Purification and characterization of a novel nitrile hydratase from Rhodococcus sp. RHA1

Sachi Okamoto, Lindsay D. Eltis

Research output: Contribution to journalArticle

38 Citations (Scopus)

Abstract

The microbial degradation of nitriles is of interest for bioremediation and green chemistry. We demonstrated that the soil bacterium Rhodococcus sp. RHA1 utilizes a range of nitriles, including acetonitrile, as growth substrates. Proteomic analysis identified 13 proteins that were more abundant in acetonitrile-grown cells, including an aliphatic amidase and a protein with no known homologue. Purification of a nitrile hydratase (NHase) from acetonitrile-grown cells identified the unknown protein as the β subunit of a two-subunit NHase. Sequence analysis revealed that the genes encoding the amidase (anhC) and the NHase (anhAB) occur in a 12.8 kbp cluster located on plasmid pRHL2. The anh gene cluster also encodes an acetyl-CoA hydrolase, transcriptional regulators, a putative cobalt transporter and a protein of unknown function. Striking features of the NHase include the amino acid sequence identity (32%) and large size (63 and 56 kDa) of the α and β subunits, as well as the enzyme's metal ion content (one cobalt, two copper and one zinc). The enzyme possessed similar specificities for acetonitrile and propionitrile (kcat/Km∼7 mM-1 s -1) followed by acrylonitrile and butyronitrile. We propose that this acetonitrile hydratase (ANHase) represents the first member of a previously unknown class of NHases.

Original languageEnglish
Pages (from-to)828-838
Number of pages11
JournalMolecular Microbiology
Volume65
Issue number3
DOIs
Publication statusPublished - Aug 1 2007

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Rhodococcus
amidase
Nitriles
Cobalt
Acetyl-CoA Hydrolase
Hydro-Lyases
Acrylonitrile
Proteins
Environmental Biodegradation
Protein Subunits
Enzymes
Multigene Family
Proteomics
Sequence Analysis
Zinc
Copper
Amino Acid Sequence
Plasmids
Soil
Metals

All Science Journal Classification (ASJC) codes

  • Molecular Biology
  • Microbiology

Cite this

Purification and characterization of a novel nitrile hydratase from Rhodococcus sp. RHA1. / Okamoto, Sachi; Eltis, Lindsay D.

In: Molecular Microbiology, Vol. 65, No. 3, 01.08.2007, p. 828-838.

Research output: Contribution to journalArticle

Okamoto, Sachi ; Eltis, Lindsay D. / Purification and characterization of a novel nitrile hydratase from Rhodococcus sp. RHA1. In: Molecular Microbiology. 2007 ; Vol. 65, No. 3. pp. 828-838.
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