• 미생물학회지
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• 제51권3호
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• pp.187-193
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• 2015

Candida (Pseudozyma로도 알려짐) antarctica lipase B(CAL-B)는 학문적으로 그리고 산업적으로 많이 활용되고 있다. CAL-B 자체에 대한 연구는 많이 진행되어온 반면, CAL-B 상동체에 관한 연구는 그리 알려진 바가 없다. 본 연구에서는 단백질 유사성 검색을 통해서 CAL-B의 상동체 탐색을 수행하였고, 6종의 단백질 서열을 찾았다. 해당하는 유전자들을 대장균에 대한 코돈 최적화를 수행하였고, 이를 바탕으로 유전자 합성을 진행하였다. 이들 유전자를 대장균 발현용 벡터에 클로닝한 후, 대장균 내에서 단백질 발현을 시도하여 이들 중 4종의 단백질이 성공적으로 발현되었다. 이들 단백질들이 가수분해 효소로서의 활성이 있는지 확인하기 위해서, 4-nitrophenyl acetate와 4-nitrophenyl butyrate를 반응기질로 하여 가수분해 반응성을 확인하였다. 이들 단백질들의 비활성(specific activity)값은 $(1.3-30){\times}10^{-2}{\mu}mol/min/mg$로 측정되었고, 이는 CAL-B의 비활성 수치보다는 다소 낮은 값에 해당하였다. (${\pm}$)-1-phenylethyl acetate의 가수분해 반응에 대한 입체선택성은 이들 상동체 효소들 중에서 Pseudozyma hubeiensis SY62에서 유래된 효소만이 CAL-B의 입체선택성과 유사함이 확인되었다.

• Asian-Australasian Journal of Animal Sciences
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• 제33권6호
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• pp.1023-1033
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• 2020

Objective: The efficiency of the knock-in process is very important to successful gene editing in domestic animals. Recently, it was reported that transient loosening of the nucleosomal folding of transcriptionally inactive chromatin might have the potential to enhance homologous recombination efficiency. The objective of this study was to determine whether histone deacetylases (HDAC) inhibitor and RAD51 recombinase (RAD51) expression were associated with increased knock-in efficiency on the β-casein (bCSN2) gene locus in mammary alveolar-large T antigen (MAC-T) cells using the transcription activator-like effector nucleases (TALEN) system. Methods: MAC-T cells were treated with HDAC inhibitors, valproic acid, trichostatin A, or sodium butyrate for 24 h, then transfected with a knock-in vector, RAD51 expression vector and TALEN to target the bCSN2 gene. After 3 days of transfection, the knock-in efficiency was confirmed by polymerase chain reaction and DNA sequencing of the target site. Results: The level of HDAC 2 protein in MAC-T cells was decreased by treatment with HDAC inhibitors. The knock-in efficiency in MAC-T cells treated with HDAC inhibitors was higher than in cells not treated with inhibitors. However, the length of the homologous arm of the knock-in vector made no difference in the knock-in efficiency. Furthermore, DNA sequencing confirmed that the precision of the knock-in was more efficient in MAC-T cells treated with sodium butyrate. Conclusion: These results indicate that chromatin modification by HDAC inhibition and RAD51 expression enhanced the homologous recombination efficiency on the bCSN2 gene locus in MAC-T cells.

• 한국물환경학회지
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• 제26권4호
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• pp.608-613
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• 2010

Electricity generation of microbial fuel cells (MFC) is greatly affected by the kind of feed substrates because substrates would change microbial community of electrochemically active bacteria (EAB) able to transfer electrons to electrode. The effect of different substrates on electricity generation and microbial community of MFC was investigated. Two-chamber MFCs fed with acetate (A-MFC), butyrate (B-MFC), propionate (P-MFC), glucose (G-MFC) and a mixture (M-MFC) of the 4 substrates (acetate : butyrate : propionate : glucose = 1 : 1 : 1 : 1 as $COD_{Cr}$ base) were operated under continuous mode. The maximum power density was found from the M-MFC ($190W/m^3$) which showed the lowest internal resistance ($89{\Omega}$). The maximum power densities of the pure substrates feed MFCs were in order of A-MFC ($25W/m^3$), P-MFC ($21W/m^3$), B-MFC ($20W/m^3$) and G-MFC ($9W/m^3$). In DGGE analysis, the microbial community structure in suspension was quite different from each others depending on feed substrates, while the community structure in the biofilm was relatively similar regardless of the substrates. This result suggests that the feed substrates would affect the microbial community of suspended growth bacteria than attached growth bacteria resulting in difference of electricity generation in MFCs.

