• Journal of Microbiology and Biotechnology
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• v.13 no.5
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• pp.717-724
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• 2003

A newly isolated Citrobacter sp. Y19 catalyzes the CO-dependent $H_2$ production (biological water-gas shift reaction) by the actions of CO dehydrogenase (CODH) and hydrogenase. Y 19 requires $O_2$ for fast growth, but its $H_2$ production activity is significantly inhibited by $O_2$. In the present study, the effect of $O_2$ on the activities of CODH ard hydrogenase was investigated quantitatively in both whole cells and broken cells, based on CO-dependent or methyl viologen (MV)-dependent $H_2$ production in addition to CO-dependent MV reduction. In crude cell extracts, CODH activity was mostly found in the soluble fraction. Inactivation of CODH and hydrogenase activities by $O_2$ followed the first-order decay kinetics, and the dependence of the rate constants on $O_2$ partial pressure could be expressed by the Michaelis-Menten equation. In whole cells, the maximum deactivation rate constants ($k_{d,max}$ of hydrogenase and CODH were quite similar: $0.07{\pm}0.03 min^{-1}\;and\;0.10{\pm}0.04 min^{-1}$, respectively. However, the first-order rate constant ($k_{d,max}/K_s$) of CODH ($0.25\;min^{-1}\;atm^{-1}$) at low $O_2$ partial pressures was about 3-fold higher than that of the hydrogenase, since the half-saturation constant ($K_s$) of CODH was about half of that of hydrogenase. In broken cells, both enzymes became significantly more sensitive to $O_2$ compared to the unbroken cells, while $k_{d,max}/K_s$ increased 37-fold for hydrogenase and 6.7-fold for CODH. When whole cells were incubated under anaerobic conditions after being exposed to air for 1 h, hydrogenase activity was recovered more than 90% in 2 h suggesting that the deactivation of hydrogenase by $O_2$ was reversible. On the contrary, CODH activity was not recovered once deactivated by $O_2$ and the only way to recover the activity was to synthesize new CODH. This study indicates that $O_2$ sensitivity of $H_2$ production activity of Citrobacter sp. Y19 is an important drawback as in other $H_2-producing$ bactria.

• Microbiology and Biotechnology Letters
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• v.41 no.4
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• pp.433-441
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• 2013

In this study, the anti-oxidative, anti-inflammatory, anti-melanogenic activities of Endlicheria anomala (Nees) Mez methanol extract (EAME) were evaluated by use of in vitro assays and cell culture model systems. The results revealed that EAME scavenges various radicals such as 1,1-diphenyl-2-picryl hydrazyl hydrogen peroxide induced reactive oxygen species, and lipopolysaccharide induced nitric oxide. Furthermore, EAME induced the expression of anti-oxidative enzymes such as heme oxygenase 1, thioredoxin reductase 1, NAD(P)H dehydrogenase 1, and their upstream transcription factor, nuclear factor-E2-related factor 2. Moreover, EAME inhibited in vitro DOPA oxidation and 3-isobutyl-1-methylxanthine induced melanogenesis in B16F10 cells. Its anti-melanogenic activity will have originated from the inhibition of tyrosinase enzyme activity and melanogenesis related protein expression. Taken together, these results provide the important new insight that E. anomala possesses various biological activities such as anti-oxidative, anti-inflammatory, and anti-melanogenic. Therefore, it might be utilized as a promising material in the fields of nutraceuticals and cosmetics.

• Korean Journal of Plant Taxonomy
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• v.42 no.4
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• pp.253-259
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• 2012

The genetic variation in populations of Eranthis byunsanensis, an endemic and rare species of Korea, was studied using starch gel electrophoresis. All five known populations were sampled for allozyme electrophoresis of nine enzymes coded by 10 loci. The overall genetic variation of E. byunsanensis population was shown to be considerably high within the populations (A = 2.4, P = 90.0, $H_E$ = 0.311). A positive $F_{IS}$ value of E. byunsanensis indicated an overall deficiency of heterozygotes, and a low $F_{ST}$ value (0.131) showed little differentiation among populations. The high genetic variation, less genetic differentiation among populations, and a significant amount of heterozygote deficiency propose the hypothesis that they have an experience of recent isolation and fragmentation of their habitat. Thus, the rate of gene flow has been drastically reduced, and the rate of inbreeding in E. byunsanensis populations has increased. Current habitats in Mai-san and Naro-do are vulnerable due to their small population size and the levels of anthropogenic activity in the region constantly threatening survival of this species. Because of the high genetic variation and low levels of differentiation among populations in E. byunsanensis, it is not issue which populations have a priority for protection, but we may concern the plan to maintain population continuously and diminish the rate of inbreeding.

