• Journal of Nutrition and Health
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• v.42 no.6
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• pp.505-515
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• 2009

Collagen is the major matrix protein in dermis and consists of proline and lysine, which are hydroxylated by prolyl hydroxylase (PH) and lysyl hydroxylase (LH) with cofactors such as vitamin C, oxygen, iron (Fe$^{2+}$), ketoglutarate and silicon. The collagen degradation is regulated by matrix metalloproteinase-1 (MMP-1), of which is the major collagen-degrading proteinase whereas tissue inhibitors of metalloproteinase-1 (TIMP-1) bind to MMP-1 thereby inhibiting MMP-1 activity. In this study, we investigated the effects of vitamin C, silicon and iron on mRNA, protein expressions of PH, LH, MMP-1 and TIMP-1. The physiological concentrations of vitamin C (0-100 $\mu$M), silicon (0-50 $\mu$M) and iron (Fe$^{2+}$：0-50 $\mu$M) were treated to human dermal fibroblast cells (HS27 cells) for 3 or 5days. The expression level of mRNA and protein was increased in not only PH but also LH when cells were incubated with vitamin C. A similar increase in LH mRNA or protein expression occurred when cells were incubated with silicon. Our results suggest that treatment of vitamin C and silicon increased mRNA and protein expression of PH and LH in human dermal fibroblast.

• Asian Pacific Journal of Cancer Prevention
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• v.16 no.10
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• pp.4369-4376
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• 2015

Background: To investigate in-vitro antagonistic effect of low-dose liquiritigenin on gemcitabine-induced capillary leak syndrome (CLS) in pancreatic adenocarcinoma via inhibiting reactive oxygen species (ROS)-mediated signalling pathways. Materials and Methods: Human pancreatic adenocarcinoma Panc-1 cells and human umbilical vein endothelial cells (HUVECs) were pre-treated using low-dose liquiritigenin for 24 h, then added into gemcitabine and incubated for 48 h. Cell viability, apoptosis rate and ROS levels of Panc-1 cells and HUVECs were respectively detected through methylthiazolyldiphenyl-tetrazoliumbromide (MTT) and flow cytometry. For HUVECs, transendothelial electrical resistance (TEER) and transcellular and paracellular leak were measured using transwell assays, then poly (ADP-ribose) polymerase 1 (PARP-1) and metal matrix proteinase-9 (MMP9) activity were assayed via kits, mRNA expressions of p53 and Rac-1 were determined through quantitative polymerase chain reaction (qPCR); The expressions of intercellular adhesion molecule 1 (ICAM-1), vascular cell adhesion molecule-1 (VCAM-1) and PARP-1 were measured via western blotting. Results: Low-dose liquiritigenin exerted no effect on gemcitabine-induced changes of cell viability, apoptosis rate and ROS levels in Panc-1 cells, but for HUVECs, liquiritigenin ($3{\mu}M$) could remarkably elevate gemcitabine-induced decrease of cell viability, transepithelial electrical resistance (TEER), pro-MMP9 level and expression of ICAM-1 and VCAM-1 (p<0.01). Meanwhile, it could also significantly decrease gemcitabine-induced increase of transcellular and paracellular leak, ROS level, PARP-1 activity, Act-MMP9 level, mRNA expressions of p53 and Rac-1, expression of PARP-1 and apoptosis rate (p<0.01). Conclusions: Low-dose liquiritigenin exerts an antagonistic effect on gemcitabine-induced leak across HUVECs via inhibiting ROS-mediated signalling pathways, but without affecting gemcitabine-induced Panc-1 cell apoptosis. Therefore, low-dose liquiritigenin might be beneficial to prevent the occurrence of gemcitabine-induced CLS in pancreatic adenocarcinoma.

• Microbiology and Biotechnology Letters
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• v.42 no.4
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• pp.393-401
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• 2014

A bacterium producing antimicrobial substance was isolated from cheonggukjang. The bacterium was identified as a strain of Bacillus subtilis by 16S rDNA sequencing and designated as Bacillus subtilis HH28. The antimicrobial substance produced from Bacillus subtilis HH28 was purified by 0-80% ammonium sulfate precipitation, DEAE-sepharose FF column chromatography, and Sephacryl S-200 HR gel chromatography. The molecular weight of the purified antimicrobial substance was estimated to be approximately 3,500 Da using Tricine sodium dodecyl sulfate-polyacrylamide gel electrophoresis and direct detection analysis. Antimicrobial substance from B. subtilis HH28 not only inhibited B. cereus, but also Listeria monocytogenes and Vibrio parahaemolyticus. The purified antimicrobial substance was stable at $40-80^{\circ}C$, and between pH 2 and 8. Antimicrobial activity of the purified substance was completely destroyed by treatment of protease, proteinase K, and pronase E, indicating that it is proteinaceous.

