Using recombinant human growth hormone as a model protein, we carried out unfolding by adding a denaturant such as urea, guanidine HCl, or SDS followed by refolding by dilution and dialysis. The objectives were to monitor the structural changes during in vitro refolding process and, based on the results, to develop a quantitative method of refolding progress assessment. The changes in surface hydrophobicity were measured by fluorescence tagging of 1-anilinonaphthalene-8-sulfonate(1,8-ANS) to the hydrophobic portions, and those in the secondary structure were monitored by using far UV-CD(circular dichroism) spectroscopy. Also, we used RP-HPLC to separate and quantify the folded and unfolded proteins to correlate the result with the structure analysis. Our results indicate the surface hydrophobicity are well correlated with the formations of the secondary structure, primarily ${\alpha}$-helices, as well as the disulfide bridges. We expect this monitoring technique can be applied in industrial fields as a means to quantitatively assess the progress of in-vitro refolding of recombinant proteins.
Vegetable oils are desirable inexpensive feedstocks for various bioproducts. The content of unsaturated fatty acids such as oleic and linoleic acids are 22% and 55% for soybean oil, 26% and 60% for corn oil, and 61% and 21% for canola oil, respectively. Keto and hydroxy fatty acids are useful industrial chemicals, used in plasticizer, surfactant, lubricant and detergent formulations because of their special chemical properties such as higher viscosity and reactivity compared with other fatty acids. In this study, a microbial isolate, Flavobacterium sp. strain DS5 (NRRL B-14859), was used to convert oleic acid to 10-ketostearic acid (10-KSA) via 10-hydroxystearic acid (10-HSA). Two bioconversion products, 10-KSA and 10-HSA, were quantitatively and qualitatively analyzed using gas chromatography, gas chromatography-mass spectrometry, and $^1H-$ and $^{13}C$-nuclear magnetic resonance. The maximum production of 10-KSA and 10-HSA in flask cultures were 3.4 g/L and 0.5 g/L, respectively. The optimum concentrations of glucose and yeast extract, addition time and volume of oleic acid for 10-KSA production were less than 20 g/L, more than 5 g/L, 18 hand 0.3 ml/50 ml, respectively.
Kim, Byung Ki;Ha, Jae Jung;Yi, Jun Koo;Oh, Don Yep;Jung, Dae Jin;Hwang, Eun Gyeong;Kim, Soo Jung;Ann, Yong Geun
The Korean Journal of Food And Nutrition
/
v.29
no.6
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pp.860-869
/
2016
This study was a report on 60 heads of Hanwoo steers(5 treatments ${\times}$ 4 heads ${\times}$ 3 repeats). Their start weights were $361.2{\pm}14.39kg$ to $380.5{\pm}27.33kg$; 12.7 to 13.6 months of age. The aim was to investigate the effect of herbal probiotic supplementation on the weight, blood composition, meat quality and immunity in beef. In case of body weight gain, con. 1 plot(183.8 kg) in the early fattening stage, T3 plot(115.1 kg) in the middle fattening stage, and T2 plot(163.3 kg) in the late fattening stage significantly increased(p<0.05). There was a statistically significant difference in the blood content of T-Cho and IgG: con. 1 plot and T1 plot in the early fattening stage, T3 plot and T2 plots in the middle fattening stage, and con. 2 plot and T1~T3 plots in the late fattening stage had higher blood content than that of other treatment plots (p<0.05). In addition, the carcass weight was highest in the T1 plot and T2 plot among the total treatment plots(p<0.05). However, no statistical significant difference was observed in the different treatments; all individual treatment plots(T1~T3 plots) were somewhat higher than the control plots(con. 1~con. 2 plots) with respect to the carcass quantity and carcass quality. Combining and analyzing all results revealed that the dietary addition of herbal probiotics for Hanwoo steers had a positive effect on the improvement of weight, amount of meat, meat quality and immunity.
Kwon, Soo Jeong;Lee, Ui Gun;Moon, Young Ja;Cho, Gab Yeon;Woo, Sun Hee;Boo, Hee Ock;Koo, Jin-Woog;Kim, Hag Hyun
Proceedings of the Korean Society of Crop Science Conference
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2017.06a
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pp.172-172
/
2017
Pollen germination and its' viability in bellflower hybrid system are of great importance. The present study was conducted to investigate the several factors underlying the pollen germination of Platycodon grandiflorum and obtain the basic data for effective artificial pollination for the production of sound specifies. The pollens of Platycodon grandiflorum started germination from one hour after planting, and the germination was actively progressed as time dependent manner. For lighting conditions, the germination of pollens under the light was faster by more than twice than that without the light. Furthermore, the germination was better in the high temperature rather than in the low temperature. The germination rate was higher in the $30^{\circ}C$. For the carbon source, the germination rate was better at the concentration of 15% regardless of the kinds. In particular, the highest value was observed with glucose. The germination rate was decreased substantially as the increasing with the higher pH. The dynamic germination of pollens was observed at the pH 5. With respect to the growth regulator, the higher concentration of NAA induced the higher the germination rate. $GA_3$ showed a good germination rate in $0.05mg{\cdot}L^{-1}$. Meanwhile, for kinetin, lower concentration increased the germination rate, unlike NAA. The higher concentrations of boric acid degraded the germination rate, and the addition of boric acid of $10mg{\cdot}L^{-1}$ demonstrated higher germination rate than the addition of other growth regulators. Notably, the addition of asparaginic acid exhibited the similar results in all test sectors regardless of concentration, whereas a little higher result was observed in the high concentration sector. Taken together, the results concluded that the boric acid was considered as one of the essential minerals that played an important role on the germination of pollens.
