• Asian-Australasian Journal of Animal Sciences
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• v.17 no.11
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• pp.1591-1598
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• 2004

To isolate the $\beta$-galactosidase producing thermophilic bacteria, samples of mud and water were collected from hot springs of avolcanic area near Golden Springs in New Zealand. Among eleven isolated strains, the strain of KNOUC112 produced the highest amounts of $\beta$-galactosidase at 40 h incubation time (0.013 unit). This strain was aerobic, asporogenic bacilli, immobile, gram negative, catalase positive, oxidase positive, and pigment producing. Optimum growth was at 70-72$^{\circ}C$, pH 7.0-7.2, and it could grow in the presence of 3% NaCl. The main fatty acids of cell components were iso-15:0 (30.26%), and iso-17:0 (31.31%). Based on morphological and biochemical properties and fatty acid composition, the strain could be identified as genus Thermus, and finally as Thermus thermophilus by phylogenetic analysis based on 16S rRNA sequence. So the strain is designated as Thermus thermophilus KNOUC112. A gene from Thermus thermophilus KNOUC112 encoding $\beta$-galactosidase was amplified by PCR using redundancy primers prepared based on the structure of $\beta$-galactosidase gene of Thermus sp. A4 and Thermus sp. strain T2, cloned and expressed in E. coli JM109 DE3. The gene of Thermus thermophilus KNOUC112 $\beta$-galactosidase(KNOUC112$\beta$-gal) consisted of a 1,938 bp open reading frame, encoding a protein of 73 kDa that was composed of 645 amino acids. KNOUC112$\beta$-gal was expressed as dimer and trimer in E. coli JM109 (DE3) via pET-5b.

• Journal of Life Science
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• v.12 no.2
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• pp.129-135
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• 2002

Several kinds of thermophilic, aerobic microorganisms (Bacillus genus), metal leaching microorganisms (Thiobacillus, T. ferooxidans), and other nondegradable chemical-degrading microorganisms (Pseudomonas genus) were utilized to study the effect on composting livestock manure. Under the Carbon-Nitrogen ratio (C/N) of 35∼40 and water content of 50∼65% conditions, the composting in the cycling drum reactor showed slower composting and lower temperature increase than that of the manual reactor. Element analysis after composting indicated relatively high levels of mineral contents with the substitutional effect of chemical fertilizer. Metal analysis before and after composting showed lower As in all, Cr in pig, Pb in cow, Hg in chicken, and Cu in cow manure compost than the regulation values. Compost maturity was ascertained by the several maturity tests. Salmonella and E. cozi detection test by SS or EMB agar plate confirmed the safety from the pathogenic microorganisms. The results suggest that the inoculation of metal and some other chemical degrading microorganisms during composting might decrease metal contamination and increase composting rate.

• Journal of Environmental Health Sciences
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• v.35 no.3
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• pp.214-219
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• 2009

This study examined the removal efficiency of the nitrogen and phosphorus in order to compensate for the combined process of ATAD(Autothermal Thermophilic Aaerobic Digestion) and EGSB(Expended Granular Sludge Bed), which are treatment methods for livestock wastewater, by introducing SBR(Sequencing Batch Reactor) as post-treatment process. The analysis on the treatment efficiency of each operation mode showed that, in the case of T-N, the treatment efficiency were 67.1% and 74.2% for Run-1 and Run-2, respectively, and in the case of T-P, the values were 71.2 and 74.1, respectively, which are indicating that the treatment efficacy is higher in the condition of Run-1, in which the time period of Anoxic and Aerobic segments were increased. In addition, the result of analyzing removal characteristics of nitrogen and phosphorus by Influx load showed that removal efficiency of nitrogen was better in the case of high influx load than in the case of low influx load. Regardless of Influx load, phosphorus showed constant influx concentration, so that removal efficiency of phosphorus was influenced littler by Influx load than that of nitrogen. This study also fed methanol as an external carbon source and performed experiment to induce denitrification under anoxic condition by using nitrate among nitrogen compounds of SBR reactor, and the results showed that intermittent feeding was more effective in Nitrogen Removal than composite feeding.

