• Title/Summary/Keyword: $H^+-ATPase$

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Antifungal Activity of Medium-Chain ($C_{6}-C_{13}$) Alkenals against, and Their Inhibitory Effect on the Plasma Membrane $H^{+}$-ATPase of Saccharomyces cerevisiae

  • Lee, Jae-Ran;Lee, Sang-Hwa;Kubo, Isao;Hong, Soon-Duck
    • Journal of Microbiology and Biotechnology
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    • v.8 no.3
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    • pp.197-202
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    • 1998
  • Aliphatic alkenals having 6 to 13 carbons were evaluated for antifungal activity against Saccharomyces cerevisiae. The activity was gradually increased with chain length, e.g., (E)-2-decenal and (E)-2-undecenal exhibited maximum potency, while (E)-2-dodecenal and (E)-2-tridecenal were completely inactive. Alkenals showed increasing inhibitory activity with chain length, as in the case of antifungal activity, towards glucose-induced medium acidification by the plasma membrane $H^+$-ATPase of S. cerevisiae. The group including (E)-2-nonenal, (E)-2-decenal, and (E)-2-undecenal exhibited maximum potency, but the potency of (E)-2-dodecenal and (E)-2-tridecenal demonstrated a sudden drop with respect to the former group. (E)-2-Nonenal revealed dose-responsive inhibition to the medium acidification and inhibited over 90% at a concentration of 1.25 mM ($175.3{\mu}g$/ml). In contrast to (E)-2-undecenal whose inhibitory efficiency increased with incubation time, inhibition by (E)-2-dodecenal was reversed with time. Of the tested alkenals, (E)-2-heptenal and (E)-2-octenal most highly inhibited ATP hydrolytic activity by the plasma membrane $H^+$ ATPase, while (E)-2-heptenal at 10 mM ($1121.8{\mu}g$/ml) showed an inhibitory efficacy of 93.2%.

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Acid Tolerance of Lactobacillus brevis Isolated from Kimchi (김치에서 분리한 Lactobacillus brevis의 내산성)

  • Lee, Kap-Sang;Shin, Yong-Seo;Lee, Cherl-Ho
    • Korean Journal of Food Science and Technology
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    • v.30 no.6
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    • pp.1399-1403
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    • 1998
  • We isolated wild lactic acid bacteria from kimchi and identified as Lactobacillus brevis by using API 50 CHL Kit, some morphological and physiological tests. In order to evaluate the acid tolerance of Lactobacillus brevis, its survivals rate, glycolysis assay, membrane permeability, and pH profiles of $H^+-ATPase$ were also determined. When Lactobacillus brevis were incubated in Lactobacilli MRS broth adjusted to various levels of pH for 2 hours, the decreases in its population at pH 4.0 and 3.0 were about 2.61 log cycles/mL and 5.89 log cycles/mL, respectively, but there was no decrease at pH 6.0 and 5.0. Glycolysis by Lactobacillus brevis had optimal pH about 6.5 and glucose degradation was reduced by 50% at a pH of 5.2. $Mg^{++}$ release from Lactobacillus brevis cells in medium with pHs of 4.0 and 3.0 was 24.3 and 71.2% of totals, respectively. The $H^+-ATPase$ of Lactobacillus brevis showed a maximal activity between pH values of approximately 6.5 to 7.0.

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Characterization of Single Stranded DNA-Dependent ATPase Activities of Deinococcus radiodurans RecA Protein (Deinococcus radiodurans RecA 단백질의 외가닥 DNA-의존성 ATPase 활성 분석)

  • Kim, Jong-Il
    • Korean Journal of Microbiology
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    • v.43 no.4
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    • pp.250-255
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    • 2007
  • The RecA protein of Deinococcus radiodurans is essential for the extreme radiation resistance of this organism. The central steps involved in recombinational DNA repair require DNA-dependent ATP hydrolysis by recA protein. Key feature of RecA protein-mediated activities is the interactions with ssDNA and dsDNA. The ssDNA is the site where RecA protein filament formation nucleates and where initiation of DNA strand exchange takes place. The effect of sequence heterogeneity of ssDNA was examined in this experiment. The rate of homopolymeric synthetic ssDNA-dependent ATP hydrolysis was constant or nearly so over a broader range of pHs. For poly(dT)-dependent ATP or dATP hydrolysis, rates were generally faster, with a broader optimum between pH 7.0 and 8.0. Activities of RecA protein were affected by the ionic environment. The ATPase activity was shown to have different sensitivity to anionic species. The presence of glutamate seemed to slimulate the hydrolytic activity. Dr RecA protein was shown to require $Mg^{2+}$ ion greater than 2 mM for binding to etheno ssDNA and the binding stoichiometry of 3 nucleotide for RecA protein monomer.

