• Title/Summary/Keyword: Common metabolite

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Improvement of Cyclosporin A Hydroxylation in Sebekia benihana by Conjugational Transfer of Streptomyces coelicolor SCO4967, a Secondary Metabolite Regulatory Gene (Sebekia benihana에서 Streptomyces coelicolor SCO4967 유전자 도입을 통한 하이드록실 사이클로스포린 A의 생전환)

  • Kim, Hyun-Bum;Lee, Mi-Jin;Han, Kyu-Boem;Kim, Eung-Soo
    • Microbiology and Biotechnology Letters
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    • v.38 no.4
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    • pp.475-480
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    • 2010
  • Actinomycetes are Gram-positive soil bacteria and one of the most important industrial microorganisms due to superior biosynthetic capabilities of many valuable secondary metabolites as well as production of various valuable bioconversion enzymes. Among them are cytochrome P450 hydroxylase (CYP), which are hemoproteins encoded by a super family of genes, are universally distributed in most of the organisms from all biological kingdoms. Actinomycetes are a rich source of soluble CYP enzymes, which play critical roles in the bioactivation and detoxification of a wide variety of metabolite biosynthesis and xenobiotic transformation. Cyclosporin A (CyA), one of the most commonly-prescribed immunosuppressive drugs, was previously reported to be hydroxylated at the position of 4th N-methyl leucine by a rare actinomycetes called Sebekia benihana, leading to display different biological activity spectrum such as loss of immunosuppressive activities yet retaining hair growth-stimulating side effect. In order to improve this regio-selective CyA hydroxylation in S. benihana, previously-identified several secondary metabolite up-regulatory genes from Streptomyces coelicolor and S. avermitilis were heterologously overexpressed in S. benihana using an $ermE^*$ promoter-containing Streptomyces integrative expression vector. Among tested, SCO4967 encoding a conserved hypothetical protein significantly stimulated region-specific CyA hydroxylation in S. benihana, implying that some common regulatory systems functioning in both biosynthesis and bioconversion of secondary metabolite might be present in different actinomycetes species.

Studies on resveratrol and its metabolite in human urine by GC/MS (GC/MS를 이용한 요 중 resveratrol과 그 대사체에 관한 연구)

  • Jung, Hyun-Joo;Paeng, Ki-Jung;Kim, Yun-Je
    • Analytical Science and Technology
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    • v.24 no.2
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    • pp.142-149
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    • 2011
  • This study was conducted to define metabolite of the resveratrol by gas chromatography- time-offlight mass spectrometric detection. From these results, we suppose that the structure of metabolite is the result of reduction of double-bond attached by two-phenyl groups. Also, validity of method for determining metabolite of resveratrol and endogenous steroids was tested. The recoveries ranged from 96.47 to 114.74%, and intraand inter-day precision ranged 11.40 - 10.87% and 1.10 - 10.93%, accuracy ranged 80.03 - 119.92% and 80.02 - 119.56%, respectively. Resveratrol and endogenous steroids had correlation coefficients above or equal to 0.996. The method was successfully validated for the determination of resveratrol and endogenous steroids. Urinary samples from volunteers dosed resveratrol were analyzed to confirm a correlation resveratrol and its metabolite. From these results, the highest level of resveratrol and its metabolite was excreted in 10 - 15 hr more slowly than common drug, and conversion rate of metabolite was higher in woman than that in man. In addition, endogenous steroids were shown same the highest level of 10 - 15 hr. For estrone and estradiol, sensitivity was relatively higher in female than in man. And there were no significant changes of excretion patterns in the other endogenous steroids. Thus, we assumed that activation of resveratrol has impact on woman than man.

Toxicological Characterization of Phthalic Acid

  • Bang, Du-Yeon;Lee, In-Kyung;Lee, Byung-Mu
    • Toxicological Research
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    • v.27 no.4
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    • pp.191-203
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    • 2011
  • There has been growing concern about the toxicity of phthalate esters. Phthalate esters are being used widely for the production of perfume, nail varnish, hairsprays and other personal/cosmetic uses. Recently, exposure to phthalates has been assessed by analyzing urine for their metabolites. The parent phthalate is rapidly metabolized to its monoester (the active metabolite) and also glucuronidated, then excreted. The objective of this study is to evaluate the toxicity of phthalic acid (PA), which is the final common metabolic form of phthalic acid esters (PAEs). The individual PA isomers are extensively employed in the synthesis of synthetic agents, for example isophthalic acid (IPA), and terephthalic acid (TPA), which have very broad applications in the preparation of phthalate ester plasticizers and components of polyester fiber, film and fabricated items. There is a broad potential for exposure by industrial workers during the manufacturing process and by the general public (via vehicle exhausts, consumer products, etc). This review suggests that PA shows in vitro and in vivo toxicity (mutagenicity, developmental toxicity, reproductive toxicity, etc.). In addition, PA seems to be a useful biomarker for multiple exposure to PAEs in humans.

