A wastewater treatment pond system was developed for treatment and recycling of dairy cattle excreta of $5\;m^1$ per day. The wastes were diluted by the water used for clearing stalls. The system was composed of three ponds in series. A submerged gas collector for the recovery of methane was installed at the bottom of secondary pond with water depth of 2.4m. This paper deals mainly with performance of methane fermentation of secondary pond which is faclutative one. The average $BOD_5$, SS, TN, and TP concentrations of influent into secondary pond were 49.1, 53.4, 48.6, and 5.3 mg/l, and those of effluent from it were 27.9, 45.7, 30.8, 3.2 mg/l respectively. Methane fermentation of 2.4-meter-deep secondary pond bottom was well established at $16^{\circ}C$ and gas garnered from the collector at that temperature was 80% methane. Literature on methane fermentation of wastewater treatment ponds shows that methane bacteria grow well around $24^{\circ}C$, the rate of daily accumulation and decomposition of sludge is approximately equal at $19^{\circ}C$, and activities of methanogenic bacteria are ceased below $14^{\circ}C$. The good methane fermentation of the pond bottom around $16^{\circ}C$, about $3^{\circ}C$ lower than $19^{\circ}C$, results from temperature stability, anaerobic condition, and neutral pH of the bottom sludge layer. It is recommended that the depth of pond water could be 2.4m. Gas from the collector during active methane fermentation was almost 83% methane, less than 17% nitrogen. Carbon dioxide was less than 1% of the gas, which indicates that carbon dioxide produced in bottom sludges was dissolved in the overlaying water column. Thus a purified methane can be collected and used as energy source. Sludge accumulation on the pond bottom for a nine month period was 1.3cm and annual sludge depth can be estimated to be 1.7cm. Design of additional pond depth of 0.3m can lead to 15 - 20 year sludge removal.
Yedam Kim;Young Seok Lee;Youngsuk Kim;Jinmyeong Song;Yeongbeen Bak;Gyulim Park;O-Mi Lee;Hong-Joo Son
Journal of Life Science
/
v.34
no.5
/
pp.304-312
/
2024
This study was conducted to characterize strain Y10, isolated from discarded chicken feathers. Strain Y10 was identified as Bacillus amyloliquefaciens through phenotypic and 16S rRNA gene analysis. B. amyloliquefaciens Y10 exhibited plant growth-promoting activities, including the production of fungal cell-degrading enzymes (cellulase, lipase, protease, and pectinase), siderophores, ammonia, and indoleacetic acid. Furthermore, strain Y10 was able to inhibit the mycelial growth of several phytopathogenic fungi. When 0.1% sucrose as a carbon source and 0.05% casein as a nitrogen source were added to the basal medium, adjusted to pH 10, and cultured at 35℃, the degradation rate of chicken feathers by strain Y10 was about two times higher than that of the basal medium, with the feathers almost completely degraded in four days. Strain Y10 also degraded various keratin substrates, including duck feathers, wool, and human nails. It was confirmed that the feather hydrolyzates prepared using strain Y10 exhibited antioxidant activities, such as 1,1-diphenyl-2-picrylhydrazyl radical scavenging activity (EC50 = 0.38 mg/ml) and superoxide dismutase-like activity (EC50 = 183.7 mg/ml). These results suggest that B. amyloliquefaciens Y10 is a potential candidate for the development of bioinoculants and feed additives applicable to the agricultural and livestock industries, as well as the microbiological treatment of keratin waste.
SOHN Heung-Sik;PARK Seong-Min;SON Byung-Yil;CHOI Hyeon-Mee;LEE Keun-Tai
Korean Journal of Fisheries and Aquatic Sciences
/
v.32
no.2
/
pp.121-126
/
1999
In order to degrade chitin by enzymatic hydrolysis, it is required from screening highly active deacetylase. To this end, we examined three fungal strains and it turned out that Mucor rouxii produced highly active deacetylase, this enzyme exhibited the highest enzymatic activity against colloidal chitin. The conditions for growing Mucor rouxii are as follows; the effective carbon source, nitrogen source, adequate initial pH, temperature and incubation time were $2\%$ glucose, $1.33\%$ yeast extract, $0.66\%$ pepton, 4.5, $25{\pm}2^{\circ}C$ and 48hr, respectively. The optimum pH and temperature for purified enzyme activity were 5.5 and $40^{\circ}C$, respectively. The purified enzyme was stable at pH ranging from 4.5 to 5.5. However, the enzyme activity was decreased to less than $50\%$ at pH blow 45 and above 7.5. At temperatures above $50^{\circ}C$, the enzyme activity was decreased remarkably. The enzyme was inhibited by LiC1, $HgCl_2$, and $BaCl_2$, but stimulated by $CaCl_2$ and $ZnC1_2$, The activity of purified enzyme was increased by L-cysteine and 2-mercaptoethanol, while decreased by O-phenanthroline, p-CMB, EDTA, and iodoacetate. The $K_m$ and the $V_{max}$ value of purified enzyme were $1.2\%$ and 59.5 U/mg, respectively. The deacetylation activity of purified enzyme was not detected at optimal reaction condition when chitin particle suspension was used.
