Cyclodextrin glucanotransferase (CGTase) of Bacillus sp. isolated from soil was purified and its enzymological characteristics were investigated. It was found that the production of CGTase reached to the maximum when the strain was cultured in the broth containing 0.1 % albumin, 2% $NH_4Cl$, 2% soluble starch and 0.2% $NH_2PO_4$ for 72 hrs at $37^{\circ}C$. The purity of CGTase was increased by 9.7 folds through purification procedures by the following column chromatography DEAE-cellulose ion exchange chromatography and Sephadex G-100, G-150 gel filtration and its specific activity was 528.0 unit/mg. The optimum pH and temperature for the CGTase activity were 8.0 and $80^{\circ}C$, respectively. The enzyme was stable in pH $8.0{\sim}11.0$ at $60{\sim}80^{\circ}C$. The activity of purified enzyme was inhibited by $Pb^{2+},\;Hg^{2+}$ and $Zn^{2+}$. When CGTase was treated with each 20.5 unit, 41 unit, 205 unit and 410 unit to investigate the transglucosylation to stevioside by purified cyclodextrin glucanotransferase, transglucosylation rate to stevioside was 74.9%, 75.7%, 68.7% and 57.9%. Brown effect was observed above the concentration amounting to 205 unit of our CGTase.
Fusarium head blight (FHB) is a severe disease problem that affects the quality and yield of barley grain. The evaluation of FHB resistance is difficult because environmental conditions greatly influence FHB infection and development. The objectives of this study were to: 1) establish an efficient screening method for selecting resistant barley to FHB, 2) compare FHB severity between the cut-spike method and pot-plant method for development of mass screening, and 3) estimate FHB resistance for barley germplasms. Barley cultivars and lines were evaluated for reaction to FHB in controlled-greenhouse condition. Spikes were spray-inoculated with a suspension $(5.0\times10^5\;macroconidia\;mL^{-1})$ of Fusarium graminearum SCK-O4 strain, and then kept in a greenhouse at $18-25^{\circ}C$ with $80-100\%$ relative humidity. Inoculation were employed at 3 different heading growth stages (heading date, three days after heading, and five days after heading). The inoculation was performed in 2 consecutive days in order to avoid escapes. The inoculated plants were maintained in the greenhouse at 4 different free moisture periods (1, 3, 5, and 7 days). The percentage of FHB severity was scored from 0 to 9 according to the rate of infected kernels per spike, and three spikes were evaluated per replication with 3 replicates. There were significant differences of FHB severity depending on the different free moisture periods, but not by the inoculation at different heading stages. The optimum evaluation point of FHB severity in the greenhouse condition was on the 7th day under free moisture condition after inoculation at the heading date. Infection level in cut-spike method highly correlated with that in pot-plant method. This suggested that cut-spike method is useful in evaluating of FHB resistance in barley. Six cultivars, such as Jinkwang, Buheung, Atahualpha 92, Chevron-b, Gobernadora-d, and MNBrite-c, were selected as resistant varieties to FHB. Correlation coefficient for the FHB severity evaluated by the pot-plant method between two seasons was 0.794, indicating the stability and accuracy of the screening method.
This study was conducted to detect the serum antibodies in the experimentally toxoplasma infected dogs and street dogs by use the of an enzyme-linked immunosorbent assay (ELISA). And this test was performed on the polystylene microplate by coating with the tachyzoites soluble antigen of T gondii (RH strain), incubated with sera diluted then, added with HPO-conjugated rabbit anti-dog IgG and o-phenylenediamine used as a substrate. Tachyzoites of T gondii harvested from mouse peritoneal cavity were purified by 30, 40 and 50% Percoll density gradient centrifugation and used as the source of antigen. The results obtained were summarized as follows; 1. The highest ratio of positive to negative (P/N ratio) was obtained at the level of $l{\mu}g/ml$ protein concentration of antigen with the 1/4000 dilution of the conjugate measured by checker-board titration. It was regarded as the optimum concentration of the antigen and conjugate. 2. Cut-off value in this IgG ELISA was 0.375 that was determined by mean absorbance (at 492nm) of IFA negative serum added with the dauble value of the standard deviation $(mean{\pm}2S.D.)$. 3. Serum ELISA IgG antibodies to T gondii in the exyerimentally infected dogs were detected firstly at the Week 3 after inoculation and the highest titer was recognized at the Week 4, 5 and 6 after inoculation. 4. Stability of the antigen absorbed in the microplates that were preserved at $4^{\circ}C$ and $-25^{\circ}C$ separately were prolonged up to 3 weeks and 10 weeks at $4^{\circ}C$ and $-25^{\circ}C$, respectively. However the reproducibility was not reliable after the preservation of 4 weeks and longer. 5. Positive rate of the specific antibodies in 312 test sera was 28.5% and there was no significant differences between the male (27.8%) and female (29.5%), respectively. 6. The IgG ELISA was proved to be a specific procedure for the detection of antibodies to canine toxoplasma infection and also evaluated as a screening test for the large scale of test samples in laboratory.
