有机物甲烷化可通过乙酸发酵型、氢营养型和甲基营养型3种途径进行,确定各个途径对甲烷生成的贡献率是其动力学研究的基本问题,稳定同位素示踪技术是研究生态体系甲烷主导生成途径的创新方法。本文综述了稳定同位素技术表征甲烷生成途径的方法进展及影响测试解析结果的因素。产甲烷过程中的稳定同位素分馏效应是影响测试结果解析的关键,甲烷菌类型、生长阶段、底物丰度、温度等是其主要影响因素;对典型产甲烷生态体系进行控制实验,积累分馏效应数据及联合应用微生态分子生物学表征方法,是提高稳定同位素技术对甲烷生成途径区分水平的可行方法。联合应用稳定同位素表征技术和微生态原位表征方法,对高浓度有机酸胁迫条件下的生物质类有机物厌氧发酵甲烷化途径的研究结果表明,面临高浓度有机酸迅速累积的环境,中温发酵条件下,优势菌种为甲烷八叠球菌,依时间次序,通过乙酸发酵和氢营养型途径利用累积的有机酸产生甲烷;而在高温条件下,有机酸则通过乙酸氧化和氢营养型甲烷化途径的串联反应被降解。运用稳定同位素表征技术揭示甲烷生成途径可为针对性的微生态调控提供依据。
Methane is biologically produced from biomass organics through three pathways, i. e. acetoclastic, hydrogenotrophic and methylotrophic methanogenesis. The methanogenic pathways can be characterized and quantified by stable isotope probing (SIP). The quantification of methanogenesis kinetics using SIP is critically determined by carbon isotope fractionation, which is affected by the types and growth phase of methanogens, substrate availability and temperature, etc. Therefore, the accurate quantification should lie on more fractionation data and the combination with molecular biology. A combination analysis of SIP and in situ microbiological methods was applied on the anaerobic digestion of biomass organics suffering from severe acids inhibition. The studies found that accumulated acids were degraded mainly by Methanosarcina firstly with acetoclastic pathway and then with hydrogenotrophic pathway under mesophilic condition, whereas acids were mainly degraded to methane by the tandem reactions of syntrophic acetate oxidation and hydrogenotrophic methanogenesis under thermophilic condition. Thereby, the microbial ecology involved in the methane production can be regulated effectively with the SIP information.