Kinetics of Enriched Chitinase as Extracellular Metabolite in Beauveria bassiana

  • Mondal, Subhoshmita;Datta, Siddhartha;Mukherjee, Alakananda;Bhattacharya, Pinaki
    • Microbiology and Biotechnology Letters
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    • v.47 no.1
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    • pp.96-104
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    • 2019
  • Beauveria bassiana, one of the most common entomopathogenic fungi, has been isolated, pre defined and characterized in-house from soil of tea cultivation area. Experiments have been performed to verify the presence of chitinase as intracellular metabolite and its release as extracellular product rendering the spores with biopesticide activity. Although there are many responsible enzymes for the pest killer action of B. bassiana, binding property of chitinase depending on presence as well as absence of serine supplemented in the media has been studied with respect to the production and kinetics. A programmed investigation conclusively indicates that the isolated spore (hyphae) of B. bassiana has been metabolically enriched with the enzyme chitinase in presence of an externally added amino acid serine with its inhibitory kinetics.

Antidepressant effect of Licium chinense Mill. and its influence on indoleamine and its metabolite of depression model rats (구기자의 항우울효과 및 indoleamine에 미치는 영향)

  • Lee Duk-Ki;Gwak Dong-Gul;Park Sun-Dong
    • Herbal Formula Science
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    • v.11 no.2
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    • pp.185-196
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    • 2003
  • Depression is very common mental disorder, so many people suffer from it, which makes the treatment of depression important. Many drugs to treat depression were developed and being prescripted. But they have a lot of side effects, so it needs to develop drugs without side effects or with less side effects. Herbal medicines have been used to treat not only physical disorder but also mental disorder and it has been reported that they have less side effects. Therefore, there is the need to discover and use herbal medicine with antidepressant effect. The purpose of this study was to reseach Antidepressant effect of Licium chinense Mill. and its influence on serotonin and its metabolite of depression model rats. We used 'forced swimming test(FST)' to know antidepressant effect of Licium chinense Mill. and HPLC to check the influence on serotonin and its metabolite(5-HIAA) of Licium chinense Mill. after rats' brains were divided into cerebral cortex, striatum, hypothalamus and hippocampus. The results were obtained as follows : In the study of antidepressant effect by 'forced swimming test(FST)' method, Licium chinense Mill. had a significant antidepressant effect. In the study of influence on serotonin and 5-HIAA by HPLC, Licium chinense Mill. mainly increased serotonin and 5-HlAA of cerebral cortex and striatum signigficantly among 4 parts of rat's brain above-mentioned. These results suggest that Licium chinense Mill. has antidepressant effect that may be related with the increase of serotonin and its metabolite as its mechanism, but more precise experiments will be need to prove their relation.

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Pharmacokinetics of a New Histone Hl Protein (p961), an Arthritis-suppressing Agent, in Rats and Rabbits (항류마치스 효과를 갖는 새로운 히스톤 H1 단백질 (p961)의 흰쥐와 토끼에 대한 약물동태)

  • 우수경;윤민혁;이재흥;권광일
    • YAKHAK HOEJI
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    • v.45 no.4
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    • pp.378-386
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    • 2001
  • A purified histone Hl protein, p961, which plays a role in mediating the condensation of DNA into chromatin, was recently proved as an arthritis-suppressing agent in the mouse CIA model. The pharmacokinetics of p961 was carried out in rats and rabbits. The rat's blood, bile and urine samples were serially collected from the femoral vein, common bile duct, and bladder respectively, after bolus i.v. injection at low (10 mg/kg) and high (30 mg/mg) doses. The rabbit's blood samples were also collected from the marginal ear vein after bolus i.v. injection at a dose 10 mg/kg. p961 and its major metabolite in the physiological samples were analyzed by reverse-phase HPLC using a Yydac C4 protein column and a multistep water-acetonitrile gradient containing 0.24% trifluoroacetic acid. The major pharmacokinetic parameters (AUC, $C_{max}$, MRT, $t_{1}$2/, $V_{ss}$ and Cl) were estimated from the time course of plasma p961 and metabolite concentrations using WinNonlin. A two-compartment model was chosen for p961 as the most appropriate pharmacokinetic model. After i.v. injection of p961 at doses of 10 mg/kg and 30 mg/kg, more than 80% of p961 was removed rapidly from the plasma within 15 min. The plasma half-life of p961 in rats and rabbits was found not to exceed 12 min. p961 (22448.9 mol wt) was rapidly cleaved to 21612 mot wt fragment and the breakdown product appeared rapidly in the circulation with no lag phase. p961 and metabolite were not detected in rat urine and bile....

