Liquid Chromatography combined with Mass Spectrometry for the Investigation of Endoglucanase Selectivity on Carboxymethyl Cellulose
2009 (English)In: Carbohydrate Research, ISSN 0008-6215, E-ISSN 1873-426X, Vol. 344, no 16, 2173-2181 p.Article in journal (Refereed) Published
Endoglucanases are useful tools in the chemical structure analysis of cellulose derivatives. However, knowledge on the endoglucanase selectivity, which is of central importance for data interpretation, is still limited. In this study, new reverse-phase liquid chromatography mass spectrometry (LC-MS) methods were developed to investigate the selectivity of the endoglucanases Cel5A, cel7B, Cel45A, and Cel74A from the filamentous fungus Trichoderma reesei. The aim was to improve the identification of the regioisomers in the complex mixtures that are obtained after enzymatic hydrolysis. Reduction followed by per-O-methylation was performed in order to improve the separation in reverse-phase LC, increase MS sensitivity, and to facilitate structure analysis by MS/MS of O-carboxymethyl glucose and cellooligosaccharides. The cellulose selective enzymes that were investigated displayed interesting differences in enzyme selectivity on CMC substrates.
Place, publisher, year, edition, pages
2009. Vol. 344, no 16, 2173-2181 p.
carboxymethyl cellulose, enzyme hydrolysis, cellulase, permethylation, liquid chromatography, mass spectrometry
IdentifiersURN: urn:nbn:se:kth:diva-9436DOI: 10.1016/j.carres.2009.08.009ISI: 000271254800009ScopusID: 2-s2.0-70349776203OAI: oai:DiVA.org:kth-9436DiVA: diva2:113968
QC 20100720. Uppdaterad från manuskript till artikel (20100720).2008-11-032008-11-032010-07-20Bibliographically approved