Rammed Aggregate Pier (RAP) method is intermediate foundation between deep and shallow foundation, and it has been built in world wide. RAP represents a relatively new method that has grown steadily over 19 years since Geopier of USA developed this revolutionary method in 1989. The investigation and research in domestic is not accomplished. In this paper, the examined details of different spacing of piles, bearing capacities, respectively, conclude with recommendations on how RAP can be used in future needs. This documentation further provides comparisons of the laboratory test results which were obtained from changing the spacing of piles, namely installed rammed aggregate pier. Laboratory model test was administered in a sand box. Strain control test was conducted to determine the bearing capacities of the piers; 20 mm, 30 mm and 40 mm RAP in diameter using drilling equipment to make holes were installed in sand at initial relative densities of 40%. By comparing different spacing of piles, in this experiment, piles are spaced structually span, form a ring shape, narrowing the distance of each other, to the center. the result shows that as diameter of pier is bigger in diameter, bearing capacity also dramatically increased due to raised stiffness. Also, as the space between each piers was closed, the settlement rate of soil was decreased significantly. From the test results, as the space between each piles were getting closer, it allows greater chances to have more resistance to deformation, and shows more improved stability of structures. After from the verification work which is continuous leads the accumulation of the site measuring data which is various, and bearing capacity and the settlement is a plan where the research will be advanced for optimum installed RAP.
The purification and characteristics of the biosynthesis enzyme of vitamin C from microorganisms related with kimchi fermentation were investigated to define vitamin C biosynthetic pathways in yeast. A yeast strain (Pichia onychis 16-4) which synthesizes vitamin C with galacturonic acid as substrate at high rate was isolated from kimchi. The enzyme $L-galactono-{\gamma}-lactone$ oxidase isolated from the yeast was purified and characterized. The specific activity of the crude enzyme was 7.26 unit/mg protein, which increased to 4,698 unit/mg protein with a chromatography of Sephacryl S-200HR; indicating a 647.1-fold level of purification. The molecular weights of the dissociated enzymes were estimated to be 31,000, 39,000, and 50,000 KD. Among the substrates tested, $L-galactono-{\gamma}-lactone$ was the most effective. The enzyme showed optimum activity ah pH 7.8 and 35c. The purified enzyme uses $O_2$ as the electron acceptor for oxidation of $L-galactono-{\gamma}-lactone$.
In order to develop a new starter for fermented milk, 1037 bacterial strains were isolated from raw milk. The strain that showed excellent acid producing and angiotensin converting enzyme (ACE) inhibitory activity (88.6%) was selected and identified as a Lactobacillus zeae based on the result of API carbohydrate fermentation pattern and 16S rDNA sequence. Lactobacillus zeae RMK354 was investigated further to study its physiological characteristics. It showed strong ACE inhibitory activity compared with commercial LAB starters tested. The optimum growth temperature of L. zeae RMK354 was $40^{\circ}C$ and it took 10 hr to reach pH 4.3 under this condition. L. zeae RMK354 showed more sensitive to penicillin-G, bacitracin, novobiocin, in a comparison of 14 different antibiotics, and showed most resistance to polymyxin B and vancomycin. It showed higher esterase and leucine arylamidase activities compared with 16 other enzymes. It was comparatively tolerant to bile juice and able to survive at pH 2 for 3 hr. It showed inhibitory activity against Salmonella Typhimurium with the rate of 60%. Based on these and previous results, L. zeae RMK354 could be an excellent starter culture for fermented milk with high level of ACE inhibitory activity.
