Lactic acid bacteria-malted vinegar: fermentation characteristics and anti-hyperlipidemic effect.

In this study, the fermentation characteristics and functional properties of lactic acid bacteria-malted vinegar (LAB-MV) were investigated during the fermentation period. Changes in the components (organic acids, free sugars, free amino acids, ß-glucan, and gamma-aminobutyric acid (GABA)) of MV (BWAF0d, BWAF10d, BWAF20d) and LAB-MV (LBWAF0d, LBWAF10d, LBWAF20d) were analyzed according to the fermentation time. The amounts of ß-glucan and GABA in LBWAF20d were greater than those in BWAF20d (122.00 µg/mL, 83.06 µg/mL and 531.00 µg/mL, 181.31 µg/mL, respectively). The ACE1 and HMG-CoA reductase inhibitory activities of LBWAF20d were 98.16% (1/20 dilution factor, DF) and 91.01% (1/25 DF), respectively. The lipid accumulation ratio and total cholesterol levels in HepG2 cells treated with LBWAF20d (1/200 DF) were reduced by 45.85% and 54.48%, respectively, compared to those in the untreated group. These results suggest that LAB-MV, which comprises barley wine manufactured from LAB and yeast, may improve hepatic lipid metabolism.

Long-term performance and bacterial community dynamics in biocovers for mitigating methane and malodorous gases.

The long-term performance of lab-scale biocovers for the simulation of engineered landfill cover soils was evaluated. Methane (CH4), trimethylamine (TMA), and dimethyl sulfide (DMS) were introduced into the biocovers as landfill gases for 134 days and the removal performance was evaluated. The biocover systems were capable of simultaneously removing methane, TMA, and DMS. Methane was mostly eliminated in the top layer of the systems, while TMA and DMS were removed in the bottom layer. Overall, the methane removal capacity and efficiency were 224.8±55.6g-CH4m-2d-1 and 66.6±12.8%, respectively, whereas 100% removal efficiencies of both TMA and DMS were achieved. Using quantitative PCR and pyrosequencing assay, the bacterial and methanotrophic communities in the top and bottom layers were analyzed along with the removal performance of landfill gases in the biocovers. The top and bottom soil layers possessed distinct communities from the original inoculum, but their structure dynamics were different from each other. While the structures of the bacterial and methanotrophic communities showed little change in the top layer, both communities in the bottom layer were considerably shifted by adding TMA and DMA. These findings provide information that can extend the understanding of full-scale biocover performance in landfills.

Inhibitory effects of sulfur compounds on methane oxidation by a methane-oxidizing consortium.

Kinetic and enzymatic inhibition experiments were performed to investigate the effects of methanethiol (MT) and hydrogen sulfide (H2S) on methane oxidation by a methane-oxidizing consortium. In the coexistence of MT and H2S, the oxidation of methane was delayed until MT and H2S were completely degraded. MT and H2S could be degraded, both with and without methane. The kinetic analysis revealed that the methane-oxidizing consortium showed a maximum methane oxidation rate (Vmax) of 3.7 mmol g-dry cell weight (DCW)(-1) h(-1) and a saturation constant (Km) of 184.1 µM. MT and H2S show competitive inhibition on methane oxidation, with inhibition values (Ki) of 1504.8 and 359.8 µM, respectively. MT was primary removed by particulate methane monooxygenases (pMMO) of the consortium, while H2S was degraded by the other microorganisms or enzymes in the consortium. DNA and mRNA transcript levels of the pmoA gene expressions were decreased to ∼10(6) and 10(3)pmoA gene copy number g-DCW(-1) after MT and H2S degradation, respectively; however, both the amount of the DNA and mRNA transcript recovered their initial levels of ∼10(7) and 10(5)pmoA gene copy number g-DCW(-1) after methane oxidation, respectively. The gene expression results indicate that the pmoA gene could be rapidly reproducible after methane oxidation. This study provides comprehensive information of kinetic interactions between methane and sulfur compounds.

Depth profiles of methane oxidation potentials and methanotrophic community in a lab-scale biocover.

