Kim, Jin-Kwang;He, Dan;Liu, Qing-Mei;Park, Hye-Yoon;Jung, Mi-Sun;Yoon, Min-Ho;Kim, Sun-Chang;Im, Wan-Taek
Journal of Microbiology and Biotechnology
/
v.23
no.4
/
pp.444-450
/
2013
A Gram-negative, strictly aerobic, non-motile, non-spore-forming, and rod-shaped bacterial strain designated FW-$6^T$ was isolated from a freshwater sample and its taxonomic position was investigated by using a polyphasic approach. Strain FW-$6^T$ grew optimally at $10-42^{\circ}C$ and at pH 7.0 on nutrient and R2A agar. Strain FW-$6^T$ displayed ${\beta}$-glucosidase activity that was responsible for its ability to transform ginsenoside $Rb_1$ (one of the dominant active components of ginseng) to Rd. On the basis of 16S rRNA gene sequence similarity, strain FW-$6^T$ was shown to belong to the family Sphingomonadaceae and was related to Novosphingobium aromaticivorans DSM $12444^T$ (98.1% sequence similarity) and N. subterraneum IFO $16086^T$ (98.0%). The G+C content of the genomic DNA was 64.4%. The major menaquinone was Q-10 and the major fatty acids were summed feature 7 (comprising $C_{18:1}{\omega}9c/{\omega}12t/{\omega}7c$), summed feature 4 (comprising $C_{16:1}{\omega}7c/iso-C_{15:0}2OH$), $C_{16:0}$, and $C_{14:0}$ 2OH. DNA and chemotaxonomic data supported the affiliation of strain FW-$6^T$ to the genus Novosphingobium. Strain FW-$6^T$ could be differentiated genotypically and phenotypically from the recognized species of the genus Novosphingobium. The isolate that has ginsenoside converting ability therefore represents a novel species, for which the name Novosphingobium ginsenosidimutans sp. nov. is proposed, with the type strain FW-$6^T$ (= KACC $16615^T$ = JCM $18202^T$).
Sikhae is a traditional Korean fermented seafood with a 7-10% salt concentration. Consumers have begun to look for low-salt food because excess salt is known to cause hypertension and gastric cancer. The quality characteristics of low-salt squid sikhae were investigated at different fermentation temperatures and periods, so as to determine its shelflife. The shelf-life of the low-salt (5%) squid sikhae at $-1^{\circ}C$ based on pH was 142 days. The functional activities of the ethanol extract of squid sikhae such as its antioxidant activity and inhibitions on ${\alpha}$-glucosidase, ${\beta}$-glucuronidase, and elastase were stronger than those of the water extract. Based on the results of sensory evaluation, the low-salted squid sikhae was very similar to fermented seafood. In conclusion, low-salt sikhae is commercially viable.
Ginsenosides have been regarded as the principal components, responsible for the pharmacological and biological activities of ginseng. Absorption of major ginsenosides at the gastrointestinal tract was extremely low, when ginseng taken orally. In order to improve the absorption and bioavailability, transformation of major ginsenosides into more active and valuable minor ginsenoside is much required. In this present study, We isolated a lactic acid bacteria Leuconostoc fallax LH3 from the Korean fermented food Kimchi, which have higher ${\beta}$-glucosidase activity. Using the ethanol precipitated curd enzyme of Leuconostoc fallax LH3, we investigated the biotransformation of ginsenoside Rd at different experimental condition to increase transformation. The maximum convertion was supported at 30 $^{\circ}C$ and decreased when temperatures increased. In order to optimize the effect of pH, the curd enzyme was mixed 20 mM sodium phosphate buffer (pH 3.5 to pH 8.0). Ginsenoside Rd was almost hydrolyzed between pH 7.0 and pH 9.0, but not hydrolyzed above pH 10.0. Ginsenoside Rd was hydrolyzed after 24 hrs incubation, but whereas the ginsenoside F2 was appeared from 36 hrs, and all ginsenoside Rd was transformed to F2 after the 60 hrs incubation. Based on this study, the curd enzyme of Leuconostoc fallax LH3 transformed the ginsenoside Rd at the 30$^{\circ}C$ and the pH optimum of 7.0 to 9.0 after the 60 hrs incubation time.
