期刊
FRONTIERS IN MICROBIOLOGY
卷 12, 期 -, 页码 -出版社
FRONTIERS MEDIA SA
DOI: 10.3389/fmicb.2021.734039
关键词
activity-based protein profiling; archaea; Thermococcus; xylan; hemicellulose degradation; glycoside hydrolases
类别
资金
- DFG-RSF Cooperation for joint German-Russian projects by the DFG [SI 642/12-1, KA 2894/6-1]
- RSF [18-44-04024]
- DFG [INST 20876/322-1 FUGG]
- Russian Science Foundation [20-14-00250]
- Russian Science Foundation [18-44-04024, 20-14-00250] Funding Source: Russian Science Foundation
This study is the first to apply activity-based protein profiling (ABPP) in Archaea, identifying two novel proteins with xylanase activity in extremophilic organisms. This expands the applicability of ABPP for glycoside hydrolase identification in hyperthermophiles.
Activity-based protein profiling (ABPP) has so far scarcely been applied in Archaea in general and, especially, in extremophilic organisms. We herein isolated a novel Thermococcus strain designated sp. strain 2319x1E derived from the same enrichment culture as the recently reported Thermococcus sp. strain 2319x1. Both strains are able to grow with xylan as the sole carbon and energy source, and for Thermococcus sp. strain 2319x1E (optimal growth at 85 degrees C, pH 6-7), the induction of xylanolytic activity in the presence of xylan was demonstrated. Since the solely sequence-based identification of xylanolytic enzymes is hardly possible, we established a complementary approach by conducting comparative full proteome analysis in combination with ABPP using alpha- or beta-glycosidase selective probes and subsequent mass spectrometry (MS)-based analysis. This complementary proteomics approach in combination with recombinant protein expression and classical enzyme characterization enabled the identification of a novel bifunctional maltose-forming alpha-amylase and deacetylase (EGDIFPOO_00674) belonging to the GH57 family and a promiscuous beta-glycosidase (EGIDFPOO_00532) with beta-xylosidase activity. We thereby further substantiated the general applicability of ABPP in archaea and expanded the ABPP repertoire for the identification of glycoside hydrolases in hyperthermophiles.
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