Proteases are enzymes that break peptide bonds between amino acids of other proteins and occupy a crucial position with respect to their applications in both physiological and commercial fields. In order to screen new source of protease, bacteria producing extracellular proteases at low temperature were isolated from deep sea water of East Sea, Korea. A bacterium showing the best growth rate and production of an extracellular protease at low temperature was designated HJ 47. The DNA sequence analysis of the 16S rRNA gene, phenotypic tests and morphology led to the placement of this organism in the genus Pseudoalteromonas. Although maximal growth was observed at $37^{\circ}C$, enzyme production per culture time was maximum at $20^{\circ}C$. At this temperature, extracellluar protease production was detected from the end of the exponential phage to stationary phase, and maximal at 15 hours after initial production. The optimum temperature and pH of the protease were found to be $35^{\circ}C$ and 8.
This study was performed to investigate the effects of microbial augmentation on the biological treatment of paper mill wastewater. Three bacteria (KN11, KN13, KN27) capable of degrading aromatic compounds and a bacterial strain (GT21) producing an extracellular cellulase were isolated from soil and wastewater by selective enrichment culture. Through morphological, physiological, and biochemical taxonomies, isolated strains of KN11, KN13, KN27, and GT21 were identified as Acinetobacter sp., Neisseria sp., Bacillus sp., and Pseudomonas sp. and named Acinetobacter sp. KN11, Neisseria sp. KN13, Bacillus sp. KN27, and Pseudomonas sp. GT21, respectively. For analysis of non-biodegradable and chemical oxygen demand (COD)-increasing matter in a paper mill wastewater, we utilized GC/MS to detect aromatic compounds and their derivatives containing several substituted functional groups. The microbial augmentation, J30 formulated with the mixture of bacteria including Acinetobacter sp. KN11, Neisseria sp. KN13, Bacillus sp. KN27, and Pseudomonas sp. GT21, was used for the treatment of paper mill wastewater. The optimum temperature and pH for COD removal of the microbial augmentation, J30, were $30^{\circ}C$ and 7.5, respectively. For evaluation of the industrial applicability of the microbial augmentation, J30 in the pilot test, treatment efficiency was examined using paper mill wastewater. The microbial augmentation, J30, showed a COD removal rate of 87%. On the basis of the above results, we designed the wastewater treatment process of the activated sludge system.
Salt accumulation in soils of greenhouse due to the massive application of nitrogen fertilizers causes salt stress on the various crops, a serious problem in domestic agriculture. Since the majority of the salinity is nitrate, the excess nitrate should be removed; therefore, a bacterial strain having high capacity of nitrate uptake and identified as Enterobacter amnigenus GG0461 was isolated from the soils of greenhouse. Optimum conditions for the bacterial growth and nitrate uptake were investigated. GG0461 was able to grow without nitrate; however, nitrate facilitated the growth. The rate of nitrate uptake increased at alkaline pH and both growth and nitrate uptake were maximal at pH 8-9. When the initial pH of culture medium was increased to pH 8 or 9, it was decreased to neutral upon bacterial growth and nitrate uptake. These results imply that the major factor mediating bacterial nitrate uptake is a nitrate/proton antiporter. The fact was supported by the effect of nitrate addition in the absence of nitrate, since the addition of nitrate greatly increased the nitrate uptake and rapidly decreased pH of media.
