新型异养硝化-好氧反硝化菌Paracoccus sp. QD-19的分离及脱氮特性研究

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

摘要/Abstract

摘要：

从沈阳市南部污水处理厂活性污泥中分离获得同时具备异养硝化和好氧反硝化能力的新型菌株，研究其脱氮特性，为改善污水厂的脱氮处理工艺奠定基础。对菌株进行形态学观察和16S rRNA基因鉴定；分别以NH₄Cl、NaNO₂、KNO₃为唯一氮源探究菌株的脱氮能力；以碳源、C/N比、pH值、温度、转速、接种量（V:V）等因素对菌株脱氮效果的影响进行研究。获得一株新型异养硝化-好氧反硝化菌株，经16S rRNA基因序列比对为副球菌属（Paracoccus），命名为Paracoccus sp. QD-19。菌株对初始氨氮浓度在300 mg/L以下的低浓度氨氮去除率能够达到100%，去除速率为8.707 mg/（L·h）且在脱氮过程中几乎没有亚硝态氮和硝态氮的积累。以亚硝态氮和硝态氮作为唯一氮源时，对此两种氮源的去除率36 h内均能达到99%，去除速率分别为4.944和5.666 mg/（L·h）。确定了去除氨氮的最佳脱氮条件：琥珀酸钠为碳源，C/N比为10，pH值为7，接种量（V:V）为1%，温度为30℃，转速为140 r/min。菌株Paracoccus sp. QD-19同时具有异养硝化和好氧反硝化的能力，并且具备不积累亚硝态氮和硝态氮的特性，为污水厂脱氮工艺的改进提供了实践基础。

关键词: 生物脱氮, 异养硝化, 同步硝化反硝化, 副球菌, 污水处理

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

张宇红, 董先博, 刘香宇, 许家琪, 徐子祾. 新型异养硝化-好氧反硝化菌Paracoccus sp. QD-19的分离及脱氮特性研究[J]. 生物技术通报, 2023, 39(3): 301-310.

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.

图/表 14

图1 菌株初筛脱氮效果

Fig. 1 Denitrification effects of strains in initial screening

图2 平板菌落形态和革兰氏染色 a：菌株QD-19平板菌落形态；b：金黄葡萄球菌革兰氏染色阳性对照；c：大肠杆菌革兰氏染色阴性对照；d：菌株QD-19革兰氏染色镜检图

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

图3 菌株QD-19的系统发育树

Fig. 3 Phylogenetic tree of the strain QD-19

图4 以氨氮为唯一氮源菌株的生长和氮的变化

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

图5 不同初始氨氮浓度下菌株的去除能力

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

表1 不同初始氨氮浓度下菌株的去除速率

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

图6 以亚硝态氮为唯一氮源菌株的生长和氮的变化

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

图7 以硝态氮为唯一氮源菌株的生长和氮的变化

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

图8 碳源对菌株生长和脱氮能力的影响

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

图9 C/N比对菌株生长和脱氮能力的影响

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

图10 pH值对菌株生长和脱氮能力的影响

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

图11 温度对菌株生长和脱氮能力的影响

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

图12 转速对菌株生长和脱氮能力的影响

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

图13 接种量对菌株生长和脱氮能力的影响

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

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