Kim, Kyung-Hee;Kim, Sang-Soo;Kim, Goo-Young;Rhim, Hyang-Shuk
Journal of Life Science
/
v.16
no.7
s.80
/
pp.1133-1140
/
2006
Human HtrA1 (High temperature requirement protein A1) is a homologue of the E. coli periplasmic serine protease HtrA. A recent study has demonstrated that HtrA1 is a serine protease involved in processing of insulin like growth factor binding protein (ICFBP), indicating that it serves as an important regulator of IGF activity. Additionally, several lines of evidence suggest a striking correlation between proteolytic activity of HtrA1 serine protease and the pathogenesis of several diseases; however, physiological roles of HtrA1 remain to be elucidated. We used the pGEX bacterial expression system to develop a simple and rapid method for purifying HtrA1, and the recombinant HtrA1 protein was utilized to investigate the optimal conditions in executing its proteolytic activity. The proteolytically active HtrA1 was purified to approximately 85% purity, although the yield of the recombinant HtrA1 protein was slightly low $460{\mu}g$ for 1 liter E. coli culture). Using in vitro endoproteolytic cleavage assay, we identified that the HtrA1 serine protease activity was dependent on the enzyme concentration and the incubation time and that the best reaction temperature was $42^{\circ}C$ instead of $37^{\circ}C$. We arbitrary defined one unit of proteolytic activity of the HtrA1 serine protease as 200nM of HtrA1 that cleaves half of $5{\mu}M\;of\;{\beta}-casein$ during 3 hr incubation at $37^{\circ}C$. Our study provides a method for generating useful reagents to investigate the molecular mechanisms by which HtrA1 serine protease activity contributes in regulating its physiological function and to identify natural substrates of HtrA1.
The objective of this research was to characterize the effect of ammonium thiosulfate(ATS) on soil urease activity and the influence of temperature and soil pH on inhibition of urea hydrolysis by ATS. The results obtained are summarized as follows. 1. Percent inhibition of urea hydrolysis ranged from 1.5~14.3% with 5% addition of ATS to urea to 7.6~20.5% with 10% inclusion at $25^{\circ}C$, which levels were lower than that of thiourea. 2. The effect of ATS on urease inhibition was greater in low temperature han high temperature. The addition of 10% ATS resulted in 62.6% and 41.7% inhibition of urea hydrolysis at $10^{\circ}C$ and $15^{\circ}C$, respectively. 3. The inhibitory effects of ATS were found rapidly or slowly at different soil pH. The inhibition of urease activity was showed after 24 hours at pH 6.0 and 6.5, while the hydrolysis of urea was inhibited by ATS after 48 hourse at pH 5.0, 7.0 and 7.5. Urea hydrolysis inhibitions after 72 hours incubation were similar at all pH tested. Therefore effects of ATS did not correlate with soil pH.
The cultural and physiological conditions for the T-2 toxin [4,15-diacetoxy-8-(3-mety1butyloxy)-12,13- epoxy-trichothec-9-en-3-01, $C_{24}H_{30}O_9$] production by Fusarium spp. were studied. Thin layer chromatography (TLC) assay and the microbiological assay uslng Rhodotomla rubra were used to quantitate tbe T- 2 toxin. Among the four strains of Fusarium spp., F tn'cinctum NRRL 3299 was best for T-2 toxin production. In solid culture, white com grit medium was best for T-2 toxm production. Temperature played a critical role in the production of T-2 toxin. T-2 toxin production was favored by long duration of low-temperature incubation. The growth and toxin production were relatively high on galactose, fructose, glucose, and sucrose media, when each was used as a sole carbon source, and relatively low on sorbitol, glycerol, and lactose media. For nitrogen sources, $NH_4^(+) and NO_3^{-}were used well as a sole nitrogen source, but $NO_2^-$ was not used. Initial pH and speed of shaker also affected the production of T-2 toxin. From temperature shifting experiment, it is clear that T-2 toxin metabolic pathway is regulated by temperature-dependent enzyme depression or enzyme induction system.
