• Title/Summary/Keyword: metabolite analysis

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Nematicidal Activity of Kojic Acid Produced by Aspergillus oryzae against Meloidogyne incognita

  • Kim, Tae Yoon;Jang, Ja Yeong;Jeon, Sun Jeong;Lee, Hye Won;Bae, Chang-Hwan;Yeo, Joo Hong;Lee, Hyang Burm;Kim, In Seon;Park, Hae Woong;Kim, Jin-Cheol
    • Journal of Microbiology and Biotechnology
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    • v.26 no.8
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    • pp.1383-1391
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    • 2016
  • The fungal strain EML-DML3PNa1 isolated from leaf of white dogwood (Cornus alba L.) showed strong nematicidal activity with juvenile mortality of 87.6% at a concentration of 20% fermentation broth filtrate at 3 days after treatment. The active fungal strain was identified as Aspergillus oryzae, which belongs to section Flavi, based on the morphological characteristics and sequence analysis of the ITS rDNA, calmodulin (CaM), and β-tubulin (BenA) genes. The strain reduced the pH value to 5.62 after 7 days of incubation. Organic acid analysis revealed the presence of citric acid (515.0 mg/kg), malic acid (506.6 mg/kg), and fumaric acid (21.7 mg/kg). The three organic acids showed moderate nematicidal activities, but the mixture of citric acid, malic acid, and fumaric acid did not exhibit the full nematicidal activity of the culture filtrate of EML- DML3PNa1. Bioassay-guided fractionation coupled with 1H- and 13C-NMR and EI-MS analyses led to identification of kojic acid as the major nematicidal metabolite. Kojic acid exhibited dose-dependent mortality and inhibited the hatchability of M. incognita, showing EC50 values of 195.2 μg/ml and 238.3 μg/ml, respectively, at 72 h post-exposure. These results suggest that A. oryzae EML-DML3PNa1 and kojic acid have potential as a biological control agent against M. incognita.

Metabolomic analysis of perilla seeds harvested from Korea and China (국내산 및 중국산 들깨 종자의 대사체 분석)

  • Gu, Suyeon;Choi, Nayoung;Son, Yejin;Park, Ji Yeong;Choi, Sung-Gil;Lee, Myoung Hee;Kim, Hyun-Jin
    • Korean Journal of Food Science and Technology
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    • v.51 no.5
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    • pp.411-419
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    • 2019
  • A comprehensive comparison of fatty acid and metabolite profiles in 8 species of perilla seeds harvested from Korea and China was carried out to understand the correlation between cultivation conditions and the quality of perilla seeds. Metabolomic studies revealed that the perilla seeds were distinct from each other, based on the partial least squares -discriminant analysis (PLS-DA) score plots; 23 compounds including metabolites, fatty acids, and phytosterols that predominantly contributed to this interspecies difference were identified. Changes in the growth environment affected these compounds. In particular, phenolic compounds were affected by rainfall and temperature. Moreover, temperature and altitude showed negative and positive correlations, respectively, to oleic acid. Although more studies on the effect of various environmental factors on cultivation of perilla plants as well as their seeds are needed, the present results provide information that would be useful in identifying perilla seeds produced in specific regions and in the cultivation of good quality perilla.

Development of the conventional crop composition database for new genetically engineered crop safety assessment (새로운 생명공학작물 안전성 평가를 위한 작물 성분 DB 구축)

  • Kim, Eun-Ha;Lee, Seong-Kon;Park, Soo-Yun;Lee, Sang-Gu;Oh, Seon-Woo
    • Journal of Plant Biotechnology
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    • v.45 no.4
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    • pp.289-298
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    • 2018
  • The Biosafety Division of the National Academy of Agricultural Science has developed a 'Crop Composition DB' that provides analytical data on commercialized crops. It can be used as a reference in the 'Comparative Evaluation by Compositional Analysis' for the safety assessment of genetically modified (GM) crops. This database provides the composition of crops cultivated in Korea, and thus upgrades the data to check the extent of changes in the compositional content depending on the cultivated area, varieties and year. The database is a compilation of data on the antioxidant, nutrient and secondary metabolite compositions of rice and capsicum grown in two or more cultivation areas for a period of more than two years. Data analysis was conducted under the guidelines of the Association of Official Analytical Chemists or methods previously reported on papers. The data was provided as average, minimum and maximum values to assess whether the statistical differences between the GM crops and comparative non-GM crops fall within the biological differences or tolerances of the existing commercial crops. The Crop Composition DB is an open-access source and is easy to access based on the query selected by the user. Moreover, functional ingredients of colored crops, such as potatoes, sweet potatoes and cauliflowers, were provided so that food information can be used and utilized by general consumers. This paper introduces the feature and usage of 'Crop Composition DB', which is a valuable tool for characterizing the composition of conventional crops.

