• Journal of Applied Biological Chemistry
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• v.61 no.3
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• pp.283-290
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• 2018

In this study, we verified the 1-1-diphenyl-2-picryl-hydrazyl radical scavenge, 2,2'-azinobis(3-ethylbenzothiazoline-6-sulfonic acid) radical scavenge, elastase, tyrosinase inhibitory effect by using the solvent fractions of Maekmoondong-tang hot water extract. As a result, the ethyl acetate fraction (MW-EA) showed the highest inhibitory activity. In cell-based assays, MW-EA treatment confirmed a 34% ($100{\mu}g/mL$) efficacy in reactive oxygen species inhibitory activity, and at the same concentration, MMPs showed more than 50% inhibition and tyrosinase inhibited 25% ($50{\mu}g/mL$). Therefore Maekmoondong-tang is considered high development potential as a material to improve the skin.

• Microbiology and Biotechnology Letters
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• v.13 no.2
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• pp.157-162
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• 1985

The cell cultures and crude extracts of Bacillus sphaericus 1593 K-5 and its mutant Spo-Dl216 were respectively bioassayed against Culex pipiens var. pollens mosquito larvae. The B. sphaeriucs 1593 K-5 showed toxic activity against the larvae. LC$_{50}$ values (cells/$m\ell$) was 2.6$\times$10$^2$. Also the LC$_{50}$ ($\mu\textrm{g}$ Protein/$m\ell$) of the crude extract was 10.26. However, B. sphaericus Spo-Dl216 didn't show toxic activity against the larvae. The soluble cytoplasmic toxin in broken B. sphaeriucs 1593k-5 cells was partially purified by gel permeation chromatography and ion exchange chromatography. Among the fractions of the gel permeation chromatography only a single fraction was found to be toxic. LC$_{50}$ values ($\mu\textrm{g}$ protein/$m\ell$) of the active fraction was 0.182. The active fraction of the gel permeation was subjected to ion exchange chromatography. Only a single fraction showed toxic activity and its LC$_{50}$ values ($\mu\textrm{g}$ protein/$m\ell$) was 0.02..02.

• The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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• v.20 no.1 s.32
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• pp.1-15
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• 2007

Objectives : RVC has long been used for a useful natural agent ameliorating inflammation related symptoms in the folk medicine recipe. This study was performed to investigate effects of RVC on the inflammation and oxidation in RAW 264.7 cells. Methods : The RVC was extracted with 80% ethanol and sequentially partitioned with solvents in order to increase polarity. With the various fractions, we determined the activities on the inflammation and oxidation in RAW 264.7 cells. Results : 1. Among the various solvent extracts of RVC, the butanol fraction showed the most powerful inhibitory ability against nitric oxide (NO) production in lipopolysaccharide (LPS)-induced RAW 264.7 cells without affecting cell viability. 2. Butanol fraction showed a oxidation inhibition effect by decreasing the DPPH and OH radicals. 3. Butanol fraction exhibited the inhibitory avilities against iNOS and COX-2. 4. Reverse transcriptase polymerase chain reaction (RT-PCR) and Westem blotting analysis revealed that the BuOH fraction provided a primary inhibitor of the iNOS protein and mRNA expression in LPS-induced RAW 264.7 cells. Among the up-regulater molecules of iNOS and COX-2, the BuOh fraction of RVC was shown the inhibitory activity of phoshporylation of c-Jun N-terminal kinase (JNK) 1/2 and threonine protein kinase (AKT), the one of the MAPKs pathway. Conclusion : Thus, the present study suggests that the response of a component of the BuOH fraction to NO generation via iNOS expression provide a important clue to elucidate anti-inflammatory and anti-oxidation mechanism of RVC.

• Korean Journal of Plant Resources
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• v.21 no.4
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• pp.265-269
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• 2008

The antioxidant activities of the extracts of butterbur (Petasites japonicus) derived from different extraction methods were investigated. SOD (superoxide dismutase)-like activity differed according to the extraction solvents, showing a greater antioxidant effect with ethanol solvent than that of water. Ethanol extracts of butterbur leaves showed higher SOD-like activity of 96.7% than those of water extracts. The contents of polyphenolic compounds were higher in water extracts than those in ethanol extracts. The highest content of polyphenolic compounds was 223mg/g dry weight for butterbur leaves. EDA of butterbur roots was 61.5% in the water extract and EDA in butterbur leaves was 34.9% in the ethanol extract. The anticancer effects with the extracts of butterbur were experimented by fractionations with different solvents. 41.9% of the growth of stomach cancer cells, SNU-719, were inhibited and also 72.7% of the growth of liver cancer cells, Hep3B, were inhibited by the butanol fractions of butterbur, while not affecting the growth of normal cell, DC 2.4.

