• 한국유기농업학회지
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• 제20권3호
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• pp.327-343
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• 2012

In order to develop a high cellulolytic direct-fed microorganism (DFM) for ruminant productivity improvement, this study isolated cellulolytic bacteria from the rumen of Holstein dairy cows, and compared their cellulolytic abilities via DM degradability, gas production and cellulolytic enzyme activities. Twenty six bacteria were isolated from colonies grown in Dehority's artificial (DA) medium with 2% agar and cultured in DA medium containing filter paper at $39^{\circ}C$ for 24h. 16s rDNA gene sequencing of four strains from isolated bacteria showed that H8, H20 and H25 strains identified as Ruminococcus flavefaciens, and H23 strain identified as Fibrobacter succinogenes. H20 strain had higher degradability of filter paper compared with others during the incubation. H8 (R. flavefaciens), H20 (R. flavefaciens), H23 (F. succinogenes), H25 (R. flavefaciens) and RF (R. flavefaciens sijpesteijn, ATCC 19208) were cultured in DA medium with filter paper as a single carbon source for 0, 1, 2, 3, 4 and 6 days without shaking at $39^{\circ}C$, respectively. Dry matter degradability rates of H20, H23 and H25 were relatively higher than those of H8 and RF since 2 d incubation. The cumulative gas production of isolated cellulolytic bacteria increased with incubation time. At every incubation time, the gas production was highest in H20 strain. The activities of carboxymethylcellulase (CMCase) and Avicelase in the culture supernatant were significantly higher in H20 strain compared with others at every incubation time (p<0.05). Therefore, although further researches are required, the present results suggest that H20 strain could be a candidate of DFM in animal feed due to high cellulolytic ability.

• 공업화학
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• 제34권3호
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• pp.327-329
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• 2023

본 연구에서는 Clostridium ljungdahlii 배양에서 유기질소공급물인 tryptone과 sodium tungstate의 배양액 내 농도가 균주 성장과 아세트산, 에탄올 생산에 대한 영향을 확인하기 위한 실험을 수행하였다. 대조군 실험의 배지 조건(0 g/L tryptone)보다 tryptone 2.5 g/L를 투입한 경우 균주 성장이 144.6% 증가하였고, 에탄올 및 아세트산 생산은 각각 8.6%와 36.7% 향상되었다. 대조군 실험에 사용된 배지의 조건(0.01 µM Na₂WO₄·2H₂O)보다 100배인 1 µM Na₂WO₄·2H₂O를 사용한 실험군에서, 균주 성장과 C2 대사산물의 총생산량에는 큰 차이가 없지만, 에탄올 생산 증가하고 아세트산 생산은 감소하여 에탄올/아세트산 생산비가 0.56로 대조군의 0.24에서 크게 증가하였다.

• Asian-Australasian Journal of Animal Sciences
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• 제18권2호
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• pp.221-225
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• 2005

An in vitro study was conducted to examine the effect of monensin or fish oil addition on bio-hydrogenation of $C_{18^-} unsaturated fatty acids and CLA production by mixed ruminal bacteria when incubated with safflower oil. Commercially manufactured concentrate (1%, w/v) with safflower oil (0.2%, w/v) were added to mixed solution (600 ml) of strained rumen fluid and McDougalls artificial saliva (control). Monensin $Rumensin^{(R)}$, 10 ppm, w/v, MO), mixed fish oil (0.02%, w/v, absorbed to 0.2 g alfalfa hay, FO) or similar amounts of monensin and fish oil (MO+FO) to MO and FO was also added into the control solution. All the culture solutions prepared were incubated in the culture jar anaerobically at $39^{\circ}C$ up to 12 h. Higher pH (p<0.047) and ammonia concentration (p<0.042) were observed from the culture solution containing MO at 12 h incubation than those from the culture solutions of control or FO. The MO supplementation increased (p<0.0001-0.007) propionate proportion of culture solution but reduced butyrate proportion at 6 h (p<0.018) and 12 h (p<0.001) of incubations. Supplementation of MO or MO+FO increased (p<0.001) the proportions of $C_{18:2}$. The MO alone reduced (p<0.022-0.025) the proportion of c9,t11-CLA compared to FO in all incubation times. The FO supplementation increased the proportion of c9,t11-CLA. An additive effect of MO to FO in the production of c9,t11-CLA was observed at 6 h incubation. In vitro supplementation of monensin reduced hydrogenation of $C_{18^-}$UFAs while fish oil supplementation increased the production of CLA.

