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Screening for distinct xylan degrading enzymes in complex shake flask fermentation supernatants
Authors:Van Gool M P  Vancsó I  Schols H A  Toth K  Szakacs G  Gruppen H
Institution:a Wageningen University, Laboratory of Food Chemistry, Bomenweg 2, 6703 HD Wageningen, The Netherlands
b Budapest University of Technology and Economics, Department of Applied Biotechnology and Food Science, 1111 Budapest, Gellert ter 4, Hungary
Abstract:The efficient degradation of complex xylans needs collaboration of many xylan degrading enzymes. Assays for xylan degrading activities based on reducing sugars or PNP substrates are not indicative for the presence of enzymes able to degrade complex xylans: They do not provide insight into the possible presence of xylanase-accessory enzymes within enzyme mixtures. A new screening method is described, by which specific xylan modifying enzymes can be detected.Fermentation supernatants of 78 different fungal soil isolates grown on wheat straw were analyzed by HPLC and MS. This strategy is powerful in recognizing xylanases, arabinoxylan hydrolases, acetyl xylan esterases and glucuronidases.No fungus produced all enzymes necessary to totally degrade the substrates tested. Some fungi produce high levels of xylanase active against linear xylan, but are unable to degrade complex xylans. Other fungi producing relative low levels of xylanase secrete many useful accessory enzyme component(s).
Keywords:Enzyme screening  Soil fungi  Xylanase  Hemicellulases  Accessory enzymes
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