Kim, Hyun-Jung;Kim, Yoon-Hee;Cho, Moon-Jung;Shin, Keum;Lee, Dong-Heub;Kim, Tae-Jong;Kim, Yeong-Suk
Journal of the Korean Wood Science and Technology
/
v.38
no.6
/
pp.547-560
/
2010
The optimum culture condition of Schizophyllum commune for the cellulase production and its enzymatic characteristics for saccharification of cellulosic biomass were analyzed. S. commune secrets ${\beta}$-1,4-xylosidase (BXL) and cellulases, including endo-${\beta}$-1,4-glucanase (EG), cellobiohydrolase (CBH), and ${\beta}$-glucosidase (BGL). The optimum reaction temperature for all cellulases was $50^{\circ}C$ and the thermostable range was $30{\sim}40^{\circ}C$C. The optimum reaction pH for all cellulases was 5.5 in a range of temperature from $0^{\circ}C$ to $55^{\circ}C$. The best nutritions for the cellulase production of S. commune among tested nutrients were 2% cellulose for the carbon source and corn steep liquor or peptone/yeast extract for the nitrogen source without vitamins. The environmental culture condition for the cellulase production was 5.5~6.0 for pH at $25{\sim}30^{\circ}C$. The enzyme activities of EG, BGL, CBH, and BXL were 3670.5, 631.9, 398.5, and 15.2 U/$m{\ell}$, respectively, after concentration forty times from the culture broth of S. commune which was grown at the optimized culture condition. Alternative filter paper unit assay showed 11 FPU/$m{\ell}$ enzyme activity. The saccharification tests using cellulase of S. commune showed the low saccharification rate on tested hardwoods but a high value of 50.5% on cellulose, respectively. The saccharification rate (50.5%) of cellulose by cellulase produced in this work is higher than 45.7% in the commercial enzyme (Celluclast 1.5L, 30 FPU/g, glucan).
The optimal culture conditions in 500$m\ell$ flask suture, 5$\ell$ jar fermenter and 2,000 $\ell$ large stale culture were investigated to maximize the production of antibiotic on Rhizoctonia solani AC4, the causal agent of vegetables damping-off, by the strain Bacillus ehimensis YJ-37. Starch (1.5%) as a carbon source, peptone (0.6%) as a nitrogen source and MgC1$_2$(0.15%) as a metal ion in the medium containing N $a_2$HP $O_4$(0.3%) showed the highest production of the antibiotic(s) in a rotary shake (200 rpm). Optimal initial pH of the culture medium, culture temperature and culture time for the antibiotic(s) production were pH 8.0, 32$^{\circ}C$ and 54hrs, respertively. Under the optimal renditions in flask culture, cell growth and antifungal activity (clear zone size) were 1.42 $\times$ 10$^{8}$ cfu/$m\ell$ (21g-DCW/ $\ell$) and 13.9 mm, respectively. In 5 $\ell$ jar fermenter (medium 3 $\ell$), optimal air flow, agitation speed and culture time for the antibiotic(s) production were 2 vvm, 200 rpm and 48hrs, respectively. Under the optimal conditions in 5 $\ell$ jar fermenter, tell growth and antifungal activity were 2.06 $\times$ 10$^{8}$ cfu/$m\ell$ (30g-DCW/ $\ell$) and 13.4 mm, respectively. Under the culture conditions of air flow (30 vvm) and agitation speed (200 rpm) at 32$^{\circ}C$ for 10 days in 2,000 $\ell$ large scale culture (medium 1,800 $\ell$, pH 8.0), cell growth and antifungal activity were 0.81$\times$10$^{8}$ cfu/$m\ell$ (12g-DCW/ $\ell$) and 8.6 mm, respectively.