• Microbiology and Biotechnology Letters
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• v.38 no.1
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• pp.84-92
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• 2010

Antibacterial compound from Bacillus subtilis MJP1 was purified using C18 Sep-Pak cartridge, ion exchange chromatography, and gel filtration chromatography. The purified antibacterial compound showed antibacterial activity against Listeria monocytogenes, Bacillus subtilis, Staphylococcus aureus subsp. aureus, and Enterococcus faecalis. The purified antibacterial compound was found to be stable at $100^{\circ}C$ for 5 min and in the pH range of 3.0~9.0, but it was unstable at pH 10.0. It was inactivated by proteinase K and pronase E, and heat treatment at $121^{\circ}C$ for 15 min, but it was stable with lipase and $\alpha$-amylase treatment, which indicated its proteineous nature. Ultra performance liquid chromatography and electrospray ionization tandem mass spectrometry analysis were used to identify the purified antibacterial compound and confirmed the existence of two peptides (3356.54 Da, 3400.5244 Da).

• Korean Journal of Fisheries and Aquatic Sciences
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• v.29 no.3
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• pp.296-308
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• 1996

Proteolytic enzymes responsible for post-mortem degradation of the fish tissues have been studied in regard with screening the proteases distributed in the fish body by reacting with the specific synthesized substrates. Activities of cathepsin L, B, H, G, and D like enzymes were detected in the muscle crude protease from the both kind of fish, dark fleshed fish (anchovy, Engraulis japonica, and gizzard-shad, Clupanodo punctatus) and white fleshed fish (seabass, Lateolabrax japonicus, and sole, Pleuronichthys cornutus), however, those of chymotrypsin, trypsin, pepsin, and peptidase like enzymes were observed 3n the viscera crude pretense from the fish. Proteolytic activities of the muscle crude protease at pH 6.0 were similar to those of the viscera crude protease at pH 8.0, but, those of the viscera crude protease at pH 8.0 were about 2 times higher than those at pH 6.0. The muscle and viscera crude protease from anchovy showed the strongest proteolytic activity among the four fish crude proteases and the proteolytic activity of the viscera crude protease was approximately 100 times higher than that of the muscle crude protease, which suggest that viscera proteases were more contributed on the development of post-mortem changes than muscle proteases. With the degradation patterns on SDS-polyacrylamide gel electrophoresis against yellowtail myofibrillar proteins, the muscle and viscera crude protease of the four fishes were primary responsible for the degradation of myosin heavy chain, and myosin light chain and actin, respectively.

• Korean Journal of Food Science and Technology
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• v.48 no.2
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• pp.165-171
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• 2016

Physicochemical properties of waxy maize flours prepared by hydrothermal and enzymatic treatments were evaluated. Waxy maize flours were hydrothermally treated using heat-moisture treatment (HMT) and annealing (ANN) and enzymatically treated using commercial enzymes (cellulase, proteinase, and pectinase). The HMT-modified waxy maize flours had low water absorption index (WAI), melting enthalpy, viscosity, and crystallinity. However, ANN-modified and enzymatically modified waxy maize flours had high WAI, melting enthalpy, and viscosity. X-ray diffraction analysis of ANN-modified and enzymatically modified waxy maize flours revealed a typical A-type pattern and displayed sharper crystalline peaks than those observed for the control groups (native waxy maize flours). In contrast, the crystallinity of HMT-modified waxy maize flours were decreased by hydrothermal treatment.

