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Human brain pyridoxal-5'-phosphate phosphatase (PLPP): protein transduction of PEP-1-PLPP into PC12 cells

  • Lee, Yeom-Pyo;Kim, Dae-Won;Lee, Min-Jung;Jeong, Min-Seop;Kim, So-Young;Lee, Sun-Hwa;Jang, Sang-Ho;Park, Jin-Seu;Kang, Tae-Cheon;Won, Moo-Ho;Cho, Sung-Woo;Kwon, Oh-Shin;Eum, Won-Sik;Choi, Soo-Young
    • BMB Reports
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    • v.41 no.5
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    • pp.408-413
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    • 2008
  • Pyridoxal-5'-phosphate phosphatase (PLPP) catalyzes the dephosphorylation of pyridoxal-5'-phosphate (PLP). A human brain PLPP gene was fused with a PEP-1 peptide and produced a genetic in-frame PEP-1-PLPP fusion protein. The purified PEP-1-PLPP fusion protein was efficiently transduced into PC12 cells in a time- and dose-dependent manner when added exogenously to culture media. Once inside the cells, the transduced PEP-1-PLPP fusion protein was stable for 36 h. The concentration of PLP was markedly decreased by the addition of exogenous PEP-1-PLPP to media pretreated with the vitamin $B_6$ precursors; pyridoxine, pyridoxal kinase and pyridoxine-5'-phosphate oxidase into cells. The results suggest that the transduction of the PEP-1-PLPP fusion protein can be one mode of PLP level regulation, and to replenish this enzyme in the various neurological disorders related to vitamin $B_6$.

Transduced PEP-1-AMPK inhibits the LPS-induced expression of COX-2 and iNOS in Raw264.7 cells

  • Shin, Min-Jea;Lee, Yeom-Pyo;Kim, Dae-Won;An, Jae-Jin;Jang, Sang-Ho;Cho, Sung-Min;Sheen, Seung-Hoon;Lee, Hae-Ran;Kweon, Hae-Yong;Kang, Seok-Woo;Lee, Kwang-Gill;Park, Jin-Seu;Eum, Won-Sik;Cho, Yong-Jun;Choi, Soo-Young
    • BMB Reports
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    • v.43 no.1
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    • pp.40-45
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    • 2010
  • AMP-activated protein kinase (AMPK) is a heterotrimeric enzyme that plays a central role in cellular metabolic stress. Modulation of nitric oxide (NO) and cyclooxygenase-2 (COX-2) is considered a promising approach for the treatment of inflammation and neuronal diseases. In this study, the AMPK gene was fused in-frame with PEP-1 peptide in a bacterial expression vector to produce a PEP-1-AMPK fusion protein. Expressed and purified PEP-1-AMPK fusion proteins were transduced efficiently into macrophage Raw 264.7 cells in a time- and dose-dependent manner. Furthermore, transduced PEP-1-AMPK fusion protein markedly inhibited LPS-induced iNOS and COX-2 expression. These results suggest that the PEP-1-AMPK fusion protein can be used for the protein therapy of COX-2 and NO-related disorders such as inflammation and neuronal diseases.

Transduced HSP27 protein protects neuronal cell death by enhancing FALS-associated SOD1 mutant activity

  • An, Jae-Jin;Lee, Yeom-Pyo;Kim, Dae-Won;Sohn, Eun-Joung;Jeong, Hoon-Jae;Kang, Hye-Won;Shin, Min-Jae;Kim, Mi-Jin;Ahn, Eun-Hee;Jang, Sang-Ho;Kang, Jung-Hoon;Kang, Tae-Cheon;Won, Moo-Ho;Kwon, Oh-Shin;Cho, Sung-Woo;Lee, Kil-Soo;Park, Jin-Seu;Eum, Won-Sik;Choi, Soo-Young
    • BMB Reports
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    • v.42 no.3
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    • pp.136-141
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    • 2009
  • Familial Amyotrophic lateral sclerosis (FALS) is a progressive neurodegenetative disorder induced by mutations of the SOD1 gene. Heat shock protein 27 (HSP27) is well-defined as a stress-inducible protein, however the its role in ALS protection has not yet been established. To investigate the role HSP27 may have in SOD1 mutant-mediated apoptosis, human SOD1 or HSP27 genes were fused with a PEP-1 peptide in a bacterial expression vector to produce a genetic in-frame fusion protein, which was then transduced into cells. We found the purified PEP-1-HSP27 fusion proteins can be transduced efficiently into neuronal cells and protect against cell death by enhancing mutant SOD1 activity. Moreover, transduced PEP-1-HSP27 efficiently prevents protein aggregation produced by oxidative stress. These results suggest that transduced HSP27 fusion protein may be explored as a potential therapeutic agent for FALS patients.

