• Korean Journal of Fisheries and Aquatic Sciences
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• v.55 no.6
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• pp.875-885
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• 2022

This study was performed to confirm the optimal extraction method for antimicrobial peptides from the Hard-shelled mussel. Extractions were performed with two processes including 1% HAc/boiling and 1% HAc/non-boiling methods and used extracts for the comparison of the antimicrobial activity, protease stability, action mechanism, AU-PAGE (acid-urea PAGE), and HPLC chromatograms. 1% HAc/boiling extract showed potent antibacterial activities both against Gram-positive and negative bacterium but 1% HAc/non-boiling extract showed antibacterial activity only against Gram-positive bacteria. Treatment of 1% HAc/boiling extract with proteases retained almost antibacterial activity against B. subtilis, but abolished significant antibacterial activity against E. coli D31. Only 1% HAc/boiling extract showed two discrete clearing antibacterial zones including slow migrating and rapid migrating zones. Both extracts showed strong DNA-binding ability but did not show bacterial membrane permeabilizing ability. In comparison of the chromatogram obtained from C₁₈ or cation-exchange HPLC, the eluted peaks from 1% HAc/boiling extract showed high hydrophobic property or absorbance compared to 1% HAc/non-boiling extract, respectively. The concentration of the purified antimicrobial peptide was also higher in 1% HAc/boiling extract than in 1% HAc/non-boiling extract. Our results suggest that the effective extraction condition for antimicrobial peptides from marine invertebrate is boiling process in a weak acetic acid solution (1%).

• Korean Journal of Fisheries and Aquatic Sciences
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• v.56 no.1
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• pp.89-98
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• 2023

We evaluated the effects of prebiotic (mannan oligosaccharides, Mos) and probiotic diet supplements on growth performance, innate immunity, antioxidant activity, and intestinal changes in the microbial flora of red seabream Pagrus major. A basal diet (Con) was formulated to meet the nutrient requirement of red seabream. The dietary starch in Con was replaced with 0.6% Mos, Lactobacillus plantarum, Bacillus subtilis, B. licheniformis and probiotic mixture (labeled as Mos, Pro-LP, Pro-BS, Pro-BL and Pro-Mix, respectively). We stocked 450 fish in 18 polypropylene tanks (400 L) in triplicate groups per dietary treatment. The fish were fed one of the diets twice (08:30, 18:30 h) a day for 63 days. Lysozyme activity was significantly higher in all the supplemented groups than that of the Con group. The immunoglobulin level of Pro-Mix, anti-protease activity of Pro-BL, and glutathione peroxidase and superoxide dismutase activity of Pro-BS, Pro-BL and Pro-Mix groups were significantly higher than those of the Con group. The ratio of total Vibrio/heterotrophic marine bacteria counts was significantly lower in Pro-LP, Pro-BL and Pro-Mix groups than that of the Con group. Therefore, dietary supplementation of Mos and probiotics to improves immune response and antioxidant enzyme activity and inhibits Vibrio bacteria in the intestine.

• Microbiology and Biotechnology Letters
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• v.37 no.2
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• pp.160-169
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• 2009

Poly-$\gamma$-glutamic acid ($\gamma$-PGA) was mixed natural flora of Bacillus subtilis, contaminated from cooked soybeans. Also, it was performed to find out the antiallergic activity by using NC/Nga mice, in vitro. The $\gamma$-PGA (PGA-HM : PGA-high molecular weight), Molecular weight 300 kDa, was decomposed and made PGA-LM (PGA-low molecular weight) which has molecular weight below 30 kDa by sonication. Therefore, it was same result between PGA-HM and PGA-LM, and reported PGA-LM as basic result. We found that PGA-LM contains antiallergic efficacy that inhibit B cells and Th2 cells activation from isolated CD4+T cells in NC/Nga atopic dermatitis model mice, and not show a cytotoxicity in the hFCs. To investigate the effects of these PGA-LM in vitro, isolation of splenic B cell and CD4+ T cells in atopic dermatitis mice were used. To elucidate the role of PGA-LM in anti-CD40+ interleukin-4 (IL-4)-mediated B-cell activation, showed that the capacity of B cells to expression IL-$1\beta$, IL-6, and TNF-$\alpha$ mRNA down-regulated, and IL-10 mRNA up-regulation by PGA-LM treatment, but it had no effect on TGF-$\beta$ expression. In addition to CD4+IFN-$\gamma$+ and CD4+CD25+foxp3+, the functions of PGA-LM in the development of the CD4+CD25+foxp3+ and CD4+IFN-$\gamma$+cells, the phenotype and functions of PGA-LM induced CD4+CD25+foxp3+, and CD4+IFN-$\gamma$+cells in CD4+T cells. These results suggested that PGA-LM could change cytokine production and generate CD4+CD25+foxp3+ Tregs in NC/Nga mice, and may be effective for immunotherapy in patients with AD.

