• 제목/요약/키워드: alkaline fermentation

검색결과 93건 처리시간 0.02초

Sodium Butyrate 처리가 대장암 세포주인 HT29 Cell의 Sphingolipid 생합성에 미치는 영향 (Effects of Sodium Butyrate on the Biosynthesis of Sphingolipids in HT29, a Human Colon Cancer Cell Line)

  • 김희숙
    • 생명과학회지
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    • 제9권2호
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    • pp.160-168
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    • 1999
  • Butyrate는 탄소사슬이 짧은 지방산 중 하나로 소화되지 않은 식이성 섬유의 혐기적 발효결과 포유동물의 위장관내에 millimolar 농도로 유지되며 대장의 상피세포에서 흡수되어 energy원으로 사용된다. 70%정도 confluent하게 자란 사람의 대장암세포주인 HT29 cell에 5mM sodium butyrate를 시간별로 처리하고 세포의 생존율, 암세포의 분화정도의 biomarker로 알려진 alkaline phosphatase 및 PLC-rl의 발현정도를 측정하였으며 sphingolipid의 생합성 및 ceramidase 활성도 측정하였다. Sodium butyrate 처리는 성장중인 HT29 cell의 부착을 저해하여 처리 1일째부터 세포수가 감소하였고 형질막 효소인 alkaline phosphatase의 발현을 증가시켰으며 PLC-${\gamma}$의 발현을 감소시켰다. 또한, 복합 sphingolipid들의 생합성을 측정한 결과, 세포성장의 저해와 함께 sphingomyelin은 2일째부터 감소하였으며, galactosyl ceramide는 1일째부터 급속히 감소하였다. 그러나 ceramide의 경우, 1일째는 처리하기 전보다 680dpm/mg protein정도 증가하였으며 2일째에는 다시 급속히 감소하였다. 또한 butyrate처리에 의하여 HT29 cell의 acid ceramidase와 neutral ceramidase활성이 저해됨을 관찰하였는데 그 결과 ceramide함량이 초기에 증가된 것으로 생각된다. 본 실험결과들로부터 HT29 cell의 sodium butyrate처리는 세포분화 또는 세포성장저해를 가져오는데 이와 함께 초기의 ceramide함량 및 alkaline phosphatase활성의 증가와 galactosylceramide함량 및 LC-rl 발현의 감소현상이 동반됨을 알 수 있다.

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분자진화 기술을 통한 Vibrio metschnikovii 유래 고활성 알칼리성 단백질 분해효소 생산균주 개발 (Strain Development for the Over-production of Alkaline Protease from Vibrio metschnikovii by Molecular Evolution)

  • 신용욱;이과수;조재형;이현환
    • 미생물학회지
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    • 제46권4호
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    • pp.383-388
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    • 2010
  • 알칼리성 단백질 분해효소 고생산 돌연변이 균주 Vibrio metschnikovii L12-23, N4-8, KS1으로부터 알칼리성 단백질 분해효소를 암호화하는 vapK (Vibrio alkaline protease K) 유전자들을 PCR에 의하여 분리한 다음 DNA shuffling, error-prone PCR 방법과 같은 분자진화 기술을 통해 고활성 단백질 분해효소를 생산하는 재조합 V. metschnikovii 균주를 제작하였다. DNA shuffling 방법을 통해 변형시킨 vapK-1 유전자와 이 유전자를 주형으로 error-prone PCR 기법을 통해 재 변형된 vapK-2 유전자를 cloning한 후 V. metschnikovii KS1 균주에 역도입하여 재조합 균주를 제조하였다. 재조합 균주들의 단백질 분해 능력을 조사한 결과 vapK-2 유전자가 2 copy 도입된 재조합 균주의 경우 야생형 균주인 V. metschnikovii RH530에 비해 43.6배 높은 단백질 분해활성을 보였으며 숙주인 V. metschnikovii KS1에 비해 약 3.9배 향상된 단백질 분해 활성을 확인할 수 있었다. 변형된 vapK-1과 vapK-2 유전자를 야생형 vapK 유전자의 염기서열을 비교 분석한 결과 단백질 분해 능력의 활성에 영향을 미치는 active site를 제외한 부분에서 변화가 일어났음을 확인 할 수 있었다. 변형된 유전자 vapK-1을 two copy를 포함한 재조합 플라스미드를 가진 V. metschnikovii KS1을 30 L fermentor로 배양 하였을 때 배양 후 35 시간에 18,000 PU/ml의 활성을 보였으며, 이는 향후 산업용 균주로서 사용될 수 있는 가능성을 제시하였다.