This study was conducted to predict the hatching time of eggs of Lycorma delicatula, to select an effective environmentally-friendly agriculture material (EFAM) and to evaluate the attraction effect of brown sticky traps for controling of Lycorma delicatula nymph and adults. Eggs hatched 55.9, 26.8, 21.6 days after incubation at 15, 20, $25^{\circ}C$ with 14L:10D condition and the hatching rates of egg were 61.9, 57.8, 30.4%, respectively. At high temperature conditions, egg development periods were shorter and the hatching rate was lower. The relationship between temperature and developmental rate was expressed by the linear equation Y=0.0028X-0.0228, $R^2$=0.9561. The low temperature threshold of eggs was $8.14^{\circ}C$ and the thermal constant required to reach larva was 355.4 DD. According to this relationship, the mean estimated hatching date was $22^{nd}$ May. The effective EFAM was natural plant extract, sophora extract, derris extract to nymph and natural plant extract, pyrethrum extract, sophora extract to adult. Among three colors of sticky trap : brown, blue and yellow, the brown sticky trap was the most attractive to nymphs and adults of L. delicatula over a 2 weeks trial period. It suggested that the brown sticky trap could be a very useful and environment-friendly control method for nymphs and adults of L. delicatula.
This experiment was carried out to study the effects of lactic acid bacteria (LAB) inoculation and addition of cell wall degrading enzymes on the fermentation characteristics and chemical compositions of Italian ryegrass silage. An inoculant LAB with or without a cell wall degrading enzyme of Acremoniumcellulase (A), or Meicellulase (M) or a mixture of both (AM), was applied to 1 kg of fresh Italian ryegrass sample. The treatments were control untreated, LAB-treated (application rate $10^5$ cfu/g fresh sample), LAB+A 0.005%, LAB + A 0.01%, LAB+A 0.02%, LAB + M 0.005%, LAB + M 0.01%, LAB + M 0.02%, LAB+AM 0.005%, LAB + AM 0.01% and LAB+AM 0.02%. The sample was ensiled into 2-L vinyl bottle silo, with 9 silages of each treatment were made (a total of 99 silages). Three silages of each treatment were incubated at 20, 30 and $40{^{\circ}C}$ for an approximately 2-months storage period. All silages were well preserved as evidenced by their low pH values (3.79-4.20) and high lactic acid concentrations (7.71-11.34% DM). The fermentation quality and chemical composition of the control untreated and the LAB-treated silages were similar, except that for volatile basic nitrogen (VBN) content was lower (p < 0.05) in the LAB-treated silages. LAB + cellulase treatments improved the fermentation quality of silages by decreasing (p < 0.01) pH values and increasing (p<0.01) lactic acid concentrations, in all of cellulase types and incubation temperatures. Increasing amount of cellulase addition resulted in further decrease (p < 0.01) of pH value and increases (p < 0.01) of lactic acid and residual water soluble carbohydrate (WSC) concentrations. LAB + cellulase treatments reduced (p<0.01) NDF, ADF, hemicellulose and cellulose contents of silages compared with both the control untreated and LAB-treated silages. LAB + cellulase treatments did not affect the silage digestibility due to fact of in vitro dry matter digestibility (IVDMD) was similar in all silages. The silages treated with cellulase A resulted in a better fermentation quality and a higher rate of cell wall reduction losses than those of the silages treated with cellulases M and AM. Incubation temperature of $30{^{\circ}C}$ seemed to be more suitable for the fermentation of Italian ryegrass silages than those of 20 and $40{^{\circ}C}$.
Kim, Young-Ho;Kong, Won-Sik;Kim, Kyung-Soo;You, Chang-Hyun;Kim, Han-Kyoung;Sung, Jae-Mo;Ryu, Young-Jin;Kim, Kwang-Ho
The Korean Journal of Mycology
/
v.26
no.2
s.85
/
pp.187-193
/
1998
The effects of medium, incubation temperature, incubation period, pH of medium and $CO_2$ ondition during mycelial growth were investigated to study the factors associated with the formation of oidia in Flammulina velutipes. Oidia formation was increased when mycelial growth was poor, while oidia formation was inhibited in optimum condition of mycelial growth. Mating type of oidia was investigated to examine the effect of oidia formation on dikaryotic strain. Di-mon matings between oidia strains and original dikaryotic strain were carried out. Monokaryotic strains derived from oidia showed only one genotype. Seventy percent among Dimon mating strains showed slow mycelial growth and low yield of fruit-body, but others showed similar or high mycelial growth and yields in comparision with original dikaryon strain. One strain from di-mon mating demonstrated some differences in isozyme band pattern.
