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2,6-Dimethoxyphenol-Based Assay for Quantitation of Polysaccharide Monooxygenase in Multienzyme Cocktails
Moscow University Chemistry Bulletin Pub Date : 2020-05-25 , DOI: 10.3103/s0027131420020157 V. D. Telitsin , M. V. Semenova , D. O. Osipov , A. V. Gusakov , A. P. Sinitsyn
中文翻译:
基于2,6-二甲氧基苯酚的定量方法测定多酶鸡尾酒中的多糖单加氧酶
更新日期:2020-05-25
Moscow University Chemistry Bulletin Pub Date : 2020-05-25 , DOI: 10.3103/s0027131420020157 V. D. Telitsin , M. V. Semenova , D. O. Osipov , A. V. Gusakov , A. P. Sinitsyn
Abstract
A rapid method for quantitation of lytic polysaccharide monooxygenase (LPMO) in multienzyme cocktails produced by mutant strains of filamentous fungi is developed. In this method, 2,6-dimethoxyphenol and hydrogen peroxide are used as cosubstrates in a nonspecific LPMO-catalyzed reaction leading to the formation of a colored product. With this method, we determine the LPMO content in the multienzyme cocktails produced by Penicillium verruculosum recombinant strains that homologously express LPMO. The results agree closely with the findings obtained using a method based on the fine chromatographic fractionation of the cocktails followed by the qualitative and quantitative evaluation of their content by electrophoresis and mass-spectrometry and protein quantitation assays in fractions collected from chromatographic separation.中文翻译:
基于2,6-二甲氧基苯酚的定量方法测定多酶鸡尾酒中的多糖单加氧酶