SCODMn 농도 및 pH 조정이 양돈분뇨의 후숙발효과정에 미치는 이화학적 영향 (Effect of SCODMn and pH Adjustment on Physicochemical Characteristics in Liquid Fertilizer Production Process Using Swine Manure)

  • 홍인기;김수량;이명규
    • 한국축산시설환경학회지
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    • 제18권sup호
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    • pp.13-20
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    • 2012
  • 연구는 양돈분뇨의 액비화 처리과정 중 농지환원 직전단계인 액비 후숙처리 단계에 있어서 $SCOD_{Mn}$ 농도 및 산도(pH)의 조정이 후숙액비의 이화학적성상에 미치는 영향을 알아보고 적절한 후숙 액비화 공정을 구축하기 위하여 수행하였으며, 실험결과를 요약하면 다음과 같다. 1. $SCOD_{Mn}$ 감소율은 알칼리발효 처리구에서 평균 29.9%, 인산중성화 처리구에서는 평균 36.9%로 인산 중성화 처리구가 비교적 높은 감소율을 보였다. 2. 30일간 처리 후 $SCOD_{Mn}$의 농도가 가장 낮은 처리구는 유입농도가 가장 낮았던 인산 중성화 처리구 T-6이다. 후숙 액비의 목표수준을 $SCOD_{Mn}$ 3,000 ppm 이하로 가정할 경우 후숙 발효조의 초기 투입농도는 5,500 ppm 이하로 조정하는 것이 적합하다고 사료된다. 3. 질소의 잔존율은 알칼리발효 처리구에서 평균 29.3%, 인산중성화 처리구에서 평균 38.9%로 인산중성화 처리구가 비교적 질소의 손실이 적은 한편, T-P의 경우 인산중성화 처리구에서 높은 농도(평균 1,473 ppm)로 유지되었다. 4. 본 연구를 통해 액비 후숙처리 시 '저인산-저질소'의 형태는 "알칼리발효 처리구", '고인산-고질소'의 형태는 "인산중성화 처리구"의 조건이 유리할 것으로 기대된다.

Effects of Environmental Conditions on Expression of Bacillus subtilis $\alpha$-Amylase in Recombinant Escherichia coli

  • Shin, Pyong-K.;Nam, Seung-H.
    • Journal of Microbiology and Biotechnology
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    • 제2권3호
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    • pp.166-173
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    • 1992
  • The expression of Bacillus subtilis $\alpha$-amylase from the phoA-amyE fusion gene in recombinant E. coli was investigated under various environmental conditions. The overexpression of cloned $\alpha$-amylase caused retardations in cell growth and synthesis of alkaline phosphatase (AP) from the chromosomal phoA gene. The change of culture temperature from $37^\circ{C}$ to $30^\circ{C}$ increased the specific activities of both $\alpha$-amylase and $\beta$-lactamase by six and two times, respectively, whereas the AP activity remained unchanged. The experiments with chlorampenicol (a translation inhibitor) suggested the enhancement of $\alpha$-amylase activity at $30^\circ{C}$, and this was partly due to the stability of $\alpha$-amylase itself. The further decrease of the temperature to $25^\circ{C}$ slowed down both the cell growth and cloned-gene expression rate. The $\alpha$-amylase activity showed a maximum at pH of 7.4 while alkaline phosphatase was most effectively produced at pH of 8.3.

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Effect of Sodium Hydroxide plus Hydrogen Peroxide Treated Mustard (Brassica campestris) Straw Based Diets on Rumen Degradation Kinetics (In sacco), Fermentation Pattern and Nutrient Utilization in Sheep

