Mining the landfill soil metagenome for denitrifying methanotrophic taxa and validation of methane oxidation in microcosm.

In the present study, the microbial community residing at different depths of the landfill was characterized to assess their roles in serving as a methane sink. Physico-chemical characterization revealed the characteristic signatures of anaerobic degradation of organic matter in the bottom soil (50-60 cm) and, active process of aerobic denitrification in the top soil (0-10 cm). This was also reflected from the higher abundance of bacterial domain in the top soil metagenome represented by dominant phyla Proteobacteria and Actinobacteria which are prime decomposers of organic matter in landfill soils. The multiple fold higher relative abundances of the two most abundant genera; Streptomyces and Intrasporangium in the top soil depicted greater denitrifying taxa in top soil than the bottom soil. Amongst the aerobic methanotrophs, the genera Methylomonas, Methylococcus, Methylocella, and Methylacidiphilum were abundantly found in the top soil metagenome that were essential for oxidizing methane generated in the landfill. On the other hand, the dominance of archaeal domain represented by Methanosarcina and Methanoculleus in the bottom soil highlighted the complete anaerobic digestion of organic components via acetoclasty, carboxydotrophy, hydrogenotrophy, methylotrophy. Functional characterization revealed a higher abundance of methane monooxygenase gene in the top soil and methyl coenzyme M reductase gene in the bottom soil that correlated with the higher relative abundance of aerobic methanotrophs in the top soil while methane generation being the active process in the highly anaerobic bottom soil in the landfill. The activity dependent abundance of endogenous microbial communities in the different zones of the landfill was further validated by microcosm studies in serum bottles which established the ability of the methanotrophic community for methane metabolism in the top soil and their potential to serve as sink for methane. The study provides a better understanding about the methanotrophs in correlation with their endogenous environment, so that these bacteria can be used in resolving the environmental issues related to methane and nitrogen management at landfill site.

Genomic characterization of denitrifying methylotrophic Pseudomonas aeruginosa strain AAK/M5 isolated from municipal solid waste landfill soil.

Municipal landfills are known for methane production and a source of nitrate pollution leading to various environmental issues. Therefore, this niche was selected for the isolation of one-carbon (C1) utilizing bacteria with denitrifying capacities using anaerobic enrichment on nitrate mineral salt medium supplemented with methanol as carbon source. Eight axenic cultures were isolated of which, isolate AAK/M5 demonstrated the highest methanol removal (73.28%) in terms of soluble chemical oxygen demand and methane removal (41.27%) at the expense of total nitrate removal of 100% and 33% respectively. The whole genome characterization with phylogenomic approach suggested that the strain AAK/M5 could be assigned to Pseudomonas aeruginosa with close neighbours as type strains DVT779, AES1M, W60856, and LES400. The circular genome annotation showed the presence of complete set of genes essential for methanol utilization and complete denitrification process. The study demonstrates the potential of P. aeruginosa strain AAK/M5 in catalysing methane oxidation thus serving as a methane sink vis-à-vis utilization of nitrate. Considering the existence of such bacteria at landfill site, the study highlights the need to develop strategies for their enrichment and designing of efficient catabolic activity for such environments.

Management of various organic fractions of municipal solid waste via recourse to VFA and biogas generation.