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Analysis of Endophytic Bacterial Communities and Investigation of Core Taxa in Apple Trees

  • Yejin Lee;Gyeongjun Cho;Da-Ran Kim;Youn-Sig Kwak
    • The Plant Pathology Journal
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    • v.39 no.4
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    • pp.397-408
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    • 2023
  • Fire blight disease, caused by Erwinia amylovora, is a devastating affliction in apple cultivation worldwide. Chemical pesticides have exhibited limited effectiveness in controlling the disease, and biological control options for treating fruit trees are limited. Therefore, a relatively large-scale survey is necessary to develop microbial agents for apple trees. Here we collected healthy apple trees from across the country to identify common and core bacterial taxa. We analyzed the endophytic bacterial communities in leaves and twigs and discovered that the twig bacterial communities were more conserved than those in the leaves, regardless of the origin of the sample. This finding indicates that specific endophytic taxa are consistently present in healthy apple trees and may be involved in vital functions such as disease prevention and growth. Furthermore, we compared the community metabolite pathway expression rates of these endophyte communities with those of E. amylovora infected apple trees and discovered that the endophyte communities in healthy apple trees not only had similar community structures but also similar metabolite pathway expression rates. Additionally, Pseudomonas and Methylobacterium-Methylorobrum were the dominant taxa in all healthy apple trees. Our findings provide valuable insights into the potential roles of endophytes in healthy apple trees and inform the development of strategies for enhancing apple growth and resilience. Moreover, the similarity in cluster structure and pathway analysis between healthy orchards was mutually reinforcing, demonstrating the power of microbiome analysis as a tool for identifying factors that contribute to plant health.

The Analysis of Common Metabolites of Organophosphorus Pesticides in Urine by Gas Chromatography/Mass Spectrometry

  • 박성수;표희수;이강진;박송자;박택규
    • Bulletin of the Korean Chemical Society
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    • v.19 no.1
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    • pp.45-50
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    • 1998
  • Most organophosphorus pesticides may be metabolized to yield some common phosphates in human or in animals, and these metabolites may be used as the exposure biomarkers to pesticides. In this study, we developed the extraction method of four phosphate metabolites from the spiked human urine in high recovery by the solid phase extraction with a reverse-phase cartridge (cyclohexyl silica) followed by the elution with methanol. The extracted urinary metabolites were derivatized with hexamethyldisilazane/trimethyl-chlorosilane/pyridine (2 : 1 : 10, v/v/v) and identified by gas chromatography/mass spectrometry. Calibration curve obtained from each metabolite standard using by GC/MS/SIM has shown good linearity and detection limits of metabolites were the range of 0.05-0.1 ㎍/㎖ in urine. Phenthoate, one of the organophosphorus pesticides, was orally administrated to rats. Four metabolites were detected in the rat urine. The results of this study may be applied to development of exposure biomarkers for monitoring of environmental pollutants.

Identification of Nandrolone and its Metabolite 5α-Estran-3β, 17α-Diol in Horse Urine after Chemical Derivatization by Liquid Chromatography Tandem Mass Spectrometry

  • Dubey, Saurabh;Beotra, Alka
    • Mass Spectrometry Letters
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    • v.8 no.4
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    • pp.90-97
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    • 2017
  • Androgenic anabolic steroids (AASs) are synthetic derivatives of testosterone with a common structure containing cyclopentanoperhydrophenanthrene nucleus. Their use enhances the muscle building capacity and is beneficial during performance. The AASs are one of the most abused group of substances in horse doping. Liquid chromatography tandem mass spectrometry ($LC/MS^n$) has been successfully applied to the detection of anabolic steroids in biological samples. However, the saturated hydroxysteroids viz: nandrolone, $5{\alpha}-estrane-3{\beta}$, $17{\alpha}-diol$ exhibit lower detection responses in electrospray ionisation (ESI) because of their poor ionisation efficiency. To overcome this limitation pre-column chemical derivatization has been introduced to enhance their detection responses in $LC-ESI-MS^n$ analysis. The aim of present study was to develop a sensitive method for identification and confirmation of nandrolone and its metabolite in horse urine incorporating pre-column derivatization using picolinic acid. The method consists of extraction of targeted steroid conjugates by solid phase extraction (SPE). The eluted steroid conjugates were hydrolysed by methanolysis and free steroids were recovered with liquid-liquid extraction. The resulting steroids were derivatized to form picolinoyl esters and identification was done using LC-ESI-MS/MS in positive ionization mode. The picolinated steroid adduct enhanced the detection levels in comparison to underivatized steroids.

Dosage Effects of Salt and pH Stresses on Saccharomyces cerevisiae as Monitored via Metabolites by Using Two Dimensional NMR Spectroscopy

  • Chae, Young Kee;Kim, Seol Hyun;Ellinger, James E.;Markley, John L.
    • Bulletin of the Korean Chemical Society
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    • v.34 no.12
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    • pp.3602-3608
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    • 2013
  • Saccharomyces cerevisiae, which is a common species of yeast, is by far the most extensively studied model of a eukaryote because although it is one of the simplest eukaryotes, its basic cellular processes resemble those of higher organisms. In addition, yeast is a commercially valuable organism for ethanol production. Since the yeast data can be extrapolated to the important aspects of higher organisms, many researchers have studied yeast metabolism under various conditions. In this report, we analyzed and compared metabolites of Saccharomyces cerevisiae under salt and pH stresses of various strengths by using two-dimensional NMR spectroscopy. A total of 31 metabolites were identified for most of the samples. The levels of many identified metabolites showed gradual or drastic increases or decreases depending on the severity of the stresses involved. The statistical analysis produced a holistic outline: pH stresses were clustered together, but salt stresses were spread out depending on the severity. This work could provide a link between the metabolite profiles and mRNA or protein profiles under representative and well studied stress conditions.