• Asian-Australasian Journal of Animal Sciences
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• 제18권3호
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• pp.326-331
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• 2005

A rumen simulated continuous culture (RSCC) system was used to study the influence of supplementation of the three different types of protein sources such as urea, casein and soy protein on rumen microbial synthesis in terms of rumen microbial synchronization. The urea treatment showed the highest pH value. Ammonia nitrogen concentration was rapidly increased after feeding and not significantly different in the urea treatment (13.53 mg/100 ml). Protozoa numbers were not significantly different for soy protein and casein treatment compared to urea treatments during incubation. The average concentration of total VFA (mMol) was not detected with significant difference among treatments, but iso-butyrate production showed the highest for soy protein treatment among treatments (p<0.001). The lowest concentration in total iso-acids (iso-butyrate and iso-valerate) production was observed in urea treatment. The soy protein treatment showed no significantly change in acetate/propionate. The amounts of dry matter (DM) out flow showed no significant difference among treatments. Organic matter (OM) flow was the highest for urea treatments and the lowest for casein treatment (p<0.03). The nitrogen flow for casein treatment was not significantly different from other treatments. The efficiency of microbial protein synthesis in terms of microbial nitrogen (MN) synthesis (g MN/kg ADOM) digested in the rumen was highest for casein treatment (58.53 g MN/kg ADOM) compared to soy protein and urea (p<0.05). This result suggests that rumen ammonia releasing rate may influence on microbial protein synthesis in the rumen.

• Asian-Australasian Journal of Animal Sciences
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• 제10권4호
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• pp.391-397
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• 1997

Six medium-framed steers, fitted with ruminal cannulas, were utilized in a $6{\times}6$ Latin square design with a $3{\times}2$ arrangement of treatments to determine the effects of sorghum hybrid and grain supplementation on nutrient digestibilities and passage rates and ruminal metabolism of silage-based diets fed to growing steers. The diets consisted of three wholes-plant silages (a high grain-containing, grain sorghum and middle-season, moderate grain-containing, and late-season, low grain-containing forage sorghums), each fed with or without 25% rolled grain sorghum. No significant interactions occurred between sorghum hybrid and grain supplementation for the digestion or passage rate criteria measured. Ruminal butyrate concentration was the only fermentation characteristic affected by a hybrid ${\times}$ grain supplementation interaction. The grain sorghum silage diets had the highest DM, OM, and ADF digestibilities; the late-season silage diets, the lowest. Digestibility of NDF tended to be highest (p < 0.10) for the grain sorghum silage, whereas starch digestibility was not affected by sorghum hybrid. Ruminal ammonia, acetate, propionate, butyrate, and total VFA concentrations were highest for the grain sorghum silage diets. Grain supplementation increased DM and OM digestibilities, but had no effect on digestibilities of NDF, ADF, and starch. Ruminal pH was decreased, but total VFA concentration and acetate : propionate ratio were not affected by grain supplementation.

• 유기물자원화
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• 제14권1호
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• pp.131-138
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• 2006

본 연구는 혐기성 발효를 이용한 수소생산과정에서 pH를 일정하게 유지한 상태에서 안정적인 수소를 생산하기 위한 최적 pH를 찾고자 하였다. 실험결과 pH 6이었을 때 최적의 조건이었으며 이때 수소생산량은 1175.87 mL/L 이고, 이론적 수소 전환율은 22.51 %였다. pH 5는 901.77 mL/L의 수소가 발생되었으며, 수소 전환율은 17.48 %, pH 7은 454.26 mL/L의 수소생산량과 8.74 %의 수소 전환율을 나타내었다. 그리고 pH 7과 8에서는 각각 452.08 mL/L와 82.25 mL/L의 저조한 수소 생산량을 나타내었다. 자당의 혐기성 발효를 통한 유기산 생성에 있어서 pH가 7과 8에서는 propionate가 주로 관찰되었으나 pH가 5와 6인 영역 에서는 butyrate의 생성이 두드러지는 결과가 나타났다.

• Applied Biological Chemistry
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• 제27권1호
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• pp.14-20
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• 1984

우리 나라에서 비교적 생산량이 많은 사과과실 3품종(Fuji, 홍옥, 국광)을 대상으로 향기성분을 분리 및 동정하고, 또한 성숙시기별로 향기성분의 함량변화를 측정하였다. 향기성분은 30종 이상 분리되었고 그 중 ester 23종, alcohol 9종 및 aldehyde 2종을 동정하였다. 성숙중 향기성분의 종류와 함량은 증가하였으며 1-butanol, isobutyl butyrate, 2-pentanol, ethyl valerate 및 hexanal 등의 함량이 많았다.