• Journal of Embryo Transfer
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• v.24 no.4
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• pp.259-263
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• 2009

Salivary lipocalin (SAL1) is a member of the lipocalin protein family that has a property to associate with many lipophilic molecules and was identified as pheromone-binding protein in pigs. Our previous study has shown that SAL1 is expressed in the uterine endometrium in a cell type- and implantation stage-specific manner and secreted into the uterine lumen in pigs. However, function of SAL1 in the uterus during pregnancy in pigs is still not known. To understand physiological function of SAL1 in the uterine endometrium during pregnancy in pigs, it needs to elucidate the ligand(s) for SAL1. Thus, to identify the ligand for SAL1 in the porcine uterus, we collected uterine luminal fluid from pigs on day 12 of pregnancy by flushing with PBS. Proteins from the uterine luminal fluid were separated by ion exchange chromatography and gel filtration. Fractions containing SAL1 protein were pooled and concentrated. Immunoblot analysis confirmed successful purification of SAL1. Then, we extracted lipids from the purified SAL1 protein and analyzed the lipids by liquid chromatography-mass spectrometry, and predicted to be steroid hormones and prostaglandins as SAL1 ligands. Results in this study showed that SAL1 protein in the uterine secretions has a small lipophilic molecule as a natural ligand. Further characterization of ligand extracted from purified SAL1 will be useful for understanding physiological function of SAL1 during pregnancy and its application to increase the pregnancy rate in pigs.

• Korean Journal of Animal Reproduction
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• v.26 no.2
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• pp.165-171
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• 2002

Follicular fluid (FF) contains an oocyte maturation inhibitor with unknown chemical properties. This study was carried out to chemically define the factor(s) inhibiting cumulus cell denudation (CD) and first polar body formation (PBF). Porcine FF (PFF) was extracted with methanol and the extract was serially separated using gel filtration on Superose 12 and Superdex columns. A Superdex fraction was derived with PITC and analyzed with an amino acid analysis column. The results obtained are as follows; PFF had an activity inhibiting both CD and PBF of porcine primary oocytes. Superdex fractions RV2.11 prepared from PFF exhibited an activity inhibiting CD and PBF. By amino acid analysis, the fraction RV2.11 appeared to be proline having the same activity inhibiting CD and PBF. In conclusion, PFF had oocyte maturation inhibitors, of which proline should inhibit CD and PBF.

• The Plant Pathology Journal
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• v.21 no.4
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• pp.301-309
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• 2005

Trichoderma species/isolates exhibited varied degree of agglutination on sclerotial (Sc) and hyphal (Hy) surface of Macrophomina phaseolina. The agglutination efficiencies on Sc and Hy ranged from $11\;to\;57\%$. Isolates of T. harzianum (Th) and T. viride (Tv) showed greater agglutination on Sc ($23-57\%$) and Hy ($16-47\%$). Different enzymes (trypsin, pepsin, proteinase k, a-chymotrypsin, lyticase and glucosidase) and inhibitors (tunicamycin, cycloheximide, brefeldin A, sodium azide, dithiothreitol and SDS) reduced the agglutination potential of conidia of Th-23/98 and Tv-25/98; however, the extent of response varied greatly in different treatments. Different fractions of Th-23/98 and Tv-25/98 exhibited haemagglutinating reaction with human blood group A, B, AB and O. Haemagglutinating activity was inhibited by different sugars and glycoproteins tested. Crude haemagglutinating protein from outer cell wall protein fraction of Th-23/98 and Tv-25/98 were eluted on Sephadex G-100 column. Initially Th-23/98 and Tv-25/98 exhibited two peaks showing no agglutination activity; however, lectin activity was detected in the third peak. Similar to crude lectin, the purified lectin also exhibited haemagglutinating activity with different erythrocyte source. SDS-PAGE analysis of partially purified lectin revealed single band with an estimated molecular mass of 55 and 52 kDa in Th-23/98 and Tv-25/98, respectively. Trypsin, chymotrypsin and b-1,3-glucanase totally inhibited lectin activity. Similarly, various pH also affected the haemagglutinating activity of Th-23/98 and Tv-25/98. From the present observations, it can be concluded that the recognition/attachment of mycoparasite (T. harzianum and T. viride) to the host surface (M. phaseolina) may be most likely due to lectin-carbohydrate interaction.

