• Title/Summary/Keyword: DJ-1

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Purification and Characterization of a New Peptidase, Bacillopeptidase DJ-2, Having Fibrinolytic Activity: Produced by Bacillus sp. DJ-2 from Doen-Jang

  • CHOI, NACK-SHICK;YOO, KI-HYUN;HAHM, JEUNG-HO;YOON, KAB-SEOG;CHANG, KYU-TAE;HYUN, BYUNG-HWA;PIL, JAE-MAENG;KIM, SEUNG-HO
    • Journal of Microbiology and Biotechnology
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    • v.15 no.1
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    • pp.72-79
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    • 2005
  • A new Bacillus peptidase, bacillopeptidase DJ-2 (bpDJ-2), with molecular mass of 42 kDa and isoelectric point (pI) of 3.5- 3.7, was purified to homogeneity from Bacillus sp. DJ-2 isolated from Doen-Jang, a traditional Korean soybean fermented food. The enzyme was identified as an extracellular serine fibrinolytic protease. The optimal conditions for the reaction were pH 9.0 and $60^{\circ}C$. The first 18 amino acid residues of the N-terminal amino acid sequence of bpDJ-2 were TDGVEWNVDQIDAPKAW, which is identical to that of bacillopeptidase F (bpf). However, based on their Nterminal amino acid sequence, molecular size, and pI, it is different from that of bpf and extracellular 90 kDa. The whole (2,541 bp, full-bpDJ-2) and mature (1,956 bp, mature-bpDJ-2) genes were cloned, and its nucleotide sequence and deduced amino acid sequence were determined. The expressed proteins, full-bpDJ-2 and mature-bpDJ-2, were detected on SDSPAGE at expected sizes of 92 and 68 kDa, respectively.

Protective effects of Tat-DJ-1 protein against streptozotocin-induced diabetes in a mice model

  • Yeo, Hyeon Ji;Yeo, Eun Ji;Shin, Min Jea;Choi, Yeon Joo;Lee, Chi Hern;Kwon, Hyeok Yil;Kim, Dae Won;Eum, Won Sik;Choi, Soo Young
    • BMB Reports
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    • v.51 no.7
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    • pp.362-367
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    • 2018
  • A major feature of type 1 diabetes mellitus (T1DM) is hyperglycemia and dysfunction of pancreatic ${\beta}$-cells. In a previous study, we have shown that Tat-DJ-1 protein inhibits pancreatic RINm5F ${\beta}$-cell death caused by oxidative stress. In this study, we examined effects of Tat-DJ-1 protein on streptozotocin (STZ)-induced diabetic mice. Wild type (WT) Tat-DJ-1 protein transduced into pancreas where it markedly inhibited pancreatic ${\beta}$-cell destruction and regulated levels of serum parameters including insulin, alkaline phosphatase (ALP), and free fatty acid (FFA) secretion. In addition, transduced WT Tat-DJ-1 protein significantly inhibited the activation of $NF-{\kappa}B$ and MAPK (ERK and p38) expression as well as expression of COX-2 and iNOS in STZ exposed pancreas. In contrast, treatment with C106A mutant Tat-DJ-1 protein showed no protective effects. Collectively, our results indicate that WT Tat-DJ-1 protein can significantly ameliorate pancreatic tissues in STZ-induced diabetes in mice.

Impaired Taste Associative Memory and Memory Enhancement by Feeding Omija in Parkinson's Disease Fly Model

  • Poudel, Seeta;Lee, Youngseok
    • Molecules and Cells
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    • v.41 no.7
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    • pp.646-652
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    • 2018
  • Neurodegeneration can result in memory loss in the central nervous system (CNS) and impairment of taste and smell in the peripheral nervous system (PNS). The neurodegeneration seen in Parkinson's disease (PD) is characterized by functional loss of dopaminergic neurons. Recent studies have also found a role for dopaminergic neurons in regulating taste memory rewards in insects. To investigate how taste memories and sugar sensitivity can be affected in PD, we utilized the $DJ-1{\beta}$ mutant fruit fly, $DJ-1{\beta}^{ex54}$, as a PD model. We performed binary choice feeding assays, electrophysiology and taste-mediated memory tests to explore the function of the $DJ-1{\beta}$ gene in terms of sugar sensitivity as well as associative taste memory. We found that PD flies exhibited an impaired ability to discriminate sucrose across a range of sugar concentrations, with normal responses at only very high concentrations of sugar. They also showed an impairment in associative taste memory. We highlight that the taste impairment and memory defect in $DJ-1{\beta}^{ex54}$ can be recovered by the expression of wild-type $DJ-1{\beta}$ gene in the dopaminergic neurons. We also emphasized the role of dopaminergic neurons in restoring taste memory function. This impaired memory property of $DJ-1{\beta}^{ex54}$ flies also allows them to be used as a model system for finding supplementary dietary foods that can improve memory function. Here we provide evidence that the associative taste memory of both control and $DJ-1{\beta}^{ex54}$ flies can be enhanced with dietary supplementation of the medicinal plant, omija.

