• Journal of Animal Science and Technology
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• v.54 no.6
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• pp.435-442
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• 2012

This study was conducted to isolate and identify the lactic acid bacteria (LAB) from spent mushroom substrates (SMS) for the effective anaerobic fermentation to utilize SMS as an animal feed and to determine the optimal medium conditions for their growth. At first, a total of 23 strains were isolated from the ensiled SMS based on the LAB counts and pH tested. Then, a total of 16 strains which rapidly produce lactate and decreased the pH, were selected for a screening test. The optical density (OD), pH, and yellow clear zone were tested for the selected 16 strains. Among the strains, KU5 strain had wider yellow clear zone and lower pH and KU13 strain had higher OD at 24 hr of incubation and wider yellow clear zone compared to other strains and control strain (Lactobacillus plantarum KCCM 12116). Accordingly, KU5 and KU13 strains were finally selected. The KU5 and KU13 were identified as Lactobacillus plantarum by the 16S rRNA sequencing. The KU5 strain was named as Lactobacillus plantarum KU5, and the KU13 strain was named as Lactobacillus plantarum KU13. Lactobacillus plantarum KU5 and Lactobacillus plantarum KU13 were registered at the National Center for Biotechnology Information (NCBI). Access number of Lactobacillus plantarum KU5 was HQ542227 and that of Lactobacillus plantarum KU13 was HQ542228. The optimal medium conditions for growth of KU5 and KU13 were soybean meal 2% and formulated feed 2%, respectively.

• Asian-Australasian Journal of Animal Sciences
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• v.27 no.12
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• pp.1726-1735
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• 2014

This study investigated the effects of acarbose addition on changes in ruminal fermentation characteristics and the composition of the ruminal bacterial community in vitro using batch cultures. Rumen fluid was collected from the rumens of three cannulated Holstein cattle fed forage ad libitum that was supplemented with 6 kg of concentrate. The batch cultures consisted of 8 mL of strained rumen fluid in 40 mL of an anaerobic buffer containing 0.49 g of corn grain, 0.21 g of soybean meal, 0.15 g of alfalfa and 0.15g of Leymus chinensis. Acarbose was added to incubation bottles to achieve final concentrations of 0.1, 0.2, and 0.4 mg/mL. After incubation for 24 h, the addition of acarbose linearly decreased (p<0.05) the total gas production and the concentrations of acetate, propionate, butyrate, total volatile fatty acids, lactate and lipopolysaccharide (LPS). It also linearly increased (p<0.05) the ratio of acetate to propionate, the concentrations of isovalerate, valerate and ammonia-nitrogen and the pH value compared with the control. Pyrosequencing of the 16S rRNA gene showed that the addition of acarbose decreased (p<0.05) the proportion of Firmicutes and Proteobacteria and increased (p<0.05) the percentage of Bacteroidetes, Fibrobacteres, and Synergistetes compared with the control. A principal coordinates analysis plot based on unweighted UniFrac values and molecular variance analysis revealed that the structure of the ruminal bacterial communities in the control was different to that of the ruminal microbiota in the acarbose group. In conclusion, acarbose addition can affect the composition of the ruminal microbial community and may be potentially useful for preventing the occurrence of ruminal acidosis and the accumulation of LPS in the rumen.

• Korean Journal of Agricultural Science
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• v.43 no.3
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• pp.387-393
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• 2016

This study was conducted to determine the fermentative characteristics of wheat bran inoculated with a starter culture of direct-fed microbes as a microbial wheat bran (DMWB) feed additive. Wheat bran was prepared with 1% (w/w, 0.5% Lactobacillus plantarum and 0.5% of Saccharomyces cerevisiae) starter culture treatment (TW) or without starter culture as a control (CW). Those were fermented under anaerobic conditions at $30^{\circ}C$ incubation for 3 days. Samples were taken at 0, 1, 2, and 3 days to analyze chemical composition, microbial growth, pH, and organic acid content. Chemical composition was not significantly different between CW and TW (p > 0.05). In TW, the number of lactic acid bacteria and yeast increased during the 3 days of fermentation (p < 0.05) and the population of lactic acid bacteria was significantly higher than in CW (p < 0.05). After 3 days, the number of yeast in TW was $7.50{\pm}0.07log\;CFU/g$, however, no yeast was detected in CW (p < 0.05). The pH values of both wheat bran samples decreased during the 3 days of fermentation (p < 0.05), and TW showed significantly lower pH than CW after 3 days of fermentation (p < 0.05). Contents of lactic acid and acetic acid increased significantly at 3rd day of fermentation in TW. However, no organic acids were generated in CW during testing period. These results suggest that 3 days of fermentation at $37^{\circ}C$ incubation after the inoculation wheat bran with starter culture makes it possible to produce a direct-feed with a high population of lactic acid bacteria at more than $10^{11}CFU/g$.

