In order to obtain the basic informations on the production of single cell protein from ethanol, 145 yeast strains utilizing ethanol as a sole carbon source were isolated from 32 soil samples in Korea. A yeast strain showing the highest cell yield among the isolated strains was selected and identified. The optimum culture condition, utilization of other carbon sources and the cultural characteristics for the selected yeast, and the chemical analysis of the yeast cell composition, and utilization of ethanol by the selected yeast were investigated. All the culture was carried out in the shaking flasks. The results obtained were as follows: 1. The selected yeast strain was identified as Debaryomyces nicotianae-SNU 72. 2. The optimum composition of the medium for the selected yeast is : Ethanol 40 ml, Urea 0.5 g, Potassium phosphate (dibasic) 0.5 g, Ammoium phosphate (monobasic) 0.15 g, Magnesium sulfate 0.05 g, Calcium chloride 0.01g, Yeast extract 0.005 g, Tap water 1000 ml. 3. The optimum pH was 5.0-5.5, the optimum temperature $30-33^{\circ}C$ and the aerobic state was unimportant. 4. Utilization of methanol, n-propanol, iso-propanol, n-butanol, iso-butanol, tert-amyl alcohol and acetic acid by the selected yeast was very weak. So substitution of the subtrate was thought to be impossible. 5. Studies on the propagation of the yeast cells showed that the lag phase of the yeast cells lasted 16 hours, and the logarithmic growth phase extended 16 to 28 hours. The specific growth rate was about $0.19\;hr^{-1}$ and the doubling time was 3.6 hours during the logarithmic growth phase. 6. As the result of the chemical analysis of the dry yeast cells, the content rate of the crude protein was 55.19 %, the content of others was similar to the average content of the yeast component. 7. After 34 hours cultivation, under the optimum culture condition investigated, the dry cell yield against the amount of the added ethanol was 53.4 % (W/V%), the dry cell yield against the amount of the utilized ethanol was 73.6 % (W/V%), the evaporation rate of ethanol was about 19.1 %.
Park, Sun-Hee;Kim, Min-Ji;Kim, Go-Eun;Park, So-Yeong;Kim, Koth-Bong-Woo-Ri;Kim, Yeon-Ji;Cho, Young-Je;Ahn, Dong-Hyun
Journal of the Korean Society of Food Science and Nutrition
/
v.46
no.8
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pp.919-928
/
2017
The immuno-enhancing effects of alginate oligosaccharides from Sargassum coreanum were investigated. The alginate oligosaccharides were produced by an alginate-degrading enzyme from S. oneidensis PKA 1008. The degraded alginate oligosaccharides were visualized by thin-layer chromatography developed using a solvent system of 1-butanol/methanol/water, 4:1:2 (v/v/v). Alginate was degraded into dimmers at 60 h. As a result, the levels of Th1 cytokine [interferon $(IFN)-{\gamma}$ and interleukin (IL)-2] and Th2 cytokine (IL-6 and IL-10) increased with increasing incubation time compared to the control in vitro. Enzymatic extract treatment promoted proliferation of splenocytes at concentrations of 100 and 200 mg/kg at 24 h in vivo. Secretion of $IFN-{\gamma}$ and IL-2 significantly increased in a dose-dependent manner at 24 h as well as induced higher production of IgG2a in serum. Natural killer cell activity was measured and tended to increase. In addition, complete blood cell counts increased in a dose-dependent manner. These results indicate that alginate oligosaccharides produced by crude enzyme from S. oneidensis PKA 1008 may have significant immune activities.