• Journal of Animal Environmental Science
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• v.9 no.1
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• pp.45-56
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• 2003

$TAO^{TM}$ System is an auto-heated thermophilic aerated digestion process using a proprietary microbe called as a Phototropic Bacteria (PTB). High metabolic activity results in heat generation, which enables to produce a pathogen-free and digested liquid fertilizer at short retention times. TAO$^{TM}$ system has been developed to reduce a manure volume and convert into the liquid fertilizer using swine manure since 1992. About 100 units have been installed and operated in Korean swine farms so far. TAO$^{TM}$ system consists of a reactor vessel and ejector-type aeration pumps and foam removers. The swine slurry manure enters into vessel with PTB and is mixed and aerated. The process is operated at detention times from 2 to 4 days and temperature of 55 to $65^{\circ}C$. Foams are occurred and broken down by foam removers to evaporate water contents. Generally, at least 30% of water content is evaporated, 99% of volatile fatty acids caused an odor are removed and pathogen destruction is excellent with fecal coliform, rotavirus and salmonella below detection limits. The effluent from TAO$^{TM}$ system, called as the "TAO EFFLUX", is screened and has superb properties as a fertilizer. Normally N-P-K contents of screened TAO Efflux are 4.7 g/L, 0.375 g/L and 2.8 g/L respectively. The fertilizer effect of TAO EFFLUX compared to chemical fertilizer has been demonstrated and studied with various crops such as rice, potato, cabbage, pumpkin, green pepper, parsley, cucumber and apple. Generally it has better fertilizer effects and excellent soil fertility improvement effects. Moreover, the TAO EFFLUX is concentrated through membrane technology without fouling problems for a cost saving of long distance transportation and a commercialization (crop nutrient commodity) to a gardening market, for example.

• Journal of Animal Science and Technology
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• v.63 no.1
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• pp.16-24
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• 2021

The suppressive effect of monensin as an ionophore-feed additive on enteric methane (CH4) emission and renewable methanogenesis were evaluated. To clarify the suppressive effect of monensin a respiratory trial with head cage was performed using Holstein-Friesian steers. Steers were offered high concentrate diets (80% concentrate and 20% hay) ad libitum with or without monensin, galacto-oligosaccharides (GOS) or L-cysteine. Steers that received monensin containing diet had significantly (p < 0.01) lower enteric CH4 emissions as well as those that received GOS containing diet (p < 0.05) compared to steers fed control diets. Thermophilic digesters at 55℃ that received manure from steers fed on monensin diets had a delay in the initial CH4 production. Monensin is a strong inhibitor of enteric methanogenesis, but has a negative impact on biogas energy production at short retention times. Effects of the activity of coprophagous insects on CH4 and nitrous oxide (N2O) emissions from cattle dung pats were assessed in anaerobic in vitro continuous gas quantification system modified to aerobic quantification device. The CH4 emission from dungs with adults of Caccobius jessoensis Harold (dung beetle) and the larvae of the fly Neomyia cornicina (Fabricius) were compared with that from control dung without insect. The cumulative CH4 emission rate from dung with dung insects decreased at 42.2% in dung beetles and 77.8% in fly larvae compared to that from control dung without insects. However, the cumulative N2O emission rate increased 23.4% in dung beetles even though it reduced 88.6% in fly larvae compared to dung without coprophagous insects. It was suggested that the antibacterial efficacy of ionophores supplemented as a growth promoter still continued even in the digested slurry, consequently, possible environmental contamination with the antibiotics might be active to put the negative impact to land ecosystem involved in greenhouse gas mitigation when the digested slurry was applied to the fields as liquid manure.