Characteristics of Cyanide Decomposition by Hydrogen Peroxide Reduction (과산화수소에 의한 시안의 분해특성)

  • 이진영;윤호성;김철주;김성돈;김준수
    • Resources Recycling
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    • v.11 no.2
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    • pp.3-13
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    • 2002
  • The characteristics of cyanide decomposition in aqueous phase by hydrogen peroxide have been explored in an effort to develop a process to recycle waste water. The self-decomposition of $H_2O$$_2$at pH 10 or below was minimal even in 90 min., with keeping about 90% of $H_2O$$_2$undissociated. On the contrary, at pH 12 only 9% of it remained during the same time. In the presence of copper catalyst at 5 g Cu/L, complete decomposition of $H_2$O$_2$was accomplished at pH 12 even in a shorter time of 40 min. The volatility of free cyanide was decisively dependent on the solution pH: the majority of free cyanide was volatilized at pH 8 or below, however, only 10% of it was volatilized at pH 10 or above. In non-catalytic cyanide decomposition, the free cyanide removal was incomplete in 300 min. even in an excessive addition of $H_2$$O_2$at a $H_2$$O_2$/CN molar ratio of 4, with leaving behind about 8% of free cyanide. On the other hand, in the presence of copper catalyst at a Cu/CN molar ratio of 0.2, the free cyanide was mostly decomposed in only 16 min. at a reducedH202/CN molar ratio of 2. Ihe efnciency of HBO2 in cyanide decomposition decreased with increasing addition of H2O2 since the seu-decomposition rate of $H_2$$O_2$increased. At the optimum $H_2$$O_2$/mo1ar ratio 0.2 of and Cu/CN molar ratio of 0.05, the free cyanide could be completely decomposed in 70 min., having a self-decomposition rate of 22 mM/min and a H$_2$$O_2$ efficiency of 57%.

Enhancement of Anti-Inflammatory Activity of PEP-1-FK506 Binding Protein by Silk Fibroin Peptide

  • Kim, Dae-Won;Hwang, Hyun-Sook;Kim, Duk-Soo;Sheen, Seung-Hoon;Heo, Dong-Hwa;Hwang, Gyo-Jun;Kang, Suk-Hyung;Kweon, Hae-Yong;Jo, You-Young;Kang, Seok-Woo;Lee, Kwang-Gill;Park, Jin-Seu;Eum, Won-Sik;Cho, Yong-Jun;Choi, Soo-Young
    • Journal of Microbiology and Biotechnology
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    • v.22 no.4
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    • pp.494-500
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    • 2012
  • Silk fibroin (SF) peptide has been traditionally used as a treatment for flatulence, spasms, and phlegm. In this study, we examined whether SF peptide enhanced the anti-inflammatory effect of PEP-1-FK506 binding protein (PEP-1-FK506BP) through comparing the anti-inflammatory activities of SF peptide and/or PEP-1-FK506BP. In the presence or absence of SF peptide, transduction levels of PEP-1-FK506BP into HaCaT cells and mice skin and anti-inflammatory activities of PEP-1-FK506BP were identified by Western blot and histological analyses. SF peptide alone effectively reduced both mice ear edema and the elevated levels of cyclooxygenase-2, interleukin-6 and $-1{\beta}$, and tumor necrosis factor-${\alpha}$, showing similar anti-inflammatory effect to that of PEP-1-FK506BP. Furthermore, co-treatment with SF peptide and PEP-1-FK506BP exhibited more enhanced anti-inflammatory effects than the samples treated with SF peptides or PEP-1-FK506BP alone, suggesting the possibility that SF peptide and PEP-1-FK506BP might interact with each other. Moreover, the transduction data demonstrated that SF peptide did not affect the transduction of PEP-1-FK506BP into HaCaT cells and mice skin, indicating that the improvement of anti-inflammatory effect of PEP-1-FK506BP was not caused by enhanced transduction of PEP-1-FK506BP. Thus, these results suggest the possibility that co-treatment with SF peptide and PEP-1-FK506BP may be exploited as a useful therapy for various inflammation-related diseases.

