Mycelial viability and cultivation characteristics of strains of Hypsizygus marmoreus after long-term storage were investigated. The experimental conditions were the storage at $4^{\circ}C$ using a slant culture technique with or without mineral oil, and in liquid nitrogen tank in the presence of 10% glycerol or 10% glycerol with 5% trehalose. The myceila of four strains of H. marmoreus were thawed at 9, 21, 33, and 45 months after beginning of the storage, and then the growth of the mycelia was measured on a PDA plate with serial transfers to new plate for the recovery. The mycelial growth data after 45 months showed that the mycelia were mostly viable but not fully active particularly when they were stored in liquid nitrogen with 10% glycerol. The growth activity could be fully recovered after second transfer to new PDA plate. Cultivation of mushroom fruiting body using the recovered mycelia also demonstrated that the storage methods employed in this work were applicable for the long-term storage of H. marmoreus.
Three hundreds thirty two bacterial colonies which were able to degrade crude oil were isolated from soil samples that were contaminated with oil in Daejeon area. Among them, one bacterial strain was selected for this study based on its higher oil degrading ability, and this selected bacterial strain was identified as Acinetobactor sp. B2 through physiological-biochemical tests and analysis of its 16S rRNA sequence. Acinetobactor sp. B2 was able to utilize various carbohydrates but did not utilize trehalose and mannitol as a sole carbon source. Acinetobactor sp. B2 showed a weak resistance to antibiotics such as kanamycin, streptomycin, tetracycline and spectinomycin, but showed a high resistance up to mg/ml unit to heavy metals such as Ba, Li, Mn, AI, Cr and Pb. The optimal growth temperature of Acinetobactor sp. B2 was $30^{\circ}C.$ The lipase produced by Acinetobactor sp. B2 was purified by ammonium sulfate precipitation, DEAE-Toyopearl 650M ion exchange chromatography and Sephadex gel filtration chromatography. Its molecular mass was about 60 kDa and condition for the optimal activity was observed at $40^{\circ}C$ and pH 10, respectively. The activation energy of lipase for the hydrolysis of pnitrophenyl palmitate was 2.7 kcal/mol in the temperature range of 4 to $37^{\circ}C,$ and the enzyme was unstable at the temperature higher than $60^{\circ}C.$ The Michaelis constant $(K_m)\;and\;V_{max}$ for p-nitrophenyl palmitate were 21.8 uM and $270.3\;{\mu}M\;min^{-1}mg^{-1},$ respectively. This enzyme was strongly inhibited by 10 mM $Cd^{2+},\;Co^{2+},\;Fe^{2+},\;Hg^{2+},$ EDTA and 2-Mercaptoethalol.
The mycelium was isolated from the fruiting body of Tricholoma matsutake collected from Mt. Namsan, Kyongju and it was named as Tricholoma matsutake DGUM 26001. For the mycelial growth of T. matsutake DGUM 26001, the complex media, yeast-malt extract medium and Czapek-Dox medium supplemented with yeast extract, were excellent. The media such as nutrient glucose medium, mushroom complex medium, and Tricholoma matsutake medium (TMM), were effective. However, There was no a mycelial growth in the media used for bacterial cultivation such as colombia medium, brain heart infusion medium, Luria-Bertani medium supplemented with glucose, and brucella medium. When carbohydrate as a carbon and energy source was supplemented in the TMM medium for the mycelial growth, starch as a polysaccharide was best. As a disaccharide, trehalose and maltose were excellent. Sorbitol, xylitol and glucose were excellent carbon sources of monosaccharose. When the mycelia were cultivated for 30 days at $24^{\circ}C$ in the TMM supplemented with 2.0% starch, the dry weight of the mycelia harvested was 8.85 g/L. When organic acid was given as a carbon source, only succinic acid was utilized. As a vitamin source, coconut water and pyridoxine were excellent. After 30 day-cultivation in the TMM medium, the dry weights with coconut water and pyridoxine were 8.65 and 8.32 g/L, respectively.