• Journal of Microbiology and Biotechnology
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• v.32 no.6
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• pp.749-760
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• 2022

α-Galactosidase is a debranching enzyme widely used in the food, feed, paper, and pharmaceuticals industries and plays an important role in hemicellulose degradation. Here, T26, an aerobic bacterial strain with thermostable α-galactosidase activity, was isolated from laboratory-preserved lignocellulolytic microbial consortium TMC7, and identified as Parageobacillus thermoglucosidasius. The α-galactosidase, called T26GAL and derived from the T26 culture supernatant, exhibited a maximum enzyme activity of 0.4976 IU/ml when cultured at 60℃ and 180 rpm for 2 days. Bioinformatics analysis revealed that the α-galactosidase T26GAL belongs to the GH36 family. Subsequently, the pET-26 vector was used for the heterologous expression of the T26 α-galactosidase gene in Escherichia coli BL21 (DE3). The optimum pH for α-galactosidase T26GAL was determined to be 8.0, while the optimum temperature was 60℃. In addition, T26GAL demonstrated a remarkable thermostability with more than 93% enzyme activity, even at a high temperature of 90℃. Furthermore, Ca²⁺ and Mg²⁺ promoted the activity of T26GAL while Zn²⁺ and Cu²⁺ inhibited it. The substrate specificity studies revealed that T26GAL efficiently degraded raffinose, stachyose, and guar gum, but not locust bean gum. This study thus facilitated the discovery of an effective heat-resistant α-galactosidase with potent industrial application. Meanwhile, as part of our research on lignocellulose degradation by a microbial consortium, the present work provides an important basis for encouraging further investigation into this enzyme complex.

• Microbiology and Biotechnology Letters
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• v.8 no.3
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• pp.155-166
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• 1980

Studies have teen undertaken to investigate the degree of microbial contamination in the subsidiary materials which have been known as an important source of microorganisms associated with spoilage of fish sausage and fish paste products. Twenty hinds of food ingredients including starch, spices and condiments, 59 samples in total collected from commercial fish sausage processing plants and supermarket in the period of July to October 1979, were examined for standard plate count, coliform and fecal coliform, mold and yeast, thermoduric microorganisms, aerobic sporeformers (mesophilic and thermophilic), anaerobic sporeformers (mesophilic and thermophilic) and sulfide spoilage anaerobes. The results obtained are summarized as follows. 1. Among the food ingredients examined, corn starch, black pepper, hot pepper, onion, garlic, ginger, beef extract and frank marked high bacterial contamination with general and sporeforming microorganisms. And bacterial content of marked samples were generally higher than that of the samples from plants. 2. The high standard plate count caused by high content of these bacteria like thermoduric, mesophilic or thermophilic sporeforming aerobes. 3. Bacterial content of food ingredients such as black pepper and beef extract being used in plants, and black pepper, hot pepper, onion and garlic from the market were exceeded the bacterial standards being enforced in Japan and U. S. A. 4. Average standard plate count was in the range of 10$^4$to 10$^{5}$ /g for black pepper, wheat flour, onion and garlic collected from plants, and 10$^{5}$ to 10$^{7}$ /g for black pepper, hot pepper, onion and garlic from market. No plate count was observed in pepper essence and coloring material. 5. Coliform organism was detected in starch, black pepper, hot pepper, onion, garlic, ginger and gluten that showed high standard plate but no fecal coliform in the samples except black pepper and hot pepper. 6. Average mold and yeast count was 140 to 460/g for corn starch, wheat flour and black pepper from plants, and 10$^3$/g for black pepper and hot pepper from market. No count was observed in the other ingredients. 7. Sulfide spoilage sporeforming anaerobes boiled for 5 min. at 10$0^{\circ}C$ and incubated at 55$^{\circ}C$ was not detected in all the samples examined.

• Journal of Animal Science and Technology
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• v.46 no.2
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• pp.261-272
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• 2004