• 한국수소및신에너지학회논문집
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• 제26권3호
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• pp.199-205
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• 2015

Organic wastes such as food waste (FW), livestock wastewater (LW), and sewage sludge (SWS) can produce hydrogen ($H_2$) by anaerobic acid fermentation. Expecially, FW which has high carbohydrate content produces $H_2$ and short chain fatty acids by indigenous $H_2$ producing microorganisms without adding inoculum, however $H_2$ production rate (HPR) and yield have to be improved to use a commercially available technology. In this study, LW was mixed to FW in different ratios (on chemical oxygen demand (COD) basis) as an auxiliary substrate. The mixture of FW and LW was pretreated at pH 2 using 6 N HCl for 12 h and then fermented at $37^{\circ}C$ for 28 h. HPR of FW, 254 mL $H_2/L/h$, was increased with the addition of LW, however, mixing ratio of LW to FW was reversely related to HPR, exhibiting HPR of 737, 733, 599, and 389 mL $H_2/L/h$ at the ratio of FW:LW=10:1, 10:2, 10:3, and 10:4 on COD basis, respectively. Maximum HPR and $H_2$ production yield of 737 $H_2/L/h$ and 1.74 mol $H_2/mol$ hexoseadded were obtained respectively at the ratio of FW:LW=10:1. Butyrate was the main organic acid produced and propionate was not detected throughout the experiment.

• 한국생물공학회:학술대회논문집
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• 한국생물공학회 2003년도 생물공학의 동향(XIII)
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• pp.351-355
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• 2003

Screening 실험을 통하여 결정된 주요인자인 sucrose, nitrogen, 배양온도로 인자의 최적수준을 결정하는 표면반응법중 하나인 Box-Behnken design을 수행하였다. 각 인자의 상관관계를 통하여 sucrose는 90 g/L, nitrogen은 41 mM, 온도는 $22^{\circ}C$가 최적수준으로 결정이 되었으며 확인실험을 통하여 대조구보다 세포생장이 증가하였을 뿐만 아니라 hGM-CSF의 생산도 약 2배 정도 증가되었음을 확인하였다. 이것은 고농도의 sucrose로 인해 배지내로의 hGM-CSF의 분비가 촉진되었기 때문이며 또한 저온으로 인해 분비된 hGM-CSF를 분해하는 protease 활성이 감소되었기 때문인 것으로 사료된다.

• Asian-Australasian Journal of Animal Sciences
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• 제20권2호
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• pp.200-207
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• 2007

In vitro rumen incubation studies were conducted to determine effects of initial pH on bacterial attachment and fiber digestion. Ruminal fluid pH was adjusted to 5.7, 6.2 and 6.7, and three major fibrolytic bacteria attached to rice straw in the mixed culture were quantified with real-time PCR. The numbers of attached and unattached Fibrobacter succinogenes, Ruminococcus flavefaciens and Ruminocococcus albus were lower (p<0.05) at initial pH of 5.7 without significant difference between those at higher initial pH. Lowering incubation media pH to 5.7 also increased bacterial numbers detached from substrate regardless of bacterial species. Dry matter digestibility, gas accumulation and total VFA production were pH-dependent. Unlike bacterial attachment, maintaining an initial pH of 6.7 increased digestion over initial pH of 6.2. After 48 h in vitro rumen fermentation, average increases in DM digestion, gas accumulation, and total VFA production at initial pH of 6.2 and 6.7 were 2.8 and 4.4, 2.0 and 3.0, and 1.2 and 1.6 times those at initial pH of 5.7, respectively. The lag time to reach above 2% DM digestibility at low initial pH was taken more times (8 h) than at high and middle initial pH (4 h). Current data clearly indicate that ruminal pH is one of the important determinants of fiber digestion, which is modulated via the effect on bacterial attachment to fiber substrates.

