UP-HILIC-MS/MS to determine the action pattern of Penicillium sp. dextranase

Yi, Lin
Sun, Xue
Du, Kenze
Ouyang, Yilan
Wu, Chengling
Xu, Naiyu
Linhardt, Robert J.
Zhang, Zhenqing
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Biology , Chemistry and chemical biology , Chemical and biological engineering , Biomedical engineering
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Full Citation
UP-HILIC-MS/MS to determine the action pattern of Penicillium sp. dextranase, L. Yi, X. Sun, K. Du, Y. Ouyang, C. Wu, N. Xu, R. J. Linhardt, Z. Zhang, Journal of the American Society of Mass Spectrometry, 26, 1174-1185 2015.
Investigation of the action pattern of enzymes acting on carbohydrates is challenging, as both the substrate and the digestion products are complex mixtures. Dextran and its enzyme-derived oligosaccharides are widely used for many industrial applications. In this work, a new method relying on ultra-performance hydrophilic interaction liquid chromatography quadrupole time-of-flight tandem mass spectrometry (UP-HILIC- Q/TOF-MS/MS) was developed to analyze a complex mixture of dextran oligosaccharide products to determine the action pattern of dextranase. No derivatization of oligosaccharides was required and the impact of the α- and β-configurations of the native oligosaccharides on the chromatographic separation was eliminated. The 1→6, 1→3, 1→4 backbone linkages and the branch linkages of these oligosaccharides were all distinguished from diagnostic ions in their MS/MS spectra, including fragments corresponding to 0,2A, 0,3A, 0,4A, B-H2O, 2,5A, and 3,5A. The sequences of the oligosaccharide products were similarly established. Thus, the complex oligosaccharide mixtures in dextran digestion products were profiled and identified using this method. The more enzyme-resistant structures in dextran were established using much less sample, labor, time, and uncertainty than in previous studies. This method provides an efficient, sensitive, and straightforward way to monitor the entire process of digestion, establish the action pattern of the dextranase from Penicillium sp., and to support the proper industrial application of dextranase.
Journal of the American Society of Mass Spectrometry, 26, 1174-1185
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The Linhardt Research Labs.
The Shirley Ann Jackson, Ph.D. Center for Biotechnology and Interdisciplinary Studies (CBIS)
The Linhardt Research Labs Online Collection
Rensselaer Polytechnic Institute, Troy, NY
Journal of the American Society for Mass Spectrometry