• Title/Summary/Keyword: U-Terminal

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Highly Efficient Biotransformation of Astragaloside IV to Cycloastragenol by Sugar-Stimulated β-Glucosidase and β-Xylosidase from Dictyoglomus thermophilum

  • Li, Qi;Wu, Tao;Zhao, Linguo;Pei, Jianjun;Wang, Zhenzhong;Xiao, Wei
    • Journal of Microbiology and Biotechnology
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    • v.29 no.12
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    • pp.1882-1893
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    • 2019
  • β-Glucosidases and β-xylosidases are two categories of enzymes that could cleave out non-reducing, terminal β-D-glucosyl and β-D-xylosyl residues with release of D-glucose and D-xylose, respectively. In this paper, two functional β-glucosidase Dth3 and β-xylosidase Xln-DT from Dictyoglomus thermophilum were heterologously expressed in E.coli BL21 (DE3). Dth3 and Xln-DT were relatively stable at 75℃ and were tolerant or even stimulated by glucose and xylose. Dth3 was highly tolerant to glucose with a Ki value of approximately 3 M. Meanwhile, it was not affected by xylose in high concentration. The activity of Xln-DT was stimulated 2.13-fold by 1 M glucose and 1.29-fold by 0.3 M xylose, respectively. Furthermore, the βglucosidase Dth3 and β-xylosidase Xln-DT showed excellent selectivity to cleave the outer C-6 and C-3 sugar moieties of ASI, which established an effective and green method to produce the more pharmacologically active CAG, an exclusive telomerase activator. We measured temperature, pH and dosage of enzyme using a single-factor experiment in ASI biotransformation. After optimization, the optimal reaction conditions were as follows: 75℃, pH 5.5, 1 U of Dth3 and 0.2 U of Xln-DT, respectively. Under the optimized conditions, 1 g/l ASI was transformed into 0.63 g/l CAG with a corresponding molar conversion of 94.5% within 3 h. This is the first report to use the purified thermostable and sugar-tolerant enzymes from Dictyoglomus thermophilum to hydrolyze ASI synergistically, which provides a specific, environment-friendly and cost-effective way to produce CAG.

Interaction of promyelocytic leukemia/p53 affects signal transducer and activator of transcription-3 activity in response to oncostatin M

  • Lim, Jiwoo;Choi, Ji Ha;Park, Eun-Mi;Choi, Youn-Hee
    • The Korean Journal of Physiology and Pharmacology
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    • v.24 no.3
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    • pp.203-212
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    • 2020
  • Promyelocytic leukemia (PML) gene, through alternative splicing of its C-terminal region, generates several PML isoforms that interact with specific partners and perform distinct functions. The PML protein is a tumor suppressor that plays an important role by interacting with various proteins. Herein, we investigated the effect of the PML isoforms on oncostatin M (OSM)-induced signal transducer and activator of transcription-3 (STAT-3) transcriptional activity. PML influenced OSM-induced STAT-3 activity in a cell type-specific manner, which was dependent on the p53 status of the cells but regardless of PML isoform. Interestingly, overexpression of PML exerted opposite effects on OSM-induced STAT-3 activity in p53 wild-type and mutant cells. Specifically, overexpression of PML in the cell lines bearing wild-type p53 (NIH3T3 and U87-MG cells) decreased OSM-induced STAT-3 transcriptional activity, whereas overexpression of PML increased OSM-induced STAT-3 transcriptional activity in mutant p53-bearing cell lines (HEK293T and U251-MG cells). When wild-type p53 cells were co-transfected with PML-IV and R273H-p53 mutant, OSM-mediated STAT-3 transcriptional activity was significantly enhanced, compared to that of cells which were transfected with PML-IV alone; however, when cells bearing mutant p53 were co-transfected with PML-IV and wild-type p53, OSM-induced STAT-3 transcriptional activity was significantly decreased, compared to that of transfected cells with PML-IV alone. In conclusion, PML acts together with wild-type or mutant p53 and influences OSM-mediated STAT-3 activity in a negative or positive manner, resulting in the aberrant activation of STAT-3 in cancer cells bearing mutant p53 probably might occur through the interaction of mutant p53 with PML.