  • Mishra, A.S.;Misra, A.K.;Tripathi, M.K.;Santra, A.;Prasad, R.;Jakhmola, R.C.
    • Asian-Australasian Journal of Animal Sciences
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    • 제17권3호
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    • pp.355-365
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    • 2004
  • Two experiments were conducted to determine the effect of alkaline hydrogen peroxide (AHP) treatment (1% NaOH+1.5% $H_2O_2$; 1 AHPMS, 2% NaOH+1.5% $H_2O_2$; 2AHPMS) on rate and extent of degradation of mustard straw (MS) in sacco in sheep, and its in vivo digestion and ruminal fermentation characteristics when fed to sheep with concentrate (200 g per sheep daily). The treatment of straw with 1 and 2% AHP increased its sodium content by 148 and 296% to that of untreated straw (UMS). There was significant decrease in NDF and hemicellulose contents of AHP treated straw and increase in cellulose and lignin contents. Phenolic acids like ferrulic, $\rho$-coumaric and o-coumaric significantly (p<0.001) reduced by AHP treatment of mustard straw. In first experiment the in sacco degradation of DM, OM and NDF was significantly (p<0.01) greater for 2 AHPMS than for UMS at all incubation periods. The disappearance of nutrient from 1 AHPMS and 2 AHPMS treated straws continue to increase up to 96 h whereas in UMS the peak disappearance was found at 48 h. By using the equation {(y=a+b) ($1-e^{-ct}$)} the degradation rates (c) for DM, OM, and NDF were significantly higher for UMS than AHP treated straws. Level of alkali (1 and 2%) had significant effect on degradation characteristics (a, b, c and $P_{0.05}$) of DM and NDF fraction of MS. However, the effect was not pronounced on OM fraction of MS. In feeding experiment, the intake of nutrients for DM, OM, cell wall constituents and energy was higher on 2 AHPMS, whereas no effect on the digestibility of these nutrients was observed. The apparent nitrogen retention was higher (p<0.05) both in 1 and 2 AHPMS groups. Water intake by animals was significantly increased due to AHP treated mustard straw feeding. Rumen liquor pH was higher in 2 AHPMS fed animals. The $NH_3-N$ of rumen liquor was not affected by feeding of AHP treated MS based diets. Total VFA concentration was significantly (p<0.01) higher in UMS fed group. The fractional out flow rate of DM was higher (p<0.05) in animals fed on 2 AHPMS diets compared to UMS and lAHPMS fed groups. The population of large holotrichs was higher (p<0.05) on AHP treated MS fed diets compared to UMS. The study indicated that treatment of mustard straw with AHP changed its chemical composition towards a better feed. The nutritive value of 2% AHP treated mustard straw was better in terms of dry matter intake and apparent nitrogen retention. The higher in sacco DM, OM and NDF disappearance however, was not confirmed by in vivo data in this study.

Alkaline Peroxide Pretreatment of Waste Lignocellulosic Sawdust for Total Reducing Sugars

  • Satish Kumar Singh;Sweety Verma;Ishan Gulati;Suman Gahlyan;Ankur Gaur;Sanjeev Maken
    • Korean Chemical Engineering Research
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    • 제61권3호
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    • pp.412-418
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    • 2023
  • The surge in the oil prices, increasing global population, climate change, and waste management problems are the major issues which have led to the development of biofuels from lignocellulosic wastes. Cellulosic or second generation (2G) bioethanol is produced from lignocellulosic biomass via pretreatment, hydrolysis, and fermentation. Pretreatment of lignocellulose is of considerable interest due to its influence on the technical, economic and environmental sustainability of cellulosic ethanol production. In this study, furniture waste sawdust was subjected to alkaline peroxide (H2O2) for the production of reducing sugars. Sawdust was pretreated at different concentrations from 1-3% H2O2 (v/v) loadings at a pH of 11.5 for a residence time of 15-240 min at 50, 75 and 90 ℃. Optimum pretreatment conditions, such as time of reaction, operating temperature, and concentration of H2O2, were varied and evaluated on the basis of the amount of total reducing sugars produced. It was found that the changes in the amount of lignin directly affected the yield of reducing sugars. A maximum of 50% reduction in the lignin composition was obtained, which yielded a maximum of 75.3% total reducing sugars yield and 3.76 g/L of glucose. At optimum pretreatment conditions of 2% H2O2 loading at 75 ℃ for 150 min, 3.46 g/L glucose concentration with a 69.26% total reducing sugars yield was obtained after 48 hr. of the hydrolysis process. Pretreatment resulted in lowering of crystallinity and distortion of the sawdust after the pretreatment, which was further confirmed by XRD and SEM results.

Impact of Lactic Acid and Hydrogen Ion on the Simultaneous Fermentation of Glucose and Xylose by the Carbon Catabolite Derepressed Lactobacillus brevis ATCC 14869

  • Jeong, Kyung Hun;Israr, Beenish;Shoemaker, Sharon P.;Mills, David A.;Kim, Jaehan
    • Journal of Microbiology and Biotechnology
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    • 제26권7호
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    • pp.1182-1189
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    • 2016
  • Lactobacillus brevis ATCC 14869 exhibited a carbon catabolite derepressed phenotype that has ability to consume fermentable sugars simultaneously with glucose. To evaluate this unusual phenotype under harsh conditions during fermentation, the effects of lactic acid and hydrogen ion concentrations on L. brevis ATCC 14869 were examined. Kinetic equations describing the relationship between specific cell growth rate and lactic acid or hydrogen ion concentration were deduced empirically. The change of substrate utilization and product formation according to lactic acid and hydrogen ion concentration in the media were quantitatively described. Although the simultaneous utilization has been observed regardless of hydrogen ion or lactic acid concentration, the preference of substrates and the formation of two-carbon products were changed significantly. In particular, acetic acid present in the medium as sodium acetate was consumed by L. brevis ATCC 14869 under extreme pH of both acid and alkaline conditions.

