Chemische Biologie, ZMB, Fakultät für Biologie, Universität Duisburg-Essen, Universitätsstr. 2, 45117 Essen, Germany.
Molekulare Enzymtechnologie und Biochemie, Biofilm Centre, ZWU, Fakultät für Chemie, Universität Duisburg-Essen, Universitätsstr. 2, 45117 Essen, Germany.
Nat Commun. 2017 May 8;8:15352. doi: 10.1038/ncomms15352.
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.
古菌的特点是在通常的极端环境中具有独特的生活方式，但由于适合的体内研究方法有限，目前对它们的深入研究受到了阻碍。在这里，我们证明了即使在这些极端条件下，基于活性的蛋白质体内分析（ABPP）也可以用于灵敏地检测活古菌中的天然或异质表达的活性酶。结合遗传工程化古菌筛选株的开发，ABPP 还可以用于（宏）基因组样品中的功能酶筛选。我们预计，我们的 ABPP 方法因此不仅可以应用于基础古菌研究，还可以用于（宏）基因组文库中新型酶的发现。
