• Title/Summary/Keyword: Fusion Activity

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Development of a Method for High throughput Screening of Antagonistic Substances against Rice Pathogens using Rice Leaf Explants (벼의 생엽절편을 이용한 병원균 억제물질의 대량 스크리닝 방법 개발)

  • Park, Sait-Byul;Lee, Choong-Hwan;Kim, Tae-Jong;Kang, Lin-Woo;Lee, Byoung-Moo;Kim, Jeong-Gu
    • Microbiology and Biotechnology Letters
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    • v.40 no.1
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    • pp.39-42
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    • 2012
  • A new method for the high throughput screening of antagonistic substances against rice pathogens using rice leaf explants was developed. This method can be used to confirm the activities of any compound or mixture suppressing rice bacterial blight (BB) before field tests. Xanthomonas oryzae pv. oryzae (Xoo) culture medium was distributed in 96 well plates with equally sized explants and the active compounds were added to the wells. The strength suppressing BB was converted into an area percent of the lesion on the rice explants. The explants under BB suppressing activity remained uninfected maintaining their actual green color, while infected explants exhibited pale yellow-colored lesions. Based on the results, this method seems to be faster and easier, dose-dependent, and can be performed all-at-once with a small amount of unspecified compounds. This method also has the potential to be applied to inspection activities for the suppression of other waterborne crop diseases.

Derepression of a Methionine Biosynthetic Gene by Utilizing a Promoter Isolated from Corynebacterium glutamicum (Corynebacterium glutamicum에서 분리된 프로모터를 이용한 메치오닌 생합성 유전자의 조절해제)

  • Park Soo-Dong;Park Ik-Hyun;Choi Jong-Soo;Kim Il-Kwon;Kim Younhee;Lee Heung-Shick
    • Korean Journal of Microbiology
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    • v.41 no.4
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    • pp.300-305
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    • 2005
  • A transcriptionally active fragment $(P_{19})$ isolated by utilizing the promoter-probe shuttle vector pSK1Cat was analyzed. By subcloning analysis, the 180 bp region $(P_{180})$ responsible for the activity was determined. Transcriptional fusion of the C. glutamicum metX gene to $P_{180}\;(P_{180}-metX)$ resulted in a 24-fold increase in MetX activity in a complex medium, while a 13-fold increase was observed with the $P_{tac}$ promoter. Additionally, the expression conferred by $P_{180}$ was not affected by methionine added to the growth medium, suggesting that the $P_{180}$ clone is useful for the deregulated expression of biosynthetic genes in C. glutamicum during amino acid fermentation. Introduction of $P_{180}-metX$ into a lysine-producing C. glutamicum resulted in the production of methionine to 0.8 g/l.

Identification and Characterization of Alternative Promoters of the Rice MAP Kinase Gene OsBWMK1

  • Koo, Sung Cheol;Choi, Man Soo;Chun, Hyun Jin;Park, Hyeong Cheol;Kang, Chang Ho;Shim, Sang In;Chung, Jong Il;Cheong, Yong Hwa;Lee, Sang Yeol;Yun, Dae-Jin;Chung, Woo Sik;Cho, Moo Je;Kim, Min Chul
    • Molecules and Cells
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    • v.27 no.4
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    • pp.467-473
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    • 2009
  • Our previous study suggested that OsBWMK1, a gene which encodes a member of the rice MAP kinase family, generates transcript variants which show distinct expression patterns in response to environmental stresses. The transcript variants are generated by alternative splicing and by use of alternative promoters. To test whether the two alternative promoters, pOsBWMK1L (promoter for the OsBWMK1L splice variant) and pOsBWMK1S (promoter for the OsBWMK1S splice variant), are biologically functional, we analyzed transgenic plants expressing GUS fusion constructs for each promoter. Both pOsBWMK1L and pOsBWMK1S are biologically active, although the activity of pOsBWMK1S is lower than that of pOsBWMK1L. Histochemical analysis revealed that pOsBWMK1L is constitutively active in most tissues at various developmental stages in rice and Arabidopsis, whereas pOsBWMK1S activity is spatially and temporally restricted. Furthermore, the expression of pOsBWMK1S::GUS was upregulated in response to hydrogen peroxide, a plant defense signaling molecule, in both plant species. These results suggest that the differential expression of OsBWMK1 splice variants is the result of alternative promoter usage and, moreover, that the mechanisms controlling OsBWMK1 gene expression are conserved in both monocot and dicot plants.

