硫氰酸盐高效降解菌群的动力学及微生物多样性研究_汪流伟.pdf

2023，45（2）DOI：10.13836/j.jjau.2023043Acta Agriculturae Universitatis Jiangxiensishttp:/江西农业大学学报 硫氰酸盐高效降解菌群的动力学及微生物多样性研究汪流伟，肖小双，安雪姣，李宁健，谢东，张庆华*（江西农业大学 生物科学与工程学院，江西 南昌 330045）摘要：【目的】硫氰酸盐（SCN-）是一种常见的金矿、纺织、印染和焦化工业污染物，具有较强毒性和化学结构稳定性，其所造成的环境污染已成为近年来亟待解决的难题。【方法】在前期构建的一组高效硫氰酸盐降解复合菌群的基础上，探究在不同初始硫氰酸盐浓度下复合菌群对硫氰酸盐降解速率及复合菌群生长速率的影响，并对其进行动力学分析；根据细菌 16S rRNA 的序列结果进行群落结构组成分析；通过 OTUs、Chao1 指数、Shannon指数和Simpson指数对原始污泥和复合菌群进行多样性分析。【结果】（1）复合菌群降解硫氰酸盐及复合菌群生长符合底物抑制模型-Haldane 方程，硫氰酸盐最佳降解速率(Vopt)与硫氰酸盐浓度（Sopt）分别为0.137 85 g/（Lh）和0.121 26 g/L、复合菌群大生长速率（Vopt）与硫氰酸盐浓度（Sopt）分别为1.278 28 g/（Lh）和2.784 68 g/L。此外，复合菌群在30 h内可完全降解4 g/L硫氰酸盐，最高可耐受15 g/L的硫氰酸盐，培养6 d，降解率可达100%，生物量增加8.65 g/L。（2）该复合菌群细菌多样性及结构与原始污泥发生了变化，细菌多样性及结构在添加硫氰酸盐定向富集驯化后发生了改变，生物多样性在减少；活性污泥中的主要菌属有 Uncultured_bacterium_f_Blastocatellaceae、Alicycliphilus、uncultured_bacterium_f_NS9_marine_group、Diaphorobacter、uncultured_bacterium_f_Mitochondria、Thiobacillus，而复合菌菌群样品中主要的菌属有uncultured_bacterium_f_Saccharimonadaceae、uncultured_bacterium_f_Chitinophagaceae、uncultured_bacterium_f_NS9_marine_group、Dokdonella、Thiobacillus。其中Thiobacillus从活性污泥中的劣势菌属（丰度大约为2.69%）变为复合菌群中的优势菌属（丰度大约为77.73%），被认为是复合菌群降解硫氰酸盐的关键菌属。【结论】研究结果初步揭示了复合菌群降解硫氰酸盐规律，并解析了其微生物演变过程，挖掘了功能微生物，为后续硫氰酸盐废水的高效处置提供一定的研究基础和理论依据。关键词：复合菌群；硫氰酸盐；废水的高效处置；降解；动力学；微生物群落结构中图分类号：Q939.9 文献标志码：A 开放科学（资源服务）标识码（OSID）：文章编号：1000-2286（2023）02-0444-09收稿日期：20220818 修回日期：20221018基金项目：国家自然科学基金项目（32160027）、江西省自然科学基金重点项目（20212ACB205003）、江西省高层次高技能领军人才项目和江西省研究生创新专项资金（YC2022-S401）Project supported by the National Natural Science Foundation of China（32160027），the Key Project of Natural Science Foundation of Jiangxi Province，China（20212ACB205003），the High-level and High-skilled Leading Talents Training Project of Jiangxi Province，China and Innovation Fund for Postgraduate Students in Jiangxi Province（YC2022-S401）作者简介：汪流伟，硕士生，orcid.org/0000-0002-7994-6485，；*通信作者：张庆华，副教授，博士，主要从事有害物的生物降解与转化研究，orcid.org/0000-0003-1067-4343，zqh_。汪流伟，肖小双，安雪姣，等.硫氰酸盐高效降解菌群的动力学及微生物多样性研究 J.江西农业大学学报，2023，45（2）：444-452.WANG L W,XIAO X S,AN X J,et al.Studies on the kinetics and microbial diversity of a microbial consortium with highly efficient thiocyanate degradation abilityJ.Acta agriculturae universitatis Jiangxiensis,2023,45(2):444-452.第 2 期汪流伟等:硫氰酸盐高效降解菌群的动力学及微生物多样性研究Studies on the Kinetics and Microbial Diversity of Microbial Consortium with Highly Efficient Thiocyanate Degradation AbilityWANG Liuwei，XIAO Xiaoshuang，AN Xuejiao，LI Ningjian，XIE Dong，ZHANG Qinghua*（College of Bioscience and Biotechnology，Jiangxi Agricultural University，Nanchang 330045，China）Abstract：Objective As a common pollutant in gold mining，textile，printing and dyeing and coking industries，Thiocyanate（SCN-）has strong toxicity and chemical structure stability，and its environmental pollution has become an urgent problem to be solved in recent years.Method Based on a group of high-efficiency thiocyanate degradation microbial community constructed in the previous stage，the effects of the microbial community on thiocyanate degradation rate and growth rate of the microbial community under different initial thiocyanate concentrations were explored.Kinetic analysis was implemented.According to the sequence results of 16S rRNA of bacteria，the community structure was analyzed.OTUs，Chao1 index，Shannon index，and Simpson index were used to analyze the diversity of raw sludge and microbial community.Result（1）The degradation of thiocyanate and growth of the microbial community conformed to the substrate inhibition model-Haldane equation.The optimum degradation rate of thiocyanate and thiocyanate concentrations were 0.137 85 g/（Lh）and 0.121 26 g/L，respectively.The maximum growth rate of microbial community and thiocyanate concentrations were 1.278 28 g/（Lh）and 2.784 68 g/L，respectively.In addition，the microbial community could completely degrade 4 g/L thiocyanate within 30 hours，the maximum tolerance of the microbial community was 15 g/L thiocyanate，and the degradation rate reached 100%after 6 days of culture.Biomass was increased by 8.65 g/L.（2）The bacterial diversity and structure of the microbial community changed from those of the original sludge，the diversity and structure of bacteria were changed after thiocyanate enrichment and acclimation，and the biodiversity decreased again.Uncultured_bacterium_f_Blastocatellaceae，Alicycliphilus，uncultured_bacterium_f_NS9_marine_group，Diaphorobacter，uncultured_bacterium_f_Mitochondria，Thiobacillus are the main genera in the original sludge samples.Dokdonella，Thiobacillus，uncultured_bacterium_f_Saccharimonadaceae，uncultured_bacterium_f_NS9_marine_group，and uncultured_bacterium_f_Chitinophagaceae are the main genera in the microbial community samples.Among them，Thiobacillus changed from the inferior genus in activated sludge（the abundance was about 2.69%）to the dominant genus in microbial community（the abundance was about 77.73%），which was considered as the key genus for thiocyanate degradation by microbial community.Conclusion The research results preliminarily revealed the rule of thiocyanate degra