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| Content Provider | PubMed Central |
|---|---|
| Author | Mcmahon, Katherine D. Dojka, Michael A. Pace, Norman R. Jenkins, David Keasling, Jay D. |
| Copyright Year | 2002 |
| Abstract | A novel polyphosphate kinase (PPK) was retrieved from an uncultivated organism in activated sludge carrying out enhanced biological phosphorus removal (EBPR). Acetate-fed laboratory-scale sequencing batch reactors were used to maintain sludge with a high phosphorus content (approximately 11% of the biomass). PCR-based clone libraries of small subunit rRNA genes and fluorescent in situ hybridization (FISH) were used to verify that the sludge was enriched in Rhodocyclus-like β-Proteobacteria known to be associated with sludges carrying out EBPR. These organisms comprised approximately 80% of total bacteria in the sludge, as assessed by FISH. Degenerate PCR primers were designed to retrieve fragments of putative ppk genes from a pure culture of Rhodocyclus tenuis and from organisms in the sludge. Four novel ppk homologs were found in the sludge, and two of these (types I and II) shared a high degree of amino acid similarity with R. tenuis PPK (86 and 87% similarity, respectively). Dot blot analysis of total RNA extracted from sludge demonstrated that the Type I ppk mRNA was present, indicating that this gene is expressed during EBPR. Inverse PCR was used to obtain the full Type I sequence from sludge DNA, and a full-length PPK was cloned, overexpressed, and purified to near homogeneity. The purified PPK has a specific activity comparable to that of other PPKs, has a requirement for Mg2+, and does not appear to operate in reverse. PPK activity was found mainly in the particulate fraction of lysed sludge microorganisms. |
| Related Links | http://dx.doi.org/10.1128/aem.68.10.4971-4978.2002 |
| Ending Page | 4978 |
| Page Count | 8 |
| Starting Page | 4971 |
| File Format | |
| ISSN | 00992240 |
| e-ISSN | 10985336 |
| Journal | Applied and Environmental Microbiology |
| Issue Number | 10 |
| Volume Number | 68 |
| Language | English |
| Publisher | American Society for Microbiology |
| Publisher Date | 2002-10-01 |
| Access Restriction | Open |
| Rights Holder | American Society for Microbiology |
| Subject Keyword | Biotechnology Food Science Ecology Applied Microbiology and Biotechnology Research in Higher Education |
| Content Type | Text |
| Resource Type | Article |
| Subject | Ecology Food Science Applied Microbiology and Biotechnology Biotechnology |
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