• Asian-Australasian Journal of Animal Sciences
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• 제13권5호
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• pp.636-640
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• 2000

Cup-plant (Silphium perfoliatum L.) has potential to produce high biomass and highly digestible forage in the wetlands where other productive forages do not grow or produce well. However, high moisture content at harvest is a considerable disadvantage of cup-plant for the production of high quality silage. This study was conducted to determine the effect of moisture content on the characteristics of cup-plant silage. Harvested cup-plant was ensiled in farm scale plastic bag silos and laboratory silos. In the plastic bag silos, first growth (FG) and regrowth (RG) cup-plant was harvested, wilted and ensiled. Dry matter content of FG and RG was 280 g/kg and 320 g/kg after 48 hr of wilting. The silage made with FG had pH 5.3 and 5.63 g/kg DM of acetate as a major volatile fatty acid. The composition of lactate, butyrate and acetate production was 1.0: 0.9: 2.3. The pH of silage made with RG was 4.5 and lactate was a major fermentation end product (16.8 g/kg DM). In the laboratory silos, wilted and unwilted first growth cup-plant material was ensiled to compare the early fermentation end products at days 2, 4, 11, and 40. Wilting increased dry matter content by 42% in the harvested material. Wilted silage showed about one unit lower pH until day 11. The contents of ammonia nitrogen and acetate were higher in un wilted silage, while that of lactate was higher in wilted silage (p<0.05). Butyrate and propionate were not detected in the wilted silage until day 40. We conclude from the results that moisture control is essential for the production of high quality cup-plant silage and high pH of cup-plant silage is due to low concentrations of fermentation end products.

• KSBB Journal
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• 제20권6호
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• pp.443-446
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• 2005

음식물 쓰레기로부터의 연속 수소 생성 실험을 다양한 수리학적 체류시간(HRT; 18, 21, 24, 30, 36, 42 h)에서 수행하였다. 음식물쓰레기는 분쇄와 알칼리 처리를 거쳐 27.0 g COD/L(average VS 4.4%)의 농도로 fed-batch 형태로 주입되었으며, 반응조 내의 pH는 $5.3{\pm}0.1$ 이상으로 유지되었다. 126일 간의 운전을 통해 유기성 폐기물로부터의 연속 수소 생성이 안정적으로 진행될 수 있음을 확인하였다. 수소 생성 효율은 HRT에 따라 변하였으며, 30 h에서 가장 높은 수치를(25.8 mL $H_2/g\;VS_{added}$, 0.36 mol $H_2/mol\;hexose_{added}$, 0.91 L $H_2/L/d$) 보였다. 대부분의 조건에서 가장 양이 많은 부산물은 노말부티르산이었으며, 수소생성이 증가함에 따라 노말부티르산의 생성이 증가하였다. 이소프로판을 역시 수소 생성과 관련이 있는 나타났다. 반면, 아세트산의 생성량은 수소생성과 반대되는 경향을 보여 수소 소모 아세트산 생성 경로로 발생되는 양이 많았다고 사료된다. 한편 산발효 효율은 $53.3{\sim}65.7%$인 것으로 나타나 기존 산 발효를 수소발효가 대체할 수 있음을 확인하였다.

• Asian-Australasian Journal of Animal Sciences
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• 제26권4호
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• pp.523-528
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• 2013

This study investigates the effects of three branched-chain amino acids (BCAA; valine, leucine, and isoleucine) on the in vitro ruminal fermentation of wheat straw using batch cultures of mixed ruminal microorganisms. BCAA were added to the buffered ruminal fluid at a concentration of 0, 2, 4, 7, or 10 mmol/L. After 72 h of anaerobic incubation, pH, volatile fatty acids (VFA), and ammonia nitrogen ($NH_3$-N) in the ruminal fluid were determined. Dry matter (DM) and neutral detergent fiber (NDF) degradability were calculated after determining the DM and NDF in the original material and in the residue after incubation. The addition of valine, leucine, or isoleucine increased the total VFA yields ($p{\leq}0.001$). However, the total VFA yields did not increase with the increase of BCAA supplement level. Total branched-chain VFA yields linearly increased as the supplemental amount of BCAA increased (p<0.001). The molar proportions of acetate and propionate decreased, whereas that of butyrate increased with the addition of valine and isoleucine (p<0.05). Moreover, the proportions of propionate and butyrate decreased (p<0.01) with the addition of leucine. Meanwhile, the molar proportions of isobutyrate were increased and linearly decreased (p<0.001) by valine and leucine, respectively. The addition of leucine or isoleucine resulted in a linear (p<0.001) increase in the molar proportions of isovalerate. The degradability of NDF achieved the maximum when valine or isoleucine was added at 2 mmol/L. The results suggest that low concentrations of BCAA (2 mmol/L) allow more efficient regulation of ruminal fermentation in vitro, as indicated by higher VFA yield and NDF degradability. Therefore, the optimum initial dose of BCAA for in vitro ruminal fermentation is 2 mmol/L.