• Korean Journal of Microbiology
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• v.51 no.4
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• pp.398-406
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• 2015

Culture-dependent ARDRA and culture-independent DGGE were employed to investigate the bacterial community associated with the marine sponge Halichondria panicea collected from Jeju Island. A total of 120 bacterial strains associated with the sponge were cultivated using modified Zobell and Marine agar media. PCR amplicons of the 16S rRNA gene from the bacterial strains were digested with the restriction enzymes HaeIII and MspI, and then assigned into different groups according to their restriction patterns. The 16S rRNA gene sequences derived from ARDRA patterns showed more than 96% similarities compared with known bacterial species, and the isolates belonged to four classes, Alphaproteobacteria, Gammaproteobacteria, Bacteroidetes, and Firmicutes, of which Alphaproteobacteria was dominant. DGGE fingerprinting of 16S rRNA genes amplified from the sponge-derived total gDNA showed 14 DGGE bands, and their sequences showed 100% similarities compared with the sequences available in GenBank. The sequences derived from DGGE bands revealed high similarity with the uncultured bacterial clones. DGGE revealed that bacterial community consisted of seven classes, including Alphaproteobacteria, Gammaproteobacteria, Acidobacteria, Actinobacteira, Bacteroidetes, Cyanobacteria, and Chloroflexi. According to both the ARDRA and DGGE methods, three classes, Alphaproteobacteria, Gammaproteobacteria, and Bacteroidetes, were commonly found in H. panicea. However, overall bacterial community in the sponge differed depending on the analysis methods. Sponge showed more various bacterial community structures in culture independent method than in culture-dependent method.

• Journal of the Korean Society of Food Science and Nutrition
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• v.45 no.1
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• pp.27-34
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• 2016

Garlic has drawn attention as a food material for its anti-oxidative and anti-inflammatory properties as well as for prevention and treatment of cancer. In order to increase efficiency, various aging methods for garlic have been attempted. In particular, thermally processed garlic is known to have higher biological activities due to its various chemical changes during heat treatment. Therefore, in this study, we investigated the anti-oxidative effects of garlic extracts aged at low temperature ($60{\sim}70^{\circ}C$). In the results, 2,2-diphenyl-1-picrylhydrazyl and 2,2-azino-bis (3-ethylbenzo-thiazoline-6-sulfonate) radical scavenging activities and ferric reducing ability of low temperature-aged garlic (LTAG) were similar to those of raw garlic. LTAG also showed decreased lipopolysaccharide (LPS)-induced production of reactive oxygen species, although there were not significant differences among samples. In addition, xanthine oxidase activity was inhibited by LTAG; the 15 days and $60^{\circ}C$ extract showed outstanding inhibition compared with the others. To understand the molecular mechanisms behind the anti-oxidative activity of LTAG, we performed quantitative real-time PCR analysis. The 30 days and $70^{\circ}C$ extract upregulated mRNA expression of antioxidant enzymes such as Cu/Zn-superoxide dismutase (SOD), Mn-SOD, glutathione peroxidase, and catalase in LPS-stimulated RAW 264.7 cells. This result indicates that LTAG can be a functional food as a nature antioxidant and antioxidant substance.

• ALGAE
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• v.32 no.4
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• pp.359-377
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• 2017

Pyropia haitanensis (T. J. Chang et B. F. Zheng) N. Kikuchi et M. Miyata is one of the most commercially useful macroalgae cultivated in southeastern China. In red algae, the biosynthesis of terpenoids through 2-C-methyl-D-erythritol 4-phosphate (MEP) pathway can produce a direct influence on the synthesis of many biologically important metabolites. In this study, two genes of cDNAs, 1-deoxy-D-xylulose-5-phosphate synthase (DXS) and 1-deoxy-D-xylulose-5-phosphate reductase (DXR), which encoding the first two rate-limiting enzymes among MEP pathway were cloned from P. haitanensis. The cDNAs of P. haitanensis DXS (PhDXS) and DXR (PhDXR) both contained complete open reading frames encoding polypeptides of 764 and 426 amino acids residues, separately. The expression analysis showed that PhDXS was significant differently expressed between leafy thallus and conchocelis as PhDXR been non-significant. Additionally, expression of PhDXR and its downstream gene geranylgeranyl diphosphate synthase were both inhibited by fosmidomycin significantly. Meanwhile, we constructed types of phylogenetic trees through different algae and higher plants DXS and DXR encoding amino acid sequences, as a result we found tree clustering consequences basically in line with the "Cavalier-Smith endosymbiotic theory." Whereupon, we speculated that in red algae, there existed only complete MEP pathway to meet needs of terpenoids synthesis for themselves; Terpenoids synthesis of red algae derivatives through mevalonate pathway came from two or more times endosymbiosis of heterotrophic eukaryotic parasitifer. This study demonstrated that PhDXS and PhDXR could play significant roles in terpenoids biosynthesis at molecular levels. Meanwhile, as nuclear genes among MEP pathway, PhDXS and PhDXR could provide a new way of thinking to research the problem of chromalveolata biological evolution.