Some characters of bacterial cellulases in goats' rumen elucidated by metagenomic DNA analysis and the role of fibronectin 3 module for endoglucanase function

  • Nguyen, Khanh Hoang Viet;Dao, Trong Khoa;Nguyen, Hong Duong;Nguyen, Khanh Hai;Nguyen, Thi Quy;Nguyen, Thuy Tien;Nguyen, Thi Mai Phuong;Truong, Nam Hai;Do, Thi Huyen
    • Animal Bioscience
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    • v.34 no.5
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    • pp.867-879
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    • 2021
  • Objective: Fibronectin 3 (FN3) and immunoglobulin like modules (Ig) are usually collocated beside modular cellulase catalytic domains. However, very few researches have investigated the role of these modules. In a previous study, we have sequenced and analyzed bacterial metagenomic DNA in Vietnamese goats' rumen and found that cellulase-producing bacteria and cellulase families were dominant. In this study, the properties of modular cellulases and the role of a FN3 in unique endoglucanase belonging to glycosyl hydorlase (GH) family 5 were determined. Methods: Based on Pfam analysis, the cellulases sequences containing FN3, Ig modules were extracted from 297 complete open reading frames (ORFs). The alkaline, thermostability, tertiary structure of deduced enzymes were predicted by AcalPred, TBI software, Phyre2 and Swiss models. Then, whole and truncated forms of a selected gene were expressed in Escherichia coli and purified by His-tag affinity column for assessment of FN3 ability to enhance enzyme activity, solubility and conformation. Results: From 297 complete ORFs coding for cellulases, 148 sequences containing FN3, Ig were identified. Mostly FN3 appeared in 90.9% beta-glucosidases belonging to glycosyl hydrolase family 3 (GH3) and situated downstream of catalytic domains. The Ig was found upstream of 100% endoglucanase GH9. Rarely FN3 was seen to be situated downstream of X domain and upstream of catalytic domain endoglucanase GH5. Whole enzyme (called XFN3GH5 based on modular structure) and truncate forms FN3, XFN3, FN3GH5, GH5 were cloned in pET22b (+) and pET22SUMO to be expressed in single and fusion forms with a small ubiquitin-related modifier partner (S). The FN3, SFN3 increased GH5 solubility in FN3GH5, SFN3GH5. The SFN3 partly served for GH5 conformation in SFN3GH5, increased modules interaction and enzyme-soluble substrate affinity to enhance SXFN3GH5, SFN3GH5 activities in mixtures. Both SFN3 and SXFN3 did not anchor enzyme on filter paper but exfoliate and separate cellulose chains on filter paper for enzyme hydrolysis. Conclusion: Based on these findings, the presence of FN3 module in certain cellulases was confirmed and it assisted for enzyme conformation and activity in both soluble and insoluble substrate.

Safety evaluation of atmospheric pressure plasma jets in in vitro and in vivo experiments

  • Lee, Ji-Yoon;Park, Shin-Young;Kim, Kyoung-Hwa;Yoon, Sung-Young;Kim, Gon-Ho;Lee, Yong-Moo;Seol, Yang-Jo
    • Journal of Periodontal and Implant Science
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    • v.51 no.3
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    • pp.213-223
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    • 2021
  • Purpose: The atmospheric pressure plasma jet (APPJ) has been introduced as an effective disinfection method for titanium surfaces due to their massive radical generation at low temperatures. Helium (He) has been widely applied as a discharge gas in APPJ due to its bactericidal effects and was proven to be effective in our previous study. This study aimed to evaluate the safety and effects of He-APPJ application at both the cell and tissue levels. Methods: Cellular-level responses were examined using human gingival fibroblasts and osteoblasts (MC3T3-E1 cells). He-APPJ was administered to the cells in the experimental group, while the control group received only He-gas treatment. Immediate cell responses and recovery after He-APPJ treatment were examined in both cell groups. The effect of He-APPJ on osteogenic differentiation was evaluated via an alkaline phosphatase activity assay. In vivo, He-APPJ treatment was administered to rat calvarial bone and the adjacent periosteum, and samples were harvested for histological examination. Results: He-APPJ treatment for 5 minutes induced irreversible effects in both human gingival fibroblasts and osteoblasts in vitro. Immediate cell detachment of human gingival fibroblasts and osteoblasts was shown regardless of treatment time. However, the detached areas in the groups treated for 1 or 3 minutes were completely repopulated within 7 days. Alkaline phosphatase activity was not influenced by 1 or 3 minutes of plasma treatment, but was significantly lower in the 5 minute-treated group (P=0.002). In vivo, He-APPJ treatment was administered to rat calvaria and periosteum for 1 or 3 minutes. No pathogenic changes occurred at 7 days after He-APPJ treatment in the He-APPJ-treated group compared to the control group (He gas only). Conclusions: Direct He-APPJ treatment for up to 3 minutes showed no harmful effects at either the cell or tissue level.

