• Title/Summary/Keyword: fermentation${\beta}$ cells

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Silybin Synergizes with Wnt3a in Activation of the Wnt/${\beta}$-catenin Signaling Pathway through Stabilization of Intracellular ${\beta}$-Catenin Protein (Silybin에 의한 Wnt/${\beta}$-catenin 신호전달체계의 활성화)

  • Kim, Tae-Yeoun;Oh, Sang-Taek
    • Microbiology and Biotechnology Letters
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    • v.40 no.1
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    • pp.50-56
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    • 2012
  • The Wnt/${\beta}$-catenin signaling pathway regulates diverse developmental processes and adult tissue homeostasis. Inappropriate regulation of this pathway has been associated with human diseases, such as cancers, osteoporosis, and Alzheimer's disease. Using a cell-based chemical screening with natural compounds, we discovered silybin, a plant flavonoid isolated from the Silybum marianum, which activated the Wnt/${\beta}$-catenin signaling pathway in a synergy with Wnt3a-conditioned medium (Wnt3a-CM). In the presence of Wnt3a-CM, silybin up-regulated ${\beta}$-catenin response transcription (CRT) in HEK293-FL reporter cells and 3T3-L1 preadipocytes through stabilization of intracellular ${\beta}$-catenin protein. Silybin and Wnt3a-CM synergistically reduced expression of important adipocyte marker genes including peroxisome-proliferator-activated $receptor{\gamma}$ ($PPAR{\gamma}$) and CAATT enhancer-binding protein ${\alpha}$ (C/$EBP{\alpha}$) in 3T3-L1 preadipocytes, accompanied by the activation of Wnt/${\beta}$-catenin signaling pathway. Taken together, our findings indicate that silybin is a small-molecule synergist of the Wnt/${\beta}$-catenin signaling pathway and can be used as a controllable reagent for investigating biological processes that involve the Wnt/${\beta}$-catenin signaling pathway.

Impeller Types and Feeding Modes Influence the Morphology and Protein Expression in the Submerged Culture of Aspergillus oryzae

  • Heo, Joo-Hyung;Vladimir Ananin;Park, Jeong-Seok;Lee, Chung-Ryul;Moon, Jun-Ok;Ohsuk Kwon;Kang, Hyun-Ah;Kim, Chul-Ho;Rhee, Sang-Ki
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.9 no.3
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    • pp.184-190
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    • 2004
  • The influences of impeller types on morphology and protein expression were investigated in a submerged culture of Aspergillus oryzae. The impeller types strongly affected mycelial morphology and protein production in batch and fed-batch fermentations. Cells that were cultured by propeller agitation grew in the form of a pellet, whereas cells that were cultured by turbine agitation grew in a freely dispersed-hyphal manner and in a clumped form. Pellet-grown cells showed high levels of protein production for both the intracellularly heterologous protein (${\beta}$-glucuronidase) and the extracellularly homologous protein (${\alpha}$-amylase). The feeding mode of the carbon source also influenced the morphological distribution and protein expression in fed-batch fermentation of A. oryzae. Pulsed-feeding mainly showed high protein expression and homogeneous distribution of pellet whereas continuous feeding resulted in less protein expression and heterogeneous distribution with pellet and dispersed-hyphae. The pellet growth with propeller agitation paralleling with the pulsed-feeding of carbon source showed a high level of protein production in the submerged fed-batch fermentation of recombinant A. oryzae.