In analyzing the operational characteristics of a rice whitening machine, the internal radial pressure of the machine was measured using strain gage equipment. Changes in cylinder and feed screw configurations, screen type, cylinder speed and counter-pressure levels were examined to determine their impact on the quality and quantity of milled rice and the performance of the machine. The results are summarized as follows: 1. The internal radial pressure in the whitening chamber varied with the surface condition of the grain being processed. During the first or second pass through the machine, pressure was relatively low, reached a maximum after two to three passes with combinations I and II, three to six with combination III and then began to fall. 2. The pitch of the feed screw and the size of the feed gate opening which determine the rate of entry of grain into the whitening chamber, appeared to be the most important factor aff-::cting the degree of radial pressure, quality and quantity of milled rice and the efficiency of the machine. Using a feed screw with a wide pitch (4.8cm), radial pressure was relatively high and head rice recovery ratio \vere quite low. In this case capacity and machine effic?iency were much higher than obtained when using a feed screw with a narrow pitch (2.3cm). Very significant responses in radial pressure, head rice recovery rates and machine capacity were observed with changes in cylinder speed and counter-pressure levels when using the wide pitch feed screw. 3. The characteristics of the screen which surrounds the whitening chamber had an important effect on whitening efficiency. The existence of small protuberances on the original screen resulted in significant increases in both machine capacity and efficiency but without a significant decrease in head rice recovery or development of excessive radial pressure. Further work is required to determine the effects of screen surface conditions and the shape of the cylinderical steel roller on the rate of bran removal, machine efficiency and recovery rates. The size of the slotted perforations 0:1 the screen affects total milled rice recovery. The opening size on the original screen was fabricated to accommodate the round shape of Japonica rice varieties but was not suitable for the more slender Indica type. Milling Indica varieties with this screen resulted in a reduction in total milled rice recovery. 4. An increase in cylinder speed from 380 to 820 rpm produced a positive effect on head rice recovery for all machine combinations at every level of counter-pressure used in the tests. Head rice recovery was considerably lower at 380rpm using a wide screw pitch when compared to the results obtained at speeds from 600 to 820 r.p.m. The effects of cylinder speed On radial pressure, capacity and machine efficiency showed contrasting results, depending on the width of the feed screw pitch. With a narrow feed screw pitch (2.3cm), a direct proportional relationship was observed bet?ween cylinder speed and both radial pressure and machine efficiency. In contrast, using a 4.8 centimeter pitch feed roller produced a series of inverse relationships between the above variables. Based on the results of this study it is recommended when milling Indica type long grain rice varieties that the cylinder speed of the original machine be increased from 500-600 rmp up to a minimum of 800 rpm to obtain a greater abrasive effect between the grain and the screen. The pitch of the feed screw should be also reduced to decr?ease the level of internal radial pressure and to obtain higher machine efficiency and increased quality of milled rice with increased cylinder speeds. Further study on the interaction between cylinder speed and feed screw pitch is recommended. 5. An increase in the counter pressure level produced a negative effect On the head rice recovery with an increase in radial pressure, capacity, and machine efficiency over all combinations and at every level of cylinder speed. 6. Head rice recovery rates were conditioned primarily by the pressure inside the whitening chamber. According to the empirical cha racteristics curve developed in this study, the relationships of head rice recovery ($Y_h$) and machine capacity ($Y_c$/TEX>) to internal radial pressure ($X_p$) followed an inverse quadratic function and a linear function respectively: $$Y_h^\Delta=\frac{1}{{1.4383-0.2951X_p^\ast+0.1425X_p^{\ast\ast}}^2} , (R^2=0.98)$$$$Y_c^\Delta=-305.83+374.37X_p^{\ast\ast}, (R^2=0.88)$$The correlation between capacity and power consumption per unit of brown rice expressed in the following exponential function: $$Y_c^\Delta=1.63Y_c^{-0.7786^\{\ast\ast}, (R^2=0.94)$$These relationships indicate that when radial pressure increases above a certain range (1. 6 to 2.0 kg/$cm^2$ based On the results of the experiment) head ricerecovery decrea?ses in a quadratic relation with a inear increase in capacity but without any decrease in power consump tion per unit of brown rice. On the other hand, if radial pressure is below the range shown above, power consumption increases dramatically with a lin?ear decrease in capacity but without significant increases in head rice recovery. During the operation of a given whitening machine, the optimum radial pressure range or the correct capacity range should be selected by controlling the feed rate and/or counter-pressure keeping in mind the condition of the grain, particulary the hardness. It was observed that the total number of passes is related to radial pessure level, feed rate and counter-pressure level. The higher theradial pressure the fewer num?ber of pass required but with decreased head rice recovery. In particular, when using high feed rates, the total number of passes should be increased to more than three by reducing the counter-pressure level to avoid decreaseases in head rice recovery (less than 65 percent head rice recovery on the basis of brown rice) at every cylinder speed. 7. A rapid rise in grain temperature seemed to have a close relationship with the pressure generated inside the whitening chamber and, subsequently with head rice reco?very rates. The higher the rate of increase, the lower were the resulting head rice recoveries.
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