To study the possible use of probiotics in fish farming, we evaluated antagonism of antibacterial strain Bacillus amyloliquefaciens H41 against the fish pathogenic bacterium Vibrio anguillarum NCMB1. The purification of growth inhibition factor produced by B. amyloliquefaciens H41 was achieved by obtaining supernatant of this bacterium. The growth inhibition factor was purified to homogeneity by 70% ammonium sulfate precipitation, DEAE-sephadex A-50 ion exchange chromatography, sephadex G-200 gel filtration column chromatography, and sephadex G-50 gel filtration column chromatography with 40.8 fold of purification and 2.9% yield. The molecular weight of the purified growth inhibition factor was 48 kDa as determined by sodium dodecyl sulfate polyacrylamide gel electrophoresis. The optimum pH and temperature for the growth inhibition factor were pH 7.5 and $30^{\circ}C$, respectively. The activity of growth inhibition factor was enhanced slightly by some metal ions, such as $Mg^{+2}$, $Mn^{+2}$, but was inhibited by the addition of $Co^{+2}$, $Hg^{+2}$, $Zn^{+2}$ and $Ag^{+2}$. NaCl stability of the growth inhibition factor was observed with 50% residual activity at 3% NaCl concentration. Toxicity test showed that the purified B. amyloliquefaciens H41 growth inhibition factor did not affect the live of Japanese flounder (Paralichthys olivaceus) and the effectiveness was 78% of residual lethality compared to commercial antibacterial agents.
CD11c and costimulatory molecules such as CD80 and CD86 express mainly in dendritic cells (DCs). In this study, we investigated the biologic effects of recombinant Fms-like tyrosine kinase-3 (Flt-3) ligand on the expression of DC surface markers, including CD11c in leukemia cell lines, such as KG-1, HL-60, NB4, and THP-1 cells. The expression of the Flt-3 receptor was found in NB4 and HL-60 cells, as well as KG-1 cells, but not in THP-1 cells. When KG-1 cells were cultured in a medium containing Flt-3 ligand or granulocyte macrophage-colony stimulating factor (GM-CSF) plus tumor necrosis factor (TNF)-$\alpha$, cell proliferation was inhibited and the expression levels of CD11c, major histocompatibility complex (MHC)-I, and MHC-II were increased in the cells. Flt-3 ligand also increased the expression level of CD11c on HL-60 and NB4 cells, but not on THP-1 cells. In comparison with CD11c expression, the expression level of CD11b on KG-1 cells, but not on NB4 and HL-60 cells, was slightly increased by Flt-3 ligand. Flt-3 ligand induced phosphorylation of extracellular signal-regulated kinase-1/2 (ERK-1/2) and p38-mitogen-activated protein kinase (p38-MAPK) in KG-1 cells, and the up-regulation of CD11c expression by Flt-3 ligand in the cells was abrogated by PD98059, an inhibitor of MEK. The results suggest that Flt-3 ligand up-regulates DC surface markers on $CD34^+$ myelomonocytic KG-1 cells, as well as promyelocytic leukemia cells, and that the differentiation of the leukemia cells into DC-like cells by Flt-3 ligand is mediated by ERK-1/2 activity.
A potent citric acid producing strain was selected by an extensive screening test of the yeasts isolated from the various sources. These experiments were conducted to identify the selected strain and investigate the cultural conditions for citric acid production. The results obtained were as fellows: 1. The selected strain of yeast was identified to Hansenula anomala var. anomala by a taxnoomic study of Lodder. 2. The optimum conditions for citric acid production in the basal medium containing 10% glucose were: temperature $30^{\circ}C$, the concentration of $CaCO_3$ 3% and the velocity of oscillation 110 oscills/min. 3. As a nitrogen source ol the basal medium $NH_4Cl(0.1%)$ was the most effective for citric acid production. Organic nitrogen sources such as peptone were adequate for growth of the strain but not for citric acid production. 4. The most effective concentration of glucose was 10% in yield ratio of citric acid from sugar. 5. The addition of defatted rape seed, defatted perilla or defatted rice bran to the medium was effective for citric acid production. When 5% extract solution of defatted rape seed was added, the citric acid production was increased as much as 40% as compared with the case of adding yeast extract(0.2%). 6. The most effective concentration of $KH_2PO_4$ and $MgSO_4{\cdot}7H_2O$ in the medium(for citric acid Production) was 0.05% and 0.025% respectively. 7. Under the optimum cultural conditions, the growth of the strain was continued up to 5 days and the increase of citric acid production was continued up to 6 days, showing the yield ratio of 46% to glucose.