Fermentation process increased antimutagenic and in vitro anticancer effects during Kochujang manufacturing (고추장 제조시 발효가 항돌연변이 및 in vitro 항암효과에 미치는 영향)

  • Kim, Ji-Young;Liu, Fang-Fang;Lim, Yaung-Iee;Park, Kun-Young
    • Food Science and Preservation
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    • v.21 no.6
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    • pp.878-884
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    • 2014
  • Increased antimutagenic and in vitro anticancer effects were observed by the fermentation process during Kochujang manufacturing. In order to confirm the increased functionality, wheat grain, first fermented wheat grains (FFWG), second fermented wheat grains (SFWG), final fermented wheat grains (FiFWG), red pepper powder (RPP), and commercial Kochujang (CK) were prepared. Kochujang manufactured with final fermented wheat grains and red pepper powder were further fermented for 15 days and 30 days. The antimutagenic effects were determined by counting the number of revertants in Salmonella Typhimurium TA100 against N-methyl-N'-nitro-nitrosoguanidine (MNNG, 1.0mg/mL). The final fermented wheat grains (52% inhibition) showed higher antimutagenic effects than the wheat grain (34%), and the commercial Kochujang showed the highest antimutagenic effects (55%). We tested the inhibitory effect on the growth of HT-29 human colon carcinoma cells and AGS human gastric adenocarcinoma cells by using MTT (3-(4,5-dimethyl-thiazol)-2,5-diphenyltetrazolium bromide) assay. The results showed that increased fermentation process continually increased the growth inhibitory effect on both cancer cells. The further fermentation for 15 days of the Kochujang product also increased inhibitory growth of the AGS cancer cells. In conclusion, the methanol extract from fermented wheat grains and commercial Kochujang showed sequentially increased antimutagenic and in vitro anticancer activity, and thus the final commercial Kochujang revealed the highest effect.

Semicontinuous Production of Blud Pigment from Gardenia Fruit by Immobilized Cells of Bacillus subtilis KS-380 Using Air Bubble Column Reactor (Air Bubble Column Reactor를 이용하여 Bacillus Subtilis KS-380의 고정화에 의한 치자로부터 청색 색소의 생산)

  • 류병호;조경자
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.19 no.5
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    • pp.395-402
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    • 1990
  • The semicontinuous production of blue pigment from gardenia fruit by immobilized cells of Bacillus subtilis KS-380 which excreted $\beta$-glucosidase was investigated in comparison with free cells, . The blue pigment produced higher productivity under the conditions of aeration of 0.2m$\ell$/min and 2mm diameter of gel beads by using 3.5% sodium alginate. Semicontinuous production by immobilized cell showed the highest productivity with replacement of fresh production medium in every 24hr for fourth fermentation cycle following the conditions of blue pigment productivity.

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Study on the variation of cellular physiology of Escherichia coli during high cell density cultivation using 2-dimensional gel electrophoresis

  • Yun, Sang-Seon;Lee, Sang-Yeop
    • 한국생물공학회:학술대회논문집
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    • 2000.04a
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    • pp.219-222
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    • 2000
  • Physiological changes of Escherichia coli during the fed-batch fermentation process were characterized in this study. Overall cellular protein samples prepared at the different stage of fermentation were separated by 2-dimensional gel electrophoresis (2-DE), and differently expressed 15 proteins, Phosphotransferase enzyme I, GroEL, Trigger factor, ${\beta}$ subunit of ATP synthase, Transcriptional regulator KDGR, Phosphoglycerate mutase 1, Inorganic pyrophosphatase, Serine Hydroxymethyl-transferase, ${\alpha}$ subunit of RNA polymerase, Elongation factor Tu, Elongation factor Ts, Tyrosine-tRNA ligase, DnaK suppressor protein, Transcriptional elongation factor, 30S ribosomal protein S6 were identified using matrix-assisted laser desorption / ionization time-of-flight mass spectrometry (MALDI-TOF MS). When bacterial cells grow to high cell density, and IPTG-inducible heterologous protein is produced, expression level of overall cellular proteins was decreased. According to their functions in the cell, identified proteins were classified into three groups, proteins involved in transport process, small-molecule metabolism, and synthesis and modification of macromolecules.

