Functional screening of a Caatinga goat (Capra hircus) rumen metagenomic library reveals a novel GH3 ?-xylosidase

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چکیده

Functional screening of metagenomic libraries is an effective approach for identification novel enzymes. A Caatinga biome goat rumen library was screened using esculin as a substrate, and gene from unknown bacterium encoding GH3 enzyme, BGL11, identified. None the BGL11 closely related genes have been previously characterized. Recombinant obtained kinetically Substrate specificity purified protein assessed seven synthetic aryl substrates. Activity towards nitrophenyl-?-D-glucopyranoside (pNPG), 4-nitrophenyl-?-D-xylopyranoside (pNPX) 4-nitrophenyl-?-D-cellobioside (pNPC) suggested that multifunctional enzyme with ?-glucosidase, ?-xylosidase, cellobiohydrolase activities. However, further testing five natural substrates revealed that, although has multiple substrate specificity, it most active xylobiose. Thus, in its native environment, likely functions extracellular ?-xylosidase acting on hemicellulose. Biochemical characterization showed optimal pH 5.6, temperature 50°C. Enzyme stability, important parameter industrial application, also investigated. At 40°C remained more than 15 hours without reduction activity, at 50°C, after 7 incubation, 60% active. The kinetic parameters K m V max xylobiose were determined to be 3.88 mM 38.53 ?mol.min -1 .mg , respectively, cat 57.79 s . In contrast BLG11, ?-xylosidases studied belong GH43 family characterized only industry, can used plant biomass deconstruction, released sugars fermented into valuable bio-products, ranging biofuel ethanol sugar substitute xylitol.

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ژورنال

عنوان ژورنال: PLOS ONE

سال: 2021

ISSN: ['1932-6203']

DOI: https://doi.org/10.1371/journal.pone.0245118