title: Cloning, expression, purification and preliminary X-ray analysis of EstN2, a novel archaeal α/β-hydrolase from Candidatus Nitrososphaera gargensis.
abstract: EstN2 is a novel α/β-hydrolase originating from the ammonia-oxidizing thaumarchaeon Candidatus Nitrososphaera gargensis. The genome of the organism was sequenced and genes conferring putative lipolytic activity were amplified and cloned into Escherichia coli as a heterologous host. Through function-based screening, esterase and lipase activity was detected. A recombinant enzyme designated EstN2 was successfully expressed, purified and crystallized. The crystals belonged to space group I2, with one molecule per asymmetric unit, and diffracted X-rays to 1.5 Å resolution.
authors.affiliation: EMBL Hamburg Outstation, c/o DESY, Notkestrasse 85, 22603 Hamburg, Germany.
authors.forename: Heidi
authors.initials: H
authors.lastname: Kaljunen
authors.affiliation: Department of Microbiology and Biotechnology, Biocenter Klein Flottbek, University of Hamburg, Ohnhorststrasse 18, 22609 Hamburg, Germany.
authors.forename: Jennifer
authors.initials: J
authors.lastname: Chow
authors.affiliation: Department of Microbiology and Biotechnology, Biocenter Klein Flottbek, University of Hamburg, Ohnhorststrasse 18, 22609 Hamburg, Germany.
authors.forename: Wolfgang R
authors.initials: WR
authors.lastname: Streit
authors.affiliation: Department of Microbiology and Biotechnology, Biocenter Klein Flottbek, University of Hamburg, Ohnhorststrasse 18, 22609 Hamburg, Germany.
authors.forename: Jochen
authors.initials: J
authors.lastname: Mueller-Dieckmann
mesh_terms: D000595:Amino Acid Sequence; D001105:Archaea / Q000201:enzymology*; D019843:Archaeal Proteins / Q000096:biosynthesis / Q000737:chemistry* / Q000302:isolation & purification; D002846:Chromatography, Affinity; D003001:Cloning, Molecular; D003460:Crystallization; D018360:Crystallography, X-Ray; D004926:Escherichia coli; D015870:Gene Expression; D006867:Hydrolases / Q000096:biosynthesis / Q000737:chemistry* / Q000302:isolation & purification; D008969:Molecular Sequence Data
chemical_list: D019843:Archaeal Proteins; D006867:Hydrolases
keywords: Candidatus Nitrososphaera gargensis; EstN2; α/β-hydrolase