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Effects of Sea Tangle and Chitosan on the Physicochemical Properties of Traditional Kochujang (다시마와 키토산을 첨가한 전통고추장의 품질특성에 관한 연구)

  • 권영미;김동한
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.31 no.6
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    • pp.977-985
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    • 2002
  • In order to improve the quality of traditional kochujang, submaterials like sea tangle and chitosan were added to kochujang and their effects on microbial characteristics, enzyme activities and physicochemical characteristics were investigated for 24 weeks of fermentation. The activities of $\alpha$,$\beta$-amylase in kochujmg were higher in sea tangle added at 2% level and chitosan added at 0.1% level. However, acidic protease activity decreased as the ratio of submaterials increased. Viable cells of yeasts in the kochujang increased rapidly for 4~8 weeks of fermentation, and bacterial counts decreased in submaterials added groups. Moisture contents of kochujang increased until 12 weeks of fermentation, but water activity decreased. As the ratio of sea tangle increased, water activity decreased. Consistency of kochujang increased after middle of fermentation, and they increased remarkably by addition of sea tangle. The degree of increase in total color difference ($\Delta$E) of sea tangle added group was lowest. The titratable acidity of kochujang decreased after 4 weeks, and they changed a little by addition of chitosan. Amino nitrogen contents of kochujang increased as mixing ratio of submaterials increased in the late period of aging. Ammonia nitrogen contents was lower in chitosan added kochujang at 24 week of fermentation. Reducing sugar contents of kochujang increased rapidly for 4~8 weeks of fermentation, and they increased as the ratio of chitosan increased. Ethanol contents of kochujang increased until 12~16 weeks of fermentation, with lower values in sea tangle added group. After 24 weeks of fermentation, the result of sensory evaluation showed that 0.1% chitosan added kochujang were more acceptable than sea tangle added kochujang in the taste, color and overall acceptability.all acceptability.

Lactic Acid Bacteria Increase Antiallergic Effect of Artemisia princeps Pampanini SS-1

  • Lee, Seung-Hoon;Shin, Yong-Wook;Bae, Eun-Ah;Lee, Bo-Mi;Min, Sung-Won;Baek, Nam-In;Chung, Hae-Gon;Kim, Nam-Jae;Kim, Dong-Hyun
    • Archives of Pharmacal Research
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    • v.29 no.9
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    • pp.752-756
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    • 2006
  • Artemisia princeps Pampanini, which is called Ssajuarissuk in Korean (SS-1), was fermented with lactic acid bacteria (LAB) and their passive cutaneous anaphylaxis reaction-inhibitory activity was investigated. Of these fermented agents, SS-1 extract fermented with Bifidobacterium infantis K-525 (F-SS-1) most effectively inhibited the release of ${\beta}$-hexosamindase from RBL-2H3 cells induced IgE. In IgE-induced RBL-2H3 cells, F-SS-1 inhibited proinflammatory cytokines IL-6 and $TNF-{\alpha}$ mRNA expression. Oral administration of SS-1 and F-SS-1 to mice inhibited passive cutaneous anaphylaxis (PCA) reaction induced by IgE and scratching behaviors induced by compound 48/80. The inhibitory activity of F-SS-1 against scratching behaviors was more effective than that of SS-1. These findings suggest that the fermentation of SS-1 with LAB can increase its antiallergic activity.

Optimization of Induction Conditions for Bacillus-derived Esterase Production by High-cell Density Fermentation of Recombinant Escherichia coli (재조합 대장균의 고농도 배양과 유도조건 최적화를 통한 Bacillus 유래 esterase의 생산)

  • Kang, Seung-Hoon;Min, Byung-Hyuk;Choi, Hong-Yeol;Kim, Dong-Il
    • Microbiology and Biotechnology Letters
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    • v.45 no.2
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    • pp.149-154
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    • 2017
  • To increase the efficiency of esterase production by Bacillus, high cell-density culture of recombinant Escherichia coli through fed batch fermentation was tested. Cells were cultured to $OD_{600}$ of 76 (35.8 g/l DCW) with dissolved oxygen level controlled to least above 30% air saturation by supplying pure oxygen. Cells were cultured to an $OD_{600}$ of 90 (42.4 g/l DCW) with glucose feeding controlled to at least 1 g/l. However, the cells reached stationary phase at the late stage of culture, despite glucose being supplied. Cells were cultured to an $OD_{600}$ of 185 (87.3 g/l DCW) by supplying additional medium with fortified yeast extract. To increase the productivity of the recombinant protein, cell growth and esterase productivity based on induction time were evaluated. Late exponential phase induction for esterase production in fed batch fermentation resulted in maximum optical density $OD_{600}$ of 190 (89 g/l DCW) and maximum esterase activity of 1745 U/l, corresponding to a 5.8-fold enhancement in esterase production, compared to the early exponential phase induction. In this study, we established fermentation methods for achieving maximum production of Bacillus-derived esterase by optimizing IPTG induction time in high-cell density culture by supplying pure oxygen and a nitrogen source.

