Isolation and characterization of 4-tert-butylphenol-utilizing Sphingobium fuliginis strains from Phragmites australis rhizosphere sediment

Appl Environ Microbiol. 2010 Oct;76(20):6733-40. doi: 10.1128/AEM.00258-10. Epub 2010 Aug 27.

Abstract

We isolated three Sphingobium fuliginis strains from Phragmites australis rhizosphere sediment that were capable of utilizing 4-tert-butylphenol as a sole carbon and energy source. These strains are the first 4-tert-butylphenol-utilizing bacteria. The strain designated TIK-1 completely degraded 1.0 mM 4-tert-butylphenol in basal salts medium within 12 h, with concomitant cell growth. We identified 4-tert-butylcatechol and 3,3-dimethyl-2-butanone as internal metabolites by gas chromatography-mass spectrometry. When 3-fluorocatechol was used as an inactivator of meta-cleavage enzymes, strain TIK-1 could not degrade 4-tert-butylcatechol and 3,3-dimethyl-2-butanone was not detected. We concluded that metabolism of 4-tert-butylphenol by strain TIK-1 is initiated by hydroxylation to 4-tert-butylcatechol, followed by a meta-cleavage pathway. Growth experiments with 20 other alkylphenols showed that 4-isopropylphenol, 4-sec-butylphenol, and 4-tert-pentylphenol, which have alkyl side chains of three to five carbon atoms with α-quaternary or α-tertiary carbons, supported cell growth but that 4-n-alkylphenols, 4-tert-octylphenol, technical nonylphenol, 2-alkylphenols, and 3-alkylphenols did not. The rate of growth on 4-tert-butylphenol was much higher than that of growth on the other alkylphenols. Degradation experiments with various alkylphenols showed that strain TIK-1 cells grown on 4-tert-butylphenol could degrade 4-alkylphenols with variously sized and branched side chains (ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, tert-butyl, n-pentyl, tert-pentyl, n-hexyl, n-heptyl, n-octyl, tert-octyl, n-nonyl, and branched nonyl) via a meta-cleavage pathway but not 2- or 3-alkylphenols. Along with the degradation of these alkylphenols, we detected methyl alkyl ketones that retained the structure of the original alkyl side chains. Strain TIK-1 may be useful in the bioremediation of environments polluted by 4-tert-butylphenol and various other 4-alkylphenols.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Butanones / isolation & purification
  • Butanones / metabolism
  • Catechols / isolation & purification
  • Catechols / metabolism
  • Culture Media / chemistry
  • DNA, Bacterial / chemistry
  • DNA, Bacterial / genetics
  • DNA, Ribosomal / chemistry
  • DNA, Ribosomal / genetics
  • Gas Chromatography-Mass Spectrometry
  • Molecular Sequence Data
  • Phenols / metabolism*
  • Plant Roots / microbiology
  • Poaceae / microbiology*
  • RNA, Ribosomal, 16S / genetics
  • Rhizosphere*
  • Sequence Analysis, DNA
  • Soil Microbiology*
  • Sphingomonadaceae / growth & development
  • Sphingomonadaceae / isolation & purification*
  • Sphingomonadaceae / metabolism*

Substances

  • Butanones
  • Catechols
  • Culture Media
  • DNA, Bacterial
  • DNA, Ribosomal
  • Phenols
  • RNA, Ribosomal, 16S
  • tert-butylcatechol
  • pinacolone
  • butylphen

Associated data

  • GENBANK/AB491315
  • GENBANK/AB491316
  • GENBANK/AB491317