Studies on the scouring of raw silk by the application of bacterial enzyme of sericinase -(Part 1.) Isolation and selection of proper bacteria and some studies of properties of this bacterial enzyme- (세균성(細菌性) Sericin 분해(分解) 효소(酵素)에 의(依)한 생사정련(生絲精練)에 관(關)한 연구(硏究) -(제1보(第1報)) 유용세균(有用細菌)의 분리(分離) 세정(選定) 및 생성(生成) Sericinase 의 효소학적(酵素學的) 성질(性質)에 대(對)하여-)

  • Ki, Woo-Kyung;Seu, Jung-Hwn
    • Applied Biological Chemistry
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    • v.12
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    • pp.75-81
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    • 1969
  • In order to obtain a method of scouring of raw silk and degumming of cocoon by the appling of bacterial enzyme of sericinase, a strain of proper bacteria was isolated and some properties of the enzyme produced by the isolated bacteria were studied and the following results were obtained. 1) Optimum pH and temperature were indicating 7.5 and $50^{\circ}C$. and on these conditions, the silk fibroin will get no modification at all. 2) Sericinase activity was inhibited by calcium ion in the free incubation but same ions reacted as an activator in the reaction with substrate. So, degumming of Tussah cocoon which contains calcium oxalate in the cocoon layer will be possible by the treatment of this enzyme. 3) This bacterial sericinase never gives any affection to the fibroin layer of the silk. 4) The factors of smooth-surface condition, damage of fibroin layer, touching, luster and degumming effect of the silk resulting by the enzyme treatment were more appropriate than the results of other scouring methods, saponin, alkali and saponin-alkali using methods. We expect to get the same scouring result that compared with the papain or pancreatin were applied.

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Studies on the glucose isomerizing enzyme. (Part IV) -On the enzymatic properties and treating conditions for high activity containing cells- (포도당(葡萄糖) 이성화(異性化) 효소(酵素)에 관(關)한 연구(硏究) (제4보(第四報)) -효소학적(酵素學的) 성질(性質)및 효소생성능(酵素生成能)에 대(對)하여-)

  • Rhee, In-Koo;Seu, Jung-Hwn
    • Applied Biological Chemistry
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    • v.12
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    • pp.69-74
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    • 1969
  • This glucose isomerizing enzyme, which Actinomyces sp. (strain, K-17) produced, was inhibited by citrate, oxalate, ethylene diamine tetraacetic acid and cysteine on the enzyme reaction. This enzyme isomerized xylose to ketose as well as glucose. The Michaelis constant of this enzyme was $7.2X\;10^{-1}M.$. on D-glucose. The enzyme activity of intact cells which were harvested in the none xylose containing medium was very weak. If these intact cells of low activity were treated in the buffered xylose solution, its activity was considerably increased. After fifteen hours at $32^{\circ}C$. on xylose treatment, the enzyme activity was increased to equilibrium and the treating condition was proper at neutral pH and in aerobic state. The adequate concentration of xylose on the treatment was about 0.5 percent.