Ji Seul Kim;Jun Ho Kim;Hyeong Chul Jeong;Eun Young Lee
The Journal of Engineering Geology
/
v.33
no.1
/
pp.15-25
/
2023
The effects of forest fires on the activity of microbial communities in topsoil and subsoil were investigated. Samples were collected from Korean forest soils comprising mainly igneous and sedimentary rocks. Analysis of beta-glucosidase, found higher microbial activity in sedimentary rocks than in igneous rocks. Enzyme activity was not observed immediately after fire, but was restored over time. The enzyme activity of subsoil was inhibited by 33~46% compared with that in the topsoil, regardless of soil damage. The effect of fire on the availability of microbial substrate was investigated using EcoPlate. The percentages of average well color development values of damaged and normal topsoil were 52.7~56.8% and 62.3~83.6%, respectively. Forest fires appear to affect the diversity and substrate availability of the subsoil microbial community by accelerating the decomposition of soil organic matter. The Shanon index, representing microbial biodiversity, was high in the topsoil of all samples; it was higher for soil microorganisms in sedimentary rocks than in igneous rocks, and higher in topsoil than in subsoil.
In order to develop the enzymatic hydrolysis system concerned with taste and flavor, strains having the high hydrolyzing activity on the soy protein were selected from some traditional Mejus. Two molds and one bacterium producing enzymes which were different in character of hydrolysis were isolated and identified. Leucine and azodye enzyme activities of both M4 and M5 were relatively high among in the isolated molds. And, leucine enzyme activity of B16 was the lowest in the isolated bacteria. These strains were isolated as microorganisms having a dissimilar hydrolysis pattern on the soy protein by enzymatic reactions. Mold M4 on the culture solid media was mycelium colors of white and its sclerotia colors were changed from white to black. According to the result of slide culture, radial conidial head, subclavate vesicle, conidia of subglobose, stipes of uncolored with smooth walls and metula and phialides were existed. Because M4 was taxonomically similar to the characteristics of Aspergillus oryzae (ahlburg) species, M4 was identified and named as Aspergillus oryzae M4.Mold M5 showed white and black mycelium on the MEA medium. Mold M5 colony exhibited grayish-green color and have long(7 mm) sporangiophores at slide culture. Sporangia became brownish-gray and the wall of larger sporangia was broken to form small collars, and smaller sporangia were fomed continually from large basal membrane. Columella is globose and hyaline, and sporangiospores are ellipsoidal of small diameter$(80\;{\mu}m)$. Because M5 was taxonomically similar to the Mucor circinelloides of zygomycetes, M5 was was identified and named as Mucor circinelloides M5. Bacteria B16 colony was opaque white, circular and lobate, and had rod shaped endospore. B16 was found positive in stain, catalase, ${\beta}-glucosidse$ and V-P tests. B16 was found to utilize D-fructose, ${\alpha}-D-glucose$, maltose, D-mannose, D-raffinose, stachyose and sucrose. By the morphological and physiological results, the characteristics of B16 was thought to correspond to that of Bacillus megaterium. However, fatty acid composition was similar to Paenibacillus marcerans, requiring further study for the definite identification. Accordingly, Bacteria B16 was provisionally classified and named as Bacillus megaterium B16.