Alcoholic Hepatotoxicity Suppression in Alcohol Fed Rats by Glutathione-enriched Yeast FF-8 Strain

  • Cha, Jae-Young;Kim, Hyeong-Soo;Kang, Sun-Chul;Cho, Young-Su
    • Food Science and Biotechnology
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    • 제18권6호
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    • pp.1411-1416
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    • 2009
  • The suppressive effects of glutathione-enriched Saccharomyces cerevisiae FF-8 strain (FF-8 GY) on alcoholinduced hepatotoxicity have been studied. FF-8 GY (256 mg/L) from the fermentation at a large scale bioreactor was used. Either of 5% FF-8 GY or 5% commercial glutathione-enriched yeast extract (GYE) with or without 30% alcohol was tested with rats for 4 weeks. FF-8 GY and GYE were found to reduce those alcohol-elevated serum alanine aminotransferase (ALT), alkaline phosphatase (ALP), and lactate dehydrogenase (LDH) activities. Blood alcohol and acetaldehyde were also decreased by FF-8 GY and GYE. Interestingly, FF-8 GY drastically increased both hepatic alcohol dehydrogenase (ADH) and acetaldehyde dehydrogenase (ALDH) activities in comparison to GYE group, thus FF-8 GY would be more effective in blood alcohol and acetaldehyde reduction. Attenuated lipid droplet accumulation in hepatocytes was observed in both FF-8 GY and GYE when alcohol stimulated the accumulation. Therefore, FF-8 GY may be useful to protect liver from alcohol-induced hepatotoxicity.

Kinetic Studies of Alkaline Protease from Bacillus licheniformis NCIM-2042

  • Bhunia, Biswanath;Basak, Bikram;Bhattacharya, Pinaki;Dey, Apurba
    • Journal of Microbiology and Biotechnology
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    • 제22권12호
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    • pp.1758-1766
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    • 2012
  • An extensive investigation was carried out to describe the kinetics of cell growth, substrate consumption, and product formation in the batch fermentation using starch as substrate. Evaluation of intrinsic kinetic parameters was carried out using a best-fit unstructured model. A nonlinear regression technique was applied for computational purpose. The Andrew's model showed a comparatively better $R^2$ value among all tested models. The values of specific growth rate (${\mu}_{max}$), saturation constant ($K_S$), inhibition constant ($K_I$), and $Y_{X/S}$ were found to be 0.109 $h^{-1}$, 11.1 g/l, 0.012 g/l, and 1.003, respectively. The Leudeking-Piret model was used to study the product formation kinetics and the process was found to be growth-associated. The growth-associated constant (${\alpha}$) for protease production was sensitive to substrate concentration. Its value was fairly constant up to a substrate concentration of 30.8 g/l, and then decreased.

Purification of Biohydrogen Produced From Palm Oil Mill Effluent Fermentation for Fuel Cell Application

  • Rohani, Rosiah;Chung, Ying Tao;Mohamad, Izzati Nadia
    • Korean Chemical Engineering Research
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    • 제57권4호
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    • pp.469-474
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    • 2019
  • Fermentation of palm oil mill effluent (POME) produces biohydrogen in a mixture at a specific set condition. This research was conducted to purify the produced mixed biohydrogen via absorption and membrane techniques. Three different solvents, methyl ethanolamine (MEA), ammonia ($NH_3$) and potassium hydroxide (KOH) solutions, were used in absorption technique. The highest $H_2$ purity was found using 1M MEA solution with 5.0 ml/s feed mixed gas flow rate at 60 minutes absorption time. Meanwhile, the purified biohydrogen using a polysulfone membrane had the highest $H_2$ purity at 2~3 bar operating pressure. Upon testing with proton exchange membrane fuel cell (PEMFC), the highest current and power produced at 100% $H_2$ were 1.66 A and 8.1 W, while the lowest were produced at 50/50 vol% $H_2/CO_2$ (0.32 A and 0.49 W). These results proved that both purification techniques have significant potential for $H_2$ purification efficiency.