The depth profiles of the CH4 oxidation potentials and the methanotrophic community were characterized in a lab-scale soil mixture biocover. The soil mixture samples were collected from the top (0-10cm), middle (10-40cm), and bottom (40-50cm) layers of the biocover where most of methane was oxidized at the top layer due to consumption of O2. Batch tests using serum bottles showed that the middle and bottom samples displayed CH4 oxidation activity under aerobic conditions, and their CH4 oxidation rates were 85 and 71% of the rate of top sample (8.40µmolgdry sample(-1)h(-1)), respectively. The numbers of methanotrophs in the middle and bottom were not significantly different from those in the top sample. There was no statistical difference in the community stability indices (diversity and evenness) among the methanotrophic communities of the three layer samples, even though the community structures were distinguished from each other. Based on microarray analysis, type I and type II methanotrophs were equally present in the top sample, while type I was more dominant than type II in the middle and bottom samples. We suggested that the qualitative difference in the community structures was probably caused by the difference in the depth profiles of the CH4 and O2 concentrations. The results for the CH4 oxidation potential, methanotrophic biomass, and community stability indices in the middle and bottom layer samples indicated that the deeper layer in the methanotrophic biocover serves as a bioresource reservoir for sustainable CH4 mitigation.

Tobermolite effects on methane removal activity and microbial community of a lab-scale soil biocover.

Three identical lab-scale biocovers were packed with an engineered soil (BC 1), tobermolite only (BC 2), and a mixture of the soil and tobermolite (BC 3), and were operated at an inlet load of 338-400 g-CH4 m(-2) d(-1) and a space velocity of 0.12 h(-1). The methane removal capacity was 293 ± 47 g-CH4 m(-2) d(-1) in steady state in the BC 3, which was significantly higher than those in the BC 1 and BC 2 (106 ± 24 and 114 ± 48 g-CH4 m(-2) d(-1), respectively). Quantitative PCR indicated that bacterial and methanotrophic densities (6.62-6.78 × 10(7) 16S rDNA gene copy number g-dry sample(-1) and 1.37-2.23 × 10(7) pmoA gene copy number g-dry sample(-1) in the BC 1 and BC 3, respectively) were significantly higher than those in the BC 2. Ribosomal tag pyrosequencing showed that methanotrophs comprised approximately 60 % of the bacterial community in the BC 2 and BC 3, while they only comprised 43 % in the BC 1. The engineered soil favored the growth of total bacteria including methanotrophs, while the presence of tobermolite enhanced the relative abundance of methanotrophs, resulting in an improved habitat for methanotrophs as well as greater methane mitigation performance in the mixture. Moreover, a batch experiment indicated that the soil and tobermolite mixture could display a stable methane oxidation level over wide temperature (20-40 °C, at least 38 µmol g-dry sample(-1) h(-1)) and pH (5-8, at least 61 µmol g-dry sample(-1) h(-1)) ranges. In conclusion, the soil and tobermolite mixture is promising for methane mitigation.

The presence of significant methylotrophic population in biological activated carbon of a full-scale drinking water plant.

Methylotrophs within biological activated carbon (BAC) systems have not received attention although they are a valuable biological resource for degradation of organic pollutants. In this study, methylotrophic populations were monitored for four consecutive seasons in BAC of an actual drinking water plant, using ribosomal tag pyrosequencing. Methylotrophs constituted up to 5.6% of the bacterial community, and the methanotrophs Methylosoma and Methylobacter were most abundant. Community comparison showed that the temperature was an important factor affecting community composition, since it had an impact on the growth of particular methylotrophic genera. These results demonstrated that BAC possesses a substantial methylotrophic activity and harbors the relevant microbes.

Comparison of RNA- and DNA-based bacterial communities in a lab-scale methane-degrading biocover.

