• Korean Journal of Microbiology
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• v.32 no.2
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• pp.147-154
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• 1994

Nuclease was secreted to the environmental media from the Escherichia coli JM107 tranformant harboring the extracellular nuclease gene of Serratia marcescens in the plasmid of pNUC4. Under the growth conditions, the amount of secreted enzyme was increased in parallel with bacterial growth conditions, the amount of secreted enzyme was increased in parallel with bacterial growth. The enzyme was purified using chromatofraphic procedures of Matrex green gel and heparin agarose affinity gel, resulted in 50-fold purification with 15% recovery of the enzyme. The apparent molecular weight of the enzyme was estimated to be 29Kda by sodium dodecylsulfate denaturing gel electrophoresis. Using the purified enzyme, polyclonal antibody was obtained from the rabbit. The specificity of the antibody was confirmed by immunoblotting and immunoprecipitaion. For the investigation of cellular distribution of the enzyme, cells were fractionated into three fractions; cytoplasm, periplasm and extracellular fluid. While more than 80% of the enzymatic activity was detected in the extracellular fluid and periplasm, a little was found in the cytoplasm, indicating that the enzyme was likely to be immediately exported to the membrane for excretion after biosynthesis. These results were confirmed again by immunocytochemistry technique using the antibody.

• Journal of Life Science
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• v.21 no.2
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• pp.194-201
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• 2011

Caudal-related homeodomain proteins play critical roles in intestine development and maintenance from Drosophila to humans. The loss or reduction of CDX1 and CDX2 are known to be associated with colon cancers. It has been well known that colorectal carcinogenesis is associated with serious oxidative stress and that catalase is decreased in colon carcinomas. However, the underlying molecular mechanisms remain elusive. Here, we report that Caudal-related homeodomain proteins positively regulate catalase expression in both Drosophila and humans. We found that Drosophila caudal heterozygotes have a decreased catalase expression and increased ROS generation in the hindgut, and that the overexpression of Caudal increases catalase promoter activity and catalase mRNA levels. We also found that CDX1 and CDX2 up-regulate catalase promoter activity and protein levels in HCT116 cells - human colorectal carcinoma cell lines. The level of catalase protein in several colorectal carcinoma cell lines was associated with CDX1 expression. These results suggest that CDX1 and CDX2 may be involved in intestinal homeostasis and tumorigenesis via regulation of catalase expression.

• Journal of Life Science
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• v.18 no.4
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• pp.537-541
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• 2008

White adipose tissue is now recognized as an important endocrine organ which secretes various signal factors and proteins termed 'adipokine'. Haptoglobin (Hp), which has been known as an acute phase protein, belongs to the adipokine. However, the function of Hp in adipose tissue remains unclear. To verify the role of Hp in preadipocytes, in this study, 3T3-L1 preadipocyte cells were stably transfected with human Hp gene and Hp-overexpressing cells were made. The Hp had no effect on cell growth of preadipocytes. By RT-PCR and Western blot analysis, the Hp inhibited gene expression of IL-6 and COX-2 and enhanced HO-1 synthesis in preadipocytes. Moreover, invasion assay showed the Hp suppressed migration of monocytes to preadipocytes. These findings suggest that the Hp may inhibit an inflammatory reaction in adipose tissue by regulating the expressions of pro-inflammatory and anti-inflammatory mediators, and by repressing monocytes/macrophages infiltration.

• Journal of Life Science
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• v.15 no.6 s.73
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• pp.884-888
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• 2005

An agar-degrading bacterium was isolated from north-eastern sea of Jeju island and cultured in marine agar 2216 media. Biochemical and morphologicl characteristics and 165 rRNA gene revealed that isolated strain was member of Agarivorans genus, and named Agarivorans sp. JA-1. Agarase was produced as growth-related and expressed regardless of agar presence. Optimal pH was 8 at 50 mM Clycine-NaOH buffer, and activity was maximum at $40^{\circ}C$E Enzymatic activity was maintained over $80\%$ at $60^{\circ}C$t and $70\%$ at $80^{\circ}C$ which is thermotolerant. Hence isolated novel Agarivorans sp. JA-1 strain and its beta-agarase could be used for the production of functional oligosaccharide from agar in solution state.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.31 no.4 s.54
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• pp.329-335
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• 2005

