• 한국축산식품학회지
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• 제35권1호
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• pp.58-70
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• 2015

This study was conducted to examine the effect of egg shell membrane hydrolysates (ESMH) on wrinkle, UV, and moisture protection for cosmetic use. ESMH were fragmented as whole ESMH (before fractioning), Fraction I (> 10 kDa), Fraction II (3-10 kDa), and Fraction III (< 3 kDa). In order to test whether fractionated ESMH can be used for functional cosmetic materials, we examined not only the level of hyaluronic acid and collagen production, but also the MMP-1 activity using a HaCaT and CCD-986Sk cell line. Our study treated each sample of fractionated ESMH with different concentrations (0.01, 0.1, 1 mg/mL). In our in vivo research, we used hairless mice that had been exposed to UV-B to induce wrinkles for 7 wk, then applied Fraction I to the treatment group for 5 wk and then tested skin thickness, minimum erythema dose and moisture content. In addition, Fraction I was high in collagen and HA biosynthesis and it was better than TGF-${\beta}$ in improving of the skin. When TNF-${\alpha}$ caused MMP-1 activity in the CCD-986Sk cells, the whole ESMH and Fraction I proved to be effective in hindering the induction of collagenase depending on the concentration, and also showed outstanding effects in the suppression of skin aging. We found that the treatment group mice's UV-B radiation-induced skin damage was largely mitigated compared to that of the non-treatment group mice. Thus, we have concluded that EMSH helps to mitigate UV-B radiation-induced wrinkles, collagen, HA, MMP-1 activity and can be used for functional cosmetic materials.

• Molecules and Cells
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• 제21권3호
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• pp.418-427
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• 2006

Ionotropic glutamate receptors (iGluRs) are ligand-gated nonselective cation channels that mediate fast excitatory neurotransmission. Although homologues of the iGluRs have been identified in higher plants, their roles are largely unknown. In this work we isolated a full-length cDNA clone (RsGluR) encoding a putative glutamate receptor from small radish. An RsGluR:mGFP fusion protein was localized to the plasma membrane. In Arabidopsis thaliana overexpressing the fulllength cDNA, glutamate treatment triggered greater $Ca^{2+}$ influx in the root cells of transgenic seedlings than in those of the wild type. Transgenic plants exhibited multiple morphological changes such as necrosis at their tips and the margins of developing leaves, dwarf stature with multiple secondary inflorescences, and retarded growth, as previously observed in transgenic Arabidopsis overexpressing AtGluR3.2 [Kim et al. (2001)]. Microarray analysis showed that jasmonic acid (JA)-responsive genes including defensins and JA-biosynthetic genes were up-regulated. RsGluR overexpression also inhibited growth of a necrotic fungal pathogen Botrytis cinerea possibly due to up-regulation of the defensins. Based on these results, we suggest that RsGluR is a glutamate-gated $Ca^{2+}$ channel located in the plasma membrane of higher plants and plays a direct or indirect role in defense against pathogen infection by triggering JA biosynthesis.

• Journal of Microbiology and Biotechnology
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• 제27권2호
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• pp.306-315
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• 2017

Metabolic engineering with a high-yielding mutant, A. terreus AN37, was performed to enhance the production of itaconic acid (IA). Reportedly, the gene cluster for IA biosynthesis is composed of four genes: reg (regulator), mtt (mitochondrial transporter), cad (cis-aconitate decarboxylase), and mfs (membrane transporter). By overexpressing each gene of the IA gene cluster in A. terreus AN37 transformed by the restriction enzyme-mediated integration method, several transformants showing high productivity of IA were successfully obtained. One of the AN37/cad transformants could produce a very high amount of IA (75 g/l) in shake-flask cultivations, showing an average of 5% higher IA titer compared with the high-yielding control strain. Notably, in the case of the mfs transformants, a maximal increase of 18.3% in IA production was observed relative to the control strain under the identical fermentation conditions. Meanwhile, the overexpression of reg and mtt genes showed no significant improvements in IA production. In summary, the overexpressed cis-aconitate decarboxylase (CAD) and putative membrane transporter (MFS) appeared to have positive influences on the enhanced IA productivity of the respective transformant. The maximal increases of 13.6~18.3% in IA productivity of the transformed strains should be noted, since the parallel mother strain used in this study is indeed a very high-performance mutant that has been obtained through intensive rational screening programs in our laboratory.

