• Title/Summary/Keyword: Synthesis of Metabolite

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Synthetic Biology Tools for Novel Secondary Metabolite Discovery in Streptomyces

  • Lee, Namil;Hwang, Soonkyu;Lee, Yongjae;Cho, Suhyung;Palsson, Bernhard;Cho, Byung-Kwan
    • Journal of Microbiology and Biotechnology
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    • v.29 no.5
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    • pp.667-686
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    • 2019
  • Streptomyces are attractive microbial cell factories that have industrial capability to produce a wide array of bioactive secondary metabolites. However, the genetic potential of the Streptomyces species has not been fully utilized because most of their secondary metabolite biosynthetic gene clusters (SM-BGCs) are silent under laboratory culture conditions. In an effort to activate SM-BGCs encoded in Streptomyces genomes, synthetic biology has emerged as a robust strategy to understand, design, and engineer the biosynthetic capability of Streptomyces secondary metabolites. In this regard, diverse synthetic biology tools have been developed for Streptomyces species with technical advances in DNA synthesis, sequencing, and editing. Here, we review recent progress in the development of synthetic biology tools for the production of novel secondary metabolites in Streptomyces, including genomic elements and genome engineering tools for Streptomyces, the heterologous gene expression strategy of designed biosynthetic gene clusters in the Streptomyces chassis strain, and future directions to expand diversity of novel secondary metabolites.

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.

Polyamines and Their Metabolites as Diagnostic Markers of Human Diseases

  • Park, Myung Hee;Igarashi, Kazuei
    • Biomolecules & Therapeutics
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    • v.21 no.1
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    • pp.1-9
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    • 2013
  • Polyamines, putrescine, spermidine and spermine, are ubiquitous in living cells and are essential for eukaryotic cell growth. These polycations interact with negatively charged molecules such as DNA, RNA, acidic proteins and phospholipids and modulate various cellular functions including macromolecular synthesis. Dysregulation of the polyamine pathway leads to pathological conditions including cancer, inflammation, stroke, renal failure and diabetes. Increase in polyamines and polyamine synthesis enzymes is often associated with tumor growth, and urinary and plasma contents of polyamines and their metabolites have been investigated as diagnostic markers for cancers. Of these, diacetylated derivatives of spermidine and spermine are elevated in the urine of cancer patients and present potential markers for early detection. Enhanced catabolism of cellular polyamines by polyamine oxidases (PAO), spermine oxidase (SMO) or acetylpolyamine oxidase (AcPAO), increases cellular oxidative stress and generates hydrogen peroxide and a reactive toxic metabolite, acrolein, which covalently incorporates into lysine residues of cellular proteins. Levels of protein-conjuagated acrolein (PC-Acro) and polyamine oxidizing enzymes were increased in the locus of brain infarction and in plasma in a mouse model of stroke and also in the plasma of stroke patients. When the combined measurements of PC-Acro, interleukin 6 (IL-6), and C-reactive protein (CRP) were evaluated, even silent brain infarction (SBI) was detected with high sensitivity and specificity. Considering that there are no reliable biochemical markers for early stage of stroke, PC-Acro and PAOs present promising markers. Thus the polyamine metabolites in plasma or urine provide useful tools in early diagnosis of cancer and stroke.

Isolation and Synthesis of an Antifungal Metabolite Derived from the Commensal Vaginal Bacterium Dermabacter vaginalis (질 내 공생세균 Dermabacter vaginalis 유래 항진균 대사체의 분리 및 합성)

  • So, Jae Seong;Kwon, Kang Mu;Lee, Munseon;Kim, Dae Keun;Hwang, In Hyun
    • Korean Journal of Pharmacognosy
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    • v.53 no.3
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    • pp.119-124
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    • 2022
  • Vaginal microbiome deeply influences vaginal health via production of messenger molecules. The healthy vaginal pH ranges between 3.5 and 4.5. However, dysbiosis of vaginal microbiome increases the pH level, leading to the incidence of vaginitis. The commensal vaginal bacterium Dermabacter vaginalis-which was isolated from the vaginal fluid of a Korean female-was incubated in acidic and neutral pH to simulate healthy and vaginitis conditions, respectively. The chemical profiles of the two different cultures were compared using HPLC. The compound showing distinctive difference between the two sets of data was presumed to be a chemical messenger, which was identified as cyclo(L-pro-L-met) by analysis of NMR, MS, and specific rotation data. Synthesis was achieved in three steps (overall yield 15%), enabling structure confirmation and antimicrobial evaluation against vaginal pathogens. Cyclo(L-pro-L-met) showed antifungal activity against Candida albicans, a major cause of vulvovaginal candidiasis.

Response of Muscle Protein Synthesis to the Infusion of Insulin-like Growth Factor-I and Fasting in Young Chickens