Mpendulo, Conference Thando;Hlatini, Vuyisa Andries;Ncobela, Cypril Ndumiso;Chimonyo, Michael
Asian-Australasian Journal of Animal Sciences
/
v.31
no.11
/
pp.1833-1839
/
2018
Objective: Incorporating high fibre ingredients into pig diets has the potential to reduce odour emissions from of pigs. The current study was carried out to determine effect of diets containing 0, 80 and 160 g/kg of each of lucerne hay, maize cobs and sunflower husks on the chemical characteristics and odours from pig slurries. Methods: Twenty eight pigs averaging $18{\pm}2.0kg$ were kept in individual cages, over four weeks. All pigs were fed ad libitum. Faeces and urine were collected, mixed in a 1:2.3 ratio (w/w), stored and fermented for 16 days in a temperature controlled room at $22^{\circ}C{\pm}2.3^{\circ}C$. The slurry was sampled twice (on day 1 and on day 16) of the fermentation period and analysed for pH, chemical oxygen demand (COD), nitrogen and short chain fatty acids (SCFA) concentration, on wet basis. All samples were tested for odour offensiveness using 18 panelists. A scale of 1 to 5 was used to rank the odour severity, (1 = not offensive, 5 = extremely offensive). Results: Slurry pH and COD varied with fibre source (p<0.05). On day 16, COD for lucerne hay, sunflower husk and maize cobs were 369, 512, and 425 (standard error of the mean = 34.2) mg of oxygen per litre. Total SCFA concentration was higher at day 16 than day 1 (p<0.05). Odour offensiveness varied with fibre source across both incubation periods (p<0.05). Sunflower husks and lucerne hay-based diets were rated as less offensive (mean rank = 2.2) than maize cob diets (mean rank of 4.3) (p<0.05). Conclusion: It was concluded that different fibre sources and incubation period influence chemical composition and odour of the slurry. There is, thereby, a need to incorporate locally available fibrous feeds in the diet of pigs because they have an economical and environmental relevance to pig management.
Proceedings of the Korean Vacuum Society Conference
/
2012.08a
/
pp.154-155
/
2012
The promise of nano-crystalites (nc) as a technological material, for applications including display backplane, and solar cells, may ultimately depend on tailoring their behavior through doping and crystallinity. Impurities can strongly modify electronic and optical properties of bulk and nc semiconductors. Highly doped dopant also effect structural properties (both grain size, crystal fraction) of nc-Si thin film. As discussed in several literatures, P atoms or radicals have the tendency to reside on the surface of nc. The P-radical segregation on the nano-grain surfaces that called self-purification may reduce the possibility of new nucleation because of the five-coordination of P. In addition, the P doping levels of ${\sim}2{\times}10^{21}\;at/cm^3$ is the solubility limitation of P in Si; the solubility of nc thin film should be smaller. Therefore, the non-activated P tends to segregate on the grain boundaries and the surface of nc. These mechanisms could prevent new nucleation on the existing grain surface. Therefore, most researches shown that highly doped nc-thin film by using conventional PECVD deposition system tended to have low crystallinity, where the formation energy of nucleation should be higher than the nc surface in the intrinsic materials. If the deposition technology that can make highly doped and simultaneously highly crystallized nc at low temperature, it can lead processes of next generation flexible devices. Recently, we are developing a novel CVD technology with a neutral particle beam (NPB) source, named as neutral beam assisted CVD (NBaCVD), which controls the energy of incident neutral particles in the range of 1~300eV in order to enhance the atomic activation and crystalline of thin films at low temperatures. During the formation of the nc-/pm-Si thin films by the NBaCVD with various process conditions, NPB energy directly controlled by the reflector bias and effectively increased crystal fraction (~80%) by uniformly distributed nc grains with 3~10 nm size. In the case of phosphorous doped Si thin films, the doping efficiency also increased as increasing the reflector bias (i.e. increasing NPB energy). At 330V of reflector bias, activation energy of the doped nc-Si thin film reduced as low as 0.001 eV. This means dopants are fully occupied as substitutional site, even though the Si thin film has nano-sized grain structure. And activated dopant