• Korean Journal of Food Science and Technology
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• v.33 no.5
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• pp.560-566
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• 2001

This study was conducted to investigate the effects of solvent extracts from medicinal plants on the fermentation of kimchi. Five microorganisms related to kimchi fermentation were selected and the antimicrobial activities of solvent fractions from medicinal plants were investigated. The chloroform fraction from the methanol extract of dandelion (Taraxacum platycarpum D.) exhibited inhibitory activity against five strains such as Lactobacilli plantarum, Lactobacilli brevis, Leuconostoc mesenteroides, Enterococcus faecalis and Saccharomyces cerevisiae. The chloroform fraction from the methanol extract of Dandelion inhibited the growth of E. faecalis and Leu. mesenteroides at the concentration of 80 mg/mL. Scanning electron micrographs of Leu. mesenteroides and E. faecalis treated with chloroform fraction 80 mg/mL exhibited morphological changes, including irregularly contracted cell surface.

• Microbiology and Biotechnology Letters
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• v.47 no.3
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• pp.449-458
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• 2019

Hepatitis B treatments using immune therapy are gaining interest because of the improvements in dendritic cell performance for antigen presentation, which induces an appropriate immune response and raises patient survival rates. This research aims to produce a significant amount of the HBcAg antigen, which can induce an immune response and have a curative effect on HBV infection. In this study, the HBV subgenotype B3 of the HBcAg gene was used, which is dominant in Indonesia. Further, Lactococcus lactis bacteria was used as the host because of its safety and tightly regulated protein expression. The codon usage for the HBcAg gene was optimized to improve protein expression in L. lactis, which is important because a codon is not random between species. The HBcAg gene is attached to a pNZ8148 plasmid and transformed into the L. lactis NZ3900 expression host. The results confirm that a positive protein band (21 kDa) in two fractions of purified HBcAg was recognized by both western blotting and dot blot hybridization, even if the HBcAg optimized codon has higher GC contents than that suggested for L. lactis expression. Overall, this research strengthens the broad use of L. lactis bacteria for any protein expression, including higher protein expression of codon optimized HBcAg gene compared to non-optimized genes. Furthermore, the improvement in the codon optimization of the HBcAg gene significantly increases the total protein expression by 10-20%, and the expression level of the codon optimized HBcAg increases 1.5 to 3.2-times that of the native HBcAg.

• Food Science of Animal Resources
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• v.41 no.4
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• pp.687-700
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• 2021

This study performed to evaluate the applicability of functional dairy food materials by comparing the calcium solubilization ability and anti-inflammatory effects of hydrolyzed casein protein. Commercial enzyme (Alcalase^®; Neutrase^®; Protamex^®; Flavourzyme^®) was added to the 10% casein solution to prepare the casein hydrolysates. Samples obtained every hour [1:200 (w/v)]. According to results of measuring the degree of hydrolysis (DH), all of four enzymatic hydrolysates increased rapidly from 30 to 40 min, and after 150 min, there were no change. Protamex^® and Neutrase^® had the highest DH compared to others enzymatic hydrolysates. After that, peptides obtained throughout a preparative liquid chromatography system. In the calcium solubility experiments, neutrase fraction (NF) 4 and NF7 showed similar activities with casein phosphopeptide (CPP). In vitro cell experiments showed that no cytotoxicity except for NF6. Also, the production of nitric oxide (NO) inhibited as the concentration of fraction samples increased. The cytokine (IL-1α, IL-6, and TNF-α) production was lower than lipopolysaccharide (+) group significantly. Therefore, the possibility of anti-inflammatory activity found in the hydrolyzed samples. According to the above experiments, NF3 and Protamex Fraction (PF) 3 selected. Amino acids selected throughout an AccQ-Tag system. As a result, 17 species of amino acids and several species of unknown amino acids identified. Both fractions had the highest content of phenylalanine. This study identified the potential of biologically active and functional peptides derived from casein that affect the food and dairy industry.