In this study, we compared the levels of methylotrophic bacterial community diversity in the leaf samples of 19 rice cultivars collected from three regions of Korea. Nineteen pink pigmented isolates showing characteristic growth on methanol were obtained. Physiological and biochemical characters of each isolate were examined according to methods described in Bergey's Manual of Systematic Bacteriology. When phylotypes were defined by performing numerical analysis of 37 characteristics, four distinct clusters were formed. The two reference strains, Methylobacterium extorquens AM1 and Methylobacterium fujisawaense KACC10744 were found to group under cluster IV and cluster III respectively. Cluster I diverged on the basis of nitrate reduction and four isolates showed tolerance upto 0.5 M NaCl concentrations. Two strains in cluster I and III were found to possess methane utilizing properties. Most of the isolates in all the four clusters utilized monosaccharides, disaccharide and polyols as carbon source. When the isolates were subjected for indole-3-acetic acid (IAA) analysis in the presence of L-tryptophan, only 8 isolates exhibited IAA production. In addition, the nitrogen source in the medium was found to influence the IAA production. Addition of $(NH_4)_2SO_4$ in the medium led to a 2 to 30 fold increase in the indole synthesis. However, $KNO_3$, $NH_4NO_3$ and $NH_4Cl$ substitution did not significantly stimulate the synthesis of IAA in the growth medium. Result of gnotobiotic root elongation assay significantly increased roots and shoots lengths, and number of lateral roots, which is mediated by IAA production in the culture medium. The rice seedlings primary roots from seeds treated with methylotrophic isolates were on average 27 to 56% longer than the roots from seeds treated with the uninoculated seeds. In addition, application of different high concentrations of authentic IAA ($400g\;mL^{-1}$) to roots of rice seedlings inhibited root growth. However, the IAA concentration from 10 to $200g\;mL^{-1}$, IAA promoted root growth of rice seedlings. These results suggest that bacterial IAA plays a major role in the development of the host plant root system.
Journal of the Korean Society of Food Science and Nutrition
/
v.42
no.7
/
pp.1125-1132
/
2013
In the kimchi manufacturing process, the starter is cultured on a large-scale and needs to be supplied at a low price to kimchi factories. However, current high costs associated with the culture of lactic acid bacteria for the starter, have led to rising kimchi prices. To solve this problem, the development of a new medium for culturing lactic acid bacteria was studied. The base materials of a this novel medium consisted of Chinese cabbage extract, a carbon source, a nitrogen source, and inorganic salts. The optimal composition of this medium was determined to be 30% Chinese cabbage extract, 2% maltose, 0.25% yeast extract, and $2{\times}$ salt stock (2% sodium acetate trihydrate, 0.8% disodium hydrogen phosphate, 0.8% sodium citrate, 0.8% ammonium sulfate, 0.04% magnesium sulfate, 0.02% manganese sulfate). The newly developed medium was named MFL (medium for lactic acid bacteria). After culture for 24 hr at $30^{\circ}C$, the CFU/mL of Leuconostoc (Leuc.) citreum GR1 in MRS and MFL was $3.41{\times}10^9$ and $7.49{\times}10^9$, respectively. The number of cells in the MFL medium was 2.2 times higher than their number in the MRS media. In a scale-up process using this optimized medium, the fermentation conditions for Leuc. citreum GR1 were tested in a 2 L working volume using a 5 L jar fermentor at $30^{\circ}C$. At an impeller speed of 50 rpm (without pH control), the viable cell count was $8.60{\times}10^9$ CFU/mL. From studies on pH-stat control fermentation, the optimal pH and regulating agent was determined to be 6.8 and NaOH, respectively. At an impeller speed of 50 rpm with pH control, the viable cell count was $11.42{\times}10^9(1.14{\times}10^{10})$ CFU/mL after cultivation for 20 hr - a value was 3.34 times higher than that obtained using the MRS media in biomass production. This MFL media is expected to have economic advantages for the cultivation of Leuc. citreum GR1 as a starter for kimchi production.