Kwon, Soo Jeong;Han, Eun Ji;Moon, Young Ja;Cho, Gab Yeon;Woo, Sun Hee;Boo, Hee Ock;Koo, Jin-Woog;Kim, Hag Hyun
Proceedings of the Korean Society of Crop Science Conference
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2017.06a
/
pp.171-171
/
2017
Propagation by crown division in Platycodon grandiflorum is too slow for producing many plants in a short time although the plants are uniform. This study was performed to enhance the mass propagation for Platycodon grandiflorum containing yellow green petals via various medium compositions and the growth regulators. The nodes containing yellow green petals were used as materials to execute the study with a variety of MS medium concentrations. The 1/4MS medium showed the best development of adventitious root, while the 1/2MS medium exhibited the potential growth. The higher the concentration of sucrose showed the better development and growth of both shoots and adventitious roots. Many adventitious roots were developed at the controlled culture medium at pH 4.8 with a tendency of suppression with higher levels of pH. Also, the cultivated node and leaf explants with the treatments of simple and combined applications with auxin and cytokinin at the 1/4MS culture medium with adding 5% of sucrose were used to identify the influences of growth regulators. The regeneration of plantlets at the 234single application showed a good result with the addition of BA $1mg{\cdot}L^{-1}$ and the development and growth of adventitious roots appeared to be good at the addition of NAA $1mg{\cdot}L^{-1}$. For the combined applications, the regeneration of plantlets and the development of adventitious roots were prosperous at the combined applications with BA $0.1mg{\cdot}L^{-1}$ and IAA $0.5mg{\cdot}L^{-1}$. The addition of IAA for the leaf explants induced a number of plantlets that showed the potential regeneration, and the highest results was obtained from the combined applications of both BA $1mg{\cdot}L^{-1}$ and IAA $2.5mg{\cdot}L^{-1}$. In addition, the development of adventitious roots showed the satisfactory results at the combined application of both BA $1mg{\cdot}L^{-1}$ and IAA $0.5mg{\cdot}L^{-1}$.
Roy, Swapan Kumar;Cho, Seong-Woo;Kwon, Soo Jeong;Kamal, Abu Hena Mostafa;Kim, Sang-Woo;Lee, Moon-Soon;Chung, Keun-Yook;Woo, Sun-Hee
Proceedings of the Korean Society of Crop Science Conference
/
2017.06a
/
pp.124-124
/
2017
Cadmium (Cd) is of particular concern because of its widespread occurrence and high toxicity and may cause serious morpho-physiological and molecular abnormalities in in plants. The present study was performed to explore Cd-induced morpho-physiological alterations and their potentiality associated mechanisms in Sorghum bicolor leaves at the protein level. Ten-day-old sorghum seedlings were exposed to different concentrations (0, 100, and $150{\mu}M$) of $CdCl_2$, and different morpho-physiological responses were recorded. The effects of Cd exposure on protein expression patterns in S. bicolor were investigated using two-dimensional gel electrophoresis (2-DE) in samples derived from the leaves of both control and Cd-treated seedlings. The observed morphological changes revealed that the plants treated with Cd displayed dramatically altered shoot lengths, fresh weights, and relative water content. In addition, the concentration of Cd was markedly increased by treatment with Cd, and the amount of Cd taken up by the shoots was significantly and directly correlated with the applied level of Cd. Using the 2-DE method, a total of 33 differentially expressed protein spots were analyzed using MALDI-TOF/TOF MS. Of these, treatment with Cd resulted in significant increases in 15 proteins and decreases in 18 proteins. Significant changes were absorbed in the levels of proteins known to be involved in carbohydrate metabolism, transcriptional regulation, translation and stress responses. Proteomic results revealed that Cd stress had an inhibitory effect on carbon fixation, ATP production and the regulation of protein synthesis. In addition, the up-regulation of glutathione S-transferase and cytochrome P450 may play a significant role in Cd-related toxicity and stress responses. Our study provides insights into the integrated molecular mechanisms involved in response to Cd and the effects of Cd on the growth and physiological characteristics of sorghum seedlings. The upregulation of these stress-related genes may be candidates for further research and use in genetic manipulation of sorghum tolerance to Cd stress.
