INVESTIGADORES
BASILE Laura Ana
artículos
Título:
Quantitative Assessment of Phenol Hydroxylase Diversity in Bioreactors Using a Functional Gene Analysis
Autor/es:
LAURA A. BASILE, LEONARDO ERIJMAN
Revista:
APPLIED MICROBIOLOGY AND BIOTECHNOLOGY
Editorial:
SPRINGER
Referencias:
Lugar: Berlin; Año: 2008 vol. 78 p. 863 - 872
ISSN:
0175-7598
Resumen:
Abstract We describe a quantitative analysis of the genetic
diversity of phenol-degrading potential in bacterial communities
from laboratory-scale activated sludge. Genomic
DNA extracted from activated sludge from two sequential
batch reactors fed with synthetic sewage plus phenol was
amplified using conserved primers for the major subunit of
the phenol hydroxylase (LmPH) gene and used to generate
clone libraries. Following phylogenetic analysis, 59 sequences
containing a 470-bp fragment clustered into six
distinct subgroups with a genetic distance of 8%, most
likely representing ecologically relevant variants of the
enzyme. Seven sets of primers were designed to target the
six clusters and used to obtain quantitative information on
the dynamics of LmPH gene diversity using real-time PCR
assays throughout 9 months of bioreactors operation. Total
LmPH gene copy number remained approximately steady
in phenol-amended and control reactors. However, a
significant increase in phenol-degrading activity in the
phenol-amended sludge was accompanied by a parallel
increase in LmPH gene diversity, suggesting that phenol
degradation in the activated sludge depends on the
combined activity of a number of redundant species.
diversity of phenol-degrading potential in bacterial communities
from laboratory-scale activated sludge. Genomic
DNA extracted from activated sludge from two sequential
batch reactors fed with synthetic sewage plus phenol was
amplified using conserved primers for the major subunit of
the phenol hydroxylase (LmPH) gene and used to generate
clone libraries. Following phylogenetic analysis, 59 sequences
containing a 470-bp fragment clustered into six
distinct subgroups with a genetic distance of 8%, most
likely representing ecologically relevant variants of the
enzyme. Seven sets of primers were designed to target the
six clusters and used to obtain quantitative information on
the dynamics of LmPH gene diversity using real-time PCR
assays throughout 9 months of bioreactors operation. Total
LmPH gene copy number remained approximately steady
in phenol-amended and control reactors. However, a
significant increase in phenol-degrading activity in the
phenol-amended sludge was accompanied by a parallel
increase in LmPH gene diversity, suggesting that phenol
degradation in the activated sludge depends on the
combined activity of a number of redundant species.
We describe a quantitative analysis of the genetic
diversity of phenol-degrading potential in bacterial communities
from laboratory-scale activated sludge. Genomic
DNA extracted from activated sludge from two sequential
batch reactors fed with synthetic sewage plus phenol was
amplified using conserved primers for the major subunit of
the phenol hydroxylase (LmPH) gene and used to generate
clone libraries. Following phylogenetic analysis, 59 sequences
containing a 470-bp fragment clustered into six
distinct subgroups with a genetic distance of 8%, most
likely representing ecologically relevant variants of the
enzyme. Seven sets of primers were designed to target the
six clusters and used to obtain quantitative information on
the dynamics of LmPH gene diversity using real-time PCR
assays throughout 9 months of bioreactors operation. Total
LmPH gene copy number remained approximately steady
in phenol-amended and control reactors. However, a
significant increase in phenol-degrading activity in the
phenol-amended sludge was accompanied by a parallel
increase in LmPH gene diversity, suggesting that phenol
degradation in the activated sludge depends on the
combined activity of a number of redundant species.