• Journal of Life Science
• /
• v.19 no.8
• /
• pp.1039-1046
• /
• 2009

The cytochrome P450s (P450s) metabolizing natural products are among the most versatile biological catalysts known in plants, but knowledge of the structural basis for their broad substrate specificity has been limited. The activity of p-coumarate 3-hydroxylase (C3H) is thought to be essential for the biosynthesis of lignin and many other phenylpropanoid pathway products in plants however, all attempts to express and purify the protein corresponding C3H gene have failed. As a result, no conditions suitable for the unambiguous assay of the enzyme are known. The detailed understanding of the mechanism and substrate-specificity of C3Hdemands a method for the production of active protein on the milligram scale. We have developed a bacterial expression and purification system for the plant C3H, which allows for the quick expression and purification of active wild-type C3H via introduction of combinational mutagenesis. The modified cytochrome P450 C3H ($C3H_{mod}$) could be purified in the absence of detergent using immobilized metal affinity chromatography and size exclusion chromatography following extraction from isolated membranes in a high salt buffer and catalytically activated. This method makes the use of isotopic labeling of C3H for NMRstudies and X-ray crystallography practical, and is also applicable to other plant cytochrome P450 proteins.

• Journal of Food Hygiene and Safety
• /
• v.38 no.3
• /
• pp.79-88
• /
• 2023

Trifludimoxazin is a triazinone herbicide that inhibits the synthesis of protoporphyrinogen oxidase (PPO). The lack of PPO damages the cell membranes, leading to plant cell death. An official analytical method for the safety management of trifludimoxazin is necessary because it is a newly registered herbicide in Korea. Therefore, this study aimed to develop a residual analysis method to detect trifludimoxazin in five representative agricultural products. The EN method was established as the final extraction method by comparing the recovery test and matrix effect with those of the QuEChERS method. Various sorbent agents were used to establish the clean-up method, and no differences were observed among them. MgSO₄ and PSA were selected as the final clean-up conditions. We used LC-MS/MS considering the selectivity and sensitivity of the target pesticide and analyzed the samples in the MRM mode. The recovery test results using the established analysis method and inter-laboratory validation showed a valid range of 73.5-100.7%, with a relative standard deviation and coefficient of variation less than 12.6% and 14.5%, respectively. Therefore, the presence of trifludimoxazin can be analyzed using a modified QuEChERS method, which is widely available in Korea to ensure the safety of residual insecticides.

• Korean Journal of Environmental Agriculture
• /
• v.14 no.3
• /
• pp.329-337
• /
• 1995

The effect of N-(4-chlorophenyl) maleimide(CPMI), plant growth regulators, and alkylating agents on gluathione(GSH) content and glutathione S-transferase(GST) activity was examined with 3-day-old etiolated sorghum(Sorghum bicolor [L.] Moench) seedlings. The GSH content and GST activity of untreated seedlings were higher in shoots than that in roots. Response of GST activity in coleoptile was significantly greater than in other tissues of sorghum seedling. In CPMI-treated seedlings, GSH content was not significantly different from that in untreated seedlings. CPM treatment resulted in 2.3-fold increase in GST activity measured with metolachlor as substrate in the coleoptile region. In contrast, change in GST activity measured with metolachlor as substrate in the coleoptile region. In contrast, change in GST activity measured with 1-chloro-2, 4-dinitrobenzene did not occur. The increase of GST activity was caused by induction of a GST isozyme, which is substrate-specific to metolachlor. Subsequently, two hypotheses related to metolachlor detoxification were evaluated on the basis of regulation of plant growth regulators and substrate induction of GST activity. In coleoptile, GST activity measured with metolachior was increased to 2.1-and 3.4-fold by both 2, 4-dichlorophenoxyacetic acid(2,4-D) and metolachlor treated at the germination stage of sorghum, respectively. Treatments of 2.4-D and metolachlor also induced isozymes exhibiting the activity toward metolachlor. One of the isozymes was co-eluted with that induced by CPMI. These results indicated that increase in GST activity by CPMI may be partially related to auxin regulation and substrate induction.

• Research in Plant Disease
• /
• v.23 no.2
• /
• pp.159-167
• /
• 2017

This study was conducted to evaluate the control efficacy of the organic farming materials on cucumber scab caused by Cladosporium cucumerinum PT1 (KACC 48094). The antifungal activities in vitro as well as the suppressive effect of 43 organic farming materials on the spore germination and germ tube growth by inoculating spore suspension on cucumber seedlings in vivo were investigated. Thirteen organic farming materials inhibited the mycelial growth of C. cucumerinum and nine of these were microbial agents. In the screening using cucumber seedlings, six organic farming materials were very effective with control efficacy value of 90%. Among them, Bacillus amyloliquefaciens M27 provided suppressive effect on both mycelial growth and spore germination against cucumber scab. Finally, nine organic farming materials were selected to test the protective and curative effects, and all chosen organic farming materials significantly suppressed disease incidence when applied in the preventive action, in comparison with the curative action. Especially, Bordeaux mixture I and III gave excellent protective control efficacy with control values of 96.7% and 73.3%, respectively, whereas its curative control effect was significant low. Among these, only Thymus quinquecostatus+Sophora extract showed curative activity, although the control value was as low as 50%. This study suggests that cucumber scab can be controlled by some organic farming materials in the farmhouses under comparatively cold and wet condition and protective treatment is more important and efficient.