Reversible Inhibitory Effect of $Hg^{2+}$ on the Microsomal $H^+$-ATPases

  • Shin, Dae-Seop;Cho, Kwang-Hyun;Kim, Young-Kee
    • Proceedings of the Korean Biophysical Society Conference
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    • 1999.06a
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    • pp.49-49
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    • 1999
  • $H^{+}$-ATPases play major roles in various cellular physiology. In order to characterize the effects of heavy metal ions on the activity of $H^{+}$-ATPases, microsomes were isolated from the roots of tomato grown hydroponically. The activity of microsomal $H^{+}$-ATPase was measured by an enzyme-coupled assay. H $g^{2+}$ inhibited the activity of microsomal $H^{+}$-ATPase as a dose-dependent manner, F $e^{3+}$ and Z $n^{2+}$ inhibited the activity although they also blocked the activities of enzymes used in the assay, and C $s^{+}$ and $Ba^{2+}$ showed no significant effect.(omitted)d)ted)d)

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Study on the Light-Induced Mitochondrial ATPase$(F_1-ATPase)$ Activated by Iron ion in Mushroom (버섯중 철이온에 활성화된 광감응성 Mitochondrial ATPase에 관한 연구)

  • Min, Tae-Jin;Lee, Mi-Ae;Park, Sang-Shin
    • The Korean Journal of Mycology
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    • v.21 no.3
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    • pp.157-164
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    • 1993
  • The effects of the iron ions on the light-induced mitochondrial $F_1-ATPase$ in Lentinus edodes was studied. This enzyme activity was stimulated by each of the ferric, ferrous and magnesium ion. Especially, the activity of the enzyme by 5.0 mM ferric ion increased up to 107% in comparision with control group(100%). In the presence of magnesium ion, each of ferric and ferrous ion increased the activity of the enzyme, particulary, coexistence of 0.1 mM magnesium and 5.0 mM ferric ion increased the activity up to 270% with magnesium ion dependence. The activity of the enzyme was stimulated up to 268% by 5.0 mM ferric ion in the presence of 0.1 mM magnesium and 0.1 mM ferrous ion. Therefore, the coexistence of ferrous ion did not affect the activity. From the above, we propose that light-induced mitochondrial $F_1-ATPase$ in Lentinus edodes is a $Mg^{2+}{\cdot}Fe^{3+}{\;}F_1-ATPase.$ The optimal pH and temperature for the enzyme were 7.5 and $66^{\circ}C$ respectively.

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Vacuolar H(+)-ATPase is not restricted to clear cells of the epididymal epithelium in cattle

  • Kim, Sung Woo;Kim, Bongki
    • Journal of Animal Science and Technology
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    • v.63 no.2
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    • pp.262-271
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    • 2021
  • Communication among epididymal epithelial cells creates the best luminal condition where spermatozoa mature, transport and are stored. Vacuolar ATPase (V-ATPase) and cytokeratin 5 (KRT5) have been used as signal indicators for clear and basal cells of the epididymal epithelium, respectively, in mice, rats, bats, and pigs; however, these two markers have not yet been described in the epididymis of bulls. Here, we examined the presence and distribution of the B1 subunit of V-ATPase (B1-VATPase) and KRT5 in the distinct regions of adult bovine epididymides, specifically, the caput, corpus, and cauda. Immunofluorescence staining and confocal microscopy showed that narrow shaped-clear cells were placed in the caput and corpus regions of the bovine epididymis; however, they were absent in the cauda epididymis. In addition, B1-VATPase was highly expressed in the cauda spermatozoa; however, it was rarely detected in the caput spermatozoa. On the other hand, KRT5-positive cells, basal cells, were maintained beneath the basal lamina and they had the traditional form with a dome-shaped morphology from the caput to cauda region of the bovine epididymis. The co-expression of B1-VATPase and KRT5 was confined to basal cells placed in the basal region of the epithelium. In summary, 1) clear cells were present with region-specific localization, 2) B1-VATPase was present in the corpus and cauda spermatozoa but absent in the caput, 3) co-expressed cells with B1-VATPase and KRT5 were present in the adult bovine epididymis, and 4) B1-VATPase was not a specific marker for clear cells in the bovine epididymis. Therefore, the perfect epididymal luminal condition created by the specific expression and localization patterns of B1-VATPase might be necessary to obtain fertilizing capacity of spermatozoa in the bovine epididymis.

Cloning and characterization of a gene encoding ABP57, a soluble auxin-binding protein

  • Lee, Keunpyo;Kim, Myung-Il;Kwon, Yu-Jihn;Kim, Minkyun;Kim, Yong-Sam;Kim, Donghern
    • Plant Biotechnology Reports
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    • v.3 no.4
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    • pp.293-299
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    • 2009
  • Auxin-binding protein 57 ($ABP_{57}$), a soluble auxin-binding protein, acts as a receptor to activate plasma membrane (PM) $H^+-ATPase$. Here, we report the cloning of abp57 and the biochemical characterization of its protein expressed in E. coli. The analysis of internal amino acid sequences of $ABP_{57}$ purified from rice shoots enabled us to search for the corresponding gene in protein DB of NCBI. Further BLAST analysis showed that rice has four abp57-like genes and maize has at least one homolog. Interestingly, Arabidopsis seems to have no homolog. Recombinant $ABP_{57}$ expressed in E. coli caused the activation of PM $H^+-ATPase$ regardless of the existence of IAA. Scatchard analysis showed that the recombinant protein has relatively low affinity to IAA as compared to natural $ABP_{57}$. These results collectively support the notion that the cloned gene is responsible for $ABP_{57}$.