• Journal of Korean Society of Environmental Engineers
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• v.36 no.12
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• pp.865-872
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• 2014

The study was conducted to evaluate the effect of addition of food waste in brown water for anaerobic hydrogen production. Batch experiment was carried out to determine appropriate food waste to brown water mixing ratio. Maximum hydrogen yield of $6.92mmol\;H_2/g\;COD_{removed}$ was obtained at 70% food waste and 30% brown water. Semi-pilot scale reactor was operated based on result of batch experiment. Semi-pilot reactor operated, mixing 70% food waste and 30% brown water showed significant increment in butyric acid concentration. B/P (Butyric to propionic acid ratio) which is considered as governing factor for hydrogen production was found high (52.64). Maximum hydrogen yield of $25.03mmol\;H_2/g\;COD_{removed}$ was obtained. Result of this study concluded that mixing of food waste to brown water at appropriate ratio assists in enhanced hydrogen fermentation.

• Transactions of the Korean hydrogen and new energy society
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• v.22 no.5
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• pp.671-677
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• 2011

A hyperthermophilic archaeon, $Thermococcus$ $onnurineus$ NA1 was studied to investigate its fermentation characteristics using various carbon sources including formate, maltose and carbon monoxide during the anaerobic batch cultivation at $80^{\circ}C$. Formate was the best carbon source for cell growth and hydrogen production among others. In the batch culture on formate, it was found that the cell concentration increased exponentially by 12 hrs of culture, after which the cell growth and formate consumption was retarded. Hydrogen production was continued more than 24 hrs although the cell growth was ceased at 18 hrs. Hydrogen production rate was directly correlated with the cell growth and formate degradation up to 18 hrs, and the average hydrogen production yield was 1.05 mole-$H_2$/mole-formate. Cell growth and hydrogen production were optimized at the initial pH 6-7, while inhibited at the initial pH lower than 5 and higher than 9.

• Journal of agriculture & life science
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• v.46 no.3
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• pp.79-85
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• 2012

This study was conducted to examine the effect of dietary n-3/n-6 fatty acid (FA) ratio on in vitro dry matter digestibility (IVDMD), fermentation indices and FA profile. Rice bran was mixed with oil sources (cotton seed oil and linseed oil) to make the diets at 0.02, 0.29 and 0.61 of dietary n-3/n-6 FA ratio. These diets (0.5g) were placed into the incubation bottles with 40 ml of anaerobic culture medium, which contained rumen fluid and Van Soest medium at 1:2 ratio. Five replicates of each diet and two blanks were incubated at $39^{\circ}C$ for 48 hours. After incubation, the incubated contents were centrifuged. The residues were freeze-dried for DMD and FA analyses. The supernatant was used for pH, $NH_3-N$ and volatile fatty acid analyses. The concentrations of lactate (p<0.001) and iso-valerate (p<0.001) decreased linearly with increasing dietary n-3/n-6 FA ratio, but acetate concentration (p=0.056) and the ratio of acetate to propionate (p=0.005) was increased linearly. The concentrations of n-3, n-6 FA and the ratio of n-3/n-6 FA in residues increased (p<0.001) linearly with increasing dietary n-3/n-6 FA ratio, but C18:1n-9 FA concentration was decreased (p<0.001) linearly. With these results, it could affect fermentation characteristics and FA profile of rumen content by dietary n-3/n-6 FA ratio.

• Asian-Australasian Journal of Animal Sciences
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• v.25 no.1
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• pp.86-91
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• 2012

An in vitro rumen batch culture study was completed to compare effects of common grasses, leguminous shrubs and non-leguminous shrubs used for livestock grazing in Australia and Ghana on $CH_4$ production and fermentation characteristics. Grass species included Andropodon gayanus, Brachiaria ruziziensis and Pennisetum purpureum. Leguminous shrub species included Cajanus cajan, Cratylia argentea, Gliricidia sepium, Leucaena leucocephala and Stylosanthes guianensis and non-leguminous shrub species included Annona senegalensis, Moringa oleifera, Securinega virosa and Vitellaria paradoxa. Leaves were harvested, dried at $55^{\circ}C$ and ground through a 1 mm screen. Serum bottles containing 500 mg of forage, modified McDougall's buffer and rumen fluid were incubated under anaerobic conditions at $39^{\circ}C$ for 24 h. Samples of each forage type were removed after 0, 2, 6, 12 and 24 h of incubation for determination of cumulative gas production. Methane production, ammonia concentration and proportions of VFA were measured at 24 h. Concentration of aNDF (g/kg DM) ranged from 671 to 713 (grasses), 377 to 590 (leguminous shrubs) and 288 to 517 (non-leguminous shrubs). After 24 h of in vitro incubation, cumulative gas, $CH_4$ production, ammonia concentration, proportion of propionate in VFA and IVDMD differed (p<0.05) within each forage type. B. ruziziensis and G. sepium produced the highest cumulative gas, IVDMD, total VFA, proportion of propionate in VFA and the lowest A:P ratios within their forage types. Consequently, these two species produced moderate $CH_4$ emissions without compromising digestion. Grazing of these two species may be a strategy to reduce $CH_4$ emissions however further assessment in in vivo trials and at different stages of maturity is recommended.

