The effects of carbon sources for exopolysaccharide production during batch cultivation of an Enterobacter sp. isolated from the composter were investigated. The highest amount of exopolysaccharide was obtained when lactose was used as carbon source. Lactose in medium was converted into glucose and galactose. Glucose was metabolized fast and was completely consumed, but about $20\%$ of lactose was accumulated as galactose. On the other hand, enzyme activity was about $350\~450$ unit with the increase of lactose concentration. Thus, it was considered that the exopolysaccharide might be produced in the course of that lactose was hydrolyzed into glucose and galactose by $\beta-galactosidase$ with respect to that enzyme activity on lactose hydrolysis was accorded to the exopolysaccharide production. When glucose and galactose were added to lactose medium, respectively, it could be considered that glucose was as a repressor and galactose was as a inducer for $\beta-galactosidase$ synthesis even though the mechanisms were not elucidated. The increase of lactose concentration was almost ineffective to the specific growth rate $(0.133\~0.151\;hr^[-1})$ but showed the difference in the biomass content. The higher carbon source concentration, the more residual sugar remained. It was assumed that the optimum lactose concentration for exopolysaccharide production was $30\~70g/L.$ On the other hand, it was considered that the nitrogen acted as growth limiting nutrients to the cell growth. In the cases of 30 and 70 g/L of the fixed carbon concentrations, the increase of the nitrogen sources concentration caused a remarkable increase within the range of $0.059\~0.225\;hr^{-1}$ and $0.141\~0.237hr^{-1}$ of the specific growth rate, respectively, while there was no significant difference in biomass.
Molecular and cultivation techniques were used to characterize the bacterial communities of biobead reactor biofilms in a sewage treatment plant to which an Aerated Up-Flow Biobead process was applied. With this biobead process, the monthly average values of various chemical parameters in the effluent were generally kept under the regulation limits of the effluent quality of the sewage treatment plant during the operation period. Most probable number (MPN) analysis revealed that the population of denitrifying bacteria was abundant in the biobead #1 reactor, denitrifying and nitrifying bacteria coexisted in the biobead #2 reactor, and nitrifying bacteria prevailed over denitrifying bacteria in the biobead #3 reactor. The results of the MPN test suggested that the biobead #2 reactor was a transition zone leading to acclimated nitrifying biofilms in the biobead #3 reactor. Phylogenetic analysis of 16S rDNA sequences cloned from biofilms showed that the biobead #1 reactor, which received a high organic loading rate, had much diverse microorganisms, whereas the biobead #2 and #3 reactors were dominated by the members of Proteobacteria. DGGE analysis with the ammonia monooxygenase (amoA) gene supported the observation from the MPN test that the biofilms of September were fully developed and specialized for nitrification in the biobead reactor #3. All of the DNA sequences of the amoA DGGE bands were very similar to the sequence of the amoA gene of Nitrosomonas species, the presence of which is typical in the biological aerated filters. The results of this study showed that organic and inorganic nutrients were efficiently removed by both denitrifying microbial populations in the anaerobic tank and heterotrophic and nitrifying bacterial biofilms well-formed in the three functional biobead reactors in the Aerated Up-Flow Biobead process.
Kim, Kwan-Chul;Lee, Hyeok-Won;Lee, Hong-Won;Choo, Soo-Jin;Yoo, Ick-Dong;Ha, Byung-Jo
Microbiology and Biotechnology Letters
/
v.42
no.2
/
pp.194-201
/
2014
Clitocybin A is a novel anti-wrinkle cosmetic agent produced by the strain from a Korean native mushroom Clitocybe aurantiaca. In this study, fermentation, extraction, and purification conditions for a large scale production of clitocybin A were optimized, and its cytotoxicity and inhibition activity on the expression of matrix metalloproteinase-1 (MMP-1) were characterized. The mass production of anti-wrinkle agent was achieved according to the 300 L fermentation process with a fed-batch cultivation using the modified yeast-maltose (YM) broth, and a total of 12.5 kg of cell mass was obtained in a 120 L culture broth for 14 days. After extraction and purification, clitocybin A was identified by HPLC. The cytotoxicity of clitocybin A was examined by the MTT assay. When assayed at 100 and 200 ${\mu}g/ml$ concentrations, clitocybin A showed no cytotoxicity, demonstrating safety. The inhibition activity of clitocybin A on the expression of MMP-1 was examined against UV irradiation. Oleanolic acid (control group) showed a relatively low MMP-1 inhibiting activity (ca. 16.7%) at 10 ${\mu}g/ml$ and showed increased cytotoxicity at higher concentrations. In contrast, clitocybin A showed no cytotoxicity at 100 ${\mu}g/ml$, and exhibited a relatively high MMP-1-inhibiting activity (33.1%). These findings indicate that clitocybin A may be a safe and effective anti-wrinkle agent for use in functional cosmetics.
