Isolation of a Novel Heterotrophic Nitrification-Aerobic Denitrification Bacterium Paracoccus sp. QD-19 and Its Characterization of Removing Nitrogen

doi:10.13560/j.cnki.biotech.bull.1985.2022-0834

Abstract

Abstract:

The study is aimed to isolate a novel strain with simultaneous heterotrophic nitrification and aerobic denitrification ability from the activated sludge of the south sewage treatment plant in Shenyang, and to evaluate the nitrogen-removing characteristics, which may lay a foundation for improving nitrogen removal process in wastewater facility. The strain Paracoccus sp. QD-19 was observed by morphological and identified by 16S rRNA gene sequencing. The nitrogen-removing capabilities of the strain were explored by NH₄Cl,NaNO₂, and KNO₃as sole nitrogen source. The effects of carbon source types, C/N ratio, pH value, temperature, shaking speed and inoculation amount（v:v）on the nitrogen removal were explored. A novel strain with heterotrophic nitrification-aerobic denitrification was obtained, identified as Paracoccus by 16S rRNA gene sequencing, and designated as that Paracoccus sp. QD-19. Results showed that 100% of ammonia nitrogen（below 300 mg/L）in wastewater was removed by the strain, and the removal rates achieved 8.707 mg/（L·h）. There was no nitrite accumulation in the nitrogen removal process. The stain Paracoccus sp. QD-19 removed 99% of nitrite or nitrate nitrogen in wastewater in 36 h, and the removal rates achieved 4.944 and 5.666 mg/（L·h）separately. The optimal conditions for ammonia nitrogen removal were: sodium succinate as carbon resource, 10（C/N ratio）, 7（pH value）, 1%（inoculation amount）, 30℃（temperature）, and 140 r/min（shaking speed）. These findings indicate that the strain Paracoccus sp. QD-19 possesses heterotrophic nitrification-aerobic denitrification capability, and the characterization without the accumulation of nitrite and nitrate nitrogen, which provides the practical basis for the improvement of nitrogen-removing process in the wastewater treatment plant.

Key words: biological denitrification, heterotrophic nitrification, simultaneous nitrification and denitrification, Paracoccus, wastewater treatment

ZHANG Yu-hong, DONG Xian-bo, LIU Xiang-yu, XU Jia-qi, XU Zi-ling. Isolation of a Novel Heterotrophic Nitrification-Aerobic Denitrification Bacterium Paracoccus sp. QD-19 and Its Characterization of Removing Nitrogen[J]. Biotechnology Bulletin, 2023, 39(3): 301-310.

Figures/Tables 14

Fig. 1 Denitrification effects of strains in initial screening

Fig. 2 Colony morphology on medium and gram stain a: Colony morphology of the strain QD-19 on medium b: The positive control for Gram staining of Staphylococcus aureus. c: The negative control for Gram staining of Escherichia coli. d: Gram staining results of the strain QD-19

Fig. 3 Phylogenetic tree of the strain QD-19

Fig. 4 Growth and nitrogen variations of strains with ammonia nitrogen as the only nitrogen source

Fig. 5 Removing ability of strains at different initial amm-onia nitrogen concentrations

Table 1 Removal rates of strains at different initial ammonia nitrogen concentrations

初始氨氮浓度Initial ammonia nitrogen concentration/（mg·L^-1）	24 h去除量Removal quantity at 24 h/（mg·L^-1）	72 h最高去除速率Maximum removal rate at 72 h/（mg·L^-1·h^-1）	72 h平均去除速率Average removal rate at 72 h（mg·L^-1·h^-1）
100	99.943	8.707	4.164
200	155.593±1.136	9.464	4.188
300	188.908±0.656	12.871	4.537±0.010
500	257.430±3.470	18.929	6.478±0.018
800	321.787±1.639	22.714	8.321±0.039
1 000	337.687±2.623	22.714	8.139±0.063

Fig. 6 Growth and nitrogen variations of strains with nit-rite nitrogen as the sole nitrogen source

Fig. 7 Growth and nitrogen variations of strains with nit-rate nitrogen as the sole nitrogen source

Fig. 8 Effects of carbon source on the growths and denitri-fication capacities of strains

Fig. 9 Effects of C/N ratio on the growth and denitrification capacity of strains

Fig. 10 Effects of pH value on the growths and denitrifica-tion capacities of strains

Fig. 11 Effects of temperature on the growth and denitri-fication capacity of strains

Fig. 12 Effects of rotational speed on the growths and denitrification capacities of strains

Fig. 13 Effects of inoculation amount on the growths and denitrification capacities of strains

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