通过富集分离的方法,从香料废水中获得一株具有异养硝化-好氧反硝化功能的菌株SVT,经菌落形态以及16S rDNA分析鉴定为陶厄氏菌Thauera butanivorans SVT.提取菌株基因组DNA并进行全基因组测序,分析和预测其氮代谢通路;以100mg/L的NH4+-N、NO3--N或NO2--N为唯一氮源研究了菌株SVT的氮代谢特征,并研究了不同单一碳源(无水乙酸钠、柠檬酸钠、琥珀酸钠、蔗糖和葡萄糖)、C/N比(1、5、10、15、30)温度(10、20、30、40和50℃)和pH(4.0、6.5、7.5、8.5、10.0)对该菌株异养硝化-好氧反硝化性能的影响.结果表明,该菌株以NH4+-N、NO3--N和NO2--N为唯一氮源时,48h内的TN去除率分别为(86.96±1.03)%、(60.52±4.70)%和(64.32±1.40)%;菌株SVT在所设置的不同碳源、C/N比、温度和pH值条件下均能够生长和异养硝化-好氧反硝化脱氮,最适条件为以无水乙酸钠为碳源,温度30℃,C/N=10,pH值为7.5.通过基因组测序,发现菌株SVT含有6种氮代谢相关功能酶的基因(napAB、nxrAB、nirBD、nirS、norBC和nosZ),结合KEGG数据库比对和单一氮源氮代谢特征结果推测其氮代谢途径包括:NH4+-N同化途径(NH4+-N→L-谷氨酸盐→L-谷氨酸)、硝化途径(NH4+-N→NO2--N→NO3--N)、反硝化途径(NO3--N→NO2--N→NO→N2O→N2)和DNRA途径(NO3--N→NO2--N→NH4+-N).研究表明菌株SVT具有高效的异养硝化-好氧反硝化能力,在废水处理中具有广阔的应用前景.
Abstract
A heterotrophic nitrifying-aerobic denitrifying bacterium, identified as Thauera butanivorans SVT, was isolated from spice wastewater. Genomic DNA of the isolate was extracted for whole-genome sequencing to analyze and predict nitrogen metabolic pathways. The nitrogen metabolism characteristics were investigated using 100mg/L NH4+-N, NO3--N, or NO2--N as sole nitrogen sources. The effects of different carbon sources (sodium acetate anhydrous, sodium citrate, sodium succinate, sucrose, and glucose), C/N ratios (1, 5, 10, 15, 30), temperatures (10°C, 20°C, 30°C, 40°C, 50°C), and pH levels (4.0, 6.5, 7.5, 8.5, 10.0) on its heterotrophic nitrification-aerobic denitrification performance were systematically investigated. Results showed total nitrogen removal rates of (86.96±1.03)%, (60.52±4.70)%, and (64.32±1.40)% within 48hours when using NH4+-N, NO3--N, and NO2--N as sole nitrogen sources, respectively. Strain SVT demonstrated growth and nitrogen removal capabilities under all tested conditions, with optimal performance achieved using sodium acetate anhydrous as carbon source at 30°C, C/N=10, and pH 7.5. Genomic analysis identified six nitrogen metabolism-related functional enzyme genes (napAB, nxrAB, nirBD, nirS, norBC, and nosZ). Combined with KEGG database alignment and single nitrogen source metabolism results, the proposed nitrogen metabolic pathways include: ammonium assimilation (NH4+-N→L-glutamate→L-glutamine), nitrification (NH4+-N→NO2--N→NO3--N), denitrification (NO3--N→NO2--N→NO→N2O→N2), and DNRA pathway (NO3--N→NO2--N→NH4+-N). This study demonstrates that strain SVT exhibits efficient heterotrophic nitrification-aerobic denitrification capabilities, showing promising potential for wastewater treatment applications.
关键词
陶厄氏菌 /
异养硝化-好氧反硝化 /
脱氮 /
基因组测序 /
氮代谢途径
Key words
Thauera /
heterotrophic nitrification-aerobic denitrification /
genomic sequencing /
nitrogen removal /
nitrogen metabolic pathway
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基金
国家自然科学基金资助项目(51579092);中国能源建设股份有限公司重大科技项目(CEEC2023-ZDYF-09)
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