• 대한의생명과학회지
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• 제11권4호
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• pp.473-479
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• 2005

Possible mechanism of decreased catechol-O-methyltransferase (COMT) activity in cholestatic rat liver was studied. Hepatic and serum COMT activities were determined from the experimental rats with common bile duct ligation (CBDL). The Michaelis-Menten constants in this hepatic enzyme were also measured. The activities of cytosolic, mitochondrial and mircosomal COMT as well as their Vmax values were found to be decreased significantly in CBDL plus taurocholic acid (TCA) injected group than in the control group, such as CBDL alone groups. However, their Km values in the experimental groups did not vary. Serum COMT activity increased slightly in the CBDL plus TCA injected group than in the control group. The above results suggest that TCA represses biosynthesis of the COMT in the liver. The elevated activity of the serum COMT is believed to be caused by the increment of membrane permeability of hepatocytes upon TCA mediated liver cell necrosis.

• 대한의생명과학회지
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• 제16권1호
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• pp.65-67
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• 2010

The endoplasmic reticulum (ER) is a multifunctional intercellular organelle in which several posttranslational modification steps occurred such as protein folding, lipid biosynthesis, calcium storage and release. Perturbations that disrupt ER homeostasis lead to the misfolding of proteins in the ER lumen and up-regulation of ER signaling pathway called the unfolded protein response (UPR). Here, we have demonstrated that ageing changes the expression of ER chaperone and associated ER membrane kinases of IRE1, ATF6 and PERK.

• 대한의생명과학회지
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• 제10권1호
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• pp.43-48
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• 2004

Changes of thiol methyltransferase (TMT) activity in cholestatic rat liver were studied. Hepatic subcellular and serum TMT activities were determined in cholestatic rat induced by common bile duct (CBD) ligation over a period 28 days. The mitochondrial and microsomal TMT activities in cholestatic rat liver were found to be significantly increased between the 1st and the 28th day after CBD ligation. The TMT activity in serum was significantly increased throughout the experiments. The Vmax values of the above hepatic TMT in cholestatic rat were significantly increased at the 7th day after CBD ligation. However, the Km values of the above hepatic enzymes did not vary in all the experimental groups. Therefore, the results indicate that the biosynthesis of TMT was increased in cholestatic rat liver. The elevated serum TMT activity is most likely caused by increased hepatocytes membrane permeability due to cholestasis mediated liver cell necrosis.

• 대한의생명과학회지
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• 제10권4호
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• pp.421-427
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• 2004

The possible mechanisms of decreased monoamine oxidase (MAO) A and B activities in cholestatic rat liver were studied. Hepatic and serum MAG activities were determined from the experimental rats with common bile duct ligation (CBDL). The Michaelis-Menten constants in these hepatic enzymes were also measured. The activities of mitochondrial MAO A and B, and mircosomal MAO B as well as their Vmax values were found to be decreased significantly in CBDL plus taurocholic acid (TCA) injected group than in the control group, such as CBDL alone groups. However, their Km values in the experimental groups did not vary. Serum MAO activity increased significantly in the CBDL plus TCA injected group than in the control group. The above results suggest that TCA represses biosynthesis of the MAO in the liver. The elevated activity of the serum MAO is believed to be caused by the increment of membrane permeability ofhepatocytes upon TCA mediated liver cell necrosis.