Characterization of ginsenoside compound K loaded ionically cross-linked carboxymethyl chitosan-calcium nanoparticles and its cytotoxic potential against prostate cancer cells

  • Zhang, Jianmei;Zhou, Jinyi;Yuan, Qiaoyun;Zhan, Changyi;Shang, Zhi;Gu, Qian;Zhang, Ji;Fu, Guangbo;Hu, Weicheng
    • Journal of Ginseng Research
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    • v.45 no.2
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    • pp.228-235
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    • 2021
  • Backgroud: Ginsenoside compound K (GK) is a major metabolite of protopanaxadiol-type ginsenosides and has remarkable anticancer activities in vitro and in vivo. This work used an ionic cross-linking method to entrap GK within O-carboxymethyl chitosan (OCMC) nanoparticles (Nps) to form GK-loaded OCMC Nps (GK-OCMC Nps), which enhance the aqueous solubility and stability of GK. Methods: The GK-OCMC Nps were characterized using several physicochemical techniques, including x-ray diffraction, transmission electron microscopy, zeta potential analysis, and particle size analysis via dynamic light scattering. GK was released from GK-OCMC Nps and was conducted using the dialysis bag diffusion method. The effects of GK and GK-OCMC Nps on PC3 cell viability were measured by using the 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide assay. Fluorescent technology based on Cy5.5-labeled probes was used to explore the cellular uptake of GK-OCMC Nps. Results: The GK-OCMC NPs had a suitable particle size and zeta potential; they were spherical with good dispersion. In vitro drug release from GK-OCMC NPs was pH dependent. Moreover, the in vitro cytotoxicity study and cellular uptake assays indicated that the GK-OCMC Nps significantly enhanced the cytotoxicity and cellular uptake of GK toward the PC3 cells. GK-OCMC Nps also significantly promoted the activities of both caspase-3 and caspase-9. Conclusion: GK-OCMC Nps are potential nanocarriers for delivering hydrophobic drugs, thereby enhancing water solubility and permeability and improving the antiproliferative effects of GK.

A possible mechanism to the antidepressant-like effects of 20 (S)-protopanaxadiol based on its target protein 14-3-3 ζ

  • Chen, Lin;Li, Ruimei;Chen, Feiyan;Zhang, Hantao;Zhu, Zhu;Xu, Shuyi;Cheng, Yao;Zhao, Yunan
    • Journal of Ginseng Research
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    • v.46 no.5
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    • pp.666-674
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    • 2022
  • Background: Ginsenosides and their metabolites have antidepressant-like effects, but the underlying mechanisms remain unclear. We previously identified 14-3-3 ζ as one of the target proteins of 20 (S)-protopanaxadiol (PPD), a fully deglycosylated ginsenoside metabolite. Methods: Corticosterone (CORT) was administered repeatedly to induce the depression model, and PPD was given concurrently. The tail suspension test (TST) and the forced swimming test (FST) were used for behavioral evaluation. All mice were sacrificed. Golgi-cox staining, GSK 3β activity assay, and Western blot analysis were performed. In vitro, the kinetic binding analysis with the Biolayer Interferometry (BLI) was used to determine the molecular interactions. Results: TST and FST both revealed that PPD reversed CORT-induced behavioral deficits. PPD also ameliorated the CORT-induced expression alterations of hippocampal Ser9 phosphorylated glycogen synthase kinase 3β (p-Ser9 GSK 3β), Ser133 phosphorylated cAMP response element-binding protein (p-Ser133 CREB), and brain-derived neurotrophic factor (BDNF). Moreover, PPD attenuated the CORT-induced increase in GSK 3β activity and decrease in dendritic spine density in the hippocampus. In vitro, 14-3-3 ζ protein specifically bound to p-Ser9 GSK 3β polypeptide. PPD promoted the binding and subsequently decreased GSK 3β activity. Conclusion: These findings demonstrated the antidepressant-like effects of PPD on the CORT-induced mouse depression model and indicated a possible target-based mechanism. The combination of PPD with the 14-3-3 ζ protein may promote the binding of 14-3-3 ζ to p-GSK 3β (Ser9) and enhance the inhibition of Ser9 phosphorylation on GSK 3β kinase activity, thereby activating the plasticity-related CREBeBDNF signaling pathway.

