Screening, cultivation, and biocatalytic performance of Rhodococcus boritolerans FW815 with strong 2,2-dimethylcyclopropanecarbonitrile hydratase activity

J Ind Microbiol Biotechnol. 2012 Mar;39(3):409-17. doi: 10.1007/s10295-011-1029-1. Epub 2011 Sep 4.

Abstract

In this work, a mild, efficient bioconversion of 2,2-dimethylcyclopropanecarbonitrile (DMCPCN) to 2,2-dimethylcyclopropanecarboxamide (DMCPCA) in distilled water system was developed. The isolate FW815 was screened using the enrichment culture technique, displaying strong DMCPCN hydratase activity, and was identified as Rhodococcus boritolerans based on morphological, physiological, biochemical tests and 16S rRNA gene sequencing. Cultivation outcomes indicated that R. boritolerans FW815 was a neutrophile, with a growth optimum of 28-32°C; its DMCPCN hydratase belonged to the Fe-type family, and was most active at 38-42°C, pH 7.0, with maximal activity of 4.51 × 10(4) U g(-1) DCW. R. boritolerans FW815 was found to be DMCPCA amidase-negative, eliminating the contamination of dimethylcyclopropanecarboxylic acid. Moreover, it displayed high activity and acceptable reusability in the non-buffered distilled water system, comparable to those in pH 7.0 phosphate buffer (50.0 mmol l(-1)).

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amidohydrolases
  • Bacterial Proteins / biosynthesis*
  • Biocatalysis
  • Carbon / metabolism
  • Cell Culture Techniques
  • Fermentation
  • Hydro-Lyases / biosynthesis*
  • Hydrogen-Ion Concentration
  • Nitrogen / metabolism
  • RNA, Ribosomal, 16S
  • Rhodococcus / genetics
  • Rhodococcus / growth & development
  • Rhodococcus / metabolism
  • Rhodococcus / physiology*

Substances

  • Bacterial Proteins
  • RNA, Ribosomal, 16S
  • Carbon
  • Amidohydrolases
  • amidase
  • 2,2-dimethylcyclopropanecarbonitrile hydratase, Rhodococcus boritolerans
  • Hydro-Lyases
  • nitrile hydratase
  • Nitrogen