• 제목/요약/키워드: pka value

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Characterization of Undaria pinnatifida Root Enzymatic Extracts Using Crude Enzyme from Shewanella oneidensis PKA 1008 and Its Anti-Inflammatory Effect

  • Xu, Xiaotong;Jeong, So-Mi;Lee, Ji-Eun;Kang, Woo-Sin;Ryu, Si-Hyeong;Kim, Kwangwook;Byun, Eui-Hong;Cho, Young-Je;Ahn, Dong-Hyun
    • Journal of Microbiology and Biotechnology
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    • 제30권1호
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    • pp.79-84
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    • 2020
  • This study investigated the characterization and functionality of Undaria pinnatifida root (UPT) extracts, degraded using a crude enzyme from Shewanella oneidensis PKA1008. To obtain the optimum degrading conditions, the UPT was mixed with alginate degrading enzymes from S. oneidensis PKA 1008 and was incubated at 30℃ for 0, 3, 6, 12, 24, and 48 h. The alginate degrading ability of these enzymes was then evaluated by measuring the reducing sugar, viscosity, pH and chromaticity. Enzymatic extract at 24 h revealed the highest alginate degrading ability and the lowest pH value. As the incubation time increased, the lightness (L ) also decreased and was measured at its lowest value, 39.84, at 12 hours. The redness and yellowness increased gradually to 10.27 at 6 h and to 63.95 at 3 h, respectively. Moreover, the alginate oligosaccharides exhibited significant anti-inflammatory activity. These results indicate that a crude enzyme from S. oneidensis PKA 1008 can be used to enhance the polysaccharide degradation of UPT and the alginate oligosaccharides may also enhance the anti-inflammatory effect.

어리연꽃 추출물이 α-MSH 유도에 의한 멜라닌 생성 억제에 미치는 영향 (Inhibitory effect of Nymphoides indica extract on α-MSH induced melanin synthesis)

  • 김동희;김유아;유재묘;박채빈;박병준;박태순
    • Journal of Applied Biological Chemistry
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    • 제60권4호
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    • pp.327-332
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    • 2017
  • 본 연구에서는 어리연꽃 추출물의 B16F10 세포에서의 미백활성을 측정하였다. 버섯유래의 tyrosinase 활성 저해능 측정결과 $1,000{\mu}g/mL$의 농도에서 42%의 저해율을 나타내었으며, B16F10 세포내의 tyrosinase, 멜라닌 생합성 저해능은 $5{\mu}g/mL$에서 26, 25%의 저해능을 나타내었다. 멜라닌 생성 관련 인자중 상위 단계인 cAMP와 PKA의 발현을 측정한 결과 농도 의존적으로 발현양이 줄어드는 것을 확인하였다. 또한 어리연꽃추출물의 미백과 관련된 tyrosinase, tyrosinase related protein(TRP) 1, TRP2, microphthalmia associated transcription factor(MITF)의 단백질 및 유전자 발현양이 감소함을 나타내었다. 본 실험을 통하여 어리연꽃 추출물이 cAMP를 효과적으로 억제함으로써, PKA의 활성을 저해 시켰으며, 이를 통해 tyrosinase 활성뿐만 아니라 TRP1, TRP2 그리고 MITF 발현을 효과적으로 저해 한다는 사실을 확인하여 기능성 미백 화장품 소재로서의 활용 가능성을 제시하고자 한다.

알쏭이 모자반(Sargassum confusum C. Agardh) 알긴산 분해 조효소 분해물의 알코올 침전에 의한 기호성 증진 효과 (Palatability-Enhancing Effect of the Alcohol Precipitate of Sargassum confusum C. Agardh Extracts Using an Alginate-degrading Crude Enzyme)

  • 나현식;김동현;이하영;유현지;박미성;우가은;조미정;안동현
    • 한국수산과학회지
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    • 제56권2호
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    • pp.204-211
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    • 2023
  • This study investigated the physicochemical properties and palatability-enhancing effects of the alcohol precipitate, in the enzymatic extracts of Sargassum confusum C. Agardh (SC), obsained using the crude enzyme of Shewanella oneidensis PKA 1008. We analyzed the oligosaccharides recovered from the alcohol precipitate using a thin-layer chromatography for SC-degrading extracts, pH, color, reducing sugar, and viscosity. Thin-layer chromatography showed that after treating with the crude enzyme for 60 h, the polysaccharides were degraded into tetramers, dimers, and trimers and pH increased in the alcohol precipitate (EtOH Sedi). In terms of color, the redness and yellowness of alcohol precipitate/supernatant (EtOH Sedi+Super) and the brightness of EtOH Sedi were the highest among enzyme treated for 0 h and 60 h, EtOH Sedi, and EtOH Sedi+Super. In the reducing sugar analysis, EtOH Sedi showed the lowest value of 13.63 ㎍/mL, and the lowest viscosity of 1.13. In terms of the sensory evaluation, EtOH Sedi+Super showed the highest value with respect to the overall preference. These results suggest that the crude enzyme of S. oneidensis PKA 1008 is effective at degrading polysaccharides, and its recovery increases the palatability of the alcohol precipitate.