Although liposome has many advantages as a pharmaceutical dosage form, its application in the industrial field has been limited because of some problems such as preparation method, reproducibility, scale-up, stability and sterilization etc. Liposomes prepared by microemulsification method had defined size, narrow size distribution, reproducibility and high entrapment efficiency. For enhancing the stability, the dry form of liposome was recommended. These types of liposome are proliposome and freeze-dried liposome. The liposome must have some properties for preparing of freeze-dried liposome; small size $(50{\sim}200\;nm)$, narrow size distribution and cryoprotectant. In this experiment, the liposomes containing 5-Fluorouracil(5-FU) and its prodrug(pentyl-5-FU-1-acetate; PFA, hexyl-5-FU-1-acetate; HFA) were made with soybean phosphatidylcholine, cholesterol, stearylamine(SA) and dicetyl phosphate(DCP) employing hydration method or microemulsification method using $Microfluidizer^{TM}$. Both or liposome types were MLV and MEL. After preparation, freeze drying and rehydration were performed. In the process of freezing, trehalose(Tr) was added as a cryoprotectant. Their evaluation methods were as follows; entrapment efficiency, mean particle size and size distribution, dissolution test, retain of entrapment efficiency and turbidity after freeze-drying. The results are summarized as belows. The entrapment efficiency of 5-FU was dependent on total lipid concentration and cholesterol content but that of PFA and HFA was decreased when cholesterol was added. When DCP and SA were added, entrapment efficiency was decreased. As the partition coefficient of drug was increased, entrapment efficiency was increased. Under the same condition, entrapment efficiency of MEL is similar to that of MLV. The mean particle size and size distribution of MEL were smaller than those of MLV. Dissolution rates of drug from both liposome types were comparatively similar. Dissolution rates of drugs with serum and liver homogenate were faster than without these material. After preparation of liposome, free drug was removed efficiency by Dowex 50W-X4. When liposome was freeze-dried and then rehydrated in the presence of Tr, characteristics of liposome were maintained well in MEL than MLV. Tr Was used successfully as a cryoprotectant in the process of freeze drying and the optimal ratio of Tr:Lipid was 4:1(g/g).
The quality characteristics of Yakju and survival rate of yeast were investigated by modifying the drying method for the cold adapted yeast strain Saccharomyces cerevisiae Y297 (SCY297). Viability and fermentation characteristics of the freeze-dried, air blast-dried, and liquid SCY297 cultures were compared after storing them at $25^{\circ}C$. In addition, 5% skimmed milk, ${\alpha}$-lactose, or trehalose was added as a protective agent for examining the effects of drying methods. During the 15-week storage period, the liquid and freeze-dried SCY297 cultures containing a protective agent showed a survival rate of 80%. However, the air blast-dried SCY297 culture showed 80% survival rate only in the skimmed milk supplemented group. Compared to the untreated cells, the acidity and amino acidity of Yakju prepared using freeze-dried or air blast-dried cultures of SCY297 increased by 2 fold and 5.7 fold respectively, while the alcohol content decreased by 5.07%. Compared to the untreated cells, the pH and amino acidity of Yakju prepared using the liquid culture of SCY297 increased by 1.5 fold and 2.5 fold respectively. Although the alcohol content decreased by 2.9%, decrease rate was lower than that observed for the freeze-dried and air blast-dried yeast cultures. Therefore, the results of this study showed that using a liquid starter culture was more advantageous than using the conventional solid culture.
These studies were carried out to identify Bacillus cereus group 1..5-] strain isolated from 5uyeong Bay. This strain was differentiated from B. cereus group using conventional, API system and fatty acid composition analysis. Colony characteristics were opague. mucoid, entire margin. convex. circular and non hemolysis on sheep blood agar plates, and were observed with central spore forming positive bacilli in a Gram stained preparation. and had no motility. The carbohydrates tested; glucose.maltose, and sucrose were assimilated but neither trehalose nor salicin were assimilated. This strain ultilized gelatin and was also inhibited by 6.5% NaCI. The results of biochemical examination were differented from B. cereus group LS-1 compared with others B. cereus group. The fatty acid composition contained major amounts of branched chain acids. iso $C_{15}$ and iso $C_{13}$ and the range of chain length was $C_{12}$ to C"$C_{17}$ and n$C_{15}$, acid was not detected. Automated fatty acid computer profile indicated "B. mycoides GC subgroup B of 0.312 similarity index." The results agreed with other research cases. On the other hand. A TB computer prolile index of API system (API 50 CHB & API 20E) identified" Doubtful profile of 99.7% B. firmus" . These results were presented with considerable discrepancies between API system and fatty acid analysis. With 67 biochemical characters. the similarity matrix of B. mycaides (KCTC 1033). B. thuringiensis (KCTC 1033). B. cereus (5-3) and B. mycoides (S-12) showed 42%. 42%. 59%, and 52%. respectively. Through the key tests and fatty acid analyses. we could notice the appearance of B. mycoides of the B. cereus group and this leads us to suspect the existence of a new biotype B. mycoides.
