High-throughput functional assessment of polysaccharide-active enzymes using matrix-assisted laser desorption/ionization-time-of-flight mass spectrometry as exemplified on plant cell wall polysaccharides.

High-throughput functional assessment of polysaccharide-active enzymes using matrix-assisted laser desorption/ionization-time-of-flight mass spectrometry as exemplified on plant cell wall polysaccharides. Research Abstract Details 

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High-throughput functional assessment of polysaccharide-active enzymes using matrix-assisted laser desorption/ionization-time-of-flight mass spectrometry as exemplified on plant cell wall polysaccharides. Abstract Text:

Edouard Leboeuf,Peter Immerzeel,Yves Gibon,Martin Steup,Markus Pauly,Edouard Leboeuf,Peter Immerzeel,Yves Gibon,Martin Steup,Markus Pauly,

Despite a wealth of sequence information on genes encoding carbohydrate-active enzymes (e.g., transferases, esterases, hydrolases), very few of these enzymes have been described in detail, particularly regarding substrate specificities. A facile and rapid method for the characterization of substrate specificities of polysaccharide-active enzymes that uses matrix-assisted laser desorption-time of flight mass spectrometry (MALDI-TOF MS) has been developed. This method has been applied to characterize a xyloglucan fucosyltransferase and a pectin methyl-esterase. Reactions were performed in liquid phase, and aliquots of the reaction mixtures were spotted on a polyvinylidene fluoride (PVDF) membrane. Reaction products were precipitated onto the membrane and cleaned by treatment with an ethanol-water mixture. Subsequently, the reaction products were hydrolyzed by specific endoglycanases, and the resulting oligosaccharides were directly analyzed onto the PVDF membrane by MALDI-TOF MS. The new method is amenable to high-throughput analysis and, thus, constitutes an emerging avenue to rapidly fill the gap in our knowledge of the specificities of polysaccharide-active enzymes.

High-throughput functional assessment of polysaccharide-active enzymes using matrix-assisted laser desorption/ionization-time-of-flight mass spectrometry as exemplified on plant cell wall polysaccharides. Publishing Authors By Initials

E Leboeuf,P Immerzeel,Y Gibon,M Steup,M Pauly,E Leboeuf,P Immerzeel,Y Gibon,M Steup,M Pauly,

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High-throughput functional assessment of polysaccharide-active enzymes using matrix-assisted laser desorption/ionization-time-of-flight mass spectrometry as exemplified on plant cell wall polysaccharides. Journal Published:

PUBLICATION TYPE: Journal Article

Journal: Analytical biochemistry

VOLUME: 373

Page Numbers: 9-17

Journal Abbreviation: Anal. Biochem.

ISSN: 0003-2697

DAY: 11

MONTH: 10

YEAR: 2007

High-throughput functional assessment of polysaccharide-active enzymes using matrix-assisted laser desorption/ionization-time-of-flight mass spectrometry as exemplified on plant cell wall polysaccharides. Information

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LANGUAGE: eng

NlmUniqueID: 370535

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Grant and Affiliation Information for High-throughput functional assessment of polysaccharide-active enzymes using matrix-assisted laser desorption/ionization-time-of-flight mass spectrometry as exemplified on plant cell wall polysaccharides.

AFFILIATION: Department of Plant Physiology, Institute of Biochemistry and Biology, University of Potsdam, D-14476 Potsdam-Golm, Germany.

Country: United States

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MEDLINETA: Anal Biochem

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