In the current study, comparisons of Oenothera Biennis seed extracts with water, ethanol, methanol, and 70% ethanol in their total polyphenolics contents, anti-oxidant, anti-neurotoxicity, anti-cancer, and immune-modulatory activities were investigated. Compared with other extracts, those concentrations of total phenolics and flavonoids were the highest in MeOH extract (31.90 mg GAE/g and 20.66 mg QE/g). The radical scavenging and reducing power activities were dose-dependently increased by treatment of O. Biennis seed water, EtOH, MeOH, and 70% EtOH extracts. Furthermore, pretreatment of water, EtOH, and MeOH extracts significantly reduced glutamate-induced cytotoxicity in HT22 hipocampal neuron cells. In the case of cancer cells, MeOH extracts showed lower $IC_{50}$ values in HepG2 ($74.21{\mu}g/ml$), A549 ($188.24{\mu}g/ml$), MCF-7 ($186.42{\mu}g/ml$), and B16 ($101.80{\mu}g/ml$) than other extracts, where those water ($101.96{\mu}g/ml$) and EtOH ($788.39{\mu}g/ml$) extracts showed the lowest $IC_{50}$ activity in HT-29 and PC-3 cells, respectively. O. Biennis seed extracts did not show any cytotoxicity in RAW 264.7 macrophages at the concentration of $1-10{\mu}g/ml$, whereas 70% EtOH extract dose-dependently enhanced nitric oxide (NO) production in RAW 264.7 cells. Overall, we evaluated that various bioactive potentials of O. Biennis seed extracts which would relate with phenolic compounds abundance, thus these can be useful to future developments as functional food ingredients and natural medicines.
Esterase EM2L8 gene isolated from deep sea sediment was expressed in Escherichia coli BL21 (DE3) and the esterase activity of the cell-free extract was assayed using p-nitrophenyl butyrate-spectrophotometric method. Its optimum temperature was $40-45^{\circ}C$ and 45% activity of the maximum activity was retained at $15^{\circ}C$. The activation energy at $15-45^{\circ}C$ was calculated to be 4.9 kcal/mol showing that esterase EM2L8 was a typical cold-adapted enzyme. Enzyme activity was maintained for 6 h and 4 weeks at $30^{\circ}C$ and $4^{\circ}C$, respectively. When each ethanol, methanol, and acetone was added to the reaction mixture to 15% concentration, enzyme activity was maintained. In the case of DMSO, enzyme activity was kept up to 40% concentration. (S)-4-Chloro-3-hydroxy butyric acid is a chiral intermediate for the synthesis of Atorvastatin, a hyperlipemia drug. When esterase EM2L8 (40 U) was added to buffer solution (1.2 mL, pH 9.0) containing ethyl-(R,S)-4-chloro-3-hydroxybutyrate (38 mM), it was hydrolyzed into 4-chloro-3-hydroxy butyric acid with a rate of $6.8\;{\mu}mole/h$. The enzyme hydrolyzed (S)-substrate more rapidly than (R)-substrate. When conversion yield was 80%, e.e.s value was 40%. When DMSO was added, hydrolysis rate increased to $10.4\;{\mu}mole/h$. The plots of conversion yield vs e.e.s in the presence or absence of DMSO were almost same, implying that the reaction enantioselectivity was not changed by the addition of DMSO. Taken together, esterase EM2L8 had high activity and stability at low temperatures as well as in various organic solvents/aqueous solutions. These properties suggested that it could be used as a biocatalyst in the synthesis of useful pharmaceuticals.
Kim, Jeong-Hwa;Cha, Youn-Jeong;Shim, Mi-Ja;Lee, Min-Woong;Lee, Tae-Soo
The Korean Journal of Mycology
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v.39
no.1
/
pp.68-77
/
2011
80% methanol and 0.9% neutral saline soluble and hot water substances (hereinafter referred to Fr. NaCl, Fr. HW and Fr. MeOH, respectively) were extracted from fruiting bodies of Grifola frondosa. In vitro cytotoxicity tests, crude polysaccharides were not cytotoxic against cancer cell lines such as Sarcoma 180 and RAW 264.7 at the concentration of 10~2000 ${\mu}g/ml$, but crude polysaccharides from Fr. NaCl was slightly toxic to HT-29 and NIH3T3 at the concentration of 2000 ${\mu}g/ml$. Intraperitoneal injection with crude polysaccharides exhibited life prolongation effect of 25.0~52.9% in mice previously inoculated with Sarcoma 180. Fr. HW increased the numbers of spleen cells by 1.3 fold at the concentration of 200 ${\mu}g/ml$ compared with control. Fr. NaCl improved the immuno-stimulating activity of B lymphocyte by increasing the alkaline phosphatase activity by 1.5 fold compared with control at the concentration of 200 ${\mu}g/ml$. 10~14 ${\mu}M$ of nitric acid were generated when Fr. NaCl was added to RAW 264.7 at the concentration of 50~500 ${\mu}g/ml$, while the control group produced 4.3 ${\mu}M$ of nitric oxide. The Fr. NaCl, Fr. HW and Fr. MeOH increased the production of TNF-${\alpha}$, IL-$1{\beta}$, Il-2 and IL-6 by more than 1.4 times compared with the control group. The Fr. of MeOH increased the numbers of peritoneal exudate cells and circulating leukocytes by 3.0 and 2.0 folds compared with the control at the concentration of 50 ${\mu}g/ml$, respectively. Therefore, the crude polysaccharides extracted from fruiting bodies of Grifola frondosa could improve antitumor activity of mice.
