suggested that an appended PA14 protein domain, unique among glycoside hydrolase family 31 members, may confer specificity for large oligosaccharide fragments of the ubiquitous plant polysaccharide xyloglucan (J. Larsbrink, A. Izumi, F. M. Ibatullin, A. Nakhai, H. J. Gilbert, G. J. Davies, H. Brumer, Biochem. J. 2011, 436, 567580). In the present study, a combination of NMR spectroscopic techniques, including saturation transfer difference (STD) and transfer NOE (TR-NOE) spectroscopy, was used to reveal extensive interactions between CjXyl31A active-site variants and xyloglucan hexa- and heptasaccharides. The data specifically indicate that the enzyme recognizes the entire cello-tetraosyl backbone of the substrate and product in positive enzyme subsites and makes further significant interactions with internal pendant a-(1?6)-linked xylosyl units. As such, the present analysis provides an important rationalization of previous kinetic data on CjXyl31A and unique insight into the role of the PA14 domain, which was not otherwise obtainable by protein crystallography.

Place, publisher, year, edition, pages
2012. Vol. 18, no 42, 13395-13404 p.
Keyword [en]
a-xylosidases, Cellvibrio japonicus, molecular recognition, mutagenesis, NMR spectroscopy, xyloglucan
National Category
Chemical Sciences
URN: urn:nbn:se:kth:diva-105659DOI: 10.1002/chem.201200488ISI: 000309597000023OAI: diva2:572828
Swedish Research Council

QC 20121129

Available from: 2012-11-29 Created: 2012-11-23 Last updated: 2013-08-26Bibliographically approved
In thesis
1. Strategies for the Discovery of Carbohydrate-Active Enzymes from Environmental Bacteria
Open this publication in new window or tab >>Strategies for the Discovery of Carbohydrate-Active Enzymes from Environmental Bacteria
2013 (English)Doctoral thesis, comprehensive summary (Other academic)
Abstract [en]

The focus of this thesis is a comparative study of approaches in discovery of carbohydrate-active enzymes (CAZymes). CAZymes synthesise, bind to, and degrade all the multitude of carbohydrates found in nature. As such, when aiming for sustainable methods to degrade plant biomass for the generation of biofuels, for which there is a strong drive in society, CAZymes are a natural source of environmentally friendly molecular tools.

In nature, microorganisms are the principal degraders of carbohydrates. Not only do they degrade plant matter in forests and aquatic habitats, but also break down the majority of carbohydrates ingested by animals. These symbiotic microorganisms, known as the microbiota, reside in animal digestive tracts in immense quantities, where one of the key nutrient sources is complex carbohydrates. Thus, microorganisms are a plentiful source of CAZymes, and strategies in the discovery of new enzymes from bacterial sources have been the basis for the work presented here, combined with biochemical characterisation of several enzymes.

Novel enzymatic activities for the glycoside hydrolase family 31 have been described as a result of the initial projects of the thesis. These later evolved into projects studying bacterial multi-gene systems for the partial or complete degradation of the heterogeneous plant polysaccharide xyloglucan. These systems contain, in addition to various hydrolytic CAZymes, necessary binding-, transport-, and regulatory proteins. The results presented here show, in detail, how very complex carbohydrates can efficiently be degraded by bacterial enzymes of industrial relevance.

Place, publisher, year, edition, pages
Stockholm: KTH Royal Institute of Technology, 2013. viii, 82 p.
TRITA-BIO-Report, ISSN 1654-2312 ; 2013:13
CAZyme discovery, xyloglucan, polysaccharide-utilisation locus, microbiota, α-xylosidase, GH31, transglucosidase, human gut
National Category
Biochemistry and Molecular Biology
Research subject
SRA - Molecular Bioscience
urn:nbn:se:kth:diva-126956 (URN)978-91-7501-834-8 (ISBN)
Public defence
2013-09-13, Oskar Klein Auditoriet, FR4, 10:00, Albanova Universitetscentrum, Roslagstullsbacken 21, Stockholm, 15:00 (English)

QC 20130826

Available from: 2013-08-26 Created: 2013-08-23 Last updated: 2016-01-26Bibliographically approved

Open Access in DiVA

No full text

Other links

Publisher's full text

Search in DiVA

By author/editor
Larsbrink, JohanBrumer, Harry
By organisation
In the same journal
Chemistry - A European Journal
Chemical Sciences

Search outside of DiVA

GoogleGoogle Scholar
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: 26 hits
ReferencesLink to record
Permanent link

Direct link