Nitrate contamination of water environments can create serious problems such as eutrophication of rivers. Conventional biological processes for nitrate removal by heterotrophic denitrification often need additional organic substrates as carbon sources and electron donors. We tried to accelerate biological denitrification by using bioelectrochemical reactor (BER) in which electrode works as an electron donor. Denitrification activity of 8 environmental samples from various sediments, soils, groundwaters, and sludges were tested to establish an efficient enrichment culture for BER. The established enrichment culture from a soil sample showed stable denitrification activity without any nitrite accumulation. Microbial community analysis by using PCR-DGGE method revealed that dominant denitrifiers in the enrichment culture were Pantoea sp., Cronobacter sakazakii, and Castellaniella defragrans. Denitrification rate (0.08 kg/m3·day) of the enrichment culture in BER with electrode poised at -0.5 V (vs Ag/AgCl) was higher than that (2.1×10-2kg/m3·day) of BER without any poised potential. This results suggested that biological denitrification would be improved by supplying potential throughout electrode in BER. Further research using BER without any organic substrate addition is needed to apply this system for bioremediation of water and wastewater contaminated by nitrate.

From sludge of S municipal wastewater treatment plant in Busan, Korea, we isolated the denitrifier DN-9 which showed the ability of denitrification under aerobic condition by the color change and gas formation in liquid culture with Giltay medium. The isolated strain was identified as Pseudomonas sp. DN-9 on the basis of the morphological, physiological, biochemical and nucleotide sequence analysis of 16S rRNA. The isolated strain, Pseudomonas sp. DN-9, has cytochrome cd1 nirS of nitrite reductase. By the co-existance of additional ammonium and nitrate ion, the strain was not affected largely on growth in SL series broth. It seemed the result of denitrification. Although Pseudomonas sp. DN-9 has a good nitrate reduction activity under aerobic condition, the activity is less than Pseudomonas stutzeri in same cultivation condition. However, Escherichia coli had little the activity of aerobic denitrification and Pseudomonas putida showed lower activity of aerobic denitrification than Pseudomonas sp. DN-9 and Pseudomonas stutzeri in this study.

The activated sludge from the aeration basin of the Su-yeong municipal wastewater treatment plant which has operated by a standard activated sludge process in Busan, Korea was investigated during April 2004 and January 2005 with several bio-indicators. The number of bacteria and fungi per gram of dry weight of MLSS were estimated to be 3.1×106~1.5×108 and 1.1×103~1.1×105 colony forming units, respectively, by the plate agar method. By cultivation-independent methods, such as 4’,6-diamidino-2-phenylindole stain and fluorescence in situ hybridization, the ratio of eubacteria to the entire biomass was evaluated by more than 80% (v/v). The ratio of ammonia-oxidizing bacteria and nitrite-oxidizing bacteria to the total eubacteria was determined to be 7.0~9.8% and 3.3~6.2% without heavy variation in spite of a period of relatively low temperature in the basin. It would be expected that the nitrification would occur or at least co-exist throughout the year in the sludge of many municipal WWTP with influents that contain the sufficient nitrogen sources although the WWTP does not have any specialized processes for the removal of nitrogen.

A species of facultative photo-organotrophic, purple, non-sulfur bacterium was isolated from the 47 point at west and south coast of Korea in September 2001. Separated 13 samples of changes with red color under 28～32 ℃, 3000 lux, anaerobe conditions for 7 days cultivated in basal medium. For pure isolation from 13 samples, we used agar-shake tube method (0.4 % agar) and separated 5 strains through 13-repetition test. EGH-24 and EGH-30 was identified as the same strain through the RAPD(Random Amplified Polymorphic DNA)-PCR of strain EGH-9, EGH-13, EGH-23, EGH-24, EGH-30. Four isolates cultivated in synthesis wastewater for wastewater biodegradation test. EGH-24 was selected with efficient wastwater treating strain. Based on the results obtained from morphology, nutrient requirements, major bacteriochlorophyll content, 16S-rDNA phylogenetic analysis, EGH-24 strain may be identified as a new strain of the genus Rhodobacter and named Rhodobacter sp. EGH-24.

The isolated strain, Rhodococcus sp. EL-GT was able to degrade high phenol concentrations up to 10 mM within 24 hours in the medium consisting of 5.3 mM KH2PO4, 95 mM Na2HPO4, 18mM NH4NO3, 1mM MgSO4·7H2O, 50μM CaCl2, 0.5μM FeCl3, initial pH8.0, temperature 30℃ in rotary shaker at 200rpm. This strain was good cell growth and phenol degradation in the alkaline pH range range, and the highest in the pH range of 7 to 9. The microorganism was able to grow at the various chlorinated phenols, benzene, toluene, and bunker-C oil. As Rhodococcus sp. EL-GT was good capable of attachment on the acryl media, it would be used as microorganism to consist of biofilm in wastewater treatment.

The research was performed to compare to the biofilm characteristics and phenol removal efficiency in RBCs(Rotating Biological Contactor) using Rhodococcus sp. EL-GT(single population) and activated sludge(mixed population) as inoculum. Both reactors showed similar tendency on variations of dry weight, thickness and dry density of biofilm. However, the growth of biofilm thickness in 3 and 4 stage of single population reactor has sustained longer than that of the mixed population reactor. Unlike the mixed population reactor, the dry density of biofilm in the single population reactor had a difference between 1, 2 stage and 3, 4 stage. The single population reactor was stably operated without the decrease of phenol removal efficiency in the range of pH 6～9 and 15mM phenol was completely degraded in these pH ranges. But in case of the mixed population reactor, the phenol degradability was dramatically decreased at over 5mM phenol concentration because of the overgrowth and detachment of its biofilm.

A biosurfactant-producing microorganism was isolated from activated sludge by enrichment culture when grown on a minimal salt medium containing n-hexadecane as a sole carbon source. This microorganism was identified as Pseudomonas sp. and it was named Pseudomonas sp. EL-G527. It's optimal culture condition is 2% n-hexadecane, 0.2% NH4NO3, 0.3% KH2PO4, 0.3% K2HPO4, 0.02% MgSO4ㆍ7H2O, 0.0025% CaCl2ㆍ6H2O, 0.0015% FeSO4ㆍ7H2O in 1ℓ distilled water and initial pH 7.0. Cultivation was initiated with a 2% inoculum obtained from starter cultures grown in 30 ㎖ of the same medium in 250 ㎖ flask. They were cultivated at 30 ℃ in reciprocal shaking incubator and the highest biosurfactant production was observed after 4 days.