• Korean Journal of Medicinal Crop Science
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• v.10 no.3
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• pp.222-229
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• 2002

Ixeris dentata root were extracted with methanol and then fractionated with n-hexane, EtOAc and BuOH to get active fractions. and their antioxidant and antimicrobial activities in each fraction were determined. Ethyl acetate fraction of Ixeris dentata root showed strong antioxidant activities, but aqueous fraction did not show any activities. But in the antimicrobial test, aqueous fraction showed strong antimicrobial activities except to Escherichia coli. especially, aqueous fraction showed the strongest activities against Hypocrea nigricans. and butanol fraction showed the strongest activities against Cladosporium herbarum. This study was performed to determine the antimutagenic and cytotoxic effect of Ixeris dentata root methanol extract on Salmonella typhimurium TA98, TA100 and cancer cell lines using ames test and cytotoxicity assay, respectively. Cancer cell lines include human lung carcinoma(A549), human breast adenocarcinoma(MCF-7) and human hepatocellular carcinoma (Hep 3B). Futher fractionations with hexane, ethyl acetate, butanol and water from methanol extract of Ixeris dentata root were performed to obtain effective fraction, methanol extracts showed 79.94% inhibitory effect on the mutagenesis induced by N' -methyl- N' -nitro-N-nitrosoguanidine(MNNG) against TA100, while 89.99% inhibition was observed on the mutagenesis induced by 4-nitroquinoline-1-oxide(4NQO) against TA98. In the meanwhile, butanol fraction showed 89.92% and 71.01% inhibitory effect on the mutagenesis induced by benzo(a)pyrene(B(a)P) against TA98 and TA100, respectively. Ethyl acetate fraction showed the strongest effect against A549, MCF-7 and Hep3B at the same concentration compared to those of other fration.

• Journal of the Korean Society of Food Science and Nutrition
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• v.39 no.12
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• pp.1745-1752
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• 2010

This study was conducted to investigate the antimicrobial activity of 70% ethanol (EtOH) extract and the five fractions of the crude extract from Sasa borealis leaves against seven food poisoning bacteria, Staphylococcus aureus, Micrococus luteus, Listeria monocytogens, Bacillus subtilis, Escherichia coli, Salmonella Typhimurium, and Pseudomonas aeruginosa. The yield of 70% EtOH extract was 11.4% and those of n-hexane, chloroform, ethyl acetate, n-butanol, and aqueous fractions were 3.0%, 1.1%, 0.6%, 1.3%, and 5.1%, respectively. The 70% EtOH extract and the four fractions except aqueous fraction demonstrated antimicrobial activity against all the seven food poisoning bacteria at a concentration of 0.5%, although it was less compared to benzoic acid. Minimal inhibitory concentration (MIC) of the 70% EtOH extract against all the food poisoning bacteria except S. aureus was $50{\mu}L$/disc. Moreover, chloroform fraction was $35{\mu}L$/disc against 3 food poisoning bacteria and $50{\mu}L$/disc against the other 4 food poisoning bacteria; ethyl acetate fraction was $50{\mu}L$/disc against all the food poisoning bacteria. In addition, n-butanol fraction was $50{\mu}L$/disc against all the food poisoning bacteria except S. aureus. Aqueous fraction, which did not show antimicrobial activity at 5%, was $200{\mu}L$/disc against only S. aureus and L. monocytogen. The 0.25%, and 0.5% of ethyl acetate fraction inhibited the growth of all the food poisoning bacteria 8 to 12 hours and 24 hours, respectively. These results indicate that the Sasa borealis leaves may be useful as a natural antimicrobial substance.

