• Korean Journal of Food Science and Technology
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• v.30 no.6
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• pp.1273-1278
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• 1998

Conditions for enzyme-protein binding assay (EPBA) were established in order to detect biotin more rapidly and reproducibly than traditional microbiological assay (MBA). EPBA with streptavidin and biotin-KLH conjugate showed cross-reactivities on biocytin, a derivative with biotin activity, at the rate of 109% $(IC_{50}=0.3\;ppb)\;and\;197%\;(IC_{50}=0.8\;ppb)$, respectively, but not on other derivatives with no biotin activities, such as desthiobiotin, diaminobiotin and 2-iminobiotin. Detection ranges of biotin by EPBA with streptavidin and biotin-KLH conjugates were $0.01{\sim}30\;ng/mL\;and\;0.01{\sim}1.0\;ng/mL(ppb)$, respectively. In the spike test with milk, fruit flake and pine-carrot juice, the correlation coefficience between MBA and EPBA with biotin-KLH conjugates was r=0.994. But MBA showed cross-reactivities both on biocytin and desthiobiotin at the rate of 80.1% and 66.7%, respectively. Detection range of biotin by MBA was $0.1{\sim}0.5\;ng/mL(ppb)$. These results strongly suggest that EPBA is efficient for biotin detection in sensitivity, detection range, cross-reactivity and time consuming.

• Food Science of Animal Resources
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• v.30 no.1
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• pp.36-41
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• 2010

The Hen's egg is widely used in many processed foods as an ingredient and is one of the most prevalent food allergens in children. To detect egg proteins in processed foods, we developed a competitive indirect enzyme-linked immunosorbent assay (ciELISA) using an anti-ovomucoid (OM) antibody, which was produced by immunization of rabbits with OM, the most heat-stable component of the egg proteins. The detection limit of this quantitative assay system was 30 ng/mL. Cross-reactivity of the anti-OM antibody toward OM, ovalbumin, skim milk, casein, whey protein isolate, and isolated soy protein was 100, 0.4, 0.2, 0.04, 0, and 0%, respectively. In the spike test of egg white powder in milk replacer, commercial sausage, and in-house sausage, the assay recoveries ($mean{\pm}SD$) were $129{\pm}13.7%$, $73.9{\pm}12.5%$, and $65.5{\pm}13.6%$, respectively. When egg white in a commercial crab meat analog and sausage was determined by ciELISA, the assay recovery was found to be 108% and 127%, respectively. The combined results of this study indicate that this novel ciELISA for OM detection could be applied for the quantification of hen's egg proteins in processed foods.

• Biomolecules & Therapeutics
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• v.30 no.5
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• pp.427-434
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• 2022

The drug repurposing strategy has been applied to the development of emergency COVID-19 therapeutic medicines. Current drug repurposing approaches have been directed against RNA polymerases and viral proteases. Recently, we found that the inhibition of the interaction between the SARS-CoV-2 structural nucleocapsid (N) and spike (S) proteins decreased viral replication. In this study, drug repurposing candidates were screened by in silico molecular docking simulation with the SARS-CoV-2 structural N protein. In the ChEMBL database, 1994 FDA-approved drugs were selected for the in silico virtual screening against the N terminal domain (NTD) of the SARS-CoV-2 N protein. The tyrosine 109 residue in the NTD of the N protein was used as the center of the ligand binding grid for the docking simulation. In plaque forming assays performed with SARS-CoV-2 infected Vero E6 cells, atovaquone, abiraterone acetate, and digoxin exhibited a tendency to reduce the size of the viral plagues without affecting the plaque numbers. Abiraterone acetate significantly decreased the accumulation of viral particles in the cell culture supernatants in a concentration-dependent manner. In addition, abiraterone acetate significantly decreased the production of N protein and S protein in the SARS-CoV-2-infected Vero E6 cells. In conclusion, abiraterone acetate has therapeutic potential to inhibit the viral replication of SARS-CoV-2.

• Journal of The Korean Society of Grassland and Forage Science
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• v.32 no.2
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• pp.149-156
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• 2012

his experiment was carried out to evaluate the effects of nitrogen top dressing levels on the growth, feed value, and anthocyanin content for developing functional feed of colored barley. A colored barley cultivar, Boanchalbori, was tested in this experiment. Nitrogen top dressing levels was six (0, 20, 40, 60, 80, 100%) and top dressing time was a regeneration time. In case of productivity, heading data was get behind and dry matter rate was significant decreased with higher nitrogen top dressing levels (p<0.05). Fresh yield, dry matter yield and TDN yield was increased with higher nitrogen top dressing levels, but not significant. In case of feed value, crude protein content was significant increased with higher nitrogen top dressing levels (p<0.05), and higher in the order of spike, whole and leaf culme of the plant. Percent NDF and ADF was decreased with higher nitrogen top dressing levels in leaf culme, but no difference in spike and whole (p<0.05), and higher in the order of leaf culme, whole and spike of the plant. TDN was increased with higher nitrogen top dressing levels in leaf culme, but no difference in spike and whole (p<0.05), and higher in the order of spike, whole and leaf culme of the plant. Total anthocyanin content was significant decreased with higher nitrogen top dressing levels in leaf culme and whole (p<0.05), and higher in the order of leaf culme, whole and spike of the plant. Specially, cyanidin-3-glucoside (C3G), delphinidin (Del), malvidin-3-glucoside (M3G) and malvidin (Mal) show a significant decrease. So there are an accumulation of anthocyanin in the culm, and standard nitrogen top dressing levels dressing on the regeneration time for produces high anthocyanin content of the colored barley.

