Identification and Denitrification Characters of a High-temperature-resistant Nitrite-denitrifying Bacterium

doi:10.13560/j.cnki.biotech.bull.1985.2019-0130

Abstract

Abstract: Due to the toxic effect of nitrite,the screening and denitrification characteristics of denitrifying bacteria suitable for removing high-concentration nitrite-nitrogen are of great significance. Enrichment culturing,preliminary screening with plates and secondary screening with flasks were used to screen denitrifying bacteria from aquaculture wastewater,and a strain Y7 was identified through the physiological and biochemical characterization and the 16S rRNA gene sequence analysis. The effects of carbon sources,nitrite concentration,pH,liquid volume,inoculum size,inoculum age and temperature to the removal rate of nitrite-nitrogen by the strain Y7 were further investigated for increasing the efficacy of removing nitrite-nitrogen. As results,strain Y7 was identified as a member of the genus Bacillus and hence designated Bacillus subtilis Y7. Results of single-factor experiment showed that strain Y7 degraded nitrite from 1 100 mg/L to zero after fermentation for 24 hours at temperature 30℃ and 170 r/min,under the conditions of the glycerol as carbon source,the pH value of the medium being 7.0,the liquid volume being 100 mL in 250 mL triangular flask and the inoculum size being 10%（OD₆₀₀=1.5）. In conclusion,strain Y7 tolerates up to 1 100 mg/L nitrite,and the high temperature of 50℃ does not affect its degradation efficiency to the nitrite-nitrogen. Therefore,strain Y7 has great potential in the treatment of high-temperature industrial wastewater containing high concentration of nitrite and summer aquaculture wastewater.

Key words: Bacillus, nitrite, denitrifying bacteria, high temperature

LI Yu-qi, MA Pei-yu, LIU Han, WANG Ling-zhi, GAO Ren-ling, LI Hui-juan. Identification and Denitrification Characters of a High-temperature-resistant Nitrite-denitrifying Bacterium[J]. Biotechnology Bulletin, 2019, 35(9): 194-201.

References

[1] Jensen FB, Hansen MN.Differential uptake and metabolism of nitrite in normoxic and hypoxic goldfish[J]. Aquatic Toxicology, 2011, 101(2):318-325.
[2] Özen H, Kamber U, Karaman M, et al.Histopathologic, biochemical and genotoxic investigations on chronic sodium nitrite toxicity mice[J]. Experimental and Toxicologic Pathology, 2014, 66(8):367-375.
[3] Guo LJ, Zhao B, An Q, et al.Characteristics of a novel aerobic denitrifying bacterium, Enterobacter cloacaestrain HNR[J]. Applied Biochemistry and Biotechnology, 2016, 178:947-959.
[4] 何腾霞, 李振轮, 杨珊. 好氧反硝化细菌及其去除亚硝酸盐的作用机理研究进展[J]. 过程工程学报, 2015, 15(1):174-180.
[5] Ji B, Yang K, Zhu L, et al.Aerobic denitrification:a review of important advances of the last 30 years[J]. Biotechnology and Bioprocess Engineering, 2015, 20:643-651.
[6] Zhao B, Cheng DY, Tan P, et al.Characterization of an aerobic denitrifier Pseudomonas stutzeri strain XL-2 to achieve efficient nitrate removal[J]. Bioresource Technology, 2018, 250:564-573.
[7] 郭端强, 刘海龙, 万亚涛, 等. 一株好氧反硝化细菌的分离鉴定及反硝化特性研究[J]. 生物技术通报, 2012, 28(10):205-209.
[8] Song ZF, An J, Fu GH, et al.Isolation and characterization of an aerobic denitrifying Bacillus sp. YX-6 from shrimp culture ponds[J]. Aquaculture, 2011, 319:188-193.
[9] Liu Y, Ai GM, Miao LL, et al.Marinobacter strain NNA5, a newly isolated and highly efficient aerobic denitrifier with zero N₂O emission[J]. Bioresource Technology, 2016, 206:9-15.
[10] Lei X, Jia YT, Chen YC, et al.Simultaneous nitrification and denitrification without nitrite accumulation by a novel isolated Ochrobactrum anthropic LJ81[J]. Bioresource Technology, 2019, 272:442-450.
[11] 张苗, 黄少斌. 高温好氧反硝化菌的分离鉴定及其反硝化性能研究[J]. 环境科学, 2011, 32(1):259-265.
[12] 赵惊鸿, 黄少斌. 一株耐高温好氧反硝化菌的筛选及特性研究[J]. 环境科学与技术, 2015, 38(1):6-10.
[13] 郝敏娜, 杨云龙, 葛启隆. 高温短程反硝化菌Brevibacillus sp. XF-03 特性及其降解动力学[J]. 生物技术通报, 2014, 30(1):177-181.
[14] Gao JQ, Gao D, Liu H, et al.Biopotentiality of high efficient aerobic denitrifier Bacillus megaterium S379 for intensive aquaculture water quality management[J]. Journal of Environmental Management, 2018, 222:104-111.
[15] 国家环保总局水和废水检测分析方法编委会. 水和废水分析监测方法[M]. 北京:中国环境科学出版社, 2002.
[16] 东秀珠, 蔡妙英. 常见细菌系统鉴定手册[M]. 北京:科学出版社, 2001.
[17] Zhao B, He YL, Huang J, et al.Heterotrophic nitrogen removal by Providencia rettgeri strain YL[J]. Journal of Industrial Microbiology and Biotechnology, 2010, 37:609-616.
[18] Zheng HY, Liu Y, Sun GD, et al.Denitrification characteristics of a marine origin psychrophilic aerobic denitrifying bacterium[J]. Journal of Environmental Sciences, 2011, 23(11):1888-1893.
[19] Chen Q, Ni J.Ammonium removal by Agrobacterium sp. LAD9 capable of heterotrophic nitrification-aerobic denitrification[J]. Journal of Bioscience and Bioengineering, 2012, 113(5):619-623.
[20] 何腾霞, 李振轮. 耐冷好氧亚硝酸盐型反硝化细菌的鉴定及脱氮特性研究[J]. 生物技术通报, 2015, 31(10):191-198.
[21] 李彦芹, 李春青, 管鹏悦, 等. 一株好氧反硝化菌的分离鉴定及脱氮条件研究[J]. 环境科学与技术, 2015, 38(7):33-37.
[22] 连红民, 邱忠平, 何昆明, 等. 一株好氧反硝化-异养硝化菌的筛选及脱氮特性研究[J]. 生物技术通报, 2015, 31(6):138-143.
[23] 傅利剑, 郭丹钊, 史春龙, 等. 碳源及碳氮比对异养反硝化微生物异养反硝化作用的影响[J]. 农村生态环境, 2005, 21(2):42-45.
[24] He TX, Li ZL, Sun Q, et al.Heterotrophic nitrification and aerobic denitrification by Pseudomonas tolaasii Y-11 without nitrite accumulation during nitrogen conversion[J]. Bioresource Technology. 2016, 200:493-499.