Analysis of quality characteristics of sugar-soaked raspberry according to storage period (저장 기간에 따른 산딸기 당 침지액의 품질특성 분석)

  • Choi, Seok-Yong;Gu, Suyeon;Ryu, Chung-Ho;Kim, Hyun-Jin
    • Journal of Applied Biological Chemistry
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    • v.65 no.1
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    • pp.7-15
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    • 2022
  • Volatile and non-volatile metabolite profiles of sugar-immersed raspberry liquid during different storage periods were analyzed and comparative analysis with their general characteristics, antioxidant activity, and sensory quality was evaluated to better understand the effect of the storage period on the quality of ugar-immersed raspberry liquid. During storage, a browning reaction occurred, resulting in a change in color and the production of volatile compounds. At the beginning of storage, sucrose was completely decomposed into fructose and glucose, and the sweetness was rapidly reduced, but the increase in succinic acid increased the sour taste. Most volatile compounds increased with an increase of the storage period, and especially, the contents of citronellol, octanoic acid, and hexanoic acid, which are known as antioxidants, showed the highest content in 10 day-sample, showing the highest antioxidant activity at this time. Although a further study on bacterial profiles and browning reaction during the storage will be needed, the results of this study showed that the quality of sugar-soaked raspberry extract was significantly affected by the storage period and can be used as basic data for commercialization of ugar-immersed raspberry liquid.

Rapid metabolic discrimination between Zoysia japonica and Zoysia sinica based on multivariate analysis of FT-IR spectroscopy (FT-IR스펙트럼 데이터의 다변량통계분석 기반 들잔디와 갯잔디의 대사체 수준 신속 식별 체계)

  • Yang, Dae-Hwa;Ahn, Myung Suk;Jeong, Ok-Cheol;Song, In-Ja;Ko, Suk-Min;Jeon, Ye-In;Kang, Hong-Gyu;Sun, Hyeon-Jin;Kwon, Yong-Ik;Kim, Suk Weon;Lee, Hyo-Yeon
    • Journal of Plant Biotechnology
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    • v.43 no.2
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    • pp.213-222
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    • 2016
  • This study aims to establish a system for the rapid discrimination of Zoysia species using metabolite fingerprinting of FT-IR spectroscopy combined with multivariate analysis. Whole cell extracts from leaves of 19 identified Zoysia japonica, 6 identified Zoysia sinica, and 38 different unidentified Zoysia species were subjected to Fourier transform infrared spectroscopy (FT-IR). PCA (principle component analysis) and PLS-DA (partial least square discriminant analysis) from FT-IR spectral data successfully divided the 25 identified turf grasses into two groups, representing good agreement with species identification using molecular markers. PC (principal component) loading values show that the $1,100{\sim}950cm^{-1}$ region of the FT-IR spectra are important for the discrimination of Zoysia species. A dendrogram based on hierarchical clustering analysis (HCA) from the PCA and PLS-DA data of turf grasses showed that turf grass samples were divided into Zoysia japonica and Zoysia sinica in a species-dependent manner. PCA and PLS-DA from FT-IR spectral data of Zoysia species identified and unidentified by molecular markers successfully divided the 49 turf grasses into Z. japonica and Z. sinica. In particular, PLS-DA and the HCA dendrogram could mostly discriminate the 47 Z. japonica grasses into two groups depending on their origins (mountainous areas and island area). Considering these results, we suggest that FT-IR fingerprinting combined with multivariate analysis could be applied to discriminate between Zoysia species as well as their geographical origins of various Zoysia species.

Rapid comparison of metabolic equivalence of standard medicinal parts from medicinal plants and their in vitro-generated adventitious roots using FT-IR spectroscopy (한약자원 품목별 표준시료와 기내 생산 부정근의 FT-IR 스펙트럼 기반 대사체 동등성 신속 비교)

  • Ahn, Myung Suk;Min, Sung Ran;Jie, Eun Yee;So, Eun Jin;Choi, So Yeon;Moon, Byeong Cheol;Kang, Young Min;Park, So-Young;Kim, Suk Weon
    • Journal of Plant Biotechnology
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    • v.42 no.3
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    • pp.257-264
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    • 2015
  • To determine whether metabolite fingerprinting for whole cell extracts based on Fourier transform infrared (FT-IR) spectroscopy can be used to discriminate and compare metabolic equivalence, standard medicinal parts from four medicinal plants (Cynanchum wilfordii Hemsley, Atractylodes japonica Koidz, Polygonum multiflorum Thunberg and Astragalus membranaceus Bunge) and their in vitro-produced adventitious roots were analyzed by FT-IR spectroscopy. The principal component analysis (PCA) and partial least square discriminant analysis (PLS-DA) from the FT-IR spectral data showed that the whole metabolic pattern from Cynanchum wilfordii was highly similar to Astragalus membranaceus. However, Atractylodes japonica and Polygonum multiflorum showed significantly different metabolic patterns. Furthermore, adventitious roots from Cynanchum wilfordii and Astragalus membranaceus also showed similar metabolic patterns compared to their standard medicinal parts. These results clearly show that mass proliferation of adventitious roots may be applied to aquire novel supply of standard medicinal parts from medicinal plants. However, the whole metabolic pattern from adventitious roots of Atractylodes japonica and Polygonum multiflorum were not similar to their standard medicinal parts. Furthermore, FT-IR spectroscopy combined with multivariate analyses established in this study may be applied as an alternative tool to discriminate the whole metabolic equivalence from several standard medicinal parts. Thus, we suggest that these metabolic discrimination systems may be applied for metabolic standardization of herbal medicinal resources.

