Fruit bodies similar to the Phellinus sp. residing on the mulberry were collected at Yang-yang in Kang-won-do province and one strain of Phellinus sp. was isolated from the fruit bodies. For mass production of the isolated mycelia in a submerged culture, the culture conditions, medium composition, and the effect of various culture systems on the mycelial growth, were investigated. The morphological characteristics of the fruit body were as follows: covered with blackish to black and rough, lower surface with yellowish-brown to dull-brown and smooth, 5-7 cm thick and hard woody. Also, the pure cultured mycelia showed yellowish-brown color, capability of purplish-brown pigment production on the PDA plate media, no-formation of clamp-connection, much binding branch, and enzyme activities such as laccase, tyrosinase and peroxidase. Therefore, pure cultured strain was identified to be Phellinus sp. In the flask culture, the optimum culture conditions for the mycelial production were obtained after cultivation of 8 days at inoculum level of 5%(v/v), media volume of 70 mL, 150 rpm, initial pH 6, and temperature of $30^{\circ}C$. Optimum medium composition from the response surface analysis were determined to be glucose 12.12 g/L, sucrose 12.12 g/L, yeast extract 11.15 g/L, malt extract 11.15 g/L, $KH_2PO_4$ 0.855 g/L and $CaCl_2$ 0.855 g/L. The production of the mycelia after 4 and 8 days of cultivation was 1.95 and 9.89 g/L, respectively. The maximum specific growth rate and productivity were $0.020\;hr^{-1}$ and 1.25 g/L/day, respectively. Among the three different culture systems for the growth of mycelia, the maximum mycelial dry weight of 7.5 g/L was obtained after cultivation of 4 days in the air-lift fermentor under aeration rate of 2.5 vvm. The maximum specific growth rate and productivity were $0.033\;hr^{-1}$ and 1.9 g/L/day, respectively, which were about 1.7 and 4.2 times higher than those of flask culture.
This study was conducted to evaluate the growth characteristics of lettuce (Lactuca sativa L.) as affected by artificial light sources and different growing media in a closed-type plant production system (CPPS). The lettuce seeds were sown in the 128-cell plug tray filled with 5 different growing media such as urethane sponge (US), rock-wool (RW), Q-plug (QP), TP-S2 (TP) and PU-7B (PU). The germination rate of lettuce seeds was examined during 12 days after sowing. On the 13 days after sowing, the lettuce seedlings were transplanted in a CPPS with temperature $25{\pm}1^{\circ}C$ and nutrient solution (EC $2.0dS{\cdot}m^{-1}$, pH 6.5) using recirculating deep floating technique system. The light sources were set with FL (fluorescent lamps) and combined RB LEDs (red : blue = 7 : 3) with $150{\pm}10{\mu}mol{\cdot}m^{-2}{\cdot}s^{-1}$ PPFD and a photoperiod of 14/10 hours (light/dark). The initial germination rate of lettuce was the highest in TP. The final germination and mean daily germination were the significantly highest in RW, QP and TP. The plant height, leaf length, leaf width, leaf area, and fresh and dry weights of shoot were the greatest in QP irradiated with RB LED. The number of leaves, fresh and dry weights of root and SPAD were the greatest in QP and TP irradiated with RB LED. The root length was the longest in TP irradiated with RB LED. Therefore, these results indicate that RB LED was effective for the growth of lettuce and it was also found that the QP and TP were effective for the germination and growth of lettuce in a CPPS. In addition, we confirmed the applicability of the newly developed growing medium TP for the lettuce production in a CPPS.
