Morris Rachel, Schauer-Gimenez Anne, Bhattad Ujwal, Kearney Colleen, Struble Craig A, Zitomer Daniel, Maki James S
Department of Biological Sciences, Marquette University, Milwaukee, WI, 53201-1881, USA.
Microb Biotechnol. 2014 Jan;7(1):77-84. doi: 10.1111/1751-7915.12094. Epub 2013 Oct 31.
Biologically produced methane (CH₄) from anaerobic digesters is a renewable alternative to fossil fuels, but digester failure can be a serious problem. Monitoring the microbial community within the digester could provide valuable information about process stability because this technology is dependent upon the metabolic processes of microorganisms. A healthy methanogenic community is critical for digester function and CH₄ production. Methanogens can be surveyed and monitored using genes and transcripts of mcrA, which encodes the α subunit of methyl coenzyme M reductase - the enzyme that catalyses the final step in methanogenesis. Using clone libraries and quantitative polymerase chain reaction, we compared the diversity and abundance of mcrA genes and transcripts in four different methanogenic hydrogen/CO₂ enrichment cultures to function, as measured by specific methanogenic activity (SMA) assays using H₂ /CO₂ . The mcrA gene copy number significantly correlated with CH₄ production rates using H₂ /CO₂ , while correlations between mcrA transcript number and SMA were not significant. The DNA and cDNA clone libraries from all enrichments were distinctive but community diversity also did not correlate with SMA. Although hydrogenotrophic methanogens dominated these enrichments, the results indicate that this methodology should be applicable to monitoring other methanogenic communities in anaerobic digesters. Ultimately, this could lead to the engineering of digester microbial communities to produce more CH₄ for use as renewable fuel.
厌氧消化器生物产生的甲烷(CH₄)是化石燃料的一种可再生替代品,但消化器故障可能是一个严重问题。监测消化器内的微生物群落可以提供有关过程稳定性的有价值信息,因为该技术依赖于微生物的代谢过程。健康的产甲烷群落对于消化器功能和CH₄产生至关重要。可以使用mcrA的基因和转录本对产甲烷菌进行调查和监测,mcrA编码甲基辅酶M还原酶的α亚基,该酶催化产甲烷作用的最后一步。我们使用克隆文库和定量聚合酶链反应,比较了四种不同的产甲烷氢/CO₂富集培养物中mcrA基因和转录本的多样性和丰度,以通过使用H₂ /CO₂的特定产甲烷活性(SMA)测定来衡量其功能。mcrA基因拷贝数与使用H₂ /CO₂的CH₄产生速率显著相关,而mcrA转录本数量与SMA之间的相关性不显著。所有富集物的DNA和cDNA克隆文库都具有独特性,但群落多样性也与SMA不相关。尽管氢营养型产甲烷菌在这些富集中占主导地位,但结果表明该方法应适用于监测厌氧消化器中的其他产甲烷群落。最终,这可能会导致对消化器微生物群落进行工程改造,以生产更多的CH₄用作可再生燃料。