Miniscale Identification and Characterization of Subtilisins from Bacillus sp. Strains

  • CHOI NACK-SHICK;JU SUNG-KYU;LEE TAE YOUNG;YOON KAB-SEOG;CHANG KYU-TAE;MAENG PIL JAE;KIM SEUNG-HO
    • Journal of Microbiology and Biotechnology
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    • v.15 no.3
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    • pp.537-543
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    • 2005
  • Subtilisin (EC 3.4.21.14) is the major extracellular alkaline serine protease of Bacillus species. Previously, we found that subtilisins did not migrate in the electrophoretic field in the Laemmili buffer system due to their high pI values (over 8.8); however, it formed a 'binding mode' at the top of the separating gel [5]. Utilizing this characteristic, four subtilisins from Bacillus sp. strains (e.g., B. subtilis 168, B. subtilis KCTC 1021, B. amyloliquefaciens KCTC 3002, and Bacillus sp. DJ-1 and DJ-4) were easily and quickly identified by an over-running electrophoretic technique with a miniscale culture supernatant (less than 20 ml) without any column chromatographic steps. Two subtilisins (DJ-l and a recombinant version) from Bacillus sp. DJ-l were characterized, and the enzymatic properties were determined by SDS-fibrin zymography and densitometric analysis. Based on this observation, the recombinant pro-subtilisin DJ-l showed the same 'binding mode,' similar to native subtilisin DJ-l. On the other hand, mature subtilisin DJ -1 without pro-peptide showed no enzymatic activity.

Pseudomonas sp. strain DJ77 균주에서 extradiol dioxygenase 를 암호화하고 있는 phnE 유전자의 염기배열

  • 김영창;신명수;윤길상;박영순;김욱현
    • Korean Journal of Microbiology
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    • v.30 no.1
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    • pp.8-14
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    • 1992
  • Nucleotide Sequence of phnE Gene Encoding Extradiol Dioxygenase fromPseudomonas sp. Strain DJ77Kim, Young-Chang'.", Myeong-Su Shin1, Kil-Sang Younl, Young-Soon Park1, andUg-Hyeon Kim'.' (Department of Microbiology, C'hungbuk National University.Cheongju 360-763, KOREA. and 'Research Center for Molecular Microbiology,Seoul National University)nal University)

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The Effect of Dangguijakyak-san on Wound Healing (당귀작약산의 창상 회복에 대한 효과)

  • Yun-Jin Lee;Chang-Hoon Woo;Young-Jun Kim;Hyeon-Ji Kim;Hee-Duk An
    • Journal of Korean Medicine Rehabilitation
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    • v.33 no.3
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    • pp.47-65
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    • 2023
  • Objectives We evaluated the wound healing effects of Dangguijakyak-san (DJ) using C57BL/6 mice that were generated open wound. Methods The study was conducted with seven C57BL/6 mice assigned to each group, divided into the normal group, control group, vitamin E group, DJ low-dose group, DJ high-dose group. We measured total polyphenol, flavonoid contents, the size of the wound, liver function, pro-inflammatory cytokine activity in serum, inflammation-related proteins, adhesion molecules and chemokine proteins, collagen-related proteins in skin tissue and histopathological changes by H&E and Masson's staining. Results DJ treatment significantly reduced the area of the wound compared to the control group. Also, inflammatory cytokines were reduced and the expression of anti-inflammatory-related factors (interleukin-4 [IL-4] and IL-10) was significantly increased in the DJ treatment group. We identified that DJ treatment inhibits both pathways of inflammation, the mitogen-activated protein kinases and nuclear factor-κB pathway. Moreover, the protein expressions of Sirt1 (sirtuin 1), MCP-1 (monocyte chemoattractant protein 1), ICAM-1 (intercellular adhesion molecule 1), and VCAM-1 (vascular cell adhesion molecule 1) were decreased by DJ administration. Also, the expression of α-smooth muscle actin and collagen type I alpha 1, collagen-related proteins, that help skin recovery was significantly increased in the DJ treatment group. Histopathologically, a relatively thin epithelial layer could be observed in the DJ administration group, as well as an increase in fibroblasts and collagen fibers. Conclusions These data suggest that DJ treatment is effective in wound healing, suppressing inflammatory proteins, increasing skin repair factors and improving histopathological changes caused by wounds.