In this study, the characteristics of alcohol fermentation using ripe Daebong persimmon juice were studied in static fermentation condition by Saccharomycess cerevisiae CS02 in an effort to develop new types of functional wine. Attempts were made to modify the ripe Daebong persimmon juice in order to find suitable conditions for alcohol fermentation. The modified ripe Daebong persimmon juice that was most suitable for alcohol fermentation contained $24^{\circ}brix$ of sugar supplemented with sucrose as a carbon source and 0.5 g/L of $(NH_4)_2HPO_4$ as a nitrogen source. After 9 days of fermentation at $25^{\circ}C$, $12.2{\pm}0.02%$ of alcohol was produced from the modified juice and its pH markedly decreased to $3.97{\pm}0.02$. The wine contained free sugar such as fructose ($0.12{\pm}0.02$ g/L), some organic acids such as malic acid ($35.92{\pm}0.24$ g/L), succinic acid ($8.12{\pm}0.03$ g/L), oxalic acid ($22.14{\pm}0.11$ g/L), and citric acid ($13.63{\pm}0.08$ g/L), as well as some flavanols and phenolic acids such as catechin gallate ($38.99{\pm}0.32$ mg/L), epicatechin gallate ($110.21{\pm}0.16$ mg/L), gallic acid ($163.88{\pm}1.11$ mg/L), epigallocatechin ($15.97{\pm}0.18$ mg/L), and tannic acid ($13.36{\pm}0.02$ mg/L). In addition, the 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical (84.25%) and $ABTS^{\cdot+}$ radical (99.65%) scavenging activities were increased significantly with a corresponding increased in the organic acid and phenolic acid contents, but decreased in the flavonoids.
Jo, Hye-Mi;Kim, Ye-Jin;Yoo, Sang-Ho;Kim, Chang-Mu;Kim, KyeWon;Park, Cheon-Seok
Korean Journal of Food Science and Technology
/
v.53
no.2
/
pp.195-203
/
2021
Aureobasidium pullulans, a black yeast, produces pullulan, a linear α-glucan composed of maltotriose repeating units linked by α(1→6)-glycosidic linkages. Pullulan can be widely used in food, cosmetic, and biotechnology industries. In this study, we isolated eight strains of A. pullulans from Forsythia koreana, Magnolia kobus DC., Spiraea prunifolia var. simpliciflora, Cornus officinalis, Cerasus, and Hippophae rhamnoides. Among them, A. pullulans MR was selected as the best pullulan producer. The effects of a carbon source, a nitrogen source, and pH on pullulan production were examined. The optimal cultivation conditions for pullulan production by A. pullulans MR were determined by response surface methodology as 15% sucrose, 0.4% soy peptone, and an initial pH of 7 at 26℃. Under these conditions, the predicted pullulan production was 47.6 g/L, which was very close to the experimental data (48.9 g/L).
The characteristics of alcohol fermentation using sweet persimmon juice were studied in static fermentation in an effort to develop new types of functional wine. The yeast strain Saccharomyces cerevisiae KCCM 12650 was selected for use in the fermentation of sweet persimmon juice. Attempts were made to modify the sweet persimmon juice in order to find suitable conditions for alcohol fermentation. The modified sweet persimmon juice (pH 4.0) that was most suitable for alcohol fermentation contained $24^{\circ}Brix$ of sugar supplemented with sucrose as a carbon source and 0.5 g/L of $(NH_4)_2HPO_4$ as a nitrogen source. After 5 days of fermentation at $25^{\circ}C$, 12.8% of alcohol was produced from the modified juice and its pH was slightly decreased to 3.9. Browning of the wine was observed during storage due to the oxidation of phenolic compounds. The initial browning of 0.08% at $OD_{420}$ after fermentation increased to 0.40 during storage for 11 weeks at room temperature. The addition of $K_2S_2O_5$ was effective in delaying the browning of the wine. The browning of the wine decreased to 0.25 at $OD_{420}$ with the addition of 200 mg/L of $K_2S_2O_5$. The wine produced in this study contained some organic acids such as malic acid (6.82% g/L) and succinic acid (1.40 g/L), some minerals such as $K^+$ (947.8 mg/L) and $Mg^{2+}$ (36.4 mg/L), as well as soluble phenolics (779 mg/L of gallic acid equivalent). Schisandra fruit was added to the sweet persimmon juice during alcohol fermentation in order to improve the sour taste and flavor. The best sensory quality (taste, flavor, and color) was obtained by adding 0.5% schisandra fruit.
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