Antioxidant activity and cell bioactivity of Sargassum macrocarpum extract (큰열매모자반(Sargassum macrocarpum) 추출물의 항산화 효과 및 세포 활성 효과)

  • Kim, Sook-hee
    • Journal of the Korea Convergence Society
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    • v.12 no.8
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    • pp.301-308
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    • 2021
  • In this study, the antioxidant and anti-inflammatory, anti-obesity properties, of Sargassum macrocarpum extracts were identified to assess the availability of Sargassum macrocarpum extracts as cosmetics and foods. To measure antioxidant activity, we conducted TPC, TFC, DPPH, ABTS, NO, FRAP. For polyphenols, 30.81±1.12 mg/g was shown. Flavonoids showed 25.72±0.94 mg/g. The DPPH experiment showed an antioxidant function of 6.746 mg ascorbic acid/g extract, the ABTS experiment showed an antioxidant function of 15.59 mg ascorbic acid/g extract, and the NO experiment showed an antioxidant function of 6.781 mg ascorbic acid/g extract. In FRAP, 1 mg of the Sargassum macrocarpum extract showed a reduction of 4.573±0.097 ㎍ of ascorbic acid. In cytotoxicity experiments, Sargassum macrocarpum extracts showed a cell survival rate of more than 80% at all concentrations, and an inflammatory inhibition of 25.95±0.85%, and an lipid accumulation inhibition of 29.75±2.35%. These results indicate that Sargassum macrocarpum extract is available as an anti-inflammatory cosmetic and anti-obesity inner beauty material. In future studies, it is necessary to study how pure substances containing Sargassum macrocarpum extract affect antioxidants, anti-inflammatory and anti-obesity

Whitening Effect of Abelmoschus esculentus on Melanoma Cells (B16F10) (B16F10 세포에서의 오크라 추출물의 미백 활성 검증)

  • Yoo, Dan-Hee;Lee, In-Chul
    • Microbiology and Biotechnology Letters
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    • v.49 no.4
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    • pp.485-492
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    • 2021
  • In this study, the whitening effect of Abelmoschus esculentus extract was investigated to confirm its applicability in cosmetics. To determine the whitening effect, the tyrosinase-inhibitory activity of Abelmoschus esculentus hot water extract (AEWE) and 70% ethanol extract (AEEE) was measured. At the final concentration of 1000 ㎍/ml, AEWE showed an inhibitory activity of 22.2% and AEEE of 32.8%. To determine the whitening effect at the cellular level, the viability of melanoma cells treated with AEWE and AEEE was evaluated using the MTT assay. At concentrations of 100 ㎍/ml or less, both AEWE- and AEEE-treated groups showed cell survival rates of >95%. Furthermore, in both AEWE- and AEEE-treated melanoma cells, the melanin content decreased in a concentration-dependent manner. The inhibitory effects of AEWE and AEEE used at 5, 10, 50, and 100 ㎍/ml on protein expression were measured by western blot, with β-actin as the positive control. At a concentration of 100 ㎍/ml, AEWE showed an inhibitory effect of 88.1%, 24.8%, 62.2%, and 42.9% on microphthalmia-associated transcription factors (MITF), tyrosinase, tyrosinase-related proteins (TRP)-1, and TRP-2 factors, respectively. At the same concentration, AEEE showed inhibitory effect of 65.3%, 58.3%, 66.2%, and 65.3% against MITF, tyrosinase, TRP-1, and TRP-2 factors, respectively. In conclusion, the whitening effects of AEWE and AEEE were verified, and their applicability as a natural ingredient in cosmetics was confirmed.

Effect of Plantaginis asiaticae Folium water extract on body fat loss in high fat-induced obese C57BL/6 mice (차전초 잎의 물 추출물이 고지방식이로 비만 유도된 C57BL/6 생쥐의 체지방 감소에 미치는 영향)

  • Kim, So Young;Jeong, Mi Jin;Kim, Yoo Jin;Lee, Un-Tak;Choo, Sung-Tae;Kim, Han-Hyuk;Kim, Mi Ryeo
    • The Korea Journal of Herbology
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    • v.33 no.2
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    • pp.59-67
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    • 2018
  • Objective : Plantago asiatica L. has been used for many years in Korea, China, and Japan for the treatment of many diseases such as anti-inflammatory, anti-asthma and diuretic. But the anti-obesity effect of Plantaginis asiaticae Folium has not been studied yet. Thus, this study aimed to demonstrate the anti-obesity efficacy of Plantaginis asiaticae Folium water extract (PAF) in high fat diet-induced obesity animal model. Method : Male, five-weeks old C57BL/6 mice were divided into 5 groups; ND (normal diet + 0.9% saline), HFD (high-fat diet + 0.9% saline), PC (high-fat diet + Garcinia cambogia 500 mg/kg), PAF 100, 300 (high-fat diet + PAF 100, 300 mg/kg. Treatments were performed daily for 6 weeks per os. We evaluated the changes of body weight, white adipose tissues weight, hepatic and fecal lipids level, pancreatic lipase activity and lipid profiles in plasma. Result : Body weights, adipose tissue weights and FER were significantly lower in mice fed the high-fat diet with PAF treatment than in mice fed the high-fat diet alone. In plasma, TG, TC, LDL-C, ATS and ALT levels significantly decreased in PAF groups compared with HFD group. Also, hepatic triglyceride (TG) levels decreased and fecal TG increased in PAF groups compared to HFD group. In addition, the activity of pancreatic lipase was decreased in the PAF groups compared to that of HFD group. Conclusion: These results suggest PAF extract is effective in preventing and improvement obesity, so it might be a potential and safe material for anti-obesity agent of weight control or functional supplements.