• 한국전자현미경학회:학술대회논문집
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• 한국현미경학회 2007년도 제38차 춘계학술대회 초록집
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• pp.75-78
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• 2007

The localization of cellulase were investigated in the capitate glandular trichomes of Pelargonium ${\times}$ fragransby a transmission electron microscopy. The secretory cells of capitate trichomes involved in biosynthesis and its secretion. Secretory material is transported to the space between the plasma membrane and cell wall, and subsequently accumulated in the secretory cavity. The splitting of secretory cell wall during the formation of secretory cavity is suggested that wall-forming enzymes, such as cellulase, may contribute to the wall separation process. Cellulase reaction product was localized in the secretory cell, the secretory cavity and in the subcuticular wall of glandular trichomes. Reaction products were present over fibrillar matrix in the secretory cavity associated with both the inner wall and the subcuticular wall.

• Journal of Microbiology and Biotechnology
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• 제10권2호
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• pp.251-257
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• 2000

Chitin synthases are identified as key enzymes of chitin biosynthesis in most of the fungi. Among them, chitin synthase II has been reported to be and essential enzyme in chitin biosynthesis, and exists as a membrane-bound form. To search and screen new antifungal agents from natural resources to inhibit chitin synthase II, the assay conditions were established using the enzyme isolated from Saccharomyces cerevisiae ECY38-38A(pAS6) that overproduces only chitin synthase II. This enzyme was activated only by partial proteolysis with trypsin. Its actibity reached the maximum at $80{\;}\mu\textrm{g}/ml$ of trypsin and was strongly stimulated by 2.0 mM $Co^{2+}$, 1.0 nM UDP-[$^{14}C$]-GicNAc, and 32 mM free-GlcNAc. Under these assay conditions, the highest chitin synthase II activity was observed by incubation at $30^{\circ}C$ for 90 min. However, and extremely narrow range of organic solvents up to as much as 25% of DMSO and 25% of MeOH was useful for determining optimal assay conditions. After a search or potent inhibitors of chitin synthase II from natural resources, prodigiosin was isolated from Serratia marcescens and purified by solvent extration and silica gel column chromatographies. The structure of prodigiosin was determined by UV, IR, Mass spectral, and NMR spectral analyses. Its molecular weight and formula were found to be 323 and $C_{20}H_{25}N_{3}O$, respectively. Prodigiosin ingibited chitin synthase II by 50% at the concentration of $115{\;}\mu\textrm{g}/ml$.

• KSBB Journal
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• 제31권4호
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• pp.228-236
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• 2016

The aim of this study was to characterize the hemolytic Aeromonas sp. MH-8 exposed to green tea catechin, epigallocatechin gallate (EGCG). Initially, the hemolytic Aeromonas sp. MH-8 was enriched and isolated from stale fish. Bactericidal effects of MH-8 exposed to EGCG ranging from 1 mg/mL to 4 mg/mL were monitored, and complete bactericidal effects were achieved within 3 h at 3 mg/mL and higher concentrations. SDS-PAGE with silver staining revealed that the amount of lipopolysaccharides increased or decreased in the strain MH-8 treated to different concentrations and exposing periods of EGCG in exponentially growing cultures. The stress shock proteins (70-kDa DnaK and 60-kDa GroEL), which might contribute to enhancing the cellular resistance to the cytotoxic effect of EGCG, were induced at different concentrations of EGCG exposed to cell culture of MH-8. Scanning electron microscopic analysis demonstrated the presence of irregular rod shapes with umbilicated surfaces for cells treated with EGCG. 2-DE of soluble protein fractions from MH-8 cultures showed 18 protein spots changed by EGCG exposure. These proteins involved in chaperons (e.g., DnaK, GroEL and trigger factor), enterotoxins (e.g., aerolysin and phospholipase C precursor), LPS synthesis (e.g., LPS biosynthesis protein and outer membrane protein A precursor), and various biosynthesis and energy metabolism were identified by peptide mass fingerprinting using MALDI-TOF. In consequence, EGCG was found to have substantial antibacterial effects against food-poisoning causing bacterium, hemolytic Aeromonas sp. MH-8. Also the results provide clues for understanding the mechanism of EGCG-induced stress and cytotoxicity on Aeromonas sp. MH-8.