The aim of this work was to establish the optimal conditions for the production of cellobiase by a marine bacterium, Cellulophaga lytica LBH-14, using response-surface methodology (RSM). The optimal conditions of rice bran, ammonium chloride, and the initial pH of the medium for cell growth were 100.0 g/l, 5.00 g/l, and 7.0, respectively, whereas those for the production of cellobiase were 91.1 g/l, 9.02 g/l, and 6.6, respectively. The optimal concentrations of $K_2HPO_4$, NaCl, $MgSO_4{\cdot}_{7H2}O$, and $(NH_4)_2SO_4$ for cell growth were 6.25, 0.62, 0.28, and 0.42 g/l, respectively, whereas those for the production of cellobiase were 4.46, 0.36, 0.27, and 0.73 g/l, respectively. The optimal temperatures for cell growth and for the production of cellobiase by C. lytica LBH-14 were 35 and $25^{\circ}C$, respectively. The maximal production of cellobiase in a 100 L bioreactor under optimized conditions in this study was 92.3 U/ml, which was 5.4 times higher than that before optimization. In this study, rice bran and ammonium chloride were developed as carbon and nitrogen sources for the production of cellobiase by C. lytica LBH-14. The time for the production of cellobiase by the marine bacterium with submerged fermentations was reduced from 7 to 3 days, which resulted in enhanced productivity of cellobiase and a decrease in its production cost. This study found that the optimal conditions for the production of cellobiase were different from those of CMCase by C. lytica LBH-14.
Kim, Joo-Hyun;Lee, Yung-Kyoon;Kim, Hae-Won;Kim, Myung-Suk;Park, Chan-Woong;Lim, Jung-Kyoo
The Korean Journal of Pharmacology
/
v.18
no.2
/
pp.79-87
/
1982
Higenamine(dl-demethylcoclaurine, dl-1-(4-hydroxybenzyl)-6,7-dihydroxy-1,2,3,4-tetrah-ydroisoquinoline hydrochloride), which has recently been isolated from Aconite root by Drs. Kosuge and Yokota, has known to be the main cardiotonic component of the Aconite root. The present study was undertaken to investigate the effects of Higenamine on the calcium binding and release and ATPase activity of fragmented cardiac sarcoplasmic reticulum under in vitro condition. The calcium binding and release of sarcoplasmic reticulum were measured by using the double-beam spectrophotometer and the calcium sensitive dye, murexide. In the presence of $10^{-4}{\sim}5{\times}10^{-3}M$ of Higenamine, the maximal calcium binding and the initial binding rate of porcine cardiac sarcoplasmic reticulum were inhibited dose dependently by up to 43%. However, the calcium release from cardiac sarcoplasmic reticulum, which was loaded with $Ca^{++}(50{\mu}M)$, was stimulated in dose dependent manner. When incubated in the medium of 20 mM Tris-maleate(pH 7.0), 100 mM KCl, 10 mM $MgCl_2,\;0.05mM\;CaCl_2\;and\;0.014{\sim}1\;mM\;Tris-ATP\;at\;30^{\circ}C$ in the presence of Higenamine $(10^{-4}{\sim}5{\times}10^{-3}M)$, both $Ca^{++}-and\;Mg^{++}-ATPase$ of sarcoplasmic reticulum were inhibited non-competitively by Higenamine and values of $K_i$ were 4.896 mM and 6.875 mM respectively. It is suggested from the above findings that the cardiotonic effects of Higenamine might be partially explained by the inhibition of calcium binding and the stimulation of calcium release from the sarcoplasimic reticulum which may increase the free intracellular calcium that is available in the contraction of the cardiac muscle fiber.
