國家衛生研究院 NHRI:Item 3990099045/5906
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    題名: Colloid-based multiplexed screening for plant biomass-degrading glycoside hydrolase activities in microbial communities
    作者: Reindl, W;Deng, K;Gladden, JM;Cheng, G;Wong, A;Singer, SW;Singh, S;Lee, JC;Yao, CH;Hazen, TC;Singh, AK;Simmons, BA;Adams, PD;Northen, TR
    貢獻者: Institute of Biotechnology and Pharmaceutical Research
    摘要: The enzymatic hydrolysis of polysaccharides into fermentable sugars is a crucial step in the conversion of biomass to lignocellulosic biofuels. An efficient hydrolysis is highly dependent on the identification and characterization of optimal glycoside hydrolases. However, existing techniques for characterizing activity are limited by the range of reaction conditions that can be used, sample complexity, and throughput. The method we present is a multiplexed approach based on nanostructure-initiator mass spectrometry (NIMS) that allows for the rapid analysis of several glycolytic activities in parallel under diverse assay conditions. By forming colloids, it was possible to perform aqueous reactions in tubes and microwell plates despite the substrate analogs' hydrophobic perfluorinated tags. This method was validated by analyzing standard enzymatic parameters (temperature, pH, and kinetics) of beta-glucosidase and beta-xylosidase in separate setups. The multiplexity of this assay system was demonstrated by the simultaneous analysis of beta-glucosidase and beta-xylosidase activities, which was then used to profile environmental samples. Enzymes secreted by microbial communities within these samples were extracted by washing the sample with buffer. Enzymatic activities were directly detected within this crude extract without any further sample pretreatment steps. The multiplexed analysis of beta-glucosidase, exo-/endoglucanase, and xylanase activities was applied for a detailed characterization of previously unknownglycoside hydrolase activities. The results show the suitability of the described method for the rapid screening of crude environmental samples under a wide range of conditions to determine enzyme activities from the microbial communities present within these samples.
    日期: 2011-08
    關聯: Energy and Environmental Science. 2011 Aug;4(8):2884-2893.
    Link to: http://dx.doi.org/10.1039/c1ee01112j
    JIF/Ranking 2023: http://gateway.webofknowledge.com/gateway/Gateway.cgi?GWVersion=2&SrcAuth=NHRI&SrcApp=NHRI_IR&KeyISSN=1754-5692&DestApp=IC2JCR
    Cited Times(WOS): https://www.webofscience.com/wos/woscc/full-record/WOS:000293213600034
    Cited Times(Scopus): http://www.scopus.com/inward/record.url?partnerID=HzOxMe3b&scp=79960999883
    顯示於類別:[李靜琪] 期刊論文

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