In order to investigate the effects of Saussurea involucrata on (relationship between) aging (and Saussurea involucrata), we examined the activities of antioxidation, in vitro MMP inhibition and UVA-induced MMP-1 expression in human dermal fibroblasts. S. involucrata showed scavenging activities radicals and reactive oxygen species (ROS) with the $IC_{50}$ values of $3.89{\mu}g/mL$ against 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical and $67.29{\mu}g/mL$ against superoxide radicals in the xanthine/xanthine oxidase system, respectively. At the concentration of $1000{\mu}g/mL$, S. involucrata showed 93.27% inhibition on lipid peroxidation of linoleic acid. S. involucrata inhibited the activities of MMP-1 in a does-dependent manner and the $IC_{50}$ value calculated from semi-log plots was $97.18{\mu}g/mL$. Also, UVA induced MMP expression in human dermal fibroblasts was reduced 42.86% by treatment with S. involucrata, and MMP-1 mRNA expression was reduced in a dose-dependent manner. Therefore S. involucrata was able to significantly inhibit MMP expression in protein and mRNA level. All these results suggested that S. involucrata might act as an anti-aging agent by antioxidation and reducing UVA-induced MMP-1 production.

• Journal of Life Science
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• v.29 no.4
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• pp.421-427
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• 2019

Mobilization of storage lipids is critical for the germination of oil seeds, as they supply carbon and energy until photosynthesis commences in cotyledons. In this study, we determined the levels of plant carnitine and associated changes in these levels from seed germination to cotyledon senescence. We also examined changes in the content of unsaturated fatty acids throughout seedling development. Carnitine levels peaked on day 3 at 14.5 nM in cotyledons and decreased sharply to 7.2 nM on day 4. On development day 3 carnitine levels were maintained at around 3 nM until day 7. The unsaturated fatty acid content dropped by half at the same time as carnitine peaked (day-3), and storage lipids were almost depleted by day 5. Thereafter, carnitine was hardly detected until the second stage of cotyledon senescence, at which stage the carnitine content was 6.8 nM, similar to that on day 4 at the time of fatty acid depletion in the cotyledons. Unsaturated fatty acids levels remained constant in green cotyledons but slightly increased in the senescing cotyledons. The latter can be explained by intracellular breakdown of membrane lipids. This is the first such discovery in developing cotyledons and may offer clues regarding other roles of the acetyl unit transport system in plants. The expression of BOU was closely associated with carnitine metabolism during seed germination and cotyledon development. The results provide support for the possibility of carbon re-routing during the glyoxylate cycle in the supply of energy for early germination and development.

• Korean Journal of Ornithology
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• v.25 no.2
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• pp.94-100
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• 2018

Bird electrocution on contact with electricity lines is well reported and is quite common in Mongolian open areas. We visited 15-kV electricity distribution pole lines in a Southern Mongolian semi-desert region three times in 2017, in April, July, and September, to assess their risks to birds. The carcasses of 45 electrocuted birds representing 12 species were identified from 250 poles (overall mortality rate of 1.12% every 10 km). The majority of these electrocuted birds were endangered Saker Falcon (n=11) and common Black Kite (n=11). The lacks of roosting or perching sites in our study sites (indeed other open areas in Mongolia) may resulted for such high rate bird electrocution. A 751-base pair (bp) cytochrome c oxidase I (COI) gene was amplified for DNA sequence-based identification of carcasses that were difficult to identify directly. Our findings revealed the high electrocution risk for birds in Mongolian open areas, especially for the birds of prey, with relatively low-efficiency electrocution mitigation approaches. The findings also indicate that there is a need for better understanding of the risk of bird electrocution, particularly in the open areas. This will contribute to the conservation of endangered species.