GA-based Two Phase Method for a Highly Reliable Network Design (높은 신뢰도의 네트워크 설계를 위한 GA 기반 두 단계 방법)

  • Jo, Jung-Bok
    • Journal of the Korea Institute of Information and Communication Engineering
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    • v.9 no.5
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    • pp.1149-1160
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    • 2005
  • Generally, the network topology design problem, which is difficult to solve with the classical method because it has exponentially increasing complexity with the augmented network size, is characterized as a kind of NP-hard combinatorial optimization problem. The problem of this research is to design the highly reliable network topology considering the connection cost and all-terminal network reliability, which can be defined as the probability that every pair of nodes can communicate with each other. In order to solve the highly reliable network topology design problem minimizing the construction cost subject to network reliability, we proposes an efficient two phase approach to design reliable network topology, i.e., the first phase employs, a genetic algorithm (GA) which uses $Pr\ddot{u}fer$ number for encoding method and backtracking Algorithm for network reliability calculation, to find the spanning tree; the second phase is a greedy method which searches the optimal network topology based on the spanning ree obtained in the first phase, with considering 2-connectivity. finally, we show some experiments to demonstrate the effectiveness and efficiency of our two phase approach.

Isolation and Molecular Analysis of Methanol Oxidation Genes in an Obligate Methylotrophic Bacterium, Metheylobacillus sp. Strain SK-5

  • Choi, Hack-Sun;Kim, Jin-Kwon;Ahn, Yeong-Hee;Koh, Moon-Joo;Kim, Si-Wouk
    • Journal of Microbiology and Biotechnology
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    • v.12 no.5
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    • pp.819-825
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    • 2002
  • Methanol dehydrogenase (MDH) is a key enzyme in the process of methanol oxidation in methylotrophic bacteria. However, information on MDH genes from genus Methylobacillus is limited. In this study, a 6.5-kb HindIII DNA fragment of Methylobacillus sp. SK-5 chromosomal DNA was isolated from the genomic library of the strain by using a degenerate oligonucleotide probe that was designed based on JV-terminal amino acid sequence of the MDH $\alpha$ subunit purified from the strain. Molecular analysis of the fragment revealed four tightly clustered genes (mxaFJGI) involved in the methanol oxidation. The first and fourth genes were very similar to mxaF (77% identity for nucleotides an 78% identity for amino acids) and mxaF (67% Identity for nucleotides and 68% Identity for amino acids) genes, respectively, from Methylovorus sp. SSI. Genes mxaF and mxaI encode $\alpha$ and $\beta$ subunits of MDH, respectively. The two subunits were identified from purified MDH from Methylobacillus sp. SK-5. A dendrogram constructed by comparison of amino acid sequences of MDH u subunits suggests that MxaF from Methylobacillus sp. SK-5 belongs to a subfamily cluster of MDH u subunits from $\beta$-subgroup Proteobacteria. The subfamily cluster is separated from the other subfamily that consists of $\beta$- and $\gamma$-subgroup Proteobacteria. This study provided information on mn genes from a methylotrophic bacterium in $\beta$-subgroup Proteobacteria, which would aid to better develop a gene probe to detect one-carbon metabolizing bacteria.

Telmisartan Inhibits TNFα-Induced Leukocyte Adhesion by Blocking ICAM-1 Expression in Astroglial Cells but Not in Endothelial Cells

  • Jang, Changhwan;Kim, Jungjin;Kwon, Youngsun;Jo, Sangmee A.
    • Biomolecules & Therapeutics
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    • v.28 no.5
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    • pp.423-430
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    • 2020
  • Telmisartan is an angiotensin-II receptor blocker and acts as a selective modulator of peroxisome proliferator-activated receptor gamma (PPARγ). Several studies have demonstrated that telmisartan ameliorates depression and memory dysfunction and reduces brain inflammation. We hypothesized that the beneficial effects of telmisartan on brain could be due to modulation of the blood-brain barrier (BBB) function. Here, we examined the effect of telmisartan on tumor necrosis factor alpha (TNF-α)-induced expression of intercellular adhesion molecule 1 (ICAM-1) which plays an important role in leukocyte transcytosis through the BBB. Telmisartan blocked TNF-α-induced ICAM-1 expression and leukocyte adhesion in U87MG human glioma cells but showed no effect on human brain microvascular endothelial cells. In U87MG cells, a PPAR antagonist, GW9662 did not block the effect of telmisartan on ICAM1 expression but rather potentiated. Moreover, GW9662 caused no change in TNF-α-induced ICAM-1 expression, suggesting no implication of PPARγ in the telmisartan effect. Further studies showed that telmisartan blocked TNF-α-induced activation of c-Jun N-terminal kinase (JNK), extracellular signal-regulated kinase 1/2 (ERK1/2), p38, and nuclear factorkappa B (NF-κB). In contrast, inhibitors of JNK, ERK1/2 and NF-κB but not p38, blocked ICAM-1 expression induced by TNF-α. Thus, our findings suggest that the beneficial effect of telmisartan is likely due to the reduction of astrocytic ICAM1 expression and leukocytes adhesion to astrocytes, and that this response was mediated by the inhibition of JNK/ERK1/2/NF-κB activation and in the PPAR-independent manner. In conclusion, this study enhances our understanding of the mechanism by which telmisartan exerts the beneficial brain function.