• Journal of Food Hygiene and Safety
• /
• v.25 no.2
• /
• pp.170-179
• /
• 2010

This study was conducted to investigate for mineral contents, total polyphenol compounds, betaine, choline and DPPH free radical scavenging activity of halophyte. The mineral concentrations of Salicornia herbacea (top part) were Na 100,006 mg/kg, K 1,385 mg/kg, Mg 6,263 mg/kg, Ca 2,750 mg/kg, Fe 90.4 mg/kg, Mn 98.9 mg/kg, Zn 33.3 mg/kg, Cu 3.4 mg/kg respectively. And Suaeda Japonica (top part) were Na 85,332 mg/kg, K 710 mg/kg, Mg 7,005 mg/kg, Ca 4,344 mg/kg, Fe 1,434.9 mg/kg, Mn 119.1 mg/kg, Zn 19.2 mg/kg, Cu 2.7 mg/kg respectively. The betaine contents of Salicornia herbacea (top part) were 15.09 mg/g and Suaeda Japonica (top part) were 14.64 mg/g. The choline contents estimated by the DBAP-choline derivatives of Salicornia herbacea (top part) were 20.9 mg/100 g, Salicornia herbacea (root) were 23.4 mg/100 g, Suaeda Japonica (top part) were 23.1 mg/100g and Suaeda Japonica (root) were 23.8 mg/100 g. Total polyphenol compounds of Salicornia herbacea (top part) were high 36.0 mg/g in growth phase. The DPPH radical scavenging activities of methanol extract Salicornia herbacea (top part) were high 90.1% in growth phase. The frozen dried powder of Salicornia herbacea (top part) 1 g was equal to Quercetin 30.26 mg, Rutin 42.65 mg, TBHQ 20.32 mg, BHA 25.86 mg, BHT 40.75 mg, Ascorbic acid 22.86 mg in DPPH radical scavenging activities.

• Korean Journal of Plant Resources
• /
• v.28 no.5
• /
• pp.568-573
• /
• 2015

Arabis glabra is a localized common rhizomatous flowering plant, This plant is often used in Korean traditional systems of medicine as a remedy for blood cleaning, detoxification, abscess, gastrospasm, arthritis, contraction and diarrhea. Generally drugs that are used for arthritis have antinociceptive and anti-inflammatory properties. However, validity of the anti-inflammatory activity has not been scientifically investigated so far. Therefore, the aim of this study was to investigate the anti-inflammatory potential of A. glabra using the ethanolic extract and its sub-fractions. To evaluate the anti-inflammatory effects, we examined the inflammatory mediators such as nitric oxide (NO) and prostaglandin E₂ (PGE₂) on RAW 264.7macrophages. Our results indicated that hexane and chloroform fraction significantly inhibited the LPS-induced NO and PGE₂ production in the cells. The hexane fraction inhibitory activity for NO tests with IC₅₀ values showed in 21.0 ㎍/㎖. The chloroform fraction inhibitory activity for PGE₂ tests with IC₅₀ values showed in 18.0 ㎍/㎖. These efficacy are expected to be able to present the potential for the development of health functional food for the prevention inflammatory diseases because it has sufficient preventive medical possibilities. Further, it is determined that it is necessary to further study the mechanism of cytokine and protein expression associated with inflammation.

• Korean Journal of Microbiology
• /
• v.50 no.4
• /
• pp.351-359
• /
• 2014

A beverage was produced by the fermentation of mixed extracts from the various fruits, vegetables, algae, and medical herbs. The physicochemical properties of the fermented beverage of plant extracts (FBPE) and microbial diversity were analyzed in cultures enriched from FBPE using 16S and 26S rRNA gene sequence analyses. The pH, acidity, $^{\circ}brix$, reducing sugar, and alcohol contents of the FBPE were determined to be the 3.48, 1.68%, 70.0, 1,026 g/L, and 3.5%, respectively. The most abundant free sugar and organic acid in the FBPE were glucose (567.83 g/L) and tartaric acid (93.68 mg/L), respectively. Lactobacillus homohiochii was the predominant species in all enriched culture samples: 100% of the species in 0B (0% sugar) and 40B (40% sugar) libraries and 95.6% of 20B library (20% sugar). Lactobacillus fructivorans was detected in the 20B library. The predominant species in the samples of enrichment cultures collected from FBPE with three different sugar concentrations were: Candida zeylanoides (45.2%) in the 0Y library (0% sugar), Candida lactis-condensi (35.7%) and C. zeylanoides (35.7%) in the 20Y library (20% sugar), and C. lactis-condensi (38.1%) in the 40Y library (40% sugar). This result may provide a useful frame of reference for further analyses of microbial population dynamics in FBPE.