Cloning of Dechlorination Genes Specifying Biodegradation of Toxic 4-Chlorobiphenyl (유독성 4-Chlorobiphenyl의 생분해를 위한 탈염소화 유전자의 클로닝)

  • Kim, Chi-Kyung;Chae, Jong-Chan;Han, Jae-Jin
    • Korean Journal of Microbiology
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    • v.32 no.2
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    • pp.126-131
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    • 1994
  • The pchABCD genes in Pseudomonas sp. DJ-12 speciyin degradation o 4-chlorobiphenyl(4CB) were cloned in Eschericia coli. The cloned cells of E. coli CU1 and CU101 showed to produce 2,3-dihydroxybiphenyl (2,3-DHBP) from 4-chlorobiphenyl by dechlorination, as Pseudomonas so. DJ-12 produced 2,3-DHBP from both biphenyl and 4CB. In particular, E. coli CU101 transformed with the recombinant plasmid of pCU101 revealed dechlorination activity to produce 2,3-DHBP from 4CB without production of 4-chlorobenzoic acid. Therefore, the pcbAB genes (2.2 kb in size) cloned from the chromosome of Pseudomonas sp. DJ-12 were found to have dechlorination activity on 4CB to produce 2,3-DHNP.

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Delayed Reduction Algorithms of DJ Graph using Path Compression (경로 압축을 이용한 DJ 그래프의 지연 감축 알고리즘)

  • Sim, Son-Kwon;Ahn, Heui-Hak
    • The KIPS Transactions:PartA
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    • v.9A no.2
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    • pp.171-180
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    • 2002
  • The effective and accurate data flow problem analysis uses the dominator tree and DJ graphs. The data flow problem solving is to safely reduce the flow graph to the dominator tree. The flow graph replaces a parse tree and used to accurately reduce either reducible or irreducible flow graph to the dominator tree. In this paper, in order to utilize Tarian's path compress algorithm, the Top node finding algorithm is suggested and the existing delay reduction algorithm is improved using Path compression. The delayed reduction a1gorithm using path compression actually compresses the pathway of the dominator tree by hoisting the node while reducing to delay the DJ graph. Realty, the suggested algorithm had hoisted nodes in 22% and had compressed path in 20%. The compressed dominator tree makes it possible to analyze the effective data flow analysis and brings the improved effect for the complexity of code optimization process with the node hoisting effect of code optimization process.

Cloning and Characterization of the Genes Responsible for Degradation of 4-Chlorobenzoic Acid (4-Chlorobenzoic Acid 분해유전자의 클로닝과 유전학적 특성)

  • 이익근;김종우;김치경
    • Korean Journal of Microbiology
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    • v.28 no.1
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    • pp.41-46
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    • 1990
  • A bacterial isolate of DJ-12 capable of degrading 4-chlorobenzoic acid (4CBA) as well as 4-chlorobiphenyl (4CB) was used in this study. Its biodegradability of 4CBA was tested and the location of the genes coding for degradation of 4CBA was investigated by the nethod of in vivo cloning. The genes were found to be existed in the plasmid of pDJ121 which is about 65kb in size and which has 9, 11, 10, and 19 restriction sites for EcoRI, HindIII, SalI, and PstI, respectively. The hybrid plasmid of pDK450 was constructed by ligation of the EcoRI fragments of pDJ121 with pKT230 as a vector. In the recombinant cells selected through transformation of the hybrid vector into Pseudomonas putida KT2440, the 4CBA-degrading genes of DJ-12 were proved to be cloned and expressed in the Pseudomonas sp.

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Protective effects of quality certified traditional Doenjang in Korea on TNF-α-induced vascular inflammation in human umbilical vein endothelial cells (혈관내피세포에서 TNF-α 자극에 의해 유도되는 혈관염증에 대한 전통식품 품질인증 된장의 효능 평가)

  • Kim, Eun-Ju;Jang, Yeon-Jeong;Kim, So-Young;Choi, Hye-Sun;Park, Shin-Young
    • Food Science and Preservation
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    • v.23 no.3
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    • pp.378-386
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    • 2016
  • Anti-atherogenic effects in tumor necrosis factor-${\alpha}$ (TNF-${\alpha}$)-stimulated human umbilical vein endothelial cells (HUVEC) are involved in the suppression of oxidative stress, cell adhesion molecules, and pro-inflammatory factors. This study investigated the vascular inflammation inhibitory activity of traditional Doenjang plays a key role in the pathogenesis and progression of atherosclerosis. The protective effects of Korean Deonjang was investigated on the expression of cell adhesion molecules (CAMs) in tumor necrosis factor (TNF)-${\alpha}$-induced human umbilical vascular endothelial cells (HUVECs). Deonjang extracts (20, 50, $100{\mu}g/mL$) decreased the expression of 20 ng/mL TNF-${\alpha}$-induced vascular cell adhesion molecule (VCAM)-1 intracellular adhesion molecule (ICAM)-1 proteins, and their corresponding mRNA levels. Nitric oxides (NO) produced by endothlial nitric oxides synthase (eNOS) dilated blood vessels, which had protective effects against platelet and leukocyte adhesion. While TNF-${\alpha}$-induced suppressed the production of nitric oxide in HUVECs, Doenjang restored NO production in HUVECs. In addition, Deonjang reduced the TNF-${\alpha}$-induced expression of inducible NO synthase (iNOS) and cyclooxygenase (COX)-2 mRNA levels. These results suggested that Doenjang can inhibited the production of cell adhesion molecules and inflammatory mediators, which could be a potential candidate for preventing atherosclerosis.