Many studies on the changes of the materials in the water-logged paddy soil have been reported, but there will be several problems to apply them on the field soil. The main differences between the method of soil packed in beaker or column tube to that of natural field furrow slice are with or without of the rice root and the effect of water percolation. On the other hand, the mechanism of the water percolation on the changes of material in the natural field furrow slice are gradually understood. The purpose of this experiment is to know the effect of the rice cultivation on the chemical and physical changes of material in the water-logged paddy soil. Obtained results are as follows. 1. The physical and chemical changes on the water-logged paddy soil in the non-planted control-plot were nearly the same as the beaker or column tube experiment, while in the planted plot, slightly altered patterns were observed. 2. The relation between the number of tillers and total cation, $Ca^{{+}{+}}$, $Mg^{{+}{+}}$, Fe and Mn in the leachate showed very high significance. T hisresult showed that the leaching of those cation was promoted by growing of the rice r- of the rice root. 3. On the other hand, the concentration of the potassium, silica and phosphorus in leachates was gradually decreased and that of $NH_4$-N could not detect after the stage of active tillering. These facts revealed that such components were absorbed by rice plant. 4. The highly significant correlation between the number of tillers and the concentration of the total cation, $Ca^{{+}{+}}$, $Mg^{{+}{+}}$, $Fe^{{+}{+}}$, Fe and Mn in the percolated water was observed except that of $Mg^{{+}{+}}$. It was also showed that the rice root promoted the leaching of those cation. 5. The very high significance in the correlation between $HCO_3{^-}$ and the number of tillers indicated that the higher activity of the rice root was, the more $HCO_3{^-}$ concentration in the leachate was increased. 6. The relationship between the $HCO_3{^-}$ and the total cation, $Ca^{{+}{+}}$, $Mg^{{+}{+}}$, $Fe^{{+}{+}}$, Fe and Mn was appeared very highly significant. $HCO_3{^-}$, the metabolite of the rice root, promoted the leaching of $Ca^{{+}{+}}$, $Mg^{{+}{+}}$, $Fe^{{+}{+}}$ and Mn. This fact might be a result that these cations were leached as the form of bicarbonate. 7. The iron in the leachate was the form of $Fe^{{+}{+}}$ and the correlation between $Fe^{{+}{+}}$ and $HCO_3{^-}$ was very highly significant. This result indicated that it seemed to be ferrous bicarbonate when it is leached out. 8. In the rhizosphere, ferrous iron was decreased gradually and the concentration of glucose was as high as 2 to 3 times in comparison with the other parts of the soil. These facts were the same as the previous reports in which rhizosphere was oxidized by the oxigen excreted from the root, and was enriched by the organic matter which was also excreted from the root and accumulated residues of the root. 9. ${\beta}$-Glucosidase and phosphatase activity in the rhizosphere was higher than that of the other parts of the soil. This facts might be attributed to the vigorous activity of microorganism in the rhizosphere where glucose concentration was high. 10. The pH in the leachate of the planted plot was lower than that of control, and the Eh on the planted soil was elevated in the last stage.
Kim, Wan-Gyu;Oh, In-Seok;Yu, Seung-Hun;Park, Jong-Seong
The Korean Journal of Mycology
/
v.12
no.3
/
pp.105-110
/
1984
Seven seed samples of corn obtained from Kangweon Provincial Office of Rural Development, Kerea were tested for seed-borne fungi, and found that all the samples tested were infected with Fusarium moniliforme to an extent of $6.0{\sim}79.5%$. Severely infected seed samples showed poor germination on blotter. Seed component plating showed that the fungus present not only in tip caps, pericarps and endosperms, but also in embryos. Heavy infection of the fungus caused severe seed rot and seedling blight in soil, but the damage was not severe and many plants grew without any symptoms when the seeds with light infection were sown in soil. However the fungus was frequently detected from inside of the stems of healthy looking seedlings. The results indicate that the fungus transmit from seed to plant systemically. In inoculation experiments, the fungus produced stem rots on corn plants of 110 days old. The cultivar of Hwangok 3 was revealed more susceptible to the fungus than that of Suweon 19.
In order to obtain a regulatory mutant strain with high cellulase activity, a newly isolated Penicillium verrculosum, strain F-3 was used as parental strain since it was proved to be an efficient cellulase producer. A number of experiments were conducted to determine the optimum conditions to in-duce mutagenesis and isolate the desirable mutant strains. Out of several restriction compounds tested, 1.5% oxgall was found to be most effective to restrict the colony size by suppressing overgrowth. Derepression of catabolites was employed as a criterion in selecting mutant strains with high cellulase productivity. Production of cellulase by Penicillium venculosum F-3 was suppressed when cultured on the media with more than 1% of glucose or glycerol. It was found that either irradiation with UV light for 19 mins or treatment with nitrosoguanidine at 200$\mu\textrm{g}$/m1 for 60 mins, induced mutagenesis at desired level, when the survival rate of the spore was 0.2% and 48%, respectively. Three mutant strains of F-3, UV-9, UV-10, and NTG-3 that had the highest cellulase productivity were finally selected, based on filter paper degradation rate, size of clearing zone on the screening plate and cellulase activity in the medium containing cellulose powder. When the mutant strains were compared with parental strain F-3, on the KC-M-W medium containing cellulose powder, the filter paper activities of UV-9, UV-10, and NTG-3 were increased by 34%, 55%, and 41%, respectively. However, the assimilation of cellobiose octaacetate by UV-9 or NTG-3 was markedly reduced. When the mutant UV-10 was grown on cellobiose octaacetate medium (CCA-4) in shaking flasks, the cellulase activities of the mutant increased by 20 to 50% compared to the parental strain. Excreation of soluble protein from the mutant also elevated up to 30%. The mutant also constitutively produced both CMCase and $\beta$-glucosidase, though at relatively low level, in the presence of glucose or cellobiose as carbon sources.
