Improvement of RNA-SIP by pyrosequencing to identify putative 4-n-nonylphenol degraders in activated sludge

O. Zemb, M. Lee, M. L. Gutierrez-Zamora, J. Hamelin, K. Coupland, Nur Hazlin Hazrin Chong, I. Taleb, M. Manefield

Research output: Contribution to journalArticle

19 Citations (Scopus)

Abstract

Nonylphenols (NP) have estrogenic potential because of their phenolic ring, but the organisms involved in the degradation of this alkylated phenol remain unidentified. Using 16S ribosomal RNA (rRNA)-based stable isotope probing (SIP) and a new method based on pyrosequencing, we identified the bacteria involved in the degradation of the aromatic ring of [U-ring- 13C] 4-n-NP in aerobic sludge. The first order degradation rate of 4-n-NP was 5.5d -1. Single strand conformation polymorphism of density-separated labeled and unlabeled 16S rRNA showed significant differences and enabled selection of four representative fractions for pyrosequencing. Nineteen phylotypes showed a significant enrichment in the heavy fraction in the labeled pulse. The relative abundances of these phylotypes were combined with the RNA concentration of each fraction to yield a simple model of the distribution of each phylotype across the gradient. This model was used to estimate the percentage of labeling for each phylotype. The sequences showing the highest labeling (11%) were closely related to Afipia sp. but represented only 2 % of the RNA in the heavy fraction of the labeled pulse. The sequences representing the largest proportion of the RNA in the heavy fraction were related to Propionibacterium acnes and Frateuria aurantia, which are known to possess enzymes for phenol degradation. The model shows that despite Afipia having the highest 13C enrichment, other species encoding phenol degradation pathways are responsible for more 13C incorporation. Last, we showed that some species represent 12% of the total RNA but contain only 1% 13C above natural abundance.

Original languageEnglish
Pages (from-to)601-610
Number of pages10
JournalWater Research
Volume46
Issue number3
DOIs
Publication statusPublished - 1 Mar 2012
Externally publishedYes

Fingerprint

Isotopes
activated sludge
stable isotope
Degradation
degradation
Phenols
phenol
Labeling
Polymorphism
Conformations
RNA
relative abundance
Bacteria
polymorphism
Enzymes
sludge
enzyme
bacterium
labelling

Keywords

  • Endocrine disruptors
  • Environmental microbiology
  • Microbial ecology
  • Nonylphenol
  • Stable isotope probing
  • Waste water

ASJC Scopus subject areas

  • Water Science and Technology
  • Waste Management and Disposal
  • Pollution
  • Ecological Modelling

Cite this

Improvement of RNA-SIP by pyrosequencing to identify putative 4-n-nonylphenol degraders in activated sludge. / Zemb, O.; Lee, M.; Gutierrez-Zamora, M. L.; Hamelin, J.; Coupland, K.; Hazrin Chong, Nur Hazlin; Taleb, I.; Manefield, M.

In: Water Research, Vol. 46, No. 3, 01.03.2012, p. 601-610.

Research output: Contribution to journalArticle

Zemb, O, Lee, M, Gutierrez-Zamora, ML, Hamelin, J, Coupland, K, Hazrin Chong, NH, Taleb, I & Manefield, M 2012, 'Improvement of RNA-SIP by pyrosequencing to identify putative 4-n-nonylphenol degraders in activated sludge', Water Research, vol. 46, no. 3, pp. 601-610. https://doi.org/10.1016/j.watres.2011.10.047
Zemb, O. ; Lee, M. ; Gutierrez-Zamora, M. L. ; Hamelin, J. ; Coupland, K. ; Hazrin Chong, Nur Hazlin ; Taleb, I. ; Manefield, M. / Improvement of RNA-SIP by pyrosequencing to identify putative 4-n-nonylphenol degraders in activated sludge. In: Water Research. 2012 ; Vol. 46, No. 3. pp. 601-610.
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AU - Hazrin Chong, Nur Hazlin

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