title: Cobalt activation of Bacillus BR449 thermostable nitrile hydratase expressed in Escherichia coli.
abstract: Expression of nitrile hydratase enzymes utilized in a new "green" process for acrylamide production has proven difficult in Escherichia coli owing to lack of a cobalt transport system to introduce the required cobalt ion into this host. We describe the expression of a thermostable nitrile hydratase from a moderate thermophile Bacillus sp. BR449 in E. coliin which the cobalt required for enzyme activation is introduced by incubation of the apoenzyme in the presence of Co++ ion at 50 degrees C, yielding active and thermostable enzyme.
authors.affiliation: Department of Microbiology, Michigan State University, East Lansing 48824, USA.
authors.forename: S H
authors.initials: SH
authors.lastname: Kim
authors.forename: R
authors.initials: R
authors.lastname: Padmakumar
authors.forename: P
authors.initials: P
authors.lastname: Oriel
mesh_terms: D020106:Acrylamide / Q000378:metabolism; D000181:Acrylonitrile / Q000378:metabolism; D001407:Bacillus / Q000201:enzymology* / Q000235:genetics; D020134:Catalytic Domain; D003035:Cobalt / Q000737:chemistry / Q000494:pharmacology*; D004789:Enzyme Activation / Q000187:drug effects; D004795:Enzyme Stability; D004926:Escherichia coli / Q000235:genetics; D015870:Gene Expression; D006836:Hydro-Lyases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D008970:Molecular Weight; D013053:Spectrophotometry; D013696:Temperature
chemical_list: D020106:Acrylamide; D003035:Cobalt; D006836:Hydro-Lyases; C048831:nitrile hydratase; D000181:Acrylonitrile