• Journal of Life Science
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• v.29 no.1
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• pp.97-104
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• 2019

Streptococcus iniae is a typical fish pathogen causing streptococcosis and it can also cause zoonotic infectious diseases. We studied S. iniae FP5228 isolated from infected olive flounder in Wando, Korea. In a study to find virulence factors in FP5228, we found that the number of live bacteria decreased dramatically in culture medium containing S. iniae FP5228 for more than 24 hr. This phenomenon was hypothesized to be related to Toxin ${\zeta}$ and Antitoxin ${\varepsilon}$ genes, components of the Toxin/ Antitoxin (TA) system on the 14 kb plasmid of FP5228. We used a protein overexpression system to identify it. The pBP1140 vector system was constructed to regulate the expression of Toxin ${\zeta}$ and Antitoxin ${\varepsilon}$ by IPTG and Arabinose. E. coli/pBP1140 strain grew slowly in early growth under toxin expression condition, and it was confirmed by microscopic observation that the strain became longer. S. iniae CK287, lacking a 14 kb plasmid of S. iniae FP5228 strain, was constructed. CK287 bacterial cells did not show rapid killing during culture, and the ability to produce biofilm was also decreased, and toxicity was weakened in cytotoxicity test and fish test. These results suggest that the TA system is involved in physiological regulation and expression of virulence factors in S. iniae FP5228.

• Journal of Life Science
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• v.31 no.5
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• pp.496-501
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• 2021

In this study, the marine agar-degrading bacterium Pseudoalteromonas sp. JH-1 was isolated, and its growth and agarase properties were investigated. Seawater was collected from the offshore of the Yonggung Temple in Busan, and agar-degrading bacteria were isolated and cultured with marine agar medium. The bacterium Pseudoalteromonas sp. JH-1 was isolated through 16S rRNA gene sequencing. The extracellularly secreted enzyme was obtained from the culture broth of Pseudoalteromonas sp. JH-1 and was used to characterize its agarase. The extracellular agarase exhibited a maximum activity of 116.6 U/l at 50℃ and pH 6.0 of 20 mM Tris-HCl buffer. Relative activities were 31, 59, 94, 100, 45, and 31% at 20, 30, 40, 50, 60, and 70℃, respectively. Relative activities were 49, 85, 100, 86, 81, and 67% at pH 4, 5, 6, 7, 8, and 9, respectively. Residual activity was more than 85% after exposure at 20, 30, and 40℃ for 2 hr, and more than 82% after exposure at 50℃ for 2 hr. Zymogram analysis confirmed that Pseudoalteromonas sp. JH-1 produced at least two agarases of 55 and 97 kDa. As the products of α-agarase and β-agarase have antioxidation, antitumor, skin-whitening, macrophage activation, and prebiotic effects, further studies are needed on the agarase of Pseudoalteromonas sp. JH-1.

• Journal of Life Science
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• v.32 no.4
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• pp.285-289
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• 2022

In this study, the growth characteristics of an agar-degrading bacterium isolated from seawater samples collected from Yeongheungdo, Incheon, and the characteristics of its agarase were analyzed. The 16S rRNA gene sequence of the isolated strain was 95% similar to that of the genus Agarivorans, and thus the isolated strain was named Agarivorans sp. HY-1. When Agarivorans sp. HY-1 was cultured in a marine broth 2216 medium at 27℃ and 250 rpm, it showed maximum growth on day 1 and showed maximum enzymatic activity on day 2. A crude enzyme solution was prepared from secreted agarase in the culture medium. The extracellular agarase of the Agarivorans sp. HY-1 strain showed maximal activity at 40℃ and pH 7.0 (20 mM Tris-HCl) with 591.91 U/l. The agarase exhibited relative activities of 64, 91, 100, 97, 89, and 60% at 20, 30, 40, 50, 60, and 70℃, respectively. At pH 5, 6, 7, and 8, the relative activities were 79, 95, 100, and 55%, respectively. Furthermore, the agarase exhibited >86% residual activity at 20, 30, and 40℃ for 2 hr and >44% residual activity at 50℃ after 2 hr. A TLC analysis confirmed that Agarivorans sp. HY-1 produced α-agarase. As the degradation products of α-agarase have anticancer and antioxidant effects, Agarivorans sp. HY-1 and its agarase may well prove useful.