• Journal of Animal Science and Technology
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• 제54권6호
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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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• 제27권12호
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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.

• 농업과학연구
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• 제43권3호
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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 $10^{11}CFU/g$.

• 대한환경공학회지
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• 제36권12호
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• pp.865-872
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• 2014

본 연구는 수소가스 생산을 위한 brown water(소변을 제외한 대변 + 대변세척수 6 L)의 혐기성 소화 시, 음식물쓰레기 혼합 효과를 평가하기 위해 실시하였다. brown water와 음식물쓰레기의 적절한 혼합 비율을 찾기 위해 회분식 실험이 수행되었고, 도출된 결과는 연속운전 세미파일럿 규모 brown water의 혐기성 소화장치의 실험에 적용되었다. 회분식 실험에서 70%의 음식물쓰레기와 30%의 brown water을 혼합하였을 때 $6.92mmol\;H_2/g\;COD_{removed}$의 최대 수소생산수율을 나타내었다. 동일한 혼합비율로 투입한 음식물쓰레기 및 brown water의 세미파일럿 규모 혐기성 소화조를 운전하였을 때, 반응조 내부에서 수소생산의 중간산물인 butyric acid의 현저한 증가를 보였다. 이 때 수소 생산의 지표인 B/P (butyrate/propionate) 비는 52.64로 나타났고, 수소생산수율은 최대 $25.03mmol\;H_2/g\;COD_{removed}$로 나타났다. 이상의 실험적 연구결과 brown water의 혐기성 수소발효에서 음식물쓰레기의 혼합은 수소발생을 촉진하기 위한 좋은 대안임을 확인하였다.

• 한국수소및신에너지학회논문집
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• 제22권5호
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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.

• 농업생명과학연구
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• 제46권3호
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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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• 제25권1호
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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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• 한국신재생에너지학회 2009년도 추계학술대회 논문집
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• pp.506-510
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• 2009

다양한 유기성 폐자원이 혐기성 소화 공정 수소 생성에 미치는 영향을 고찰하기 위해 혼합균주를 사용하여 혐기 발효 회분식 형태 연구하였다. 폐자원은 부패 과일 폐수로 각각 사과, 배, 옥수수가루 및 황산염이 함유된 유기폐수 (황산염 농도 <= 1000 mg/L)를 이용하였다. 최대 수소 생성(547.1 mL)은 부패된 사과 폐수에서 나타났으며, 유입 농도는 132.2 g COD/L이다. 황산염 함유 폐수 적용결과, 황산염 농도에 관계없이 수소가 발생하였고, 잔존황산염은 pH 5.3-5.5에서 96-98 %로 나타나 황산염 환원은 발생되지 않았다. 모든 실험 조건에서 유기산은 HBu > HAc > HPr로 발생하였다. 본 연구결과 다양한 유기 폐자원에서 발효공정을 통해 수소 발생이 가능한 것으로 사료된다.

• Journal of Microbiology and Biotechnology
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• 제20권11호
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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.

• 한국미생물·생명공학회지
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• 제13권4호
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• pp.397-402
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• 1985

C. thermosaccharolyticum을 이용하여 xylose, glucose, cellobiose등을 각각 단일기질 및 혼합기질로 배양한 결과는 다음과 같다. 기질의 종류에 관계없이 에탄올의 생산양상은 Leudeking-Piret모델에 따랐으며 q$_{p}$=$\alpha$ $\mu$＋$\beta$의 식으로 나타낼 수 있었다. 단일당에서의 배양결과 glucose, xylose, cellobiose 각 당에서의 비증식속도는 0.363 h$^{-1}$, 0.242 h$^{-1}$, 0.144 h$^{-1}$로 균의 증식은 glucose를 기질로 사용하였을 때가 가장 좋았다. 에탄올 생산성은 xylose에서 가장 좋았으며, 각 당에서의 에탄올수율은 0.38g ethanol/g xylose, 0.34 g ethanol/g cellobiose, 0.242g ethano1/g glucose 이었다. glucose에서의 배양결과, 배양 12시간이후 균의 자기소화현상이 현저히 일어났으며, 다른 당에서와는 달리 에탄올 생산양상은 $\alpha$값이 음수를 나타내므로써, 균의 생육이 증가하면 오히려 에탄을 비생산속도는 감소하는 경향을 보였다. 서로 다른 당을 혼합하여 기질로 사용할 경우, 각 당의 이용특성은 이들을 단독기질로 배양하였을 때의 소비양상과 유사하여 각 당은 서로 영향을 미치지 않고, 독립적으로 자화되는 것으로 생각된다. 혼합당에서의 기질소비는 glucose, xylose, cellobiose순으로 소비되었으며, 또한 에탄을 생산성은 glucose가 첨가된 경우 glucose 농도가 낮고(5g/l), glucose를 기질로 하여 균이 왕성하게 증식한 상태에서 xylose나 cellobiose를 이용하므로써 glucose만을 단독기질로 배양한 결과보다 좋았다.