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Cloning, expression, purification and preliminary X-ray analysis of EstN2, a novel archaeal ?/?-hydrolase from Candidatus Nitrososphaera gargensis.

ABSTRACT: EstN2 is a novel ?/?-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.

SUBMITTER: Kaljunen H

PROVIDER: S-EPMC4188087 | biostudies-literature | 2014 Oct

REPOSITORIES: biostudies-literature

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Cloning, expression, purification and preliminary X-ray analysis of EstN2, a novel archaeal α/β-hydrolase from Candidatus Nitrososphaera gargensis.

Kaljunen Heidi H Chow Jennifer J Streit Wolfgang R WR Mueller-Dieckmann Jochen J

Acta crystallographica. Section F, Structural biology communications 20140925 Pt 10

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 o ...[more]

PMID: 25286947

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