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Studies on the application of bacterial sericinase -(Part II) Appling on the unwinding for mulberry cocoon- (세균성(細菌性) Sericin 분해효소(分解酵素)의 이용(利用)에 관(關)한 연구(硏究) -(제II보(第II報)) Sericinase에 의(依)한 가잠견(家蠶繭)의 해서(解舒)에 관(對)하여-)

  • Seu, Jung-Hwn;Song, Bang-Ho;Han, Yung-Gu;Kho, Yung-Hee
    • Applied Biological Chemistry
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    • v.13 no.1
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    • pp.87-92
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    • 1970
  • Sericin hydrolyzing enzyme, produced by the selected bacteria, S4-1-1, was studied and following properties were obtained. 1. The activity of this bacterial sericinase was not decreased for 30 days of storage at $5^{\circ}C$. But at $20^{\circ}C$, for 30 hrs. was the maximum period to keep the initial activity of this enzyme. 2. This bacterial enzyme gave only sericinase activity but never indicated fibroin hydrolyzing activity. 3. The chelating reagent of EDTA and Ag or Hg ions were classified as strung inhibitors but Cu and Cd ions were indicated as moderate inhibitors to this enzyme action. 4. This enzyme was not inhibited by the surface active agent, Peretex-N, but strongly activated by this agent at low concentration. In the other hand, by the application of this enzyme to the unwinding works on the mulberry cocoon, the following results were also obtained. 1. On the weight and length, nonbreaking Length, size, colour and unwinding ratio of have, the enzyme appling method was superior to generally used cooking method. 2. The tested results of strength and elongation, bouchon, haririness loops, neatness and evenness of have were also indicated spuerior properties.

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The Enhancing Effect of Atopic Dermatitis by cosmetics containing estern medical herbs (한방소재 화장품의 아토피 피부염 개선 효과)

  • Kang, Shin-Jyung;Kim, Ae-Jung;Lee, Yeon-Hee;Lee, Myoung-Sook;Joung, Kyung-Hee;Cho, Nam-Ji
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.10 no.11
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    • pp.3500-3505
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    • 2009
  • Sixteen moderate atopic dermatitis patients were treated with applying toner and cream using estern medical herbs[Estern medical complex(Radix Glycyrrhizae, Radix Angelicae Dahuricae, Rhizoma Ligustici Chuanxiong, Fructus ponciri Seu Aurantii Immaturus, Radix Rehmanniae Praeparata, Radix Puerariae, Rhizoma Corydalis), Cypress, Green tea, Radix Astragali, Red ginseng, Syrup ferment, Apple ferment filtrate] on the atopic area 2-3 times per day for 6 weeks. After 6 weeks of study, the result was a decrease in SCORAD index($26.9{\pm}11.8$ to $14.9{\pm}9.7$(p<0.05)), IgE($641.9{\pm}1294.6$ to $565.8{\pm}1076.8$), Eosinophil count($246.7{\pm}203.5$ to $203.3{\pm}130.7$(p<0.05)) and TEWL($16.7{\pm}5.1$ to $15.4{\pm}8.7$(p<0.05)). And skin hydration was increased($36.6{\pm}10.0$ to $44.0{\pm}10.3$(p<0.01)) maintaining skin pH level. The patient;s and physician's global assessment also improved. Therefore, estern medical herbs may play a role in treatment of atopic dermatitis.

Studies on the Utilization of Persimmons -(Part 1) Effect of Carton Dioxide Treatment on the Removal of Astringency- (감의 이용(利用)에 관(關)한 연구(硏究) -제1보(第一報) 탄산(炭酸)가스처리(處理)가 탈삽(脫澁)에 미치는 영향(影響)-)

  • Seu, Ohn-Soo;Sohn, Tae-Hwa
    • Applied Biological Chemistry
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    • v.19 no.2
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    • pp.93-98
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    • 1976
  • The experiment was made to investigate concentrations of $CO_2\;and\;O_2$ in the removal of astringency of persimmon fruits and to study the changes of total, reducing sugar and hardness during the removal of astringency. 1. During the removal of astringency of persimmon fruits, channel of tannin content showed the rapid decrease in high concentration of $CO_2$ and the gradual decrease in low concentration of $CO_2$. 2. Concentration of oxygen did not show effect on the removal of astringency at high concentration of $CO_2$ but at low concentration of $CO_2$ in Chungdo-Bansi. 3. Optimal concentration of the removal of astringency was $60{\sim}70%,\;CO_2$ and $6{\sim}8%\;O_2$ in Chungdo-Bansi and Sagoksi 4. In changes of sugar content total sugar was decreased and reducing sugar was increased.

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