Fermentation Increases Antidiabetic Effects of Acanthopanax Senticosusbhpark@chonbuk.ac.kr (발효에 의한 오가피의 항당뇨 활성 촉진)

  • Ham, Seong-Ho;Lim, Byung-Lak;Yu, Jia-hua;Ka, Sun-O;Park, Byung-Hyun
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.22 no.2
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    • pp.340-345
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    • 2008
  • Extract of Acanthopanax senticosus has recently been demonstrated to possess significant antidiabetic potential, in accordance with the traditional use of this plant as an antidiabetic natural health product. The present study evaluated the effects of fermented extract (FE) of this plant on glucose-stimulated insulin secretion, glucose uptake, and streptozotocin-induced type 1 diabetes model. A 3 h pretreatment with FE prevented $IL-1{\beta}$ and $IFN-{\gamma}$ toxicity in isolated rat islets. However, it did not affect insulin-stimulated glucose uptake in C2C12 myotubes. In addition, pretreatment of mice with FE blocked the destruction of streptozotocin-induced islets and the development of type 1 diabetes. FE reduced blood glucose level, increased insulin secretion, and improved glucose tolerance in streptozotocin-treated mice, whereas nonfermented extract (NFE) had moderate effects. Immunohistochemical staining for insulin clearly showed that pretreatment with FE blocked the STZ-induced islets destruction and restored the number of islet cells that secreted insulin to the level of the control. Although the active principles and their mechanisms of action remain to be identified, FE may nevertheless represent a novel complementary therapy and a source of novel therapeutic agents against type 1 diabetes mellitus.

Effect of Combined Use of various Anti-microbial Materials on Brewing of Low Salted Kochujang (저식염 고추장 양조시 항균물질 혼합첨가의 영향)

  • Park, Seon-Ju;Kim, Dong-Han
    • Applied Biological Chemistry
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    • v.50 no.4
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    • pp.287-294
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    • 2007
  • Effect of combined use of anti-microbial materials, such as ethanol, mustard and chitosan, on the quality of low salted kochujang was investigated during fermentation at $20^{\circ}C$ for 12 weeks. Viable cells of yeast increased remarkably during fermentation, but increasing ratio was significantly low in ethanol-mustard added kochujang. Activity of ${\beta}-amylase$ was high in anti-microbial material added kochujang, whereas ${\alpha}-amylase$ and protease activities were low in those groups. Water activity decreased during fermentation with being low in the control kochujang prepared with normal-salt without anti-microbial materials. Hunter L-, a- and b-values of kochujang increased during fermentation, and the degree of increase in total color difference $({\Delta}E)$ was low in ethanol added kochujang. Titratable acidity of kochujang was decreased in anti-microbial materials added group at late aging period, and oxidation-reduction potential was low in the control kochujang. Total sugar and reducing sugar contents of kochujang were high in ethanol-mustard added kochujang. Ethanol contents of kochujang increased at late aging period, with high values in ethanol-chitosan added kochujang. Amino nitrogen content increased during middle of fermentation, and ammonia nitrogen content of kochujang decreased in ethanol-mustard-chitosan added group during fermentation. After 12 weeks fermentation, sensory results showed that ethanol or ethanol-mustard added kochujang were the highest in color and flavor with the highest overall acceptability.