concentration is recorded as high as up to 1020 #/$cm^3$ at very low process temperature (< $80^{\circ}C$) process without any post annealing. Theoretical solubility for the higher dopant concentration in Si thin film for order of 1020 #/$cm^3$ can be done only high temperature process or post annealing over $650^{\circ}C$. In general, as decreasing the grain size, the dopant binding energy increases as ratio of 1 of diameter of grain and the dopant hardly be activated. The highly doped nc-Si thin film by low-temperature NBaCVD process had smaller average grain size under 10 nm (measured by GIWAXS, GISAXS and TEM analysis), but achieved very higher activation of phosphorous dopant; NB energy sufficiently transports its energy to doping and crystallization even though without supplying additional thermal energy. TEM image shows that incubation layer does not formed between nc-Si film and SiO2 under later and highly crystallized nc-Si film is constructed with uniformly distributed nano-grains in polymorphous tissues. The nucleation should be start at the first layer on the SiO2 later, but it hardly growth to be cone-shaped micro-size grains. The nc-grain evenly embedded pm-Si thin film can be formatted by competition of the nucleation and the crystal growing, which depend on the NPB energies. In the evaluation of the light soaking degradation of photoconductivity, while conventional intrinsic and n-type doped a-Si thin films appeared typical degradation of photoconductivity, all of the nc-Si thin films processed by the NBaCVD show only a few % of degradation of it. From FTIR and RAMAN spectra, the energetic hydrogen NB atoms passivate nano-grain boundaries during the NBaCVD process because of the high diffusivity and chemical potential of hydrogen atoms.
Oh, Youn-Lee;Jang, Kab-Yeul;Kong, Won-Sik;Shin, Pyung-Gyun;Kim, Eun-Sun;Oh, Min ji;Choi, In-Geol
Journal of Mushroom
/
v.13
no.3
/
pp.243-249
/
2015
Agaricus bitorquis is an edible white mushroom of the genus that is cultivated at high temperature($25{\pm}1^{\circ}C$) unlike A. bisporus is cultivate at $16{\pm}2^{\circ}C$. Unlike Agaricus bisporus, an edible white A. bitorquis mushroom is cultivated at high temperature ($25{\pm}1^{\circ}C$). Most farmers cultivate this mushroom for a long cultivation period in Korea. For this reason, we made heterokayons to develop a new cultivar that generate fruitbodies for short cultivation period. Over one hundred SSIs(single spores isolates) were collected from selected A. bitorquis ASI1151 and ASI1349 strains. Seventy-three SSIs were germinated on CDA(compost dextrose agar) media after 20 days (minimum) or 83 days (maximum) incubation under different media condition. The mycelial growth rate of germinated SSIs was different. 9 homokaryons in ASI 1151 and 11 homokaryons in ASI 1349 from SSIs were selected by OPN-02 primer in RAPD analaysis. Also this primer was used to select heterokaryon that cross among each homokaryon with compatible locus. Therefore 44 compatible matings were confirmed of 99 crossed lines.
Nguyen, Bao Hung;Chu, Hyeonjin;Kim, Won-Il;Hwang, Injun;Kim, Hyun-Ju;Kim, Hwangyong;Ryu, Kyoungyul;Kim, Se-Ri
Journal of Food Hygiene and Safety
/
v.34
no.6
/
pp.542-550
/
2019
To detect Escherichia coli from agri-food and production environments, a device based on IoT (internet of things) technology that can check test results in real time on a mobile phone has been developed. The efficiency of the developed device, which combines an incubator equipped with a UV lamp, a high-resolution camera and software to detect E. coli in the field, was evaluated by measuring the device's temperature, detection limit, and detection time. The device showed a difference between its programmed temperature setting and actual temperature of about 1.0℃. In a detection limit test performed with a single-colony inoculation, a color change to yellow and a florescent signal were detected after 12 and 15 h incubations, respectively. The incubation time also decreased along with increasing bacteria levels. When applying the developed method and device to various samples, including utensils, gloves, irrigation water, seeds, and vegetables, detection rates of E. coli using the device were higher than those of the Korean Food Code method. These results show that the developed protocol and device can efficiently detect E. coli from agri-food production environments and vegetables.
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