A hybrid anaerobic solid-liquid system was used for anaerobic digestion of organic fraction of municipal solid waste (OFMSW) consisting of mixed food + fruit waste and vegetable waste. Hydrolysis and acidogenesis potential of the above wastes were evaluated with the aim of producing value-added products in the form of volatile fatty acids (VFAs) and biogas recovery. Efficient hydrolysis and acidogenesis of mixed food + fruit waste was observed at a hydraulic retention time (HRT) of 1-3 d with a five-fold increase in soluble chemical oxygen demand (SCOD) followed by VFA production consisting of 50-75% acetic acid. Longer time was required for hydrolysis of vegetable waste with optimum hydrolysis and SCOD generation at 9 d HRT followed by VFA synthesis consisting of 45% acetic acid. Higher inoculum:substrate ratios resulted in improved hydrolysis and acidogenesis rates for vegetable waste in shorter time of 6 d with higher VFA production and increase in acetic acid content to 70%. When acidogenic leachate was fed into methanogenic reactors, detectable biogas production was observed after 25 d with 37-53% SCOD removal from leachate from mixed food + fruit waste and methane production of 0.066-0.1 L g(-1) SCOD removed and methane content of 38%. Though biogas yield from acidogenic leachate from vegetable waste was lower, nearly 94% volatile solids (VS) removal was observed in the reactors thereby providing methane yield of 0.13-0.21 L g(-1) VS consumed. Thus, the study provided a method for generation of value-added products from an otherwise misplaced resource in the form of OFMSW.

Annotating Multiple Prebiotic Synthesizing Capabilities Through Whole Genome Sequencing of Fusarium Strain HFK-74.

In the present study, seven fungal isolates from effluent treatment plants were screened for the production of prebiotic fructooligosaccharide synthesizing enzymes with the highest activity of fructofuranosidase (17.52 U/mL) and fructosyl transferase (18.92 U/mL) in strain HKF-74. Mining of genome sequence of strain revealed the annotation of genes providing multiple carbohydrate metabolizing capacities, such as amylases (AMY1), beta-galactosidase (BGAL), beta-xylosidase (Xyl), ß-fructofuranosidase (ScrB), fructosyltransferase (FTF), and maltose hydrolases (malH). The annotated genes were further supported by ß-galactosidase (15.90 U/mL), xylanase (17.91 U/mL), and α-amylase (14.05 U/mL) activities for synthesis of galactooligosaccharides, xylooligosaccarides, and maltooligosaccharides, respectively. In addition to genes encoding prebiotic synthesizing enzymes, four biosynthetic gene clusters (BGCs) including Type I polyketide synthase (PKS), non-ribosomal peptide synthetase (NRPS), NRPS-like, and terpene were also predicted in strain HKF-74. This was significant considering their potential role in pharmaceutical and therapeutic applications as well as in virulence. Accurate taxonomic assignment of strain HKF-74 by in silico genomic comparison indicated its closest identity to type strains Fusarium verticillioides NRRL 20984, and 7600. The average nucleotide identity (ANI) of strain HKF-74 with these strains was 92.5% which was close to the species threshold cut-off value (95-96%) while the DNA-DNA hybridization (DDH) value was 83-84% which was greater than both, species delineating (79-80%), and also sub-species delineating (70%) boundaries. Our findings provide a foundation for further research into the use of Fusarium strains and their prebiotic synthesizing enzymes for the development of novel prebiotic supplements.

Recent trends in the biotechnology of functional non-digestible oligosaccharides with prebiotic potential.

Prebiotics as a part of dietary nutrition can play a crucial role in structuring the composition and metabolic function of intestinal microbiota and can thus help in managing a clinical scenario by preventing diseases and/or improving health. Among the different prebiotics, non-digestible carbohydrates are molecules that selectively enrich a typical class of bacteria with probiotic potential. This review summarizes the current knowledge about the different aspects of prebiotics, such as its production, characterization and purification by various techniques, and its link to novel product development at an industrial scale for wide-scale use in diverse range of health management applications. Furthermore, the path to effective valorization of agricultural residues in prebiotic production has been elucidated. This review also discusses the recent developments in application of genomic tools in the area of prebiotics for providing new insights into the taxonomic characterization of gut microorganisms, and exploring their functional metabolic pathways for enzyme synthesis. However, the information regarding the cumulative effect of prebiotics with beneficial bacteria, their colonization and its direct influence through altered metabolic profile is still getting established. The future of this area lies in the designing of clinical condition specific functional foods taking into consideration the host genotypes, thus facilitating the creation of balanced and required metabolome and enabling to maintain the healthy status of the host.