Inhibitory effect of ethanolic extract of Abeliophyllum distichum leaf on 3T3-L1 adipocyte differentiation

  • Thomas, Shalom Sara;Eom, Ji;Sung, Nak-Yun;Kim, Dong-Sub;Cha, Youn-Soo;Kim, Kyung-Ah
    • Nutrition Research and Practice
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    • v.15 no.5
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    • pp.555-567
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    • 2021
  • BACKGROUND/OBJECTIVES: Abeliophyllum distichum is a plant endemic to Korea, containing several beneficial natural compounds. This study investigated the effect of A. distichum leaf extract (ALE) on adipocyte differentiation. MATERIALS/METHODS: The cytotoxic effect of ALE was analyzed using cell viability assay. 3T3-L1 preadipocytes were differentiated using induction media in the presence or absence of ALE. Lipid accumulation was confirmed using Oil Red O staining. The mRNA expression of adipogenic markers was measured using RT-PCR, and the protein expressions of mitogen-activated protein kinase (MAPK) and peroxisome proliferator-activated receptor gamma (PPAR𝛾) were measured using western blot. Cell proliferation was measured by calculating the incorporation of Bromodeoxyuridine (BrdU) into DNA. RESULTS: ALE reduced lipid accumulation in differentiated adipocytes, as indicated by Oil Red O staining and triglyceride assays. Treatment with ALE decreased the gene expression of adipogenic markers such as Ppar𝛾, CCAAT/enhancer binding protein alpha (C/ebp𝛼), lipoprotein lipase, adipocyte protein-2, acetyl-CoA carboxylase, and fatty acid synthase. Also, the protein expression of PPAR𝛄 was reduced by ALE. Treating the cells with ALE at different time points revealed that the inhibitory effect of ALE on adipogenesis is higher in the early period treatment than in the terminal period. Furthermore, ALE inhibited adipocyte differentiation by reducing the early phase of adipogenesis and mitotic clonal expansion. This was indicated by the lower number of cells in the Synthesis phase of the cell cycle (labeled using BrdU assay) and a decrease in the expression of early adipogenic transcription factors such as C/ebp𝛽 and C/ebp𝛿. ALE suppressed the phosphorylation of MAPK, confirming that the effect of ALE was through the suppression of early phase of adipogenesis. CONCLUSIONS: Altogether, the results of the present study revealed that ALE inhibits lipid accumulation and may be a potential agent for managing obesity.

Population Dynamics of Mabled sole Limanda yokohamae($G{\"{U}}NTHER$) in Tokyo Bay, Japan (동경만산 문치가자미Limanda yokohamae($G{\"{U}}NTHER$)의 자원양 변동의 해석)

  • PARK Jong-Soo;SIMIZU Mako-to
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.24 no.1
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    • pp.1-8
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    • 1991
  • Population dynamics of Matted sole Limanda yokohamae($G{\"{U}}NTHER$) in Tokyo bay, Japan has been studied by virtual population analysis (VPA) for multi cohort and experimental fishing. Based on the biological data, the present parameters of the Limanda yekohamae stock at the Tokyo bay, Japan were estimated as follows: natural mortality coefficient(M) were 0.313 for male and 0.250 for female, terminal fishing mortality coefficient(F) were 2.190 for male, and 0.798 for female, rate of exploitation(E) was $30\%\;to\;50\%$. From the result of virtual population analysis for multi cohort, the population size were estimated from 3,5000,000 to 9,200,000 fishes, according to the result of experimental fishing, estimated stock size were 2,400,000 to 8,700,000 fishes. Stock size difference of the two methods were about two times in 1987, however, other years has been showed from 0.8 to 1.5 times. Both method has been showed same increase and decrease tendency of the c. p. u. e. and catches. From the isopleth diagram plot by Beverton and Holt's yield per recruit, the catches could be increase two times for female, 1.3 times for male than present aspects by the fishing management. And further, as reducing fishing effort, extension of mesh size and rising the length at first caputre, are reasonable in order to manage the stock at the optimum level.