Bio-secure culture of olive flounder Paralichthys olivaceus in the IBK (Intensive Bioproduction Korean) recirculating system with dry pellet was tested for 6 months. The IBK system consists of 12 rearing tanks, 6 sedimentation tanks. 4-sectioned submerged biofilter chamber and channels. The size of each rearing tank was 3m in diameter and 1m in depth. The size of each biofilter chamber was $3.1\times3.3\times2.0$ m (D) and was filled with corrugated plastic plates as a biofilter medium. Total surface area of the biofilter was 3,789.7 $m^2$ Water was circulated by one of two vertical axial pump and circulating rate was about 34 times per day. A UV light sterilizer was used to treat inlet sea water with the flow rate of 4 ton/hr. All fish were treated with 150 ppm formalin 3 times with 5 day interval before stocking. It took 60 days for 'conditioning' the biofilter with the stocking density of 4.5 kg of fish $m^2$. The concentrations of ammonium-nitrogen, nitrite-nitrogen and nitrate-nitrogen in the system remained at the range of 0.096-0.315 mg/L, 0.015-0.504 mg/L, and 2.530-39.517 mg/L, respectively. Water temperature fluctuated from 17.5 to 25.1$^{\circ}C$ and salinity was from 30.1 to 33.5 ppt during rearing period. The fish grew from the average weight of 615.2 g to 1,201.1 g for 180 days. Initial and final fish densities were 8.4 and 15.9$kg/m^2$, Survival rate was 97.1 %. Neither parasites nor noticeable diseases was observed during the raring period even Vibrio spp. were detected from some fish in the system.
Kim, Yi-Joon;Cao, Wa;Lee, Yu-Jeong;Lee, Sang-Un;Jeong, Jeong-Han;Lee, Jin-Woo
Journal of Life Science
/
v.22
no.10
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pp.1295-1306
/
2012
A microorganism producing carboxymethylcellulase (CMCase) was isolated from seawater and identified as Bacillus atrophaeus. This species was designated as B. atrophaeus LBH-18 based on its evolutionary distance and the phylogenetic tree resulting from 16S rDNA sequencing and the neighbor-joining method. The optimal conditions for rice bran (68.1 g/l), peptone (9.1 g/l), and initial pH (7.0) of the medium for cell growth was determined by Design Expert Software based on the response surface method; conditions for production of CMCase were 55.2 g/l, 6.6 g/l, and 7.1, respectively. The optimal temperature for cell growth and the production of CMCase by B. atrophaeus LBH-18 was $30^{\circ}C$. The optimal conditions of agitation speed and aeration rate for cell growth in a 7-l bioreactor were 324 rpm and 0.9 vvm, respectively, whereas those for production of CMCase were 343 rpm and 0.6 vvm, respectively. The optimal inner pressure for cell growth and production of CMCase in a 100-l bioreactor was 0.06 MPa. Maximal production of CMCase under optimal conditions in a 100-l bioreactor was 127.5 U/ml, which was 1.32 times higher than that without an inner pressure. In this study, rice bran was developed as a carbon source for industrial scale production of CMCase by B. atrophaeus LBH-18. Reduced time for the production of CMCase from 7 to 10 days to 3 days by using a bacterial strain with submerged fermentation also resulted in increased productivity of CMCase and a decrease in its production cost.
The aim of this study was to isolate and identify marine bacterium with anti-methicillin-resistant Staphylococcus aureus (MRSA) activity, and to purify the anti-MRSA compound, as well as to determine its activity and synergistic effects. Among the marine bacteria isolated in this study, the YJ-1 isolate had the strongest anti-MRSA activity. The YJ-1 isolate was identified on the basis of its biochemical characteristics and an analysis of 16S rRNA gene sequences. The YJ-1 isolate showed over 99.2% homology with Pseudomonas stutzeri, and was designated as a Pseudomonas sp. YJ-1. The optimal culture conditions were $25^{\circ}C$ and initial pH 7.0. For the purification of the anti-MRSA compounds, the YJ-1 was cultured in Pa PES-II medium, and the culture filtrates were extracted by ethyl acetate, hexane, and 80% MeOH. The 80% MeOH fraction was separated by a $C_{18}$ ODS column, silica gel chromatography and a reverse phase HPLC, to yield three anti-MRSA agents, the MR1, MR2, and MR3 compounds. When the MR1 compound of $250{\mu}g\;mL^{-1}$ concentration was applied to the MRSA cells, over 95% of bacterial cells was killed within 48 hr. Compared with vancomycin and ampicillin, the MR1 compound showed significant anti-MRSA activity. In addition, the anti-MRSA activity was increased by dose and time dependent manners. Furthermore, the combination of an MR1 compound with vancomycin produced a more rapid decrease in the MRSA cells than did the MR1 compound alone. Taken together, our results suggest that the Pseudomonas sp. YJ-1 and its anti-MRSA compounds could be employed as a natural antibacterial agent in MRSA infections.
