REVIEW: Recent Developments in the Biochemistry and Ecology of Enhanced Biological Phosphorus Removal

G. J. J. Kortstee^1*, K. J. Appeldoorn², C. F. C. Bonting¹, E. W. J. van Niel¹, and H. W. van Veen³

¹Laboratory of Microbiology, Department of Biomolecular Sciences, Wageningen University, Hesselink van Suchtelenweg 4, 6703 CT, Wageningen, The Netherlands; fax: +31-317-483829; E-mail: Gerard.Kortstee@Algemeen.MICR.WAU.NL

²BKH Consulting Engineers, Poortweg 10, 6212 PA, Delft, The Netherlands

³Department of Microbiology, University of Groningen, Kerklaan 30, 9751 NN, Haren, The Netherlands

^* To whom correspondence should be addressed.

Received December 27, 1999

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Most of the genes encoding the enzymes involved in polyP synthesis and degradation and in phosphate transport have been studied in various Gram-negative bacteria. Progress has also been made in studying the biochemical mechanisms underlying the process of enhanced biological phosphorus removal (EBPR), in particular in lab-scale systems fed with acetate or acetate plus glucose as the sole carbon and energy sources. By applying ¹³C-NMR, previous models concerning anaerobic carbon metabolism have been advanced and the role of glycogen in providing reducing equivalents in EBPR is definitely demonstrated. The role of the citric acid cycle in supplying reducing equivalents for the conversion of acetyl-CoA into poly-beta-hydroxybutyrate and poly-beta-hydroxyvalerate has been discussed. An incomplete citric acid cycle has been proposed to provide a small part of the reducing equivalents. Polyphosphate:AMP phosphotransferase and polyphosphatase were readily detectable in EBPR sludge fed with acetate plus glucose, but polyphosphate kinase remained undetected. In a lab-scale EBPR system, fed for several months with only acetate as carbon source, a Rhodocyclus-like bacterium (R6) was highly enriched and is therefore probably responsible for EBPR in systems fed with acetate only. This R6-type bacterium was however also present in other EBPR sludges (but to a lesser extent), and may therefore play an important role in EBPR in general. This organism accumulates polyhydroxyalkanoates anaerobically and polyP under aerobic conditions. Unlike members of the genus Rhodocyclus, bacterium R6 cannot grow phototrophically. Therefore a provisional new genus Candidatus and species Accumulibacter phosphatis was proposed.

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KEY WORDS: inorganic polyphosphates, enzymes, enhanced biological phosphorus removal, bacteria, ecology

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