• Journal of Microbiology and Biotechnology
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• 제10권1호
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• pp.1-7
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• 2000

Bioconversion of fumarate to succinate was anaerobically conduced in a synthetic medium containing glycerol as a hydrogen donor and fumarate as a hydrogen acceptor. We investigated the effects of pH, carbon and nitrogen sources, conversion substrate, and other culture conditions on the production of succinate using a nwely isoloated Enterococcus facalis PKY1. Addition of a variety of carbonates to the medium significantly increasd the rates of production of succinate. The production of succinate and cell growth were relatively satisfactory in the pH range of 7.0-7.6. By using glycerol as a hydrogen donor, high purity succinate was produced with few byproducts. Yeast extract as a sole nitrogen source was the most effective for producing succinalte. As a result, the optimum condition of biconversion was obtained at a medium containing 20g/I glycerol, 50 g/l fumarate, 15 g/l yeast extract, 10 g/l $K_2HPO_4$, 1 g/I NaCl, 50ppm $MgCl_2{\cdot}6H_2O$, 10ppm $FeSo_4{\cdot}7H_2O$, and 5 g/I $Na_2CO_3$ at pH 7.0-7.6. Under the optimum condition, a succinate concentration of 153 g/I was produced in 36 h. The total volumetric production rate and the molar yield of succinate were 4.3 g/l/h and 85%, respectively.

• 한국식품과학회지
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• 제23권2호
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• pp.167-174
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• 1991

토양으로부터 생물고분자의 생산균주로 분리한 알칼리 내성 Bacillus sp.의 생물고분자 생산을 위한 최적 배양조건을 검토하였고, 반응 표면분석법에 의한 생산 최적화를 수행하였다. 최대의 생물고분자 생산은 soluble starch 8%, yeast extract 0.75%, $NaNO_3$ 0.1%, $MgSO_4\;7H_2O$ 0.05% 및 $Na_2CO_3$ 1%(pH 10)의 조성배지에서 배양온도 $30^{\circ}C$로 36시간 이상 진탕배양하였을 때 얻어졌으며, 이때의 생물고분자 생산량은 약 44g/l(대당수율로는 약 55%)이었다. C/N비, 온도 및 pH를 독립변수로 한 반응 표면분석에 의한 생물고분자 생산의 최적조건은 C/N비 15.16, 배양온도 $34.62^{\circ}C$ 및 pH 9.50의 정상점에서 얻어졌으며, 이 조건에서 생물고분자의 생산량은 66.84g/l로 추정 되었다.

• Journal of Microbiology and Biotechnology
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• 제23권10호
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• pp.1454-1459
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• 2013

The changes in prodiginines productions caused by pH shock culture of Streptomyces coelicolor strains were estimated. In Streptomyces coelicolor M511, undecylprodiginine and streptorubin B productions increased 1.8-fold (37.22 mg/g) and 2.5-fold (18.61 mg/g), respectively, by pH shock (from 7.2 to 4.0). In contrast, this resulted in the significantly decreased prodigignines production in the redP deletion mutant SJM1; 3.7-fold for undecylprodiginine, 4.4-fold for streptorubin B, 5.2-fold for methylundecylprodiginine, and 6.4-fold for methyldodecylundecylprodiginine, respectively. RT-PCR analyses showed that, during pH shock, expression of redD, the transcription activator gene, was increased while the expression of fabH, the decarboxylative condensation enzyme gene in fatty acid biosynthesis, was decreased in both strains. The enhanced redD expression was in good accordance with the increased total prodiginines production of M511. However, for SJM1 mutant, the decrease of fabH expression occurred more strikingly, such that it became almost completely turned off during acidic pH shock culture. Therefore, a down-regulation of fabH was considered to be the cause of decreased amount of total prodiginines produced, although redD expression was high in SJM1 mutant.

• 한국미생물·생명공학회지
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• 제23권6호
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• pp.647-651
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• 1995

A tylosin-hyperproductive mutant of S. fradiae MNU20 was isolated among 3500 strains obtaincd from either MNNG- or UV-treated Streptomyces fradiae NRRL2702. The composition of optimal medium for tylosin production was formulated as followed as: 4 g soluble starch, 1 g glucose, 1 g corn steep liquor, 7.5 ml soy bean oil, 0.2 g KH$_{2}$PO$_{4}$, 1 g Na$_{2}$S$_{2}$O$_{3}$$\cdot $5H$_{2}$O, 2 g CaCO$_{3}$, 2 g NaCl, 0.001 g CoCl$_{2}$$\cdot $6H$_{2}$O in 1 liter of distilled water. With the optimal medium, S. fradiae MNU20 was able to produce 159 mg tylosin (g biomass)$^{-1}$, indicating that tylosin productivity of Streptomyces fradiae NRRL2702 was increased 14 times higher by mutation. When the effect of valine, succinate, and natural zeolite on tylosin production was investigated by using the optimal medium, these substances essentially enhanced tylosin production and their addition time during culture period appeared to be critical for the increase of tylosin production.