This experiment was carried out to investigate the effects of limited nutrient supply during 21 days before and after pollination stage on the growth, fruit quality and nutrient uptake of muskmelon in rockwool culture. Muskmelon, cv. Earl's Favorite seeds sowed on rockwool cube and transplanted on rockwool slab($90\times15\times7.5cm$) when 2 to 3 true leaf appeared on Sep. 6, 1991. Three kinds of nutrient composition recommended by Shizuoka university, combinated with the composition of Otsuka house A and composition Shizuoka III. One half of calcium nitrate(Ca(NO$_3$)$_2$.4$H_2O$) for limiting nitrogen supply during 21 days was treated and then fertigated the nutrient composition recommended by Shizuoka university up to harvest time. Trickling nozzles(Netafim Co. Israel) were used for fertigation of nutrient solution and noncirculating system was employed. Temperature was maintained $18^{\circ}C$ in night but 23 to $25^{\circ}C$ for 10 days after pollination for softening the fruit. The drainage ratio of nutrient solution was adjusted 20 to 30 percent. Fertigated and drained amount, and the pH and EC of nutrient solution were recorded. The concentrations of mineral elements including N, P, K, Ca, and Mg were analyzed and compared among treatments. In both autumn and winter cultivation, the limitation of nutrient supply by adjustment of nutrient composition(NO$_3$-N : 8meㆍ$\ell^{-1}$) caused the nutrient deficiency in muskmelon plant due to the limited nutrient supply. After pollination nutrient limitation by the lowering the nitrate retarded the over thickening of upper leaves of muskmelon but plant height and fresh weight of fruit were higher in the plot of nonlimited nutrient supply. The phenomena were attributed to the differences of the amount of nutrient uptake due to the limited time of nutrient solution, duration of nutrient supply and concentration of nutrient solution. These results suggested that increasing nutrient supply in the pollination stage was favorable for better appearance of fruit and improving fruit quality. Further trials would be required for the incre-ment of sugar degree of muskmelon grown in rockwool.
Streptomyces spp. isolated from turfgrass rhizosphere and tested for their response to large-patch control fungicides. The tested fungicides were actually used in golf course or turfgrass cultivation to prevent large-patch disease. Tolerance to 3 triazole group of the strains was the highest to the PR fungicide, and following the SR fungicide, whereas the isolated strains were no tolerance to HR fungicide. Tolerances to three kind of Strobilurin group were similar for the all of the tested Streptomyces spp.. Growth and sporulation of the all strain was normal in CB and AP fungicide treatments. However no spore formulated in double concentration. Strains, tolerance to acetanilide fungicides, appeared that KT fungicide tolerance was higher than MK fungicide. The selected strains showed strong tolerance against AT fungicide but have no tolerance to ATR fungicides. In conclusion, the bacterial strains showed tolerance against 1 carbamate, 1 organophosphate and 1 cyanopyrrole group, while have no tolerance against two mixture formulations (1 Quinone + Strobilurin and 1 Imidazole + Triazole).
Some factors of wounding methods, solidifying agents, origin of leaf explants, cone. of acetosyring-one, and MES affecting regeneration and transformation by Agrobacterium tumefaciens were investigated to establish an efficient transformation system of apple. Wounding by cutting the leaves merely showed the tendency of regeneration and transformation with higher efficiency compared with that of wounding by non-traumatic forcep when carrying out co-cultivation for three days after bacterial inoculation. While examining the solidifying agents of medium with the combination of agar (A)+Gelrit $e^{ }$ (G) in g. $L^{-1}$ , the higher concentration of Gelrit $e^{ }$ increased the efficiency of regeneration. However, there was no difference in the efficiency of transformation from the treatments of 2.5 G, 3.5 A+1.2 G, and 7.0 G. The origin of leaf explants showed no difference statistically in the efficiency of regeneration and transformation, but that from the shoots of proliferation medium showed the tendency with higher efficiency. The concentration of above 0.1 mM acetosyringon had an increase in the efficiency of regeneration and transformation and the concentration of 0.15 mM had the highest efficiency of transformation in the treatment of acetosyringone with different concentration. There was no effect of MES on regeneration and transformation.ion.
To develop a selection system for regenerating plants from transformed tissues, effects of four antibiotics (kanamycin, hygromycin, carbenicillin, cefotaxime) and herbicide (phosphinotricin) on shoot regeneration from cotyledon and hypocotyl explants of Chinese cabbage (Brassica campestris L. ssp. pekinensis) were studied. For cotyledon, shoot induction was not significantly affected by kanamycin at $1mg{\cdot}L^{-1}$, but the number of shoots formed was significantly reduced at $2mg{\cdot}L^{-1}$, and no shoots were regenerated from any explants at $6mg{\cdot}L^{-1}$ or higher. Hypocotyl explants showed similar result as cotyledon. Kanamycin at $7mg{\cdot}L^{-1}$ may be adequate for selecting Chinese cabbage transformants. Hygromycin at $4mg{\cdot}L^{-1}$ or higher completely inhibited the growth and shoot regeneration of Chinese cabbage explants. Therefore, resistance gene to hygromycin may also be used as a selective marker for Chinese cabbage transformation. Carbenicillin and cefotaxime, the cephalosporin type of antibiotics, had little effect on shoot regeneration of Chinese cabbage explants. Since carbenicillin and cefotaxime have low toxicity to Chinese cabbage, they are suitable for use in tissue culture to eliminate Agrobacterium in transformation experiments after co-cultivation. Shoot regeneration from cotyledon and hypocotyl explants was significantly reduced in presence of $1mg{\cdot}L^{-1}$ phosphinotricin (PPT) and completely inhibited by $2mg{\cdot}L^{-1}$ or higher. PPT, same as antibiotics, may also be used to select transformed cells. Since Chinese cabbage is known to be recalcitrant to in vitro shoot regeneration compared to other Brassica species, even though lower levels of selectable markers result in more transformants but simultaneously allow more untransformed escapes to develop, lower levels of antibiotics and herbicides could be successfully used as a selectable marker to reduce selection pressure.
