Insights into the microbial community structure of anaerobic digestion of municipal solid waste landfill leachate for methane production by adaptive thermophilic granular sludge

BACKGROUND: The amount of municipal solid waste (MSW) gradually increased along with the rapid development of modern cities. A large amount of landfill leachate are generated with excessive chemical oxygen demand (COD), which create a great deal of pressure on the environment-friendly treatment process. Anaerobic digestion is an ideal technique to solve the above problem. RESULTS: A thermophilic granular sludge was successfully adapted for anaerobic digestion of MSW leachate (from an aging large-scale landfill) for methane production. The COD degradation efficiency improved by 81.8%, while the methane production rate reached 117.3 mL CH4/(g VS d), which was 2.34-fold more than the control condition. The bacterial and archaeal communities involved in the process were revealed by 16S rRNA gene high-throughput pyrosequencing. The richness of the bacterial community decreased in the process of thermophilic granular sludge, while the archaeal community structure presented a reverse phenomenon. The bacterial genus, Methanosaeta was the most abundant during the mesophilic process, while Methanobacterium, Methanoculleus, Methanosaeta and Methanosarcina were more evenly distributed. The more balanced community distribution between hydrogenotrophic and acetotrophic methanogens implied a closer interaction between the microbes, which further contributed to higher methane productivity. The detailed relationship between the key functional communities and anaerobic digestion performances were demonstrated via the multivariate canonical correspondence analysis. Conclusions: With the assistance of adaptive thermophilic granular sludge, microbial community structure was more evenly distributed, while both of COD degradation rate and methane production was improved during anaerobic digestion of MSW landfill leachate.

A mini-IRES sequence for stringent selection of high producers.

Internal Ribosome Entry Site (IRES) sequences have been widely used to link the expression of two independent proteins on the same mRNA transcript. Genes encoding fluorescent proteins or drug-resistance enzymes are usually placed downstream of IRES, serving as expression indicators or selection markers. In biological applications where the upstream gene-of-interest is to be expressed at extremely high levels, it is often desirable to purposely reduce IRES downstream gene expression to economize the cellular resources and/or to generate more stringent selection pressure. Here we describe a miniature IRES mutant sequence (IRESmut3) with dramatically diminished co-translational efficiency to fulfill these purposes.