• Title/Summary/Keyword: alkaline fermentation

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

  • 김희숙
    • Journal of Life Science
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    • v.9 no.2
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    • pp.160-168
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    • 1999
  • Butyrate is one of the short-chain fatty acids that are present in the colon of mammals in millimolar concentration as a result of microbial anaerobic fermentation of dietary fiber, undigested starch, and proteins. In this study, sodium butyrate was examined in HT29 cell, human colonic cancer cell line, on cell viability, alkaline phosphatase activity, PLC-${\gamma}$1 expression and complex sphingolipid biosynthesis. Treatment with butyrate showed that the decrease of cell adhesion and viability was time-dependent. Sodium butyrate also induced to increase the activity of alkaline phosphatase which is a differentiation marker enzyme and decrease the expression of PLC-${\gamma}$1. Biosynthesis of sphingomyelin and galactosylceramide by butyrate treatment were decreased so fast but ceramide was increased 680dpm/mg protein% more than untreated group on first day and then decreased fast. In addition, acid ceramidase and neutral ceramidase activity were inhibited early stage by sodium butyrate. These results suggest that sodium butyrate causes cell differentiation or cell growth arrest of HT29 cell accompanied by early increase of ceramide content and alkaline phosphatase activity and decrease of galactosylceramide content and PLC-r1 expression.

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

  • Shin, Yong-Uk;Lee, Gwa-Soo;Jo, Jae-Hyung;Lee, Hyune-Hwan
    • Korean Journal of Microbiology
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    • v.46 no.4
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    • pp.383-388
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    • 2010
  • Alkaline protease-overproducing strains of Vibrio metschnikovii were developed by using the molecular evolution from the classical mutants V. metschnikovii L12-23, N4-8, and KS1. Each vapK (Vibrio alkaline protease K) was obtained from the genomic DNAs of mutants by PCR to carry out the DNA shuffling. The modified vapK-1 obtained by DNA shuffling was used again as a template for the error-prone PCR to make the vapK-2. Both genes were cloned in the plasmid pKF3 to construct the recombinant plasmids which have one or two copies of the modified genes. The recombinant plasmids were back-transformed to V. metschnikovii KS1 to construct recombinant V. metschnikovii that expresses the alkaline protease. About 3.9-fold more protease activity was measured in the strain which has the plasmid containing two copies of vapK-2 when compared to strain KS1. When compared to wild type V. metschnikovii RH530, 43-fold more activity was achieved. Comparison of amino acids among vapK, vapK-1, and vapK-2 revealed that the active sites was highly conserved and not changed. However, many amino acids except the active sites were changed. These results suggested that the changes in amino acids might play an important role in the increase of protease activity by allowing the easy access of substrate to active sites of the protease. The fermentation of alkaline protease from the V. metschnikovii KS1 harboring the plasmid that contains two copies of vapK-1 showed the possibility of this strain to be used as industrial producer.

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

  • Hong, In-Gi;Kim, Soo-Ryang;Lee, Myung-Gyu
    • Journal of Animal Environmental Science
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    • v.18 no.sup
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    • pp.13-20
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    • 2012
  • This research investigated the effect of $SCOD_{Mn}$ concentrations and pH adjustment at the stage before land application, namely 2nd-aeration treatment stage of liquid fertilizer in the liquid fertilizer treatment process of swine manure on the physicochemical compositions of 2nd-aeration treated liquid fertilizer. The liquid fertilizer used in this research is the alkaline fermented liquid fertilizer of swine manure more than pH 9.0 through aeration treatment (Alkaline fermentation treatment group). About the alkaline liquid fertilizer, phosphate neutralization treatment was conducted with phosphoric acid and it was a phosphate neutralization treatment group. In 2nd-aeration treatment of liquid fertilizer for 30 days, each group was divided into alkaline treatment groups (T-1, T-2, and T-3) and phosphate neutralization treatment groups (T-4, T-5, and T-6) according to early $SCOD_{Mn}$ concentrations. The research results are as follows. 1. As for $SCOD_{Mn}$ reduction rate, the average 29.9% in alkaline treatment groups and the average 36.9% in phosphate neutralization treatment groups were shown and so the relatively high reduction rate was shown in phosphate neutralization treatment groups. 2. After finishing the experiment, the group of the lowest $SCOD_{Mn}$ concentrations was the phosphate neutralization treatment group, T-6 with the lowest inflow concentrations. In case the final goal level of 2nd-aeration treated liquid fertilizer is assumed as concentrations less than $SCOD_{Mn}$ 3,000 ppm, it would be desired that inflow concentrations of 2nd-aeration treatment groups are adjusted less than $SCOD_{Mn}$ 5,500 ppm. 3. As for the persistence rate of nitrogen, the average 29.3% in alkaline treatment groups and the average 38.9% in phosphate neutralization treatment groups were shown and so phosphate neutralization treatment groups showed the relatively low loss rate of nitrogen, meanwhile, in the case of T-P, phosphate neutralization treatment groups maintained high concentrations (average 1,473 ppm). 4. In the event of 2nd-aeration treatment of liquid fertilizer, "alkaline fermentation treatment" condition in 'low phosphate-low nitrogen' type and "phosphate neutralization treatment" condition in 'high phosphate-high nitrogen' type are expected to be favorable.

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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    • v.2 no.3
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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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    • v.17 no.3
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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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    • v.61 no.3
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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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    • v.26 no.7
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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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    • v.18 no.6
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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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    • v.22 no.12
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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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    • v.57 no.4
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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.