Chang, Bo Yoon;Han, Ji Hye;Cha, Bum-Suk;Ann, Sung-Ho;Kim, Sung Yeon
Journal of Food Hygiene and Safety
/
v.30
no.3
/
pp.295-301
/
2015
The functions of probiotics, particularly Lactic acid bacteria, have been studied in a range of human diseases, including cancer, infectious diseases, gastrointestinal disorders, and allergies. Among the many benefits associated with the consumption of probiotics, modulation of immune activity has received the most attention. This study aimed at investigating the improved immune stimulatory and stability of L. plantarum when cultivated on modified basal media supplemented with the Undaria pinnatifida co-cultured with L. plantarum. An in vitro test showed that U. pinnatifida media cultured L. plantarum is strong enough to survive in the gastric juice (gastric and bile acid). Mouse macrophage-derived cell lines RAW 264.7 was used to measured immune stimulating activity of L. plantarum. When U. pinnatifida media cultured by L. plantarum was NO and $TNF-{\alpha}$ production is significantly increased compared to basal media cultured L. plantarum. These results show that U. pinnatifida could be applied for a component for cultivation of L. plantarum. This optimized U. pinnatifida medium can be used the improving of stability and immune function on production of probiotics.
Growers in plug seedling production think that root media in which rockwool is a component has given rise to several environmental problems. Therefore, the demand for new materials as a substitute for rockwool has been increased. This study examined the possibility of cultivation of tomato plug seedlings using a newly developed growing medium with phenolic foam. Plug seeds of tomato cultivar 'Madison' were sown in four pellet-type growing media: Grodan rockwool (GRW), UR rockwool (URW), phenolic foam LC (LC) or phenolic foam LC-lite (LC-lite). Then, the seedlings were transplanted to the four cube-type growing media 19 days after sowing. Seeds were germinated in a growth chamber ($25{\pm}2^{\circ}C$, 80% relative humidity, and dark) for 4 days and then the seedlings were grown with a nutrient solution supplied by an overhead irrigation system in a greenhouse. Plant height, number of leaves, leaf area, and fresh or dry weight of tomato seedlings were the greatest for the seedlings transplanted to URW cube media after being grown on LC-lite pellets. Root grade was the greatest for the seedlings transplanted on LC or LC-lite cube media after being grown on LC pellets. Chemical properties of all media tested for tomato growth were maintained within a stable range, while physical properties of URW showed high values in container capacity, air space, and total porosity. These results demonstrated that the phenolic foam media were effective for seedling growth and can substitute for rockwool as a root medium.
Kim, Yu-Jin;Jang, Seo-Jung;Park, Jung-Min;Kim, Chang-Uk;Park, Young-Seo
Food Engineering Progress
/
v.14
no.1
/
pp.65-74
/
2010
Culture conditions of L. plantarum TJ-LP-002, the garlic resistant strain isolated from pakimchi (green onion kimchi), were investigated for the use of feed additives. Acetic acid, citric acid, lactic acid, and tartaric acid were detected in the culture supernatant, and especially the concentrations of lactic acid and acetic acid significantly increased during cultivation. The antimicrobial activity of L. plantarum TJ-LP-002 was not affected by proteases, calatase or cellulase, which showed that the antimicrobial activity might be due to the production of acids rather than proteinaceous antimicrobial substances. L. plantarum TJ-LP-002 was resistant to neomycin sulfate, spectinomycin dihydrochloride, and lincomycin hydrochloride, sensitive to streptomycin sulfate, and intermediate resistant to ampicillin trihydrate, chloramphenicol, erythromycin, tetracycline hydrochloride, and kanamycin sulfate. The optimum initial pH of medium, fermentation temperature and time for the cell growth and antibacterial activity were pH 7.0, 30${^{\circ}C}$ and 24hr, respectively. The optimal composition of culture medium for the cell growth and antimicrobial activity was 3%(w/v) glucose as a carbon source, 3%(w/v) yeast extract as a nitrogen source, and manganese sulfate and ammonium citrate as inorganic salts. The combinatorial supplementation of these inorganic salts, rather than sole addition as an inorganic salt, resulted in better antibacterial activity.
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