NH3和H2S降解功能菌的分离鉴定及降解特性研究

doi:10.13560/j.cnki.biotech.bull.1985.2022-1202

摘要/Abstract

摘要：

为了减少厨余垃圾堆放时恶臭气体的排放，本研究利用富集驯化、定性初筛和定量复筛法从厨余垃圾中分离筛选到2株对NH₃和H₂S均具有高效降解作用的菌株CN5和CS2。通过16S rDNA序列分析，分别鉴定为肠球菌属（Enterococcuss sp.）和假单胞菌属（Pseudomonas sp.）。通过单因素实验，确定两株菌的最佳除臭条件为:培养温度为35℃、pH为7、碳氮比25∶1、碳源为蔗糖、氮源为氯化铵，对NH₃和H₂S的降解率均能达到85%左右。菌株CN5和CS2在2∶3的混合状态下生长状况最好，pH值稳定，OD₆₀₀最大值为1.342。通过氨氮含量和硫酸盐含量测定实验发现，最终氨氮含量为50.5 mg/L，菌剂对氨氮降解率为85.3%；硫酸盐含量为302.7 mg/L，菌剂对硫酸盐生成率为89.2%。结果表明，从厨余垃圾中筛选得到的2株菌对NH₃和H₂S具有良好的降解能力，且两菌株相互之间具有协同作用。

关键词: NH₃降解菌, H₂S降解菌, 筛选, 鉴定, 优化

Abstract:

In order to reduce the emission of malodorous gas during the stacking of kitchen waste, two strains CN5 and CS2 with high degradation of NH₃ and H₂S were isolated and screened from kitchen waste by enrichment and domestication, qualitative primary screening and quantitative secondary screening methods. They were identified as Enterococcus sp. and Pseudomonas sp. by 16S rDNA sequence analysis. The optimal deodorizing conditions of microbial deodorant were determined by single factor experiment: culture temperature was 35℃, pH was 7, carbon nitrogen ratio was 25∶1, carbon source was sucrose, nitrogen source was ammonium chloride, and the degradation rate of NH₃ and H₂S could be maintained at about 85%. Strains CN5 and CS2 grew best in the mixed state of 2∶3, and the maximum value of OD₆₀₀ was 1.342. The determination of ammonia nitrogen content and sulfate content, it was found that the final ammonia nitrogen content was 50.5 mg/L, and the degradation rate of bacteria to ammonia nitrogen was 85.3%. The sulfate content was 302.7 mg/L, and the sulfate production rate of bacterial agent was 89.2%. The results showed that the two strains screened from the kitchen waste had good ability to degrading NH₃ and H₂S, and the two strains had synergistic effect with each other.

Key words: NH₃ degrading bacteria, H₂S degrading bacteria, screening, identification, optimization

江海溶, 崔若琪, 王玥, 白淼, 张明露, 任连海. NH₃和H₂S降解功能菌的分离鉴定及降解特性研究[J]. 生物技术通报, 2023, 39(9): 246-254.

JIANG Hai-rong, CUI Ruo-qi, WANG Yue BAI, Miao ZHANG, Ming-lu , REN Lian-hai. Isolation, Identification and Degradation Characteristics of Functional Bacteria for NH₃ and H₂S Degradation[J]. Biotechnology Bulletin, 2023, 39(9): 246-254.

图/表 10

表1 除臭条件优化实验设计

Table 1 Experiment design of optimal deodorizing conditions

测试项目Test item	接种量Inoculation amount/%
测试项目Test item	5	10	15	20	—	—	—
pH	6.0	6.5	7.0	7.5	8.0	8.5	—
C/N	1∶15	1∶20	1∶25	1∶30	1∶35	—	—
碳源 Carbon source	无水葡萄糖 Anhydrous glucose	柠檬酸钠 Sodium citrate	乙酸钠 Sodium acetate	蔗糖 Sucrose	丁二酸钠 Sodium succinate	—	—
氮源 Nitrogen source	尿素 Carbamide	氯化铵 Ammonium chloride	硫酸铵 Ammonium sulphate	酵母提取物 Yeast extract	蛋白胨 Peptone	硝酸钾 Potassium nitrate	亚硝酸 Nitrous acid

表2 筛选结果及其降解率

Table 2 Screening results and degradation rates

NH₃降解菌株编号NH₃ degrading strain	降解率Degradation rate/%	H₂S降解菌株编号H₂S degrading strain	降解率Degradation rate/%
CN10	71.72	CS2	70.66
CN5	77.75	CS4	62.30
PN1	65.75	CS6	66.90
PN3	67.77	CS7	60.32

图1 菌株对氨氮（A）和硫酸盐（B）的转化情况

Fig. 1 Transformation of ammonia nitrogen（A）and sulfate（B）by strains

图2 菌株的显微镜形态特征（10×100）

Fig. 2 Morphological characteristics of strains（10×100） under microscope

图3 菌株CN5和CS2的系统发育进化树

Fig. 3 Phylogenetic tree derived from 16S rDNA sequences of the CN5 and CS2 strains

图4 菌株在不同温度培养时的OD600值 A:菌株CN5的OD600值；B: 菌株CS2的OD600值

Fig. 4 OD600 value of strains cultured at different temperatures A: OD600 value of strain CN5; B: OD600 value of strain CS2

图5 菌株CN5和菌株CS2在不同培养条件下降解率的变化

Fig. 5 Changes in degradation rates of strain CN5 and strain CS2 under different culture conditions

图6 两菌株不同混合比例下OD600的变化

Fig. 6 Changes of OD600 under different mixing ratios of two strains

图7 两菌株不同混合比例下pH的变化

Fig. 7 Changes of pH under different mixing ratios of two strains

图8 两株菌混合条件下氨氮和硫酸盐离子浓度变化曲线

Fig. 8 Variation curve of ammonium nitrogen and sulfate ion concentration under mixed conditions of two strains

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NH₃和H₂S降解功能菌的分离鉴定及降解特性研究

Isolation, Identification and Degradation Characteristics of Functional Bacteria for NH₃ and H₂S Degradation

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