• Microbiology and Biotechnology Letters
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• v.26 no.6
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• pp.523-528
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• 1998

We isolated microorganism secreting antimicrobial substance from tomato and identified as Enterococcus faecium. This substance was completely inactivated by pretense treatment and retained activity after catalase treatment. This result indicated that the antimicrobial activity of this substance was due to proteinaceous substance known as bacteriocin. The bacteriocin inhibited growth of Gram positive bacteria, such as Listeria monocytogenes, Leuconostoc mesenteroides, Lactobacillus plantarum, Streptococcus agalactiae, Streptococcus pyrogenes, and Gram negative bacteria, such as Pseudomonas aeruginosa. Purification of the bacteriocin was achieved by ethanol precipitation, ion exchange chromatography on CM Sepharose CL-6B, and gel filtration on Sephacryl S-100 HR. After these purification steps, the specific activity of the bacteriocin was increased 35.8 fold compared with culture broth. Purified bacteriocin was shown single band on SDS-PAGE and molecular weight was estimated 51 kDa. The residual activity of this bacteriocin was 3.3% at 10$0^{\circ}C$ for 60 min, and this bacteriocin was stable at pH 2~7.

• Food Science and Preservation
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• v.19 no.5
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• pp.757-766
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• 2012

The SSH100-10 bacterial strain, which exhibits strong antifungal (anti-mold and anti-yeast) activity, was isolated from traditional korean soysauce aged 100 years. The strain was identified as Bacillus velezensis based on Gram-staining, the biochemical properties and 16S rRNA gene sequence determination. B. velezensis SSH100-10 showed strong proteinase activity and NaCl tolerance, but did not produce enterotoxin. Two-antifungal compounds from B. velezensis SSH100-10 were purified using SPE, preparative HPLC, and reverse phase-HPLC. The purified antifungal compounds were identified as $C_{14}$ and $C_{15}$ iturin through MALDI-TOF-MS and amino acid composition analysis. The stability characteristics of the antifungal compounds after temperature, pH, and enzyme treatments suggested that B. velezensis SSH100-10 produced more than two antifungal compounds; pH-stable $C_{14}$ iturin A and $C_{15}$ iturin A, and unidentified pH-unstable compounds. The results suggested that B. velezensis SSH100-10 can be used in soybean fermentation as a starter. Moreover it has potential as a biopreservative in the food and feed industry and as a biocontrol agent in the field of agriculture.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.32 no.2
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• pp.127-133
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• 1999

The purpose of this study was to determine the optimum hydrolysis conditions for the production of enzymatic hydrolysate from tuna boiled extract (TBE) using membrane (molecular weight cut off 10,000Da) reator. The tuna pyloric caeca crude enzyme (TPCCE) was identified as the most suitable enzymes for the hydrolysis of TBE. The optimum hydrolysis conditions of TBE in the batch reactor were $40^{\circ}C$, pH 9 and substrate to TPCCE ratio 50 (w/w). For 6hr under the above conditions, $70\%$ of the total amount of initial TBE was hydrolysed. The optimum hydrolysis conditions of TBE in the membrane reactor were $40^{\circ}C$, pH 9, enzyme 0,1 g/$\ell$, volume 1$\ell$ and substrate to enzyme ratio 100(w/w). The degree of hydrolysis of TBE was above $60\%$ for 3 hr. The TBE hydrolysate were prepared with $5\%$ TBE solution under the optimum hydrolytic conditions in the membrane reactor

• Microbiology and Biotechnology Letters
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• v.35 no.4
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• pp.325-333
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• 2007

One bacterium with high proteinase production and spore-forming ability was isolated from korean traditional soybean paste(doenjang). The isolated strain was identified as Bacillus subtilis, based on gram-staining, biochemical properties and l6S rRNA gene sequencing, and designated as B. subtilis DJI. Its growth rate was very fast, and it reached its stationary phase within 9 h, and then started to form spores. Bacterial-koji and doenjang were prepared using B. subtilis DJI. Chemical components of the doenjang were determined after 2 months of aging period: amino nitrogen 507 mg%, crude protein 14.3%, crude fat 4.8% and water 54.9%. The composition of total and free amino acids and their ratios of doenjang were changed during the aging period. Among total amino acids in DJI doenjang, glutamic acid, aspartic acid, leucine and arginine were the major amino acids. The fibrinolytic activities of DJI doenjang and traditional doenjangs were 909.7 units/ml and $363.3{\sim}618.6\;units/ml$, respectively. Flavor compounds of DJI doenjang and traditional doenjang were extracted by SDE(simultaneous steam distillation and extraction), and analyzed by GC/MS; DJI doenjang possessed the typically favorable flavor compounds in traditional korean doenjang, with reduced off-flavor compounds.