• Korean Journal of Plant Resources
• /
• v.35 no.5
• /
• pp.565-573
• /
• 2022

Gingival inflammation is one of the main causes that can be related to various periodontal diseases. Human gingival fibroblast (HGF) is the major constituent in periodontal connective tissue and secretes various inflammatory mediators, such as nitric oxide (NO) and prostaglandin E₂ (PGE₂), upon lipopolysaccharide stimulation. This study is aimed at investigating the anti-inflammatory and antioxidative activities of Lotus Root extract (LRE) in Porphyromonas gingivalis derived lipopolysaccharide (LPS-PG)-stimulated HGF-1 cells. The concentration of NO and PGE₂, as well as their responsible enzymes, inducible NO synthase (iNOS), and cyclooxygenase-2 (COX-2), was analyzed by Griess reaction, ELISA, and western blot analysis. LPS-PG sharply elevated the production and protein expression of inflammatory mediators, which were significantly attenuated by LRE treatment in a dose-dependent manner. LRE treatment also suppressed activation of Toll-like receptor 4 (TLR4)/myeloid differentiation primary response gene 88 (MyD88) and nuclear factor-κB (NF-κB) in LPS-PG-stimulated HGF-1 cells. In addition, one of phase II enzyme, NAD(P)H quinone dehydrogenase (NQO)-1, and its transcription factor, Nuclear factor erythroid 2-related factor 2 (Nrf2), were significantly induced by LRE treatment. Consequently, these results suggest that LRE ameliorates LPS-PG-induced inflammatory responses by attenuating TLR4/MyD88-mediated NF-κB, and activating NQO-1/Nrf2 antioxidant response element signaling pathways in HGF-1 cells.

• Journal of Plant Biotechnology
• /
• v.45 no.3
• /
• pp.250-256
• /
• 2018

To determine whether metabolite fingerprinting for whole cell extracts based on Fourier transform infrared spectroscopy (FT-IR) can be used to discriminate and compare metabolic equivalence, standard medicinal parts of Cynanchum wilfordii (Maxim.) Hemsl. and their adventitious roots were subjected to FT-IR. The principal component analysis (PCA) and partial least square discriminant analysis (PLS-DA) from FT-IR spectral data showed that whole metabolic pattern from the adventitious root of Cynanchum wilfordii was highly similar to its standard medicinal parts. These results clearly showed that mass proliferation of adventitious roots could be applied for the novel supply of standard medicinal parts of medicinal plants. Furthermore, FT-IR spectroscopy combined with multivariate analysis established in this study could be applied as an alternative tool for discriminating of whole metabolic equivalence from standard medicinal parts. Thus, it is proposed that these metabolic discrimination systems from the adventitious root of Cynanchum wilfordii could be applied for metabolic standardization of in vitro grown Cynanchum wilfordii.

• Research in Plant Disease
• /
• v.14 no.3
• /
• pp.187-192
• /
• 2008

Five environment-friendly farm materials including $Chitomate^{(R)}$, $Diegyun^{(R)}$, IC-$66D^{(R)}$, Gold $Bordo^{(R)}$, and $Biospot^{(R)}$ were examined for their growth inhibition effect of the 7 fungal pathogens of grape in vitro. $Diegyun^{(R)}$, being composed of natural ingredients which are extracted from a plant, was the most effective in suppression of mycelial growth of the fungi. $Diegyun^{(R)}$ inhibited the mycelial growth of all of fungi over 75% at $2,500{\mu}g{\cdot}mL^{-1}$ on potato dextrose agar(PDA) except Colletotrichum gloeosporioides 04-159. Growth inhibition effect of $Chitomate^{(R)}$, being composed of the chitosan, varied depending on the fungal pathogens on PDA. It inhibited the mycelial growth of the Botrytis cinerea 06-063 at the rate of 75.8% at $40,000{\mu}g{\cdot}mL^{-1}$ on PDA while it inhibited the mycelial growth of the C. gloeosporioides 04-159 at the rate of 6.5%. IC-$66D^{(R)}$ and Gold $Bordo^{(R)}$ are two different formula of the Bordeaux mixture, showed different control effects on mycelial growth inhibition. Except of Acremonium sp. the growth inhibition of IC-$66D^{(R)}$ was a little higher than Gold $Bordo^{(R)}$. $Biospot^{(R)}$, a chlorine formula, showed the strongest growth inhibition on C. gloeosporioides 04-159 among the farm materials used. Inhibition of spore germination of $Chitomate^{(R)}$, $Biospot^{(R)}$ and Gold $Bordo^{(R)}$ was higher than mycelial growth inhibition for Pseudocercospora vitis 04-152. The results suggest that the different types of environment-friendly farm materials are needed for different disease control in organic grape farm.