1. Isolation and identification of amylase-producing bacteria. The powerful strain A-12 and S-8 were respectively isolated from air and soil after screening a large number of amylase-producing bacteria. Their bacterial characteristics have been investigated and it has been found that all characteristics of strain A-12 and S-8 are similar to Bac. subtilis of Bergey's manual except for the acid formation from a few carbohydrates and the citrate utilization, i.e., the strain A-12 shows negative in the citrate utilization, and the acid formation from arabinose and xylose, S-8 shows negative in the acid formation from xylose. 2. Amylase production by Liquid cultures with solid materials. Several conditions for amylase production by strain A-12 in stationary cultures have been studied. The results obtained are as follows. (1) The optimum conditions are:temperature $35^{\circ}C$, initial pH 6.5 to 7.0 and incubation time 3 to 4 days. (2) The amylase production is not affected by the preservation period of the stock cultures. (3) Among the various solid material, the defatted soy bean is found to be the best for t1e amylase production. However, the alkali treatment of the defatted soy bean gives no effect contrary to the cage of defatted rape seed. The addition of soluble starch to the alkali extract of defatted soy bean shows the increased amylase production. (4) Up to 1% addition of ethanol to carbon dificient media gives the improved amylase production, whereas the above effect is not found in the case of carbon rich media. (5) The amylase production can be increased 2.5 times when 10% of defatted soy bean is admixed to cheaply available wheat bran. (6) The excellent effect is found for amylase production when 20% of wheat bran is admixed to defatted dry milk which is a poor medium. The activity is found to be $D^{40^{\circ}}_{30'}$ 7,000(L.S.V. 1,800) in 10% medium. (7) No significant effect is observed due to the addition of various inorganic salts. 3. Amylase production by solid cultures. Several conditions for amylase production by strain A-12 in wheat bran cultures have been studied and the results obtained are as follows. (1) The optimum conditions: are temperature $33^{\circ}C$, incubation lime 2 days, water content added 150 to 175% and the thickness of the medium 1.5cm, The activity is found to be $D^{40^{\circ}}_{30'}$ 36,000(L.S.V. 15,000) (2) No significant effect is found in the case of the additions of various organic and inorganic substances.
Journal of the Korean Society for Marine Environment & Energy
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v.13
no.4
/
pp.288-295
/
2010
Metal exposure experiments using polychaete (Perinereis nuntia) as a bio-indicator of trace metals contamination were conducted to evaluate the bioaccumulation and the biomarkers responses such as metallothionein-like protein (MTLPs) and glutathione S-transferase (GST) which was simultaneously exposed to Cadmium (Cd) and Copper (Cu). Cu and Cd concentrations in polychaete were enhanced with increasing exposure time and their concentrations of aqueous medium. Initial accumulation of Cd was higher than that of Cu. Our results showed that the bioaccumulation of Cu and Cd were prohibited, especially at higher Cu levels, suggesting the different cellular uptake mechanisms when Cu and Cd are co-exist. Net accumulation rate of Cu was declined with exposure time but it did not show any significant change for Cd. Although the highest MTLPs concentration was observed at 6 hr of exposure time, it did not show any significant change related to exposure times and metals concentrations. An increase of GST activity tended to increase as a function of exposure time and metals concentrations. And GST activities in P. nuntia have similar tendency with bioconcentration factors in high concentration of Cu (treatment group IV) at post 24 h of exposure. Our results provide new information of the bioaccumulation and biomarker responses to understand the effects of co-existing contaminants (Cu and Cd) using polychaete. Further studies are required to elucidate the bioaccumulation and biomarkers responses for various contaminants.
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