• Journal of Microbiology and Biotechnology
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• v.31 no.8
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• pp.1123-1133
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• 2021

Biodegradation is the key process involved in natural lignocellulose biotransformation and utilization. Microbial consortia represent promising candidates for applications in lignocellulose conversion strategies for biofuel production; however, cooperation among the enzymes and the labor division of microbes in the microbial consortia remains unclear. In this study, metagenomic analysis was performed to reveal the community structure and extremozyme systems of a lignocellulolytic microbial consortium, TMC7. The taxonomic affiliation of TMC7 metagenome included members of the genera Ruminiclostridium (42.85%), Thermoanaerobacterium (18.41%), Geobacillus (10.44%), unclassified_f__Bacillaceae (7.48%), Aeribacillus (2.65%), Symbiobacterium (2.47%), Desulfotomaculum (2.33%), Caldibacillus (1.56%), Clostridium (1.26%), and others (10.55%). The carbohydrate-active enzyme annotation revealed that TMC7 encoded a broad array of enzymes responsible for cellulose and hemicellulose degradation. Ten glycoside hydrolases (GHs) endoglucanase, 4 GHs exoglucanase, and 6 GHs β-glucosidase were identified for cellulose degradation; 6 GHs endo-β-1,4-xylanase, 9 GHs β-xylosidase, and 3 GHs β-mannanase were identified for degradation of the hemicellulose main chain; 6 GHs arabinofuranosidase, 2 GHs α-mannosidase, 11 GHs galactosidase, 3 GHs α-rhamnosidase, and 4 GHs α-fucosidase were identified as xylan debranching enzymes. Furthermore, by introducing a factor named as the contribution coefficient, we found that Ruminiclostridium and Thermoanaerobacterium may be the dominant contributors, whereas Symbiobacterium and Desulfotomaculum may serve as "sugar cheaters" in lignocellulose degradation by TMC7. Our findings provide mechanistic profiles of an array of enzymes that degrade complex lignocellulosic biomass in the microbial consortium TMC7 and provide a promising approach for studying the potential contribution of microbes in microbial consortia.

• Journal of Microbiology and Biotechnology
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• v.31 no.4
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• pp.570-583
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• 2021

Pyrococcus furiosus α-amylase can hydrolyze α-1,4 linkages in starch and related carbohydrates under hyperthermophilic condition (~ 100℃), showing great potential in a wide range of industrial applications, while its relatively low productivity from heterologous hosts has limited the industrial applications. Bacillus subtilis, a gram-positive bacterium, has been widely used in industrial production for its non-pathogenic and powerful secretory characteristics. This study was conducted to increase production of P. furiosus α-amylase in B. subtilis through three strategies. Initial experiments showed that co-expression of P. furiosus molecular chaperone peptidyl-prolyl cis-trans isomerase through genomic integration mode, using a CRISPR/Cas9 system, increased soluble amylase production. Therefore, considering that native P. furiosus α-amylase is produced within a hyperthermophilic environment and is highly thermostable, heat treatment of intact culture at 90℃ for 15 min was performed, thereby greatly increasing soluble amylase production. After optimization of the culture conditions (nitrogen source, carbon source, metal ion, temperature and pH), experiments in a 3-L fermenter yielded a soluble activity of 3,806.7 U/ml, which was 3.3- and 28.2-fold those of a control without heat treatment (1,155.1 U/ml) and an empty expression vector control (135.1 U/ml), respectively. This represents the highest P. furiosus α-amylase production reported to date and should promote innovation in the starch liquefaction process and related industrial productions. Meanwhile, heat treatment, which may promote folding of aggregated P. furiosus α-amylase into a soluble, active form through the transfer of kinetic energy, may be of general benefit when producing proteins from thermophilic archaea.

• Journal of Animal Environmental Science
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• v.15 no.2
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• pp.155-160
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• 2009

This study was carried out to investigate the feasibility of Multi Process of Aerobic Digestion (MPAD) for recycling of swine manure slurry as fertilizer. MPAD consisted of three kinds of difference process which are thermophilic aerobic oxidation (TAO) system, lime solidification system, and reverse osmosis (R/O) membrane system. TAO system was studied well previously for decade. The chemical composition of the lime-treated solid fertilizer was as like that organic matter 17.4%, moisture 34.1%, N 0.9%, P 1.7%, K 0.3%, Ca 12.7%, and which was expected to be useful as acid soil amendment material. The concentrated liquid material produced by R/O membrane system was also expected as a good fertilizer for crops production and soil fertility improvement.