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Production of manganese peroxidase from Pleurotus ostreatus using a rotary draft tube bioreactor (RTB) and characterization of its activity (회전식 통풍관 생물반응기 사용에 따른 느타리균의 manganese peroxidase 생산 및 특성)

  • Ha, Hyo-Cheol
    • Journal of Mushroom
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    • v.19 no.4
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    • pp.316-321
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    • 2021
  • Ligninolytic enzymes were produced by Pleurotus ostreatus No.42, cultivated in a new kind of bioreactor that has a rotating draft tube with a helical ribbon. Maximum laccase (Lac) production (about 8,200 U/bioreactor) was reached after 3 days of incubation, then production decreased. Production of manganese peroxidase (MnP) in this fermenter reached a maximum level of about 8,400 U/bioreactor after 6 days of incubation. Lignin peroxidase (LiP) was not detected under these growth conditions. These results indicate that the rotary draft tube bioreactor (RTB) is compatible with large scale production of ligninolytic enzymes. MnP produced under these fermentation conditions was purified via a multistep process that included chromatography on Sepharose CL-6B, prep grade Superdex 75, and Mono-Q. This major isoenzyme was confirmed to have an apparent molecular weight of 36,400 by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), and its isoelectric point (IEF) was determined to be 3.95. N-terminal sequencing of the major isoenzyme from this fermentation was identical to that reported for an MnP3 isoenzyme isolated under different cultivation conditions, including stationary and shaking culture.

Cadms/SynCAMs/Necls/TSLCs Interact with Multi-PDZ Domain Protein MUPP1 (Cadms/SynCAMs/Necls/TSLCs와 multi-PDZ domain protein MUPP1 단백질의 결합)

  • Jang, Won Hee;Jeong, Young Joo;Choi, Sun Hee;Kim, Sang-Jin;Urm, Sang-Hwa;Moon, Il Soo;Seog, Dae-Hyun
    • Journal of Life Science
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    • v.24 no.12
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    • pp.1276-1283
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    • 2014
  • Cell adhesion molecules determine the cell-cell binding and the interactions between cells and extracellular signals. Cell-cell junctional complexes, which maintain the structural integrity of tissues, consist of more than 50 proteins including multi-PDZ domain protein 1 (MUPP1). MUPP1 contains 13 postsynaptic density-95/disks large/zonula occludens-1 (PDZ) domains and serves a scaffolding function for transmembrane proteins and cytoskeletal proteins or signaling proteins, but the mechanism how MUPP1 links and stabilizes the juxtamembrane proteins has not yet been elucidated. We used the yeast two-hybrid system to identify proteins that interact with PDZ domains of MUPP1. We found an interaction between MUPP1 and cell adhesion molecule 1 (Cadm1, also known as SynCAM1, Necl-2, or TSLC1). Cadm1 bound to the second PDZ domain of MUPP1. The carboxyl (C)-terminal end of Cadm1 has a type II PDZ-association motif (-Y-F-I) which was essential for the interaction with MUPP1 in the yeast two-hybrid assay. MUPP1 also bound to the C-terminal cytoplasmic tail region of other Cadm family members (Cadm2, Cadm3, and Cadm4). In addition, these protein-protein interactions were observed in the glutathione S-transferase (GST) pull-down assay and by co-immunoprecipitation. Anti-MUPP1 antibody co-immunoprecipitated Cadm1 and Cadm4 with MUPP1 from mouse brain extracts. These results suggest that MUPP1 could mediate interaction between Cadms and cytoskeletal proteins.

Effects of a Palliative Care Program based on Home Care Nursing (가정간호기반 완화케어프로그램의 효과)

  • Hwang, Moon-Sook;Ryu, Ho-Sihn
    • Journal of Korean Academy of Nursing
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    • v.39 no.4
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    • pp.528-538
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    • 2009
  • Purpose: This study was done to develop and test a palliative care program based on home care nursing. Methods: A quasi-experimental design was employed. Changes in the variables were evaluated to test effects of the developed program. Participants were patients with terminal cancer and their families receiving home care nursing from six hospitals (experimental group: 24 and control group: 22). Data collection was conducted from February to October, 2006. Chi-square test, Fisher's exact test, t-test, Mann-Whitney U test and repeated measures ANOVA were used to analyse the data. Results: Hypothesis 1, the experimental group receiving this program will experience less pain (severe, average, weak pain) than the control group, was supported. Hypothesis 2, the experimental group will have less symptom experience than the control group, was supported. Hypothesis 3, the experimental group will have higher QOL than the control group, was supported and the last hypothesis 4, family burden in the experimental group will be less than the control group, was supported. Conclusion: The home care nursing based palliative program developed in this study was found to be an effective program to reduce patient pain and symptom experience, to improve patient QOL and to decrease family burden.