$\beta$-Glucosidase of Cellulomonas sp. CS1-1 in cellular compartment was localized with cell-bound form while Avicelase and carboxymethylcellulase (CMCase) were appeared with extracellular enzyme. Cell growth on cellulose or CMC minimal broth was increased by glucose addition. $\beta$-Glucosidase production on cellobiose or CMC minimal broth was repressed by the addition of glucose. However, on CMC minimal broth, the enzyme production was specially stimulated by cellobiose addition. $\beta$-Glucosidase production was also induced by CMC, starcth and maltose compared with glycerol, arabinose, xylose and trehalose. From the above results, it was concluded that glucose effect on $\beta$-glucosidase biosynthesis showed catabolite repression, but enzyme production was induced by cellobiose, CMC, and starch, indicating that $\beta$-glucosidase is inducible enzyme. Yeast extract stimulated $\beta$-glucosidase production more than peptone and ammonium sulfate. $\beta$-Glucosidase activity was increased with 50mM MgCl$_2$in 10mM potassium phosphate buffer (pH 7.0). Optimum conditions for enzyme activities were pH 6.0 and 42$^{\circ}C$, Km value of $\beta$-glucosidase for p-nitrophenyl-$\beta$-D-glucosidase was 0.256mM and Ki for $\beta$-D(+)-glucose was 9.0mM.
Previously, we isolated plant growth promoting rhizobacterium (PGPR) Bacillus licheniformis K11 which could produce auxin, cellulase and siderophore. The siderophore of B. licheniformis K11 $(siderophore_{K11})$ was determined to be a catechol type siderophore which is produced generally by Bacillus spp. B. licheniformis K11 could produce the siderophore most highly after 96 h of incubation under nutrient broth at $20^{\circ}C$ with initial pH 9.0. For the production of the $siderophore_{K11}$, trehalose and $NH_4Cl$ were the best carbon and nitrogen sources in Davis minimal medium, respectively. The $siderophore_{K11}$ was Produced in M9 medium (pH 9.0) after 4 days at $20^{\circ}C$, and purified from culture broth of B. licheniformis K11 by using Amberlite XAD-2, Sephadex LH-20 column chromatography, and reversed-phase HPLC. The $siderophore_{K11}$ had the biocontrol activity against spore germination of P. capsici and F. oxysporum on potato dextrose agar (PDA). The results indicate that the $siderophore_{K11}$ is an antifungal mechanism of B. licheniformis K11 against phytopathogenic fungi.
Kim, Hong-Kyu;Yang, Euy-Seog;Park, Gi-Moon;Kim, Gwan-Hou;Kim, Hyun-Ho;Lee, Ka-Soon
Journal of the Korean Society of Food Science and Nutrition
/
v.40
no.8
/
pp.1179-1183
/
2011
This study was conducted to investigate the quality characteristics, antioxidant activity, and ${\alpha}$-glucoamylase inhibitory activity of Dahyang, a Chungnam Agricultural Research & Extension Service's newly bred cultivar of brown button mushroom. Total phenolic compound contents of Dahyang and the no. 705 mushroom were 189${\pm}$12 mg% and 168${\pm}$8 mg%, respectively. The major free sugars in Dahyang were mannitol (3.11%), xylose (0.12%), and trehalose (0.08%). ${\beta}$-Glucan content was 28.34% in Dahyang and 26.55% in the no. 705 mushroom, respectively. Electron donating ability by DPPH in Dahyang and the no. 705 mushroom was 52.14% and 45.27% for the water extract, and 57.81% and 46.93% for the 80% ethanol extract, respectively. ${\alpha}$-Glucoamylase inhibitory activity in a 10 mg/mL concentration of water extract were was 33.25% in Dahyang and 29.22% in the no. 705 mushroom, respectively.
This study analyzed components of the leaves of Elaeagnus multiflora as part of studies on the nutritional and functional materials of fruits and leaves of this plant. The moisture content of the leaves was 71.6% and the carbohydrate, crude protein, lipid and ash contents were 24.1, 1.4, 0.4 and 2.5%, respectively. Concentrations of reducing sugars, soluble proteins and polyphenols were 460.0, 503.3 and 805.6 mg/100 g, respectively. Fructose was the dominant free sugar, and arabinose, maltose, glucose, and a small amount of trehalose were also detected. Malic acid was the main organic acid in E. multiflora leaves, and acetic acid, citric acid, lactic acid, and succinic acid were also present. E. multiflora leaves were high in K, Ca and Mg. Of hydrolyzed amino acids, alanine was present at the highest concentration (112.0 mg/100 g), with threonine, leucine, valine and phenylalanine being the next most common. Glutamic acid and ornithine were the dominant free amino acid and amino acid derivative, respectively.
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