• Journal of Korea Soil Environment Society
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• v.4 no.3
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• pp.57-65
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• 1999

Generation of gaseous ammonia has been a major problem in composting facilities. Microbial inocula. KMT-199(brand name: CompoBac$^{TM}$). was developed in INBI0NET CORPORATION and tested in the field for its ammonia reducing capability. When KMT-199 was applied. a ten-fold increase of mesophilic and thermophilic microorganisms was observed during the early stage of composting process. Also. the temperature and pH of early stage compost increased at a higher rate when compared to control. KMT-199 treated compost reached highest temperature of $75^{\circ}C$at day 9, indicating treatment could shift the maximum composting temperature to 3 days earlier The highest temperature also reached $3^{\circ}C$ higher than the control. The pH of compost gradually increased during composting. KMT-199 treated compost reached a plateau of pH 9.32 at day 15 after treatment, and then slowly decreased thereafter. On the other hand. pH of the control steadily increased until day 38 of composting. 29% reduction of gaseous ammonia generation during composting was observed compared to that of the control. KMT-199 amended compost resulted in a higher germination rate of radish seeds than the control. These results indicate that application of microbial inocula facilitates degradation of organic materials, including ammonia during the composting process.

• Journal of Animal Environmental Science
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• v.15 no.3
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• pp.271-280
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• 2009

The best way to treat livestock manure is to recycle as much as possible. The composting of livestock manure is a safe and economical treatment process. This study was carried out to investigate decomposition effect of pig manure by adding fermented compost. The fermented compost was added in pig manure mixed with sawdust as an inoculator, and the mixture was fed to composting reactor. Supplemental levels of fermented compost on the pig manure mixed with sawdust were regulated at 5, 10, 15 and 20% (V/V) respectively. The results were as follows ; 1. In all cases, PH range was between 7.6~9.05 during composting period. 2. The highest temperature and the long duration of thermophilic stage were observed in control treatment. 3. The number of microorganism reached at maximum on day 4, which recorded the highest temperature 4. Compost pile mixed with 10% of inoculator (fermented compost) showed the highest C/N ratio reduction.

• Journal of Environmental Health Sciences
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• v.28 no.5
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• pp.77-85
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• 2002

The objective of this study was to evaluate the performances of the ASBR under critical conditions of solid-liquid separation, caused by extremely high solids concentration, for wider application of the ASBR to various wastes. The ASBRs and completely-mixed daily-fed control runs were operated using a municipal mixed sludge at 35$^{\circ}C$ and 55$^{\circ}C$. Conversion of completely-mixed daily-fed reactor to sequencing batch mode and changes in HRT of all ASBRs were easily achieved without adverse effect, regardless of digestion temperature. Solids accumulation was remarkable in the ASBRs, and directly affected by settleable solids concentration of the feed sludge. Noticeable difference in solids-liquid separation was that flotation thickening occurred in the mesophilic ASBRs, while gravity thickening was a predominant solid-liquid separation process in the thermophilic ASBRS. Solids profiles at the end of thickening step dramatically changed at solid-liquid interface, and slight difference in solids concentrations was observed within thickened sludge bed. Organics removals based on subnatant or supernatant after thickening always exceeded 80% in all reactors. Thickened sludge volume and gas production of the ASBRs affected mutually. Gas production increased as thickened sludge accumulated, and continuous gas evolution during thickening could cause thickened sludge to expand or resuspend. Thickened sludge volume exceeding a predetermined withdrawal level resulted in loss of organic solids as well as biomass during withdrawal step, leading to decrease in gas production ind SRT. Such an adverse mutual effect was significant in gravity thickening, while it was not sensitive in flotation thickening. Changes in organic loading had no significant effect on organic removals and gas production after build-up of solids in the ASBRs.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.27 no.6
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• pp.675-681
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• 1994

[ $F_0$ ]-values of the canned tuna in cottonseed oil (CTCO) were investigated under different sterilizing conditions to optimize the energy consumption and microbiological safety. The $F_0$-values were measured using a microcomputer based technique. The exact cold point was not the volumetric center of the cans, and it was located in the center of meat mass in can which had ca. $6\%$ of head space. Location of the test cans in retort showed no remarkable influence on the $F_0$-values when the cans were jumble loaded. The process time before sterilization should be shortened as much as possible to prevent the contamination of microorganisms. Thermophilic spore forming bacteria found from raw and precooked tuna were Bacillus subtilis, Bacillus cereus and Bacillus pasteurii, and the most heat resistant was Bacillus subtilis. The rational $F_0$-value for the CTCO obtained from the preservation test was regarded as 6min.