• BMB Reports
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• v.44 no.7
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• pp.484-489
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• 2011

Annexin-1 (ANX1) is an anti-inflammatory protein as well as an important modulator in inflammation. However, the precise action of ANX1 remains unclear. To elucidate the protective effects of ANX1 on lipopolysaccharide (LPS)-induced murine macrophage Raw 264.7 cells, we constructed a cell-permeable Tat-ANX1 protein. The transduced Tat-ANX1 protein markedly inhibited the expression of cyclooxygenase-2, production of prostaglandin $E_2$, and generation of pro-inflammatory cytokines in the cells. Furthermore, transduced Tat-ANX1 protein caused a significant reduction in the activation of nuclear factor-kappa B (NF-${\kappa}B$) and mitogen-activated protein kinase (MAPK). The results indicate that Tat-ANX1 inhibits the production of inflammatory response cytokines and enzymes by blocking NF-${\kappa}B$ and MAPK. Therefore, Tat-ANX1 protein may be useful as a therapeutic agent against various inflammatory diseases.

• Journal of Life Science
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• v.30 no.5
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• pp.483-490
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• 2020

The ubiquitin system uses ligases and deubiquitinases (DUBs) to regulate ubiquitin position on protein substrates and is involved in many biological processes which determine stability, activity, and interaction of the target substrate. DUBs are classified in six groups according to catalytic domain, namely ubiquitin-specific proteases (USPs); ubiquitin C-terminal hydrolases (UCHs); ovarian tumor proteases (OTUs); Machado Joseph Disease proteases (MJDs); motif interacting with Ub (MIU)-containing novel DUB family (MINDY); and Jab1/MPN/MOV34 metalloenzymes (JAMMs). Otubain 1 (OTUB1) is a DUB in the OTU family which possesses both canonical and non-canonical activity and can regulate multiple cellular signaling pathways. In this review, we describe the function of OTUB1 through regulation of its canonical and non-canonical activities in multiple specifically cancer-associated pathways. The canonical activity of OTUB1 inhibits protein ubiquitination by cleaving Lys48 linkages while its non-canonical activity prevents ubiquitin transfer onto target proteins through binding to E2-conjugating enzymes, resulting in the induction of protein deubiquitination. OTUB1 can therefore canonically and non-canonically promote tumor cell proliferation, invasion, and drug resistance through regulating FOXM1, ERα, KRAS, p53, and mTORC1. Moreover, clinical research has demonstrated that OTUB1 overexpresses with high metastasis in many tumor types including breast, ovarian, esophageal squamous, and glioma. Therefore, OTUB1 has been suggested as a diagnosis marker and potential therapeutic target for oncotherapy.

• Journal of Life Science
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• v.22 no.5
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• pp.627-633
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• 2012

$Microbulbifer$ sp. AJ-3 was used to acquire the degrading products from $Capsosiphon$ $fulvescens$ (DPCF), which were investigated to determine its physiological activities. A crude enzyme extract from $Microbulbifer$ sp. AJ-3 hydrolyzes polysaccharide substrates such as agar, agarose, alginic acid, fucoidan, laminaran, starch, and chitin. Among them, agarose, laminaran, and alginic acid showed higher activities, especially alginic acid. The antioxidant activity of DPCF was measured by using 2,2-diphenyl-1-picryl-hydrazyl (DPPH) and superoxide dismutase (SOD)-like activities and were about 32% and 93% at 2 mg/ml, respectively. In addition, the nitrite-scavenging activity of DPCF was about 82%, 53%, and 12% at pH levels of 1.2, 3.0, and 6.0, respectively. To determine the influence of DPCF on alcohol metabolism, the generating activity of reduced-nicotinamide adenine dinucleotide (NADH) by alcohol dehydrogenase (ADH) was measured. The facilitating rate of ADH activity by DPCF was 130% at 2 mg/ml. The tyrosinase inhibitory activity of DPCF was slightly increased in a dose-dependent manner and was about 28% at 2 mg/ml. These results indicated that the enzymatic products from DPCF have a strong antioxidant, nitrite scavenging, and alcohol metabolizing activity.

• Microbiology and Biotechnology Letters
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• v.18 no.1
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• pp.81-88
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• 1990

For the selection of powerful antagonistic bacterium for biological control of soilborne Fusarium solani causing root rot of many important crops, the best YPL-1 strain was selected among 300 strains of bacteria isolated from rhizosphere in ginseng root rot-suppressive soil. The strain was identified to be a species to Pseudomonas stutzeri. With in vitro fungal inhibition tests, antagonistic substance of P. stutzeri YPL-1 against F. solani was presumed to be heat unstable, macromolecular substances such as protein. Also, it was shown that antifungal activity of P. stutzeri YPL-1 increased in proportion to its chitinase production. P. stutzeri YPL-M122 (chi-, lam -) which was deprived of the productivity of chitinase and laminarinase by NTG mutagenesis had lost antifungal activity, completely. And P. stutzeri YPL-MI53 (chi-) had only 4.1% of its antifungal activity. P. stutzeri YPL-1 was not able to produce any extracellular siderophore in iron-deficent minimal medium. It is confident that the antifungal mechanism of P. stutzeri YPL-1 for biocontrol of F. solani depends on lysis rather than antibiosis :the mechanism of lysis appears to involve enzymatic degradation of the cell will components of F. solani by hydrolytic enzymes of more chitinase and less laminarinase.