Technology Competitiveness in the AI-Edutech Field: Using Patent Indice and Hurdle Negative Binomial Model (특허 자료를 활용한 AI-에듀테크 분야 국가 간 기술 경쟁력 분석: 특허 통계 지표와 허들 음이항 모델의 활용)

  • Ilyong Ji;Hyun-young Bae
    • Journal of Industrial Convergence
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    • v.22 no.8
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    • pp.1-17
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    • 2024
  • Recently, interest in edutech has been focused on its fusion with AI technology, and the market in this field is expanding. This study aims to analyze the technological competitiveness and key technological areas of major countries in the AI-edutech field. Additionally, considering that AI-edutech is a convergence of AI technology and edutech, the study seeks to examine the path dependence of AI-edutech in each country to determine whether they are based on existing AI technologies or edutech. To this end, AI-edutech patents were collected and competitiveness was analyzed using patent activity, patent impact, and market acquisition indicators. Path dependence for each country was analyzed using the hurdle negative binomial regression model. The analysis results indicate that the major countries in the AI-edutech field are China, South Korea, the United States, India, and Japan. In terms of patent activity, China had the highest level, followed by South Korea. In terms of patent impact and market securing power, the United States was high in both aspects, Japan had high market securing power, and South Korea had high patent influence. The results of the hurdle negative binomial analysis presented unique findings. The logit part results indicated that the possession of existing AI and edutech did not positively affect the emergence of current AI-edutech, but the count part results showed a positive influence. This suggests that, overall, it is difficult to assert that current AI-edutechs are based on past AI and edutechs. However, once some AI-edutechs based on existing AI and edutechs emerge, they are influenced by the existing technologies. These findings provide implications for future research and technological strategies in this field.

The Effect of Environmental Factors on the Hydrolysis Characteristics of Lipase (환경인자가 리파제의 가수분해 특성에 미치는 영향)

  • Park, Geon-Gyu;Kim, Eun-Gi;Heo, Byeong-Gi
    • KSBB Journal
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    • v.14 no.4
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    • pp.511-516
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    • 1999
  • The effects of environmental and compositon factors, such as reaction time, metal ions, pH, agitation speed, the weight ratio of water to oil, and the weight of enzyme, on the hydrolysis of oils by Lipase-OF were investigated. In case of oils with low melting point, the optimum temperature of hydrolysis were the enzyme activity was maximum was 37$^{\circ}C$. However, when the melting temperature was higher than 4$0^{\circ}C$, the optimum temperature was around the fusion temperature. The activity of Lipase-OF decreased very rapidly with increase of temperature in the range of higher than 45$^{\circ}C$ and the activity perished above $65^{\circ}C$. The effect of agitation speed was investigated from 150 to 650 rpm. The hydrolysis of oils increased as the agitation speed increased up to 350 rpm, but it did not increase any more above 350 rpm. The weight ratio of water to oil was changed from 1 : 9 to 9 : 1 for the investigation of the effect on the hydrolusis. The weight ratio for maximum hydrolysis was 1 : 1. $Ca^{2+}\;and\;Mg^{2+}$ among various metal ions had some effect on the stimulation of hydrolysis. The optimum concentration of the ions was about 100ppm at which the hydrolysis increased, compared with that of distilled water, by 2 to 3%. The Optimum pH of Lipase-OF was 7. The hydrolysis decreased as the pH decreased as the pH decreased and also decreased as the pH increased. The content of enzyme affected the hydrolysis of oil. The hydrolysis increased with the content of Lipase-OF in the range of less than 0.013 wt% of substrate. However, the increase of hydrolysis with the content of Lipase-OF ceased above 0.013 wt%. The experiments investigating the effect of environmental and composition factors on the hydrolysis of oils showed that the optimum temperature was 37$^{\circ}C$, the pH 7, the concentration of $Ca^{2+}\;or\;Mg^{2+}$ 100 ppm, the agitation speed 350 rpm, the weight ratio of water to oil 1 : 1, and the content of Lipase-OF 0.013 wt% of substrate.

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