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Nat Commun. 2017 May 8;8:15352. doi: 10.1038/ncomms15352.

Activity-based protein profiling as a robust method for enzyme identification and screening in extremophilic Archaea.

Author information

1
Chemische Biologie, ZMB, Fakultät für Biologie, Universität Duisburg-Essen, Universitätsstr. 2, 45117 Essen, Germany.
2
Molekulare Enzymtechnologie und Biochemie, Biofilm Centre, ZWU, Fakultät für Chemie, Universität Duisburg-Essen, Universitätsstr. 2, 45117 Essen, Germany.
3
Molecular Biology of Archaea, Institute of Biology II-Microbiology, University of Freiburg, Schänzlestr. 1, 79104 Freiburg, Germany.
4
Leibniz-Institut für Analytische Wissenschaften-ISAS e.V., Bunsen-Kirchhoff-Str. 11, 44139 Dortmund, Germany.

Abstract

Archaea are characterized by a unique life style in often environmental extremes but their thorough investigation is currently hampered by a limited set of suitable in vivo research methodologies. Here, we demonstrate that in vivo activity-based protein profiling (ABPP) may be used to sensitively detect either native or heterogeneously expressed active enzymes in living archaea even under these extreme conditions. In combination with the development of a genetically engineered archaeal screening strain, ABPP can furthermore be used in functional enzyme screenings from (meta)genome samples. We anticipate that our ABPP approach may therefore find application in basic archaeal research but also in the discovery of novel enzymes from (meta)genome libraries.

PMID:
28480883
PMCID:
PMC5424146
DOI:
10.1038/ncomms15352
[Indexed for MEDLINE]
Free PMC Article

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