This study was conducted to investigate indices affecting composts maturity for swine manure compost produced in a commercial composting facility with air-forced from the bottom. The composting was made of swine manure mixed with puffing rice hull(6: 4) and turned by escalating agitator twice a day. Composting samples were collected periodically during a 45-d composting cycle at that system, showing that indices of Ammonium-N to Nitrate-N ratio were sensitive indicators of composting quality. Pile temperature maintained more than 62$^{\circ}C$ and water contents decreased about 20% for 25days of composting. A great variety and high numbers of aerobic thermophilic heterotropic microbes playing critical roles in stability of composts have been examined in the final composts, sbowing that they were detected $10^8$ to $10^{10}$ $CFUg^{-1}$ in mesophilic bacteria, $10^3$ - $10^4$ in fungi and $10^6$ - $10^8$ in actinomycetes, respectively. The results of this study for detennining a factor affecting compost stability evaluations based on composting steps were as follows; 1. Ammonium-N concentrations were highest at the beginning of composting, reaching approximately 421mg/kg. However Ammonium-N concentrations were lower during curing, reaching approximately l04mg/kg just after 45 day. The ratio between $NH_4-N$ and $NO_3-N$ was above II at the beginning of composting and less than 2 at the final step(45 day). 2. Seed germination Index was dependent upon the compost phytotoxicity and its nutrition. The phytotocity caused the GI to low during the period of active composting(till 25 days of composting time) depending on the value of the undiluted. After 25 days of composting time, the GI was dependent upon compost nutrition. The Gennination index of the final step was calculated at over 80 without regard to treatments. 3. E4: E6 ratio in humic acid of composts was correlatively decreased from 8.86 to 6.76 during the period of active composting. After 25 days of composting time, the E4: E6 was consistently decreased from 6.76 to 4.67($r^2$ of total composting period was 0.95). 4. Water soluble carbon had a tendency to increase from 0.54% to 0.78%during the period of active composting. After 25 days of composting time, it was consistently decreased from 0.78% to 0.42%. Water soluble nitrogen increased from 0.22% to 0.32% during the period of 15 days after initial composting while decreased from 0.32% to 0.21% after 15days of composting. In consequence, the correlation coefficient($r^2$) between water soluble carbon and water soluble nitrogen was 0.12 during the period of active composting mule was 0.50 after 25 days of composting time

• Journal of Animal Environmental Science
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• v.10 no.3
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• pp.169-174
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• 2004

The present study investigated the possibility of applying the fermented swine liquid manure handled with TAO system to hydroponic cultivation. The fermented swine liquid manure (FLM) used for the study was in low concentration of phosphoric acid (P). Hence, when doing hydroponic cultivation, the concentration of P and K was supplemented. The concentrations of N, P, and K in compensated FLMs were as fellows; (a) N 58mg/1, P 0.5mg/1 and K 31 mg/1 for FLM-1, (b) N 58mg/1, P 31 mg/l and K 39mg/1 for FLM-2, and (c) N 58mg/1, P 31 mg/1 and K 61 mg/1 for FLM-3, respectively. For hydroponic cultivation, it was possible to use diluted solution by 100 times through experiment of germination index. According to the experiment using lettuce, the compensated FLMs with P and K were superior in leaf width, yield and dry weight to FLM without any treatment. Therefore, it is necessary to supplement some nutrition before applying. In conclusion, it is highly estimated that the fermented swine liquid manure can be ap-plied to hydroponic cultivation. It is also necessary to conduct further researches related to nutritional compensation depending on the vegetables.

• Korean Journal of Food Science and Technology
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• v.36 no.1
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• pp.105-110
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• 2004

Microorganism (strain DF 218) producing thermostable pretense was isolated from Korean soil and compost. It was Gram-positive, rod-shaped, aerobic, and spore-forming with yellowish white colony color, Temperature range for growth at pH 6.5 was $30-65^{\circ}C$, with optimum growth at $60^{\circ}C$. pH range for growth at $60^{\circ}C$ was 5-7 with optimum of 6.5, which indicates strain DF 218 to be thermophilic. The 16S rDNA sequence of strain DF 218 had 95% sequence similarity with that of Bacillus flexus. Based on physiological properties and phylogenetic analysis, we proposed the isolated strain as Bacillus sp. DF 218. Pretense was produced aerobically at $60^{\circ}C$ for 32 hr in a medium (pH 6.5) containing 1% each trypton, glucose, and NaCl. Its molecular weight was estimated as 61 kDa, with optimum temperature and pH of $60^{\circ}C$ and 7.5, respectively.