• The Korean Journal of Food And Nutrition
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• v.17 no.3
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• pp.237-245
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• 2004

The physicochemical and physiological activities of domestic garlic from 3 different areas (Namhae, Jeju and Uiseong) were analyzed. The contents of moisture, ash, crude protein and crude fiber in garlic were little different in 3 kinds of area. Total sugar and water soluble phenolic compounds were higher in garlic from Namhae. The free sugars found in garlic were fructose, sucrose and lactose. Five kinds of organic acids were determined. Malonic acid and citric acid contents in garlic from Namhae were 23.7${\pm}$1.16 mg% and 22.1${\pm}$0.82 mg%, respectively. Total mineral content of garlic samples were in a range of 7112.6～9067.3 mg%, the potassium content showed the highest concentration (4117.3${\pm}$7.19～5175.3${\pm}$9.61 mg%). The electron donating abilities in 0.2% of garlic from Namhae and Uiseong showed 46.2${\pm}$1.25% and 37.0${\pm}$1.l6%, respectively. The nitrite scavenging effect was measured at different conditions (pH 1.2 and 4.2). The nitrite scavenging effects were higher at pH 1.2, and reached more than 95% by adding 0.2% and 0.1 % of garlic juice at pH 1.2. Addition 0.02～0.001 % garlic juice in showed the SOD-like activities. Its activity of garlic from Namhae. was a range of 6.0${\pm}$0.37～14.4～0.69%. It was found that 0.2% and 0.1% garlic showed strong antimicrobial action against growth of all the tested bacteria. Antimicrobial action. was showed 74.7${\pm}$0.70% and 51.7${\pm}$1.l1% on Sal. typhimurium in 0.2% of garlic from Namhae and Uiseong and 28.6${\pm}$0.90% on B. subtilis in garlic from Jeju.

• The Korean Journal of Food And Nutrition
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• v.25 no.3
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• pp.505-512
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• 2012

This study evaluated the quality characteristics of rice-doenjang prepared with Aspergillus oryzae MD05 and Bacillus subtilis HJ18-4. The physicochemical characteristics of rice-doenjang prepared with three types of meju, such as grain type(Rice 1), disc shape(Rice 2), and brick shape(Rice 3), were compared. We determined amino and ammonia nitrogen contents, protease and amylase activities, reducing sugar content, and total aerobic counts and lactic acid bacteria counts. The amino nitrogen and reducing sugar contents in all meju types gradually increased during fermentation. The amino nitrogen contents reached 151.5~200.0 mg%. Rice 3 were had the highest levels among the 4 types of rice-doenjang. After 42 days of fermentation, amylase and protease activity of Rice 3 were 366.57 and 94.26 unit/g, respectively, which were the highest levels among the 4 types of rice-doenjang. The total aerobic bacteria counts were not changed during 42-day fermentation. This results suggest that high quality characteristics of brick-shaped meju(Rice 3) were better than those of the other meju preparations.

• Journal of Life Science
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• v.23 no.1
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• pp.15-23
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• 2013

AprE51 from Bacillus amyloliquefaciens CH51 is a 27 kDa subtilisin-like protease with fibrinolytic activity. AprE51-6 showing increased catalytic activity was produced previously. To enhance the thermostability of AprE51-6, 2 residues, Gly-166 and Asn-218 based on B. subtilis subtilisin E were mutated by site-directed mutagenesis. The results of the mutational analysis showed that substitution of arginine for Gly-166 (AprE51-7) increased the fibrinolytic activity 1.8-fold. An N218S mutant (AprE51-8) also increased the fibrinolytic activity up to 4.5-fold in a fibrin plate assay. Purified AprE51-7 and AprE51-8 mutants had a 1.9- and a 2.5-fold higher $k_{cat}$, respectively, and a 2.1-1.9-fold lower $K_m$, respectively. This resulted in a 3.8- and a 4.7-fold increase in catalytic efficiency ($k_{cat}/K_m$), respectively, relative to that of wild-type AprE51. AprE51-8 had a broader pH range than AprE51-6 and nattokinase, especially at an alkaline pH value. In addition, AprE51-8 showed higher thermostability than AprE51-6 at $60^{\circ}C$. The half-lives of AprE51-7 and AprE51-8 at $50^{\circ}C$ were 21.5 and 27.3 min, respectively, which are 2.0 and 2.6 times longer, respectively, than that of the wild-type AprE51.