• Journal of Ginseng Research
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• v.47 no.1
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• pp.123-132
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• 2023

Background: Pseudotyped virus systems that incorporate viral proteins have been widely employed for the rapid determination of the effectiveness and neutralizing activity of drug and vaccine candidates in biosafety level 2 facilities. We report an efficient method for producing severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) pseudovirus with dual luciferase and fluorescent protein reporters. Moreover, using the established method, we also aimed to investigate whether Korean Red Ginseng (KRG), a valuable Korean herbal medicine, can attenuate infectivity of the pseudotyped virus. Methods: A pseudovirus of SARS-CoV-2 (SARS-2pv) was constructed and efficiently produced using lentivirus vector systems available in the public domain by the introduction of critical mutations in the cytoplasmic tail of the spike protein. KRG extract was dose-dependently treated to Calu-3 cells during SARS2-pv treatment to evaluate the protective activity against SARS-CoV-2. Results: The use of Calu-3 cells or the expression of angiotensin-converting enzyme 2 (ACE2) in HEK293T cells enabled SARS-2pv infection of host cells. Coexpression of transmembrane protease serine subtype 2 (TMPRSS2), which is the activator of spike protein, with ACE2 dramatically elevated luciferase activity, confirming the importance of the TMPRSS2-mediated pathway during SARS-CoV-2 entry. Our pseudovirus assay also revealed that KRG elicited resistance to SARS-CoV-2 infection in lung cells, suggesting its beneficial health effect. Conclusion: The method demonstrated the production of SARS-2pv for the analysis of vaccine or drug candidates. When KRG was assessed by the method, it protected host cells from coronavirus infection. Further studies will be followed for demonstrating this potential benefit.

• Genomics & Informatics
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• v.21 no.2
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• pp.17.1-17.12
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• 2023

Coronavirus has left severe health impacts on the human population, globally. Still a significant number of cases are reported daily as no specific medications are available for its effective treatment. The presence of the CD147 receptor (human basigin) on the host cell facilitates the severe acute respiratory disease coronavirus 2 (SARS-CoV-2) infection. Therefore, the drugs that efficiently alter the formation of CD147 and spike protein complex could be the right drug candidate to inhibit the replication of SARS-CoV-2. Hence, an e-Pharmacophore model was developed based on the receptor-ligand cavity of CD147 protein which was further mapped against pre-existing drugs of coronavirus disease treatment. A total of seven drugs were found to be suited as pharmacophores out of 11 drugs screened which was further docked with CD147 protein using CDOCKER of Biovia discovery studio. The active site sphere of the prepared protein was 101.44, 87.84, and 97.17 along with the radius being 15.33 and the root-mean-square deviation value obtained was 0.73 Å. The protein minimization energy was calculated to be -30,328.81547 kcal/mol. The docking results showed ritonavir as the best fit as it demonstrated a higher CDOCKER energy (-57.30) with correspond to CDOCKER interaction energy (-53.38). However, authors further suggest in vitro studies to understand the potential activity of the ritonavir.

• Proceedings of the Korean Society of Crop Science Conference
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• 2022.10a
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• pp.63-63
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• 2022

Wheat flour can be categorized into bread, all-purpose, cake flour according to its protein content. Since optimal wheat flour protein content is different for each end use, it is necessary to diversify the nitrogen fertilizer methods depending on the end use and cultivar. Optimal wheat flour protein content of soft wheat (for cake flour) is lowest (<=10%) among all end use, it is necessary to develop nitrogen fertilizer methods for high yield and low protein content. In order to analyze the yield and quality changes of soft wheat as nitrogen fertilizer amount and splitting timing, soft wheat cultivar 'goso' was sown on paddy soil in jeunju, Republic of Korea ('21.10). the amount of nitrogen fertilizer was divided into 4 levels by adjusting 2kg/10a increments from 5.1 to ll.lkg/lOa, and in the N 7.1 and 9.1 kg/1 Oa(standard) treatment, N amount divided into sowing date:regrowing stage=3:7,4:6(standard), 5:5. In regrowing stage, Tiller number and N fertilizer amount at sowing date showed a correlation; y=-121.14x²+792.66x-525.41 (R²=0.77^*, y: Tiller number/m², x: N amount at sowing date(kg/10a)). Tiller number in regrowing stage was the highest when the nitrogen fertilizer amount at sowing date was 3.23kg/10a. spike number per m² was the highest when N fertilizer was divided into sowing date:regrowing stage=3:7(N amount: 9.1kg/10a). If N fertilizer amount was fixed, grain yield was also the highest when N fertilizer was divided into sowing date :regrowing stage=3:7. Also, N amount at sowing date and grain yield showed no correlation, but N amount at regrowing stage and grain yield showed significant correlation. As N amount increased, protein content also showed a tendency to increase.