  • Kita, K.;Shibata, T.;Aman Yaman, M.;Nagao, K.;Okumura, J.
    • Asian-Australasian Journal of Animal Sciences
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    • v.15 no.12
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    • pp.1760-1764
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    • 2002
  • In order to elucidate the physiological function of circulating IGF-I on muscle protein synthesis in the chicken under malnutritional conditions, we administrated recombinant chicken IGF-I using a osmotic mini pump to fasted young chickens and measured the rate of muscle protein synthesis and plasma metabolite. The pumps delivered IGF-I at the rate of $22{\mu}g/d\{300{\mu}g{\cdot}(kg\;body\;weight{\cdot}d)^{-1}\}$. Fractional rate of protein synthesis in the muscle was measured using a large dose injection of L-[$2,6-^3H$]phenylalanine. Constant infusion of chicken IGF-I did not affect plasma glucose level. Significant interaction between dietary treatment and IGF-I infusion was observed in plasma NEFA and total cholesterol concentrations. When chicks were fasted, IGF-I infusion decreased plasma NEFA and total cholesterol concentrations. On the other hand, IGF-I administration did not affect plasma levels of both metabolites. Fasting reduced plasma triglyceride concentration significantly. IGF-I infusion also decreased the level of plasma triglyceride. Plasma IGF-I concentration of young chickens was halved by fasting for 1 d. IGF-I infusion using an osmotic minipump for 1 d increased plasma IGF-I concentration in fasted chicks to the level of fed chicks. Fasting decreased body weight and the loss of body weight was significantly ameliorated by IGF-I infusion. There was a significant interaction between dietary treatment and IGF-I infusion in the fractional rate of breast muscle protein synthesis. There was no effect of IGF-I infusion on muscle protein synthesis in fed chicks. Muscle protein synthesis reduced by fasting was ameliorated by IGF-I infusion, but did not reach to the level of fed control. Muscle weight of fasted chicks infused with IGF-I was similar to fasted birds without IGF-I infusion, which suggests that muscle protein degradation would be increased by IGF-I infusion as well as protein synthesis in fasted chicks.

Bioprocess Considerations for Production of Secondary Metabolites by Plant Cell Suspension Cultures

  • Chattopadhyay, Saurabh;Farkya, Sunita;Srivastava, Ashok K.;Bisaria, Virendra
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.7 no.3
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    • pp.138-149
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    • 2002
  • Plant cell culture provides a viable alternative over whole plant cultivation for the production of secondary metabolites. In order to successfully cultivate the plant cells at large scale, several engineering parameters such as, cell aggregation, mixing, aeration, and shear sensitivity are taken into account for selection of a suitable bioreactor. The media ingredients, their concentrations and the environmental factors are optimized for maximal synthesis of a desired metabolite. Increased productivity in a bioreactor can be achieved by selection of a proper cultivation strategy (batch, fed-batch, two-stage etc.), feeding of metabolic precursors and extraction of intracellular metabolites. Proper understanding and rigorous analysis of these parameters would pave the way towards the successful commercialization of plant cell bioprocesses.

Regulation of Cell Growth and Tylosin Biosynthesis through Flux Control of Metabolic Intermediate in Streptomyces fradiae (Streptomyces fradiae에서 대사중간산물 이용속도에 의한 균체 성장과 tylosin 생합성의 조절)

  • 강현아;이계준
    • Korean Journal of Microbiology
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    • v.25 no.3
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    • pp.189-197
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    • 1987
  • The aim of the present study was to investigate the effect of glutamate on the biosynthesis of tylosin. Activities of enzymes involved in the metabolic pathway of glutamate to form tylactone, an essential precursor of tylosin, were determined using Streptomyces fradiae grown at different concentration of glutamate. As results, it was found that cell growth and tylactone formation was controlled by the metabolic flux of oxaloacetate. It was clear that cell growth was favored by the activities of citrate synthase and aspartate aminotransferase, while the tylactone synthesis was stimulated by the activity of methylmalonyl-CoA carboxyltransferase. Therefore it was concluded that channelling of oxaloacetate was a point for favoring either cell growth or tylosin biosynthesis.

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Draft Genome Sequence of the Neodothiora populina-Like Yeast Strain JAF-11, Which Produces the Biosurfactant myo-Inositol Lipids

  • Jeong-Seon Kim;Parthiban Subramanian;Seunghwan Kim;Jun Heo;Bong-Sik Yun;Yiseul Kim
    • Microbiology and Biotechnology Letters
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    • v.51 no.3
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    • pp.328-331
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    • 2023
  • Genomic information of biotechnologically and industrially important microorganisms provides the basis for understanding their metabolic potential. Here, we report the draft genome sequence of the Neodothiora populina-like yeast strain JAF-11 capable of producing biosurfactant myo-inositol lipids. The draft genome contained genes associated with secondary metabolite biosynthesis, including transport and metabolism of lipids, which are a major component of fungal surfactants. Identification of myo-inositol and acyl chain synthesis genes in the draft genome corresponded to the specific biosurfactant produced by strain JAF-11. Further experimental studies could help to elucidate the genes responsible for the production of biosurfactant by strain JAF-11.

Synthesis of Benzoxazole and Bezothiazole-linked TZD Analogs as PPARν Specific Ligands

  • Kim, Hae-Sung;Park, So-Yeon;Raok Jeon
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 2003.11a
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    • pp.117-117
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    • 2003
  • PPARs (peroxisome proliferator activated receptors) are member of nuclear hormone receptors superfamily. Activations of PPARs upon binding with ligands modulate glucose metabolite, differentiation of adipocyte, inflammation response, and so on. Thiazolidinedione analog is one of potential antidiabetic drug that binds and activates PPARν selectively and enhances insulin sensitivity. In an effort to develop novel and effective antidiabetic thiazolidindione analogs, syntheses of benzoxazole and benzothiazole-linked thiazolidinedione analogs were performed via coupling reaction of benzoxazolylalkylaminoethanol with hydroxybenzylthiazolidinedione to develop novel and effective antidiabetic thiazolidindiones. All compounds were evaluated their biological potency by PPARν transactivation assay and revealed the similar potency with Troglitazone. However, lengthening of N-alkyl substituent did not seem to be beneficial for the activity.

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