Physiological Changes of Saccharomyces cerevisiae by High Voltage Pulsed Electric Field Treatments (고전압 펄스 전기장 처리에 의한 Saccharomyces cerevisiae의 생리적 변화)

  • Park, Hee Ran;Yoon, So Jung;Park, Han-Sul;Shin, Jung-Kue
    • Korean Journal of Food Science and Technology
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    • v.45 no.5
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    • pp.590-597
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    • 2013
  • High voltage pulsed electric fields (PEF) treatment is a promising non-thermal processing technology that can replace or partially substitute for thermal processes. The aim of this research was to investigate the microbial inactivation mechanisms by PEF treatment in terms of physiological changes to Saccharomyces cerevisiae. PEF was applied at the electric field strength of 50 kV/cm, treatment time of 56 ${\mu}s$ and temperature of $40^{\circ}C$. The microbial cells treated with PEF showed loss of salt tolerance on the cell membrane and collapse of the relative pH gradient on in-out of cells. Cell death or injury resulted from the breakdown of homeostasis, decreased $H^+$-ATPase activity, and loss of glycolysis activity.

Acid Stress Response of Lactobacillus rhamnosus GG in Commercial Yogurt (발효유제품에서 Lactobacillus rhamnosus GG의 생육 특성)

  • Bang, Miseon;Jeong, Anna;Park, Dong-June;Lim, Kwang-Sei;Oh, Sejong
    • Journal of Dairy Science and Biotechnology
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    • v.33 no.1
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    • pp.83-91
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    • 2015
  • Yogurt is a product of the acidic fermentation of milk, which affects the survival of lactic acid bacteria (LAB). The aim of this present study was to examine the survival and acid stress response of Lactobacillus rhamnosus GG to low pH environment. The survival of LAB in commercial yogurt was measured during long-term storage. The enumeration of viable cells of LAB was determined at 15-day intervals over 52-weeks at $5^{\circ}C$. L. acidophilus, L. casei, and Bifidobacterium spp. showed low viability. However, L. rhamnosus GG exhibited excellent survival throughout the refrigerated storage period. At the end of 52-weeks, L. rhamnosus GG survived 7.0 log10 CFU/mL. $F_0F_1$ ATPase activity in L. rhamnosus GG at pH 4.5 was also evaluated. The ATPase activities of the membranes were higher when exposed at pH 4.5 for 24 h. The survival of L. rhamnosus GG was attributable to the induction in $F_0F_1$ ATPase activity. In addition, the mRNA expression levels of acid stress-inducible genes at low pH were investigated by qRT-PCR. clpC and clpE genes were up-regulated after 1 h, and atpA and dnaK genes were up-regulated after 24 h of incubation at pH 4.5. These genes could enhance the survival of L. rhamnosus GG in the acidic condition. Thus, the modulation of the enzymes or genes to assist the viability of LAB in the low pH environment is thought to be important.

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Isolation of an acid-tolerant Leuconostoc mesenteroides LM3 from Kimchi (김치에서 산내성을 가진 Leuconostoc mesenteroides LM3의 분리)

  • 사금희;백상규;윤혜선;강경희;정진국;김일섭;문혜연;김사열;유춘발
    • Journal of Life Science
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    • v.12 no.6
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    • pp.812-820
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    • 2002
  • In order to understand stress response of Leuconostoc mesenteroides against lactic acid, a new Leuconostoc sp. which has acid tolerance was isolated from various Kimchi samples. And it identified as Leuconostoc mesenteroides LM3 by comparing its fatty acid composition with reference strain. Its growth pattern was investigated by adding a given concentration of lactic acid at the lag phase to the stationary phase. In the DeMan, Rogosa, Sharpe (MRS) media containing over 0.4% (final v/v) lactic acid, this strain slowed slowly After exposure of the stationary phase cells to 4% of lactic acid for 60 min, this strain could survive, whereas a reference strain, Leuconostoc mesenteroides KCTC3505, showed no survival. And changes of trehalose concentration, the activity of trehalase and ATPase in the growth of Leuconostoc mesenteroides LM3 after addition of 0.6% (final v/v) lactic acid were investigated : After exposure to lartic acid, trehalose concentration in this strain was increased in comparison with no treatment, but its trehalase activity was not changed. And its ATPase activity was constant, and intracellular pH was almost constant. This result meant Leuconostoc mesenteroides LM3 should have a tolerance against lactic acid. It remains to further study the mechanism of this acid tolerance.