This study was carried out to investigate into the effect of light-emitting diode (LED) for the light quality as a light source on the broccoli seed germination and the physiological activity of vegetable sprouts. We have also germinated seeds of the broccoli and applied LED as a light quality such as blue, green, red, white, yellow and red + blue color lights to their sprouts for 14 hours and kept dark for 10 hours at the temperature of $25^{\circ}C$ (day)/$18^{\circ}C$ (night). Broccoli sprouts were extracted by methanol and their physiological activities were examined. All broccoli seeds were germinated at 3 days after seeding regardless of the light color. Total sprout fresh weight were mostly became highest by 0.389g (10 plants) at 8 days after seeding when their sprouts were grown under blue color light. Total phenol compound contents in broccoli sprouts were extremely increased by $83.0\;mg{\cdot}L^{-1}$ under the white light, and total flavonoid contents were most much more by $72.6\;mg{\cdot}L^{-1}$ under the blue light. DPPH radical scavenging activity at $2,000\;mg{\cdot}L^{-1}$ were most highest by 93.5% in broccoli sprouts grown under the white light. Nitrite radical scavenging activity at the concentration of $500\;mg{\cdot}L^{-1}$ in sprout extracts were the most increased by 66.9% under the yellow light, and tyrosinase inhibition activity at $2,000\;mg{\cdot}L^{-1}$ in sprout extracts were by 14.5% under red light.
The indigenous Saccharomyces cerevisiae strains S13 and D8 were isolated at the microbial succession stage during spontaneous fermentation of Campbell Early wine as a resistant to potassium metabisulfite and a high sugar concentration. In this study, the fermentation characteristics of Campbell Early wine were investigated and compared with those of S. cerevisiae W-3, an industrial wine yeast. Alcohol production by the two strains was delayed at the initial fermentation stage, but increased fast when the fermentation continued. After the fermentation, the S13 and D8 wines contained 12.6% and 13.2% (v/v) alcohol, respectively, which were significantly higher than the alcohol content of the W-3 wine (12%, v/v). No marked differences were observed in the residual soluble solid content and the pH. However, the S13 and D8 wines showed high levels of total acid content, including malic and lactic acids. Especially, the lactic acid content was 8.9-fold in the S13 wine and six-fold in the D8 wine, compared with that of the W-3 wine. The two strains produced a higher level of acetaldehyde and a lower amount of methanol in the wine than the W-3 strain. The iso-Butanol content was lower in the two indigenous yeast wines with similar levels of n-propanol and iso-amyl alcohol contents than that in the W-3 wine. In the sensory evaluation, the S13 and D8 wines had higher scores for their color, flavor, taste and overall preferences than the W-3 wine. Especially, the S13 and D8 wines had much higher scores than the W-3 wine for flavor and color, respectively.
A bacteriocin-producing bacterium identified as Bacillus licheniformis was isolated from soybean sauce. Antibacterial activity was confirmed by paper disc diffusion method, using Micrococcus luteus as a test organism. The bacteriocin also showed antibacterial activities against Bacillus sphaericus, Lactobacillus bulgaricus, Lactobacillus planiarum, Paenibacillus polymyxa, and Pediococcus dextrinicus. Optimal culture conditions for the production of bacteriocin was attained by growing the cells in an MRS medium at a pH of 6.5~ 7.0 and a temperature of 37$^\circ$C for 36$\sim$48 hr. Solvents such as chloroform, ethanol, acetone, and acetonitrile had little effect on bacteriocin activity. However, about 50% of bacteriocin activity diminished with treatment of methanol and isopropanol at the final concentration of 50% at 25$^\circ$C for 1 hr. It was stable against a pH variation range from 3.0 and 7.0, but the activity reduced to 50% at a pH range from 9.0 to 11.0. It's activity was not affected by heat treatment at 100$^\circ$C for 30 min and 50% of activity was retained after heat treatment at 100$^\circ$C for 60 min, showing high thermostability. The bacteriocin was purified to a homogeneity through ammonium sulfate precipitation, SP-Sepharose ion-exchange chromatography, and reverse-phase high-performance liquid chromatography (HPLC). The entire purification protocol led to a 75-fold increase in specific activity and a 13.5% yield of bacteriocin activity. The molecular weight of purified bacteriocin was estimated to be about 2.5 kDa by tricine-SDS-PAGE.