• Journal of Ginseng Research
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• v.47 no.2
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• pp.329-336
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• 2023

Background: Panax ginseng Meyer is a medicinal plant well-known for its antiviral activities against various viruses, but its antiviral effect on coronavirus has not yet been studied thoroughly. The antiviral activity of Korean Red Ginseng (KRG) and ten ginsenosides against Human coronavirus OC43 (HCoV-OC43) was investigated in vitro. Methods: The antiviral response and mechanism of action of KRG extract and ginsenoside Rc, Re, Rf, Rg1, Rg2-20 (R) and -20 (S), Rg3-20 (R) and -20 (S), and Rh2-20 (R) and -20 (S), against the human coronavirus strain OC43 were investigated by using plaque assay, time of addition assay, real-time PCR, and FACS analysis. Results: Virus plaque formation was reduced in KRG extract-treated and HCoV-OC43-infected HCT-8 cells. KRG extract decreased the viral proteins (Nucleocapsid protein and Spike protein) and mRNA (N and M gene) expression, while increased the expression of interferon genes. Conclusion: KRG extract exhibits antiviral activity by enhancing the expression of interferons and can be used in treating infections caused by HCoV-OC43.

• Proceedings of the Korean Society of Crop Science Conference
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• 2022.10a
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• pp.62-62
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• 2022

Among annual precipitation in Korea (1306.3 mm), 54% of it falls intensively in summer, and only about 12.4% falls in April and May, when the water requirement of wheat is the highest. Korean wheat also could be damaged by soil water excess stress as frozen soil thaws after winter (late Feb-Mar). This study was conducted to evaluate effect of soil water stress on yield and quality of Korean wheat cultivar 'Saegeumgang'. Soil water treatments consisted of 4 treatments; water excess treatment in tilling stage (3.23-3.30), drought treatment in ripening stage (Apr-Jun), irrigation treatment in ripening stage (5.10) and standard condition. There was no significant difference between the treatment conditions for culm length, and the number of spike number was the highest in the order of irrigation in the ripening period (951)> standard cultivation (876)> excess water treatment in the tilling stage (752)> drought treatment in the ripening stage (767/m²). Test weight and Thousand grain weight were 548g/L and 22. lg respectively, which were lower than other treatments, and there was no significant difference between the other treatments. Abortive grain was 5.4kg/10a which was lower than other treatment, and there was no significant difference between the other treatment than other treatments. In drought treatment, protein content was 11.9% which is the highest among all treatments, and SDS-sedimentation value was 27.2ml under drought treatment which was very low compared to other treatments. Therefore, wheat yield and spike number were decreased in excess water condition at tilling stage and drought condition at ripening stage. Furthermore, wheat quality became deteriorate in drought condition at ripening stage.

• Journal of Microbiology and Biotechnology
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• v.32 no.10
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• pp.1335-1343
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• 2022

COVID-19 is an emerging disease that poses a severe threat to global public health. As such, there is an urgent demand for vaccines against SARS-CoV-2, the virus that causes COVID-19. Here, we describe a virus-like nanoparticle candidate vaccine against SARS-CoV-2 produced by an E. coli expression system. The fusion protein of a truncated ORF2-encoded protein of aa 439~608 (p170) from hepatitis E virus CCJD-517 and the receptor-binding domain of the spike protein from SARS-CoV-2 were expressed, purified and characterized. The antigenicity and immunogenicity of p170-RBD were evaluated in vitro and in Kunming mice. Our investigation revealed that p170-RBD self-assembled into approximately 24 nm virus-like particles, which could bind to serum from vaccinated people (p < 0.001) and receptors on cells. Immunization with p170-RBD induced the titer of IgG antibody vaccine increased from 14 days post-immunization and was significantly enhanced after a booster immunization at 28 dpi, ultimately reaching a peak level on 42 dpi with a titer of 4.97 log₁₀. Pseudovirus neutralization tests showed that the candidate vaccine induced a strong neutralizing antibody response in mice. In this research, we demonstrated that p170-RBD possesses strong antigenicity and immunogenicity and could be a potential candidate for use in future SARS-CoV-2 vaccine development.