耐盐异养硝化-好氧反硝化菌Bacillus megatherium N07的分离及脱氮特性

doi:10.13560/j.cnki.biotech.bull.1985.2021-1111

摘要/Abstract

摘要：

基于异养硝化-好氧反硝化菌的生物脱氮技术可有效去除污水中的氮素。高盐废水处理过程中存在抑制微生物生长、脱氮效率低等诸多短板。为解决高盐废水难脱氮问题,从农药厂化工污水中筛选出一株耐受8%（W/V）高盐浓度的异养硝化-好氧反硝化菌株,命名为N07。在不同盐度和不同溶氧条件下测定了菌株的氮素去除率,并探究了单因素环境因子对菌株脱氮性能的影响。结果表明：菌株N07鉴定为巨大芽孢杆菌（Bacillus megatherium）;能耐受8%盐度且繁殖旺盛,对氨氮和亚硝态氮去除率最高达到73.61%和83.56%,属于中度嗜盐菌;该菌在有氧条件下表现出高效的反硝化能力,12 h对硝态氮和亚硝态氮去除率为11.87 mg/（L· h）和8.26 mg/（L· h）;菌株在碳源为蔗糖、C/N为25、pH为7、转速160 r/min、温度为28℃时,硝化能力最佳。Bacillus megatheriumN07因能在高盐分环境降解氮素的特性,为异养硝化-好氧反硝化菌在沿海养殖业等高盐含氮污水处理的实际应用提供参考。

关键词: 异养硝化好氧反硝化, 生物脱氮, 高盐废水, 巨大芽孢杆菌

Abstract:

Biological nitrogen-removing technology based on heterotrophic nitrification and aerobic denitrification bacteria may effectively remove nitrogen in sewage. There are many shortcomings in the treatment of high salt wastewater,such as inhibiting microbial growth and low nitrogen removal efficiency. In order to solve the problem of difficult nitrogen removal from high salt wastewater,a strain of heterotrophic nitrification and aerobic denitrification with high salt concentration of 8%（W/V）was selected from chemical wastewater of agricultural pharmaceutical factory and named N07. The denitrification rate of the strain was measured under different salinity and dissolved oxygen conditions,and the effects of single environmental factors on the denitrification performance of the strain was investigated. The results showed that strain N07 was identified as Bacillus megatherium. It tolerated 8% salinity and reproduced vigorously. The removal rates of ammonia nitrogen and nitrite nitrogen were up to 73.61% and 83.56%,belonging to moderate halophilic bacteria. The bacterium showed high denitrification ability under aerobic conditions,with nitrate and nitrite removal rates of 11.87 mg/（L· h）and 8.26 mg/（L· h）at 12 h. The best nitrification capacity was obtained when carbon source was sucrose,C/N was 25,pH was 7,rotation speed was 160 r/min and temperature was 28℃. B. megatherium N07 can degrade nitrogen in high salinity environment,which provides reference for the practical application of heterotrophic nitrification and aerobic denitrification bacteria in the treatment of high salinity nitrogen wastewater such as coastal aquaculture industry.

Key words: heterotrophic nitrification aerobic denitrification, biological nitrogen removal, high salt wastewater, Bacillus megatherium

高宇轩, 靳静晨, 徐利杉, 高雅娟, 张闻天, 李晨晨, 张国伟, 靳永胜. 耐盐异养硝化-好氧反硝化菌Bacillus megatherium N07的分离及脱氮特性[J]. 生物技术通报, 2022, 38(7): 247-257.

GAO Yu-xuan, JIN Jing-chen, XU Li-shan, GAO Ya-Juan, ZHANG Wen-tian, LI Chen-chen, ZHANG Guo-wei, JIN Yong-sheng. Isolation of Halophilic Heterotrophic Nitrification-aerobic Denitrification Bacterium Bacillus megatherium N07 and Its Denitrification Characteristics[J]. Biotechnology Bulletin, 2022, 38(7): 247-257.

图/表 12

表1 三株反硝化菌脱氮性能比较

Table 1 Comparison of nitrogen removal performance of three denitrification strains

编号No.	氨氮Ammonia nitrogen		总氮Total nitrogen
编号No.	去除率 Removal rate/%	降解速率 Degradation rate/（mg·（L·h）^-1）	去除率 Removal rate/%	降解速率 Degradation rate/（mg·（L·h）^-1）
N01	55.53	4.63	33.78	2.81
N04	50.21	4.16	24.77	2.06
N07	70.95	5.91	63.84	5.33

图1 菌株N07的形态学鉴定 A：固体培养基的菌落形态;B：革兰氏染色结果;C：SEM扫描电镜菌体形态

Fig. 1 Morphological identification of strain N07 A：Colony morphology of solid medium. B：Gram staining results. C：SEM cell morphology

图2 菌株N07的系统发育树

Fig. 2 Phylogenetic tree of strain N07

图3 不同盐度下菌株N07的生长情况 A：培养基以氨氮为唯一氮源;B：培养基以亚硝态氮为唯一氮源。图中误差线表示标准差,不同字母表示存在显著性差异,P <0.05（n=3）,下同

Fig.3 Growth of strain N07 under different salinities A：Ammonia nitrogen was the only nitrogen source in the medium. B：Nitrite nitrogen was the only nitrogen source in the medium. The error line in the figure represents the standard deviation,different letters indicated significant difference,P <0.05（n=3）. The same below

图4 不同盐度下菌株N07的脱氮效果 A：培养基以氨氮为唯一氮源;B：培养基以亚硝态氮为唯一氮源

Fig. 4 Nitrogen removal effect of strain N07 at different salinities A：Ammonia nitrogen was the only nitrogen source in the medium. B：Nitrite nitrogen was the only nitrogen source in the medium

图5 不同溶氧条件下菌株N07降解硝态氮的性能 A：好氧条件;B：微厌氧条件

Fig. 5 Degradation performance of strain N07 under different DO A：Aerobic conditions;B：Micro-anaerobic conditions

图6 不同溶氧条件下菌株N07降解亚硝态氮的性能 A：好氧条件;B：微厌氧条件

Fig. 6 Degradation of nitrite nitrogen by strain N07 under different DO A：Aerobic conditions;B：Micro-anaerobic conditions

图7 碳源对菌株N07脱氮能力的影响

Fig.7 Effect of carbon source on the nitrification capacity of strain N07

图8 碳氮比对菌株N07脱氮能力的影响

Fig. 8 Effects of C/N ratio on the denitrification capacity of strain N07

图9 初始pH值对菌株N07脱氮能力的影响

Fig.9 Effect of initial pH value on the denitrification capacity of strain N07

图10 转速对菌株N07脱氮能力的影响

Fig.10 Effect of rotation speed on the denitrification capacity of strain N07

图11 温度对菌株N07脱氮能力的影响

Fig.11 Effect of temperature on the denitrification capacity of strain N07

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