Purification and Characterization of Protein Phosphatase 2C from Rat Liver

  • Oh, Joung-Sook;Hwang, In-Seong;Choi, Myung-Un
    • BMB Reports
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    • 제30권3호
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    • pp.222-228
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    • 1997
  • Protein phosphatase 2C (PP2C) is one of the four major serine/threonine phosphatases which is dependent on $Mg^{2+}$ for its activity. PP2C was purified from rat liver cytosol and its characteristics were investigated. The substrate employed for routine assay was $[^{32}P]casein$ phosphorylated by PKA. The purification process involved DEAE chromatography, ammonium sulfate fractionation, phenyl sepharose chromatography, sephacryl 5-200 gel filtration, and histone agarose chromatography. The SDS-PAGE of PP2C showed one major single protein band at a position corresponding to a molecular mass of 43 kd and the purification fold was 637. The enzyme showed a pH optimum of 8 and $K_M$ value was $1.9\;{\mu}M$. However, when the substrate was changed to $[^{32}P]histone$, the pH optimum was shifted to 7 and $K_M$ value was $2.3\;{\mu}M.\;Mg^{2+}$ was essential to the enzyme activity and okadaic acid did not exert any inhibitory effect on the enzyme. To examine residue in the active site of PP2C effects of some protein-modifying reagents were tested.

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Rous Sarcoma Virus에 의해 형질전환된 NRK 세포의 형태변화를 유도하는 활성물질 MT 1154의 분리와 생물학적 활성 (Purification and Biological Activities of MT 1155 Inducing Morphological Change of Rous Sarcoma Virus-Transformed Normal Rat Kidney Cell)

  • 안종석;박문수;박찬선;윤병대;민태익;안순철;오원근;이현선;윤병대
    • 한국미생물·생명공학회지
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    • 제21권1호
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    • pp.59-65
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    • 1993
  • 방선균 분리주 GMT 1153로부터 ts/NRK 세포의 형태를 변형된 세포형태에서 정상세포의 형태로 전환하는 활성물질을 분리, 정제하였다. 활성물질 MT 1155는 UV, IR, 1H-NMR, 13C-NMR, mass,원소분석 등의 기기분석을 통하여 항진균성 항암물질인 toyocamycin으로 동정되었다. MT 1155는 ts/NRK 세포의 형태전환에 대한 활성외에도 PKA 효소활성 저해와 CTLL 세포에 대한 세포독성이 있으나 K562 소포형성 억제 및 PKC 효소활성에 대한 저해효과도 없었다.

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이부프로펜의 피페라진 유도체에 대한 약리활성연구 (Studies of Pharmacological Activity on the Piperazine Derivatives of Ibuprofen)

  • 조한진;나성범;남상철;박목순;지웅길
    • 약학회지
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    • 제34권2호
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    • pp.126-132
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    • 1990
  • To enhance the activity of ibuprofen, amides of ibuprofen, 1-piperazinyl-2-(4-isobutylphenyl)propionamide(Ibu-P.A.) and 1-(4-methylpiperazinyl)-2-(4-isobutylphenyl)propionamide (Ibu-M.P.), were synthesized and the pharmaceutical properties and the pharmacological activities of the amides were studied. The lipid:water partition coefficients and pKa values were examined in vitro, and the antiinflammatory effect, analgesic effects, acute toxicity, and intestinal absorption were studied for the amides and compared with ibuprofen in vivo. The results are summarized as belows; 1) The lipid:water partition coefficients of Ibu-M.P. were higher than those of ibuprofen. 2) The calculated pKa values of ibuprofen and Ibu-M.P. were 5.49 and 8.66, respectively. 3) The antiinflammatory effects of ibuprofen, Ibu-P.A., and Ibu-M.P. were same intensity, but the duration of the effects of Ibu-P.A. and Ibu-M.P. were longer than that of ibuprofen. 4) The analgesic effect of Ibu-M.P. was more potent than those of ibuprofen and Ibu-P.A. in the acetic acid-induced writhing test. 5) The $LD_{50}$ was 495 mg/kg for ibuprofen, 187 mg/kg for Ibu-M.P., and over 1250 mg/kg for Ibu-P.A.. 6) The absorption rate constants(k) and half-life($t_{1/2}$) were 0.74($hr^{-1}$) and 0.94(hr) for ibuprofen, and 0.72 ($hr^{-1}$) and 0.96 (hr) respectively for Ibu-M.P..