Kang, Dong Hyeon;Han, Eun Hye;Jin, Changbae;Kim, Hyoung Ja
Journal of the Korean Society of Food Science and Nutrition
/
v.45
no.11
/
pp.1610-1616
/
2016
This study aimed to establish an optimal extraction process and high performance liquid chromatography-ultraviolet (HPLC-UV) analytical method for determination of 3,5-dicaffeoylquinic acid (3,5-DCQA) as a part of materials standardization for the development of a xanthine oxidase inhibitor as a health functional food. The quantitative determination method of 3,5-DCQA as a marker compound was optimized by HPLC analysis using a Luna RP-18 column, and the correlation coefficient for the calibration curve showed good linearity of more than 0.9999 using a gradient eluent of water (1% acetic acid) and methanol as the mobile phase at a flow rate of 1.0 mL/min and a detection wavelength of 320 nm. The HPLC-UV method was applied successfully to quantification of the marker compound (3,5-DCQA) in Aster glehni extracts after validation of the method with linearity, accuracy, and precision. Ethanolic extracts of A. glehni (AGEs) were evaluated by reflux extraction at 70 and $80^{\circ}C$ with 30, 50, 70, and 80% ethanol for 3, 4, 5, and 6 h, respectively. Among AGEs, 70% AGE at $70^{\circ}C$ showed the highest content of 3,5-DCQA of $52.59{\pm}3.45mg/100g$ A. glehni. Furthermore, AGEs were analyzed for their inhibitory activities on uric acid production by the xanthine/xanthine oxidase system. The 70% AGE at $70^{\circ}C$ showed the most potent inhibitory activity with $IC_{50}$ values of $77.01{\pm}3.13{\sim}89.96{\pm}3.08{\mu}g/mL$. The results suggest that standardization of 3,5-DCQA in AGEs using HPLC-UV analysis would be an acceptable method for the development of health functional foods.
Cladosporium fulvum, Aspergillus ochraceus, Aspergillus terreus and N-1 (unidentified species) were cultured on the artificial media containing sucrose as a carbon source at 20, 25 and $30^{\circ}C$ for 10 to 12 days. The lipids in the felts were extracted with chloroform-methanol mixture and the class composition and fatty acids of the lipid were determined. The summarized results are as follows 1. The average felts produced by each species per 100 ml of media were $3.82{\pm}0.30g$ for Cl. fulvum, $2.62{\pm}0.23g$ for Asp. ochraceus, $4.24{\pm}0.25g$ for Asp. terreus and $4.62{\pm}0.10g$ for N-1. Their crude fat contents $27.5{\pm}1.61%,\;50.47{\pm}1.00%,\;46.6{\pm}1.59%$ and 33.78 % and the fat coefficient 6.92, 8.88, 13.01 and 10.28, respectively. 2. The lipids produced by these species were mainly composed of triglyceride and the next free fatty acid in Cl. fulvum and N-1 and phospholipid Asp. ochraceus and Asp. terreus. 3. The major fatty acids of the lipids were in order of oleic, palmitic, linoleic and stearic acids in Asp. ochraceus, Asp. terreus and Cl. fulvum and linoleic, palmitic, oleic and stearic acid in N-1. The total percentage contents of these major fatty acids were over 98 % the former and over 95 % the latter. 4. The constituent fatty acids of the lipid were changed depending on the incubation temperature but hardly found a certain tendency except linoleic acid which was higher at lower temperature. 5. The total percentages of unsaturated fatty acids in the lipids were $50{\sim}60%$ and comparatively higher at lower incubation temperature.
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