Yu, Je Hyeok;Kwon, Soo Jeong;Cho, Seong-Woo;So, Yoon-Sup;Jung, Hyun-Jin;Chung, Jong-Wook;Ma, Kyung-Ho;Son, Eun-Ho;Lee, Sok-Young;Woo, Sun-Hee
Proceedings of the Korean Society of Crop Science Conference
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2017.06a
/
pp.76-76
/
2017
Buckwheat has gained scientists concern due to its nutritional and medicinal values in recent years, and many important bioactive compounds. The present study was performed to investigate the influence of the varietal performance and environmental conditions on the content of rutin and quercetin in Tartary buckwheat germplasm. A total of 44 foreign Tartary buckwheat germplasms were examined and compared their contents based on the collected countries, seed shape, and seed color. The highest number of germplasm (16) was found in ranged from 600 mg/100 g to 800 mg/100 g of rutin content. In case of quercetin content, the highest number of germplasm (19) was observed in ranged from 5 mg/100 g to 10 mg/100 g. However, the rutin content showed the highest value (1326.6 mg/100 g) from CBU408 and quercetin content showed the highest value (22.5 mg/100 g) from CBU456. The number of germplasm showed the highest value (23) in ranged from 3000 mg/100 g to 4000 mg/100 g of rutin content. The number of germplasm is the highest value (30) in ranged from 100 mg/100 g to 200 mg/100 g of quercetin content. When rutin and quercetin content in seed was compared according to collected countries, seed shape, and seed color respectively, no association was appeared with rutin and quercetin content. In sprout of whole Tartary buckwheat germplasm, mean of rutin content and quercetin content was 3362.9 mg/100 g and 14.2 mg/100 g respectively. In conclusion, the results observed from the present study suggest that highly rutin germplasm could be used more efficiently via breeding to develop a highly rutin content variety for using in sprout. No correlation was appeared in the case of quercetin. However, the quercetin content in sprout was 4~90 times more than seed. So, sprout could be used more efficiently than seed in the case of quercetin. Our results suggest that the varietal variation and qualitative characteristic differences of rutin and bioactive compounds may provide important nutrient sources and industrial application of Tartary buckwheat.
This study was carried out to examine a molecular marker system for parentage test in Jeju Black cattle (JBC). Based on the preliminarily studies, we finally selected for construction of a novel genetic marker system for molecular traceability, identity test, breed certification, and parentage test in JBC and its related industrial populations. The genetic marker system had eight MS markers, five indel markers, and two single nucleotide polymorphisms (SNPs; g.G299T and g.del310G) within MC1R gene which is critical to verify the breed specific genotypes for coat color of JBC differing from those of exotic black cattle breeds such as Holstein and Angus. The results showed lower level of a combined non-exclusion probability for second parent (NE-P2) of $4.1202{\times}10^{-4}$ than those previously recommended by International Society of Animal Genetics (ISAG) of $5.000{\times}10^{-4}$ for parentage, and a combined non-exclusion probability for sib identity (NE-SI) of $2.679{\times}10^{-5}$. Parentage analysis has been successfully identified the JBC offspring in the indigenous population and cattle farms used the certified AI semens for production using the JBC-derived offspring for commercial beef. This combined molecular marker system will be helpful to supply genetic information for parentage test and traceability and to develop the molecular breeding system for improvement of animal productivity in JBC population.
An analytical method of nine synthetic food colors in Korea, Japan, Joint FAO/WHO Expert Committee of Food Additives (JECFA), and USA were compared. Contents of water insoluble matter in synthetic food colors tested were not different by general test methods of various organs. Contents of chloride and sulfate salts in Food Green No.3, Food Red No.3, Food Blue No.2, and Food Yellow No.4 slightly differed in various methods, and but up to the standard, and general test methods of JECFA and USA needed far more time. For the measurement of arsenic contents in food colors, colorimetric method in Korea and Japan, and silver diethyldithiocarbamate colorimetric method in USA and JECFA were used, but the standards of them were different. Content of heavy metals was up to the standard, but the methods were used colorimetric method in Korea, atomic absorption method in Japan, and both methods in JECFA and USA.
Center for Food Standard Evaluation, Korea Food and Drug AdministrationThis study was performed to compare analytical methods of nine synthetic food colors and six food color lakes in Korea, Japan, Joint FAO/WHO Expert Committee of Food Additives (JECFA), and USA. The experimental protocol of this study consists of three parts: titration method with titanium chloride, gravimetric and spectrophotometric method. To measure the total contents of food colors, Korea and Japan used titration method with titanium chloride, USA used the average value of titration method with titanium chloride equipped with Kipp generator and spectrophotometric method. Also, JECFA used titration method with titanium chloride equipped with KiPP generator. However, All the low organizations used gravimetric method to measure the total content of coloring matter on Food Red No.3. Although all organizations use various methods for analysis of coloring matters, total contents of coloring matter on food colors tested fell into the standard showing $85.08-96.40\%$ in synthetic food colors and $10.00-36.86\%$ in food color lakes.
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