The purpose of this study was to improve the quality characteristics of soy ice cream supplemented with oligosaccharide, and to test its blood glucose lowering effect. Boiled soybean powder was compared to parched soybean powder and to milk, as an ingredient. The soybean powder base was prepared by incubating with fructooligosaccharide (FOS) and apple juice, along with Lactobacillus acidophilus and L. bulgaricus at $30-40^{circ}C$ for 24 hr. With the fermentation process, the fishy smell of the soybean was removed and the taste improved. The overrun and melt-down values of the boiled soybean ice cream were significantly higher than those of the parched soybean ice cream, although they were significantly lower than those of the milk ice cream. The sensory characteristics of the soy ice cream prepared with the fermented base of boiled soybeans were significantly improved, as compared to those of the ice cream made using parched soybeans, but they were not significantly different from those of the milk ice cream. The blood glucose level at 120 min after ingestion of the ice cream prepared with FOS and the fermented base of boiled soybean powder was significantly lower than that occurring with the milk ice cream made with sugar.
This study was conducted to analyze the microbial community and propionic acid production ability of natural microflora in the rice cakes. Genetic analysis of natural microflora in Jorangyi rice cake was performed to select propionic acid - producing bacteria. Selected propionic acid-producing bacteria were cultivated in TSB (tryptic soy broth) supplemented with glucose, and growth characteristics were analyzed by temperature and production of propionic acid was analyzed by gas chromatography (GC-FID). Linearity, detection limit, quantitative limit, and recovery rate were measured to verify propionic acid assay. A total of 98 microbial strains were detected from microflora of Joraengyi rice cake that grew after expiration of shelf life. Lactobacillus casei group accounted for 50.48% and Lactobacillus buchneri was 29.60%. Propionic acid - producing bacteria were Propionibacterium thoenii, P. cyclohexanicum, Propionibacterium_uc, P. jensenii, and P. freudenreichii. Natural bacteria and Lactobacillus spp. did not produce propionic acid during 14 days but P. cyclohexanicum, P. freudenreichii subsp. Shermanii, P. thoenii and P. jesenii produced $263.47{\mu}g/mL$, $338.90{\mu}g/mL$, $325.43{\mu}g/mL$ and $222.17{\mu}g/mL$ during 4 days and 2,462.02 and 2,904.78, 2,220.64, $3,519.17{\mu}g/mL$ during 14 days. As a result of this study, it was affirmed that the natural microflora of Joraengyi rice cake during storage can produce propionic acid from natural sources even if a high concentration of propionic acid is not intentionally added. Because of characteristics of rice cake composed of starch and glucose. This study will be used as a recognition criterion to detect natural preservatives such as propionic acid in starchy foods such as rice cakes and as reference standard safety management data.
Ham, Seung-Hee;Choi, Nack-Shick;Moon, Ja-Young;Baek, Sun-Hwa;Lee, Song-Min;Kang, Dae-Ook
Journal of Life Science
/
v.27
no.2
/
pp.202-210
/
2017
As an effort to find a potential biopreservative, we isolated bacterial strains producing bacteriocin from fermented foods. A strain was finally selected and characteristics of the bacteriocin were investigated. The selected strain was identified as Bacillus subtilis E9-1 based on the 16S rRNA gene analysis. The culture supernatant of B. subtilis E9-1 showed antimicrobial activity against Gram-positive bacteria. Subtilisin A, ${\alpha}$-chymotrypsin, trypsin and proteinase K inactivated the antimicrobial activity, which means its proteinaceous nature, a bacteriocin. The bacteriocin activity was fully retained at the pH range from 2.0 to 8.0 and stable at up to $100^{\circ}C$ for 60 min. Solvents such as ethanol, isopropanol and methanol had no effect on the antimicrobial activity at the concentration of 100% but acetone and acetonitrile reduced the activity at up to 100% concentration. Cell growth of four indicator strains was dramatically decreased in dose-dependent manner. Listeria monocytogenes was the most sensitive, but Enterococcus faecium was the most resistant. Bacillus cereus and Staphylococcus aureus showed the medium sensitivity. The bacteriocin showed its antimicrobial activity against B. cereus and L. monocytogenes via bactericidal action. The number of viable cells of L. monocytogenes started to reduce after addition of bacteriocin to the minced beef. The bacteriocin was purified through acetone concentration, gel filtration chromatography and RP-HPLC. The whole purification step led to a 6.82 fold increase in the specific activity and 6% yield of bacteriocin activity. The molecular weight of the purified bacteriocin was determined to be 3.3 kDa by MALDI-TOF/TOF mass spectrometry.
Red spotted grouper, Epinephelus akaara is distributed in the south and west coasts of Korea. The natural stocks of the fish are decreasing sharply year by uear because of reckless overfishing. This research was carried out to understand general informations on maturation, sex composition and sex-reversals of the fish. Annual fishing uields of red spotted grouper in the castal area of Byonsan Peninsular of Kora decreased over 10% from 1992 to 1994. The main fishing season was from May to July with fishing gear of Hand-lines. Gonadosomatic index (GSI) and condition factor were highest on early and late July, respectively, thus main spawning reriod was assumed from late July to early August. The relationship between total length (X) and body weight (Y) for wild adults was represented as a regression, Y=$0.0169X^{2.9705}$, ($r^2$=0.96). Frequency of sex of wild red spotted gouper showed that the number of female below 38cm in total length was more than that of male, and hermaphrodite mainly occurred from 28cm to 32cm in total length the frequency of male and female were almost same. Also hermaphrodite occurred mainly between 25~29cm. Sex reversal ration of the adults reared in a tank for a year with different sexual compositions revealted that the frequency of female reversed from male was more than that of male reversed from female at 1:1 and 1:2 stocking densities of female and male, respectively. Also, about 20% of female was reversed to male when all females were reared. And the size of the fish reversed to male was larger than that of non-reversed female.