• Journal of Life Science
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• v.27 no.11
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• pp.1276-1289
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• 2017

This study isolated and purified the antimicrobial peptide McSSP-31 from an acidified hemocyte extract of a Mytilus coruscus. The antimicrobial peptide was purified by using a $C_{18}$ reversed-phase high-performance liquid chromatography (HPLC). The peptide was determined to be 3330.549 Da by matrix assisted-laser desorption ionization time-of-flight mass spectrophotometer (MALDI-TOF/MS). The N-terminus of a 14 amino-acid sequence was identified as P-S-P-T-R-R-S-T-S-R-S-K-S-R by Edman degradation method. The acquired sequence showed a 93% similarity with the sperm-specific protein Phi-1, which is from M. californianus. The identified open-reading frame (ORF) of peptide was 306 bp encoding 101 amino acids, which was analyzed by rapid amplification of cDNA ends (RACE), cloning and sequencing analysis. We compared the full sequence with other known proteins that reveal the sperm-specific protein Phi-1 (93.5%) of M. californianus. Synthesized antimicrobial peptide (McSSP-31) showed antibacterial activity against gram-positive bacteria including B. subtilis, S. mutans, S. aureus and gram-negative bacteria including E. coli, K. pneumoniae, P. mirabilis, P. aeruginosa and fungi, C. albicans. Also, synthesized peptide showed strong antibacterial activity against antibiotic-resistant strains, including S. aureus. The cytotoxicity of the peptide was determined by using the HUVEC human cell line. The peptide did not exhibit any significant cytotoxic effects on the normal human cell line, and it had very low hemolytic activity with flounder hemoglobin. The results demonstrated that peptide purified from the hemocyte of a M. coruscus exhibits antibacterial activity against various bacteria and has the potential to be an alternative antibiotic agent.

• Journal of the Korean Society of Food Science and Nutrition
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• v.43 no.7
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• pp.989-998
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• 2014

We investigated the characteristics and biological activities of vinegars added with different levels (0%, 0.5%, 1%, 2%, and 3%) of young leaves of Akebia quinata. During alcohol fermentation, alcohol and total acidity contents of vinegars increased. During acid fermentation, total acidity and amino acid contents increased. Vinegar added with 3% A. quinata leaf showed the highest total sensory score. The contents of total polyphenols, flavonoids, and tannin significantly increased during fermentation according to the amount of A. quinata leaf. After 22 days of fermentation, total polyphenol, total flavonoid, and tannin contents of vinegar added with 3% A. quinata were 4,079.08 mg GAE/100 g, 2,927.08 mg CE/100 g, and 3,618.00 mg TAE/100 g, respectively. ABTS radical scavenging activity of vinegar added with 3% A. quinata was 79.63%. Anti-cancer activities of vinegar added with 3% A. quinata were 48.65% and 52.90% against MCF-7 and HepG2 cells, respectively. Vinegar added with 3% A. quinata showed anti-bacterial activities against Bacillus cereus, Shigella flexneri, Salmonella enterica, Bacillus subtilis, and Klebsiella pneumoniae. Our results demonstrate that the biological activities of vinegar added with 3% A. quinata leaf (22 days of fermentation) were excellent, and their enhanced total polyphenol, flavonoid, and tannin contents were associated with antioxidant, anti-cancer and anti-microbial activities. Thus, A. quinata can be used as a functional material in vinegar and other foods.