• Journal of Animal Environmental Science
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• v.17 no.sup
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• pp.1-6
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• 2011

Korean Government announced 'The Roadmap to realize a low carbon green society on year 2020' on July 12, 2011 in order to mitigate greenhouse gas (GHG) emissions. Non-energy category of Food, Agriculture, Forestry and Fishery (FAFF) should mitigate 1,349 kilo $CO_2$-equivalent ($CO_2$-eq.) tonnes which is 7.1% of Business-As-Usual on year 2020. The mitigation from animal manure treatment system (AMTS) comprises ca. 45% of the total mitigated amount of Non-energy category of FAFF. Hence, the precise evaluation of GHG emissions from AMTS is important to find effective mitigation measures. Life cycle assessment was used to evaluate GHG emissions from AMTS. The most GHG emitter was a composting/liquid fertilizer/activated sludge system (1,649.45 kg $CO_2$-eq./head/year) and the least GHG emitter was a activated liquid fertilizer system (1,024.46 kg $CO_2$-eq./head/year). Thermophilic oxic process showed the highest ratio (34.9%) of GHG emissions by the use of electricity to total GHG emissions from systems. Energy efficiency should be considered to mitigate GHG emissions from AMTS.

• Journal of Mushroom
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• v.18 no.3
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• pp.274-279
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• 2020

This study was conducted to investigate the characteristics of the medium used on the composting step, comparing the excavator agitator with the self-propelled turner. The temperature of the outdoor composting medium tended to increase rapidly after flipping in the turner. The late composting medium temperature was maintained at the excavator treatment area (farm practice), and the late composting effect progressed. During the field composting stage, various microorganisms such as Bacillus spp., Actinomycetes, fluorescent Pseudomonas spp., and filamentous fungi were distributed in the medium, and the density of aerobic bacteria involved in the decomposition of the medium was increased. Under high-temperature composting conditions, blue fungi, and mesophilic actinomycetes were inhibited or killed. Thermophilic actinomycetes, which play an important role in decomposing organic matter, showed higher densities than those observed in farm practices in the self-propelled turner process. The length of rice straw was slightly shorter when the self-propelled turner was used, and the water content did not show any significant difference between treatments. The a and b values tended to increase as the inverter was turned over. The CN ratio of the composting broth was lowered from 23.1 to 16.2 for the 5th turnover in the context of farming practices, and from 23.3 to 16.9 in the context of the self-propelled turner. The yield of each treatment was increased by 20% in 1 period, 28% in 2 periods, and 26% in 3 periods; the overall yield was 23%.

• Asian-Australasian Journal of Animal Sciences
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• v.18 no.8
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• pp.1199-1208
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• 2005

Abatement of greenhouse gas emitted from ruminants and promotion of biogas energy from animal effluent were comprehensively examined in each anaerobic fermentation reactor and animal experiments. Moreover, the energy conversion efficiency of biomass energy to power generation were evaluated with a gas engine generator or proton exchange membrane fuel cell (PEMFC). To mitigate safely rumen methanogenesis with nutritional manipulation the suppressing effects of some strains of lactic acid bacteria and yeast, bacteriocin, $\beta$1-4 galactooligosaccharide, plant extracts (Yucca schidigera and Quillaja saponarea), L-cysteine and/or nitrate on rumen methane emission were compared with antibiotics. For in vitro trials, cumulative methane production was evaluated using the continuous fermented gas qualification system inoculated with the strained rumen fluid from rumen fistulated Holstein cows. For in vivo, four sequential ventilated head cages equipped with a fully automated gas analyzing system were used to examine the manipulating effects of $\beta$1-4 galactooligosaccharide, lactic acid bacteria (Leuconostoc mesenteroides subsp. mesenteroides), yeast (Trichosporon serticeum), nisin and Yucca schidigera and/or nitrate on rumen methanogenesis. Furthermore, biogas energy recycled from animal effluent was evaluated with anaerobic bioreactors. Utilization of recycled energy as fuel for a co-generator and fuel cell was tested in the thermophilic biogas plant system. From the results of in vitro and in vivo trials, nitrate was shown to be a strong methane suppressor, although nitrate per se is hazardous. L-cysteine could remove this risk. $\beta$1-4 galactooligosaccharide, Candida kefyr, nisin, Yucca schidigera and Quillaja saponarea are thought to possibly control methanogenesis in the rumen. It is possible to simulate the available energy recycled through animal effluent from feed energy resources by making total energy balance sheets of the process from feed energy to recycled energy.