Gemcitabine-induced Cell Death in Lung Cancer Cells : the Role of p53 (폐암 세포에서 Gemcitabine에 의한 세포 사멸과 p53의 역할)

  • Kim, Doh-Hyung;Bae, Gang-U;Yong, Wha-Shim;Choi, Eun-Kyung;Kim, Youn-Seup;Park, Jae-Seuk;Jee, Young-Koo;Lee, Kye-Young
    • Tuberculosis and Respiratory Diseases
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    • v.53 no.3
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    • pp.275-284
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    • 2002
  • Background : Gemcitabine is a new anti-cancer agent for treating non-small cell lung cancer. Functioning as an antimetabolite, it induces anti-cancer effects by suppressing DNA synthesis after being incorporated into the DNA as a cytosine arabinoside analogue. When Gemcitabine is incorporated into the DNA, the p53 gene may be activated by induction of the DNA defect. However, there are a few studies on the molecular mechanisms of Gemcitabine-induced cell death. This study examined the role of p53 in Gemcitabine-induced cell death. Methods : A549 and NCl-H358 lung cancer cells were used in this study. The cell viability test was done using a MTT assay at Gemcitabine concentrations of 10nM, 100nM, 1uM, 10uM and 100uM. A FACScan analysis with propium iodide staining was used for the cell cycle analysis. Western blot analysis was done to investigate the extent of p53 activation. For the functional knock-out of p53, stable A549-E6 cells and H358-E6 cells were transfected pLXSN-16E6SD which is over expresses the human papilloma virus E6 protein that constantly degrades p53 protein. The functional knock out of p53 was confirmed by Western blot analysis after treatment with a DNA damaging agent, doxorubicine. Results : Gemcitabine exhibited cell toxicity in dose-dependent fashion. The cell cycle analysis resulted in an S phase arrest. Western blot analysis significant p53 activation in time-dependent manner. Gemcitabine-induced cytotoxicity was reduced by 20-30% in the A549-E6 cells and the 30-40% in H358-E6 cells when compared with the A549-neo and H358-neo control cells. Conclusion : Gemcitabine induces an S phase arrest, as expected for the anti-metabolite, and activates the p53 gene, Furthermore, p53 might play an important role in Gemcitabine-induced cell death. Further investigation into the molecular mechanisms on how Gemcitabine activates the p53 gene and its signaling pathway are recommended.

The Possible Mechanisms Involved in Citrinin Elimination by Cryptococcus podzolicus Y3 and the Effects of Extrinsic Factors on the Degradation of Citrinin

  • Zhang, Xiaoyun;Lin, Zhen;Apaliya, Maurice Tibiru;Gu, Xiangyu;Zheng, Xiangfeng;Zhao, Lina;Abdelhai, Mandour Haydar;Zhang, Hongyin;Hu, Weicheng
    • Journal of Microbiology and Biotechnology
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    • v.27 no.12
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    • pp.2119-2128
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    • 2017
  • Citrinin (CIT) is a toxic secondary metabolite produced by fungi belonging to the Penicillium, Aspergillus, and Monascus spp. This toxin has been detected in many agricultural products. In this study, a strain Y3 with the ability to eliminate CIT was screened and identified as Cryptococcus podzolicus, based on the sequence analysis of the internal transcribed spacer region. Neither uptake of CIT by cells nor adsorption by cell wall was involved in CIT elimination by Cryptococcus podzolicus Y3. The extracellular metabolites of Cryptococcus podzolicus Y3 stimulated by CIT or not showed no degradation for CIT. It indicated that CIT elimination was attributed to the degradation of intracellular enzyme(s). The degradation of CIT by C. podzolicus Y3 was dependent on the type of media, yeast concentration, temperature, pH, and initial concentration of CIT. Most of the CIT was degraded by C. podzolicus Y3 in NYDB medium at 42 h but not in PDB medium. The degradation rate of CIT was the highest (94%) when the concentration of C. podzolicus Y3 was $1{\times}10^8cells/ml$. The quantity of CIT degradation was highest at $28^{\circ}C$, and there was no degradation observed at 3$5^{\circ}C$. The study also showed that acidic condition (pH 4.0) was the most favorable for CIT degradation by C. podzolicus Y3. The degradation rate of CIT increased to 98% as the concentration of CIT was increased to $20{\mu}g/ml$. The toxicity of CIT degradation product(s) toward HEK293 was much lower than that of CIT.