This study was carried out to investigate the physiological activities of the ethanol extract from Gymnopilus spectabilis mycelium (EGM) and of the supernatant obtained from fermentation broth (SGB). The contents of polysaccharides, phenol compounds and total ${\beta}-glucans$ of EGM were found to be 80.14%, 3.5 mg/ml and 5.91%, respectively and those for SGB were 78.68%, 3.32 mg/ml and 3.28%, respectively. Both EGM and SGB exhibited dose-dependent nitrate-scavenging abilities at pH 1.2. In addition, both EGM and SGB on the autoxidation rate of the linoleic acid demonstrated powerful antioxidant activities at 1 mg/ml level. With respect to fibrolytic activity, EGM showed 1,180 unit/g, which was the same activity as streptokinase, while SGB was 1,011 unit/g. The angiotensin converting enzyme inhibition activity of EMG determined by both the normal and pretreatment methods were estimated to be 8.2% and 10.2%, respectively. However, SGB showed no corresponding activity. The growth inhibitory effects of EGM on AGS, A549, HeLa and NCTC cells were over 58.88%, respectively. And the growth inhibitory effects of the SGB on HeLa and NCTC cells were 44.92 and 76.76%, respectively. Also, EGM and SGB activated the components of the alternative complement pathway from 51 and 62% at the concentration of 100 mg/ml, The xanthine oxidase inhibition activities of EGM and SGB (1 mg/ml) were 9.53 and 16.92%, respectively.
The present study performed the isolation of cytosolic ascorbate peroxidase (APX) isozymes and analyzed the pattern of their activity development and also investigated the change in some other enzyme activities related to the ascorbate-glutathione pathway from the senescing wheat leaves. The aim of this work is to examine the possibility that in the cytoplasm of wheat leaves the ascorbate-glutathione pathway p!ays a significant role in relation to leaf senescence involving an $H_2O_2$ accumulation and then to show the effect of benzyladenine (BA) on that pathway. During the leaf senescence characterized by increases in ChI breakdown and H202 accumulation under the 4-day dark incubation of matured leaf segments; i) no significant increase of total cytosolic APX was observed, ii) a dehydroascorbate reductase (DHAR) activity was decreased rapidly, iii) a slight increase of glutathione reductase (GR) activity occurred. In the BA-treated leaves; however, i) the total activity of APX increased conspicuously, ii) the decrease of DHAR activity was relatively inhibited, iii) the GR activity increase was more enhanced, and iv) the decrease of ascorbate content and the increase of H202 content were retarded as compared with those of control leaves. Three isozymes of cytosolic APX were found by using a native-electrophoretic gel in senescing wheat leaves and two of them occurred with major activity. In the developmental patterns of cytosolic APX isozymes, only two isozyme bands ("a" and "b") appeared with almost constant activity through 4 days of incubation in the control leaves, while one additional weak isozyme band ("c") and a little increase of "b" isozyme activity were detected in the BA-treated leaves. EspeciaUy, the development of "a" isozyme activity increased remarkably compared with that of control leaves. The increased capacity for peroxide scavenging due to the enhanced activity of all 3 enzymes (APX, DHAR, GR) participating in the ascorbate-glutathione pathway in BA-treated leaves suggested that this pathway might playa significant role in the processes related to the wheat leaf senescence.scence.
In this study, we evaluated the antidiabetic effect of a submerged culture of Ceriporia lacerata mycelium (CL01) on hematological indices, as well as protein and mRNA expression of the insulin-signaling pathway, in db/db mice. After CL01 was administrated for 4 weeks, blood glucose levels decreased consistently, and plasma insulin and c-peptide levels each decreased by roughly 55.8%, 40% of those in the negative control (p<0.05). With regard to HOMA-IR, an insulin resistance index, insulin resistance of the CL01-fed group improved over that of the negative control group by about 62% (p<0.05). In addition, we demonstrated that the protein expression levels of pIR, pAkt, pAMPK, and GLUT4 and the mRNA expression levels of Akt2, IRS1, and GLUT4 in the muscle cells of db/db mice increased in the CL01-fed group compared to the corresponding levels in the control group. These results demonstrate that CL01 affects glucose metabolism, upregulates protein and gene expression in the insulin-signaling pathway, and decreases blood glucose levels effectively by improving insulin sensitivity. More than 90% of those who suffer from type 2 diabetes are more likely to suffer from hyperinsulinemia, hypertension, obesity, and other comorbidities because of insulin resistance. Therefore, it is possible that CL01 intake could be used as a fundamental treatment for type 2 diabetes by lowering insulin resistance, and these results may prove be useful as basic evidence for further research into the mechanisms of a cure for type 2 diabetes.
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