Methanotrophs must become established and active in a landfill biocover for successful methane oxidation. A lab-scale biocover with a soil mixture was operated for removal of methane and nonmethane volatile organic compounds, such as dimethyl sulfide (DMS), benzene (B), and toluene (T). The methane elimination capacity was 211±40 g m(-2) d(-1) at inlet loads of 330-516 g m(-2) d(-1). DMS, B, and T were completely removed at the bottom layer (40-50 cm) with inlet loads of 221.6±92.2, 99.6±19.5, and 23.4±4.9 mg m(-2) d(-1), respectively. The bacterial community was examined based on DNA and RNA using ribosomal tag pyrosequencing. Interestingly, methanotrophs comprised 80% of the active community (RNA) while 29% of the counterpart (DNA). Types I and II methanotrophs equally contributed to methane oxidation, and Methylobacter, Methylocaldum, and Methylocystis were dominant in both communities. The DNA vs. RNA comparison suggests that DNA-based analysis alone can lead to a significant underestimation of active members.

Reduction of perchlorate by salt tolerant bacterial consortia.

Two perchlorate-reducing bacterial consortia (PRBC) were obtained by enrichment cultures from polluted marine sediments. Non-salt-tolerant PRBC (N-PRBC) was enriched without the addition of NaCl, and salt tolerant-PRBC (ST-PRBC) was enriched with 30 g-NaCl L(-1). Although the perchlorate reduction rates decreased with increasing NaCl concentration, ST-PRBC (resp., N-PRBC) could reduce perchlorate until 75 g-NaCl L(-1) (resp., 30 g-NaCl L(-1)). The reduction yield (1.34±0.05 mg-perchlorate per mg-acetate) and maximum perchlorate reduction rate (86 mg-perchlorateL(-1) h(-1)) of ST-PRBC was higher than those (1.16±0.03 mg-perchlorate per mg-acetate and 48 mg-perchlorate L(-1) h(-1)) of N-PRBC. Kinetic analysis showed that NaCl acted as an uncompetitive inhibitor against both PRBCs. The inhibition constants were 25 and 41 mg-NaCl L(-1) for N-PRBC and ST-PRBC, respectively.

Characterization of methane oxidation by a methanotroph isolated from a landfill cover soil, South Korea.

A methane-oxidizing bacterium was isolated from the enriched culture of a landfill cover soil. The closest relative of the isolate, designated M6, is Methylocystis sp. Based on a kinetic analysis, the maximum specific methane oxidation rate and saturation constant were 4.93 mmol·g--dry cell weight--1·h⁻¹ and 23 microM, respectively. This was the first time a kinetic analysis was performed using pure methanotrophic culture. The methane oxidation by M6 was investigated in the presence of aromatic (m- and p-xylene and ethylbenzene) or sulfur (hydrogen sulfide, dimethyl sulfide, methanthiol) compounds. The methane oxidation was inhibited by the presence of aromatic or sulfur compounds.

Earthworm cast as a promising filter bed material and its methanotrophic contribution to methane removal.

The use of biocovers is a promising strategy toward mitigating CH(4) emission from smaller and/or older landfills. In this study, a filter bed material consisting of a mixture of earthworm cast and rice paddy soil in a biocover was evaluated. Although the CH(4) oxidation rate of the enriched paddy soil was 4.9 microg g-dry soil(-1) h(-1), it was enhanced to 25.1 microg g-dry soil(-1) h(-1) by adding an earthworm cast with a 3:7 ratio of earthworm cast:soil (wet weight). CO(2) was found as the final oxidation product of CH(4), and the mole ratio of CO(2) production to CH(4) consumption was 0.27. At a moisture content range of 15-40% and a temperature range of 20-40 degrees C, the CH(4) oxidation rates of the enriched mixture were more than 57% of the maximum rate obtained at 25% moisture content and 25 degrees C. By denaturing gradient gel electrophoresis analysis employing primers for the universal bacterial 16S rRNA gene, and terminal-restriction fragment length polymorphism analysis using primers for the pmoA gene, the bacterial and methanotrophic communities in the enriched mixture were mainly originate from paddy soil and earthworm cast, respectively. Both type I (mainly Methylocaldum) and type II methanotrophs (mainly Methylocystis) played important roles in CH(4) oxidation in the enriched mixture.