Determination of glycoside hydrolase specificities during hydrolysis of plant cell walls using glycome profiling
| Autor: | Maryam Mirzai, Michael G. Hahn, Johnnie A. Walker, Emily T. Beebe, Lai F. Bergeman, Sivakumar Pattathil, Kai Deng, Brian G. Fox, Trent R. Northen |
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| Rok vydání: | 2017 |
| Předmět: |
0106 biological sciences
0301 basic medicine Xyloglucanase Glycome profiling Management Monitoring Policy and Law Biology Polysaccharide 01 natural sciences Applied Microbiology and Biotechnology Industrial Biotechnology Cell wall 03 medical and health sciences Hydrolysis Enzymatic hydrolysis Glycoside hydrolase 2. Zero hunger chemistry.chemical_classification Xylanase Renewable Energy Sustainability and the Environment Research Nanostructure-initiator mass spectrometry Chemical Engineering Oligosaccharide Glycome 030104 developmental biology General Energy Enzyme Biochemistry chemistry Enzyme specificity 010606 plant biology & botany Biotechnology |
| Zdroj: | Walker, JA; Pattathil, S; Bergeman, LF; Beebe, ET; Deng, K; Mirzai, M; et al.(2017). Determination of glycoside hydrolase specificities during hydrolysis of plant cell walls using glycome profiling. Biotechnology for Biofuels, 10(1). doi: 10.1186/s13068-017-0703-6. Lawrence Berkeley National Laboratory: Retrieved from: http://www.escholarship.org/uc/item/0z0563wg Biotechnology for biofuels, vol 10, iss 1 Biotechnology for Biofuels |
| ISSN: | 1754-6834 |
| Popis: | Background Glycoside hydrolases (GHs) are enzymes that hydrolyze polysaccharides into simple sugars. To better understand the specificity of enzyme hydrolysis within the complex matrix of polysaccharides found in the plant cell wall, we studied the reactions of individual enzymes using glycome profiling, where a comprehensive collection of cell wall glycan-directed monoclonal antibodies are used to detect polysaccharide epitopes remaining in the walls after enzyme treatment and quantitative nanostructure initiator mass spectrometry (oxime-NIMS) to determine soluble sugar products of their reactions. Results Single, purified enzymes from the GH5_4, GH10, and GH11 families of glycoside hydrolases hydrolyzed hemicelluloses as evidenced by the loss of specific epitopes from the glycome profiles in enzyme-treated plant biomass. The glycome profiling data were further substantiated by oxime-NIMS, which identified hexose products from hydrolysis of cellulose, and pentose-only and mixed hexose-pentose products from the hydrolysis of hemicelluloses. The GH10 enzyme proved to be reactive with the broadest diversity of xylose-backbone polysaccharide epitopes, but was incapable of reacting with glucose-backbone polysaccharides. In contrast, the GH5 and GH11 enzymes studied here showed the ability to react with both glucose- and xylose-backbone polysaccharides. Conclusions The identification of enzyme specificity for a wide diversity of polysaccharide structures provided by glycome profiling, and the correlated identification of soluble oligosaccharide hydrolysis products provided by oxime-NIMS, offers a unique combination to understand the hydrolytic capabilities and constraints of individual enzymes as they interact with plant biomass. Electronic supplementary material The online version of this article (doi:10.1186/s13068-017-0703-6) contains supplementary material, which is available to authorized users. |
| Databáze: | OpenAIRE |
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