• 한국신재생에너지학회:학술대회논문집
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• 2009.11a
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• pp.506-510
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• 2009

The effects of various organic wastes on anaerobic fermentative hydrogen production were studied using enriched mixed microflora in batch tests. Rotten fruit, corn powder and organic wastewater enriched with sulfate (up to 1,000 mg/L) were used for experiments. Maximum hydrogen production (547.1 mL) was observed from rotten apple with initial substrate concentration of 132.2 g COD/L. The experimental result on sulfate enriched organic wastewater indicated that hydrogen production is not adversely influenced by relatively high sulfate concentration. Residual sulfate content remained at 96-98 % after 75 hours of reaction, which showed that no major sulfate reduction was occurred at pH 5.3-5.5 in the reactor. The volatile fatty acid (VFA) fractions produced during the reaction was in the order of butyrate > acetate > propionate in all experiments. The results of this study would be useful for controlling the conditions on fermentative hydrogen production using different feedstocks.

• Journal of Microbiology and Biotechnology
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• v.20 no.11
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• pp.1529-1533
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• 2010

The inulinase gene (INU1) from Kluyveromyces marxianus NCYC2887 was overexpressed by using the GAL10 promotor in a ${\Delta}ga180$ strain of Saccharomyces cerevisiae. The inulinase gene lacking the original signal sequence was fused in-frame to a mating factor ${\alpha}$ signal sequence for secretory expression. Use of the ${\Delta}ga180$ strain allowed for the galactose-free induction of inulinase expression using a glucose-only medium. Shake-flask cultivation in YPD medium produced 34.6 U/ml of the recombinant inulinase, which was approximately 13-fold higher than that produced by K. marxianus NCYC2887. It was found that the use of the ${\Delta}ga180$ strain improved the expression of inulinase in the recombinant S. cerevisiae in both aerobic and anaerobic conditions by about 2.9- and 1.7-fold, respectively. A 5-l fed-batch fermentation using YPD medium was performed under aerobic condition with glucose feeding, which resulted in the inulinase production of 31.7 U/ml at the $OD_{600}$ of 67. Ethanol fermentation of dried powder of Jerusalem artichoke, an inulin-rich biomass, was also performed using the recombinant S. cerevisiae expressing INU1 and K. marxianus NCYC2887. Fermentation in a 5-l scale fermentor was carried out at an aeration rate of 0.2 vvm, an agitation rate of 300 rpm, and with the pH controlled at 5.0. The temperature was maintained at $30^{\circ}C$ and $37^{\circ}C$, respectively, for the recombinant S. cerevisiae and K. marxianus. The maximum productivities of ethanol were 59.0 and 53.5 g/l, respectively.

• Microbiology and Biotechnology Letters
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• v.13 no.4
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• pp.397-402
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• 1985

The fermentation of various sugars by C. thermosaccharolyticum was examined under pH controlled, anaerobic condition. The kinetic model for Product formation at various sugars was the combination of growth and non-growth associated mode. In the utilization of a single sugar, glucose was the best carbon source for growth. The specific growth rate of glucose, xylose and cellobiose were 0.363 h$^{-1}$, 0.242 h$^{-1}$ and 0.144 h$^{-1}$ respectively. The production of ethanol from glucose showed a negatively growth associated mode, so the higher growth rate decreased the productivity of ethanol. The maximum concentrations of the produced ethanol were 2.42 g/l, 3.76 g/l, and 3.4 g/l on glucose, xylose, and cellobiose. No glucose was detected during cellobiose fermentation. Sequential utilization of sugars was observed in the mixtures of glucose, xylose and cellobiose. It preferred glucose, followed by xylose and then cellobiose. The presence of other sugars had little or no effect on the rate of another sugar utilization. Cell lysis at the end of fermentation occured more slowly in the mixtures of sugars than a single sugar.