Kim, Wha-Sun;Kim, Sang-Soon;Park, Yong-Kon;Seog, Ho-Moon
Korean Journal of Food Science and Technology
/
v.23
no.5
/
pp.554-560
/
1991
The physicochemical properties of Korean yam starches (D. aimadoimo, D. batatas and D. japonica) were investigated. The mean granular size of starches were 23.5 μm for D. aimadoimo, 23.9 μm for D. batatas and 18.2 μm for D. japonica. Amylose content, blue value and water binding capacity was $29{\sim}33%,\;0.42{\sim}0.51%\;and\;109.9{\sim}118.3%$, respectively. The optical transmittance of 0.3% (dry basis) yam starch suspensions were increased at $70{\sim}75^{\circ}C$ and D. japonica showed typical two-step transmittance curve. The swelling power and solubility patterns increased over $60^{\circ}C$, and D. aimadoimo was the highest values. Amylogram patterns of 5% (dry basis) yam starch suspensions, determined by Brabender amylograph, were similar to that of yam flours and the viscosity of D. aimadoimo had 630 BU, which was about 5 times higher than 130 BU for D. batatas and D. japonica. Observation under scanning electron microscope lefted marks of resistance to glucoamylase because these surfaces were similar to the natural granules. In rates of solubiliazation by dimethyl sulfoxide, D. aimadoimo showed the highest value. (3-Amylolysis limits of yam starches and their amylose were $71.8%{\sim}75.5%\;and\;90.2{\sim}92.1%$, respectively. Gel filtration patterns of debranched amylopectin by pullulanase were divided into 3 peaks. The weight ratios of peak III to peak II in yam starches were $2.15%{\sim}2.42%$.
Null mutants generated by targeted gene replacement are frequently used to reveal function of the genes in fungi. However, targeted gene deletions may be difficult to obtain or it may not be applicable, such as in the case of redundant or lethal genes. Constitutive expression system could be an alternative to avoid these difficulties and to provide new platform in fungal functional genomics research. Here we developed a novel platform for functional analysis genes in Magnaporthe oryzae by constitutive expression under a strong promoter. Employing a binary vector (pGOF1), carrying $EF1{\beta}$ promoter, we generated a total of 4,432 transformants by Agrobacterium tumefaciens-mediated transformation. We have analyzed a subset of 54 transformants that have the vector inserted in the promoter region of individual genes, at distances ranging from 44 to 1,479 bp. These transformants showed increased transcript levels of the genes that are found immediately adjacent to the vector, compared to those of wild type. Ten transformants showed higher levels of expression relative to the wild type not only in mycelial stage but also during infection-related development. Two transformants that T-DNA was inserted in the promotor regions of putative lethal genes, MoRPT4 and MoDBP5, showed decreased conidiation and pathogenicity, respectively. We also characterized two transformants that T-DNA was inserted in functionally redundant genes encoding alpha-glucosidase and alpha-mannosidase. These transformants also showed decreased mycelial growth and pathogenicity, implying successful application of this platform in functional analysis of the genes. Our data also demonstrated that comparative phenotypic analysis under over-expression and suppression of gene expression could prove a highly efficient system for functional analysis of the genes. Our over-expressed transformants library would be a valuable resource for functional characterization of the redundant or lethal genes in M. oryzae and this system may be applicable in other fungi.
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