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Gonadotropin-releasing Hormone and Its Receptor as a Therapeutic Concept in the Progression of Epithelial Ovarian Cancer

  • Kim, Ki-Yon;Choi, Kyung-Chul
    • 한국수정란이식학회지
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    • 제24권1호
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    • pp.1-14
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    • 2009
  • Ovarian cancer is a significant cause of cancer-related death in women, but the main biological causes remain open questions. Hormonal factors have been considered to be an important determinant causing ovarian cancer. Recent studies have shown that gonadotropin-releasing hormone (GnRH)-I and its analogs have clinically therapeutic value in the treatment of ovarian cancer. In addition, numerous studies have shown that the potential of GnRH-II in normal reproductive system or reproductive disorder. GnRH-I receptors have been detected in approximately 80% of ovarian cancer biopsy specimens as well as normal ovarian epithelial cells and immortalized ovarian surface epithelium cells. GnRH-II receptors have also been found to be more widely expressed than GnRH-I receptors in mammals, suggesting that GnRH receptors may have additional functions in reproductive system including ovarian cancer. The signal transduction pathway following the binding of GnRH to GnRH receptor has been extensively studied. The activation of protein kinase A/C (PKA/PKC) pathway is involved in the GnRH-I induced anti-proliferative effect in ovarian cancer cells. In addition, GnRH-I induced mitogen-activated protein kinase (MAPK) activation plays a role in anti-proliferative effect and apoptosis in ovarian cancer cells and the activation of transcriptional factors related to cellular responses. However, the role of GnRH-I and II receptors, there are discrepancies between previous reports. In this review, the role of GnRH in ovarian cancer and the mechanisms to induce anti-proliferation were evaluated.

Chronic persistent post-surgical pain following staging laparotomy for carcinoma of ovary and its relationship to signal transduction genes

  • Saxena, Ashok Kumar;Chilkoti, Geetanjali T;Chopra, Anand K;Banerjee, Basu Dev;Sharma, Tusha
    • The Korean Journal of Pain
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    • 제29권4호
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    • pp.239-248
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    • 2016
  • Background: The present study was undertaken to evaluate the incidence of chronic persistent post-surgical pain (CPPP) and the role of signal transduction genes in patients undergoing staging laparotomy for carcinoma ovary. Methods: The present observational study was undertaken following institutional ethical committee approval and informed consent from all the participants. A total 21 patients of ASA grade I to III with age 20-70 years, scheduled for elective staging laparotomy for carcinoma ovary were included. Patients were excluded if had other causes of pain, cognitive dysfunction or chronic neurological disorders. Statistical analysis of pool data was done using SPSS version-17. For various scales like GPE, PDQ, NPSI, the visual analogue scale (VAS), global perceived effect (GPE), the pain DETECT questionnaire (PDQ), and neuropathic pain symptoms inventory (NPSI), one factor repaeted measure ANOVA applied with simple contrast with baseline as on post-operative day 1 (considered as reference and compared with subsequent time-interval), and the P values were adjusted according to "Bonferroni adjustments". In patients with CPPP, the ${\Delta}ct$ values of mRNA expressions of genes at the end of postoperative day 90 were compared with the baseline control values by one factor repeated ANOVA. P value < 0.005 significant. Results: The present study demonstrates 38.1% (8 out of 21 patients) incidence of CPPP. The functional status and quality of life as were observed to be significantly diminished in all patients with chronic pain. An up-regulation in the mRNA expression of signal transduction and a positive correlation was noted between the mRNA expression of signal transduction genes and VAS score in all patients with CPPP at the end of postoperative day 90. Conclusions: The reported incidence of CPPP in patients with carcinoma ovary was 38.1%. An up-regulation and positive correlation between mRNA expression of signal transduction genes and VAS score depicts its potential role in the pathogenesis of CPPP.