• Title/Summary/Keyword: Secondary metabolites

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Enhanced Secondary Metabolite Biosynthesis by Abiotic Elicitor in Transformed Plant Root System

  • Jeong, Gwi-Taek;Hwang, Baik;Woo, Je-Chang;Park, Don-Hee
    • 한국생물공학회:학술대회논문집
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    • 2005.10a
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    • pp.417-420
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    • 2005
  • Plants generally produce secondary metabolites in nature as a defense mechanism against pathogenic and insect attack. In this study, we applied several abiotic elicitors in order to enhance growth and ginseng saponin biosynthesis in the hairy roots of P. ginseng. Generally, elicitor treatments were found to inhibit the growth of the hairy roots, although simultaneously enhancing ginseng saponin biosynthesis. The addition of selenium at inoculum time did not significantly affect ginseng saponin biosynthesis. However, when 0.5 mM selenium was added as an elicitor after 21 days of culture, ginseng saponin content and productivity increased to about 1.31 and 1.33 times control levels, respectively. These results suggest that processing time for the generation of ginseng saponin in a hairy root culture can be reduced via the application of an elicitor.

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Genetic and Functional Analyses of the DKxanthene Biosynthetic Gene Cluster from Myxococcus stipitatus DSM 14675

  • Hyun, Hyesook;Lee, Sunjin;Lee, Jong Suk;Cho, Kyungyun
    • Journal of Microbiology and Biotechnology
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    • v.28 no.7
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    • pp.1068-1077
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    • 2018
  • DKxanthenes are a class of yellow secondary metabolites produced by myxobacterial genera Myxococcus and Stigmatella. We identified a putative 49.5 kb DKxanthene biosynthetic gene cluster from Myxococcus stipitatus DSM 14675 by genomic sequence and mutational analyses. The cluster consisted of 15 genes (MYSTI_06004-MYSTI_06018) encoding polyketide synthases, non-ribosomal peptide synthases, and proteins with unknown functions. Disruption of the genes by plasmid insertion resulted in defects in the production of yellow pigments. High-performance liquid chromatography and liquid chromatography-tandem mass spectrometry analyses indicated that the yellow pigments produced by M. stipitatus DSM 14675 might be novel DKxanthene derivatives. M. stipitatus did not require DKxanthenes for the formation of heat-resistant viable spores, unlike Myxococcus xanthus. Furthermore, DKxanthenes showed growth inhibitory activity against the fungi Aspergillus niger, Candida albicans, and Rhizopus stolonifer.

Fish Oil Enriched Diet-Induced in vivo Lipid Peroxidation and Increased Excretion of Urinary Lipophilic Lipid Metabolites in Rats

  • Kim, Song-Suk
    • Nutritional Sciences
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    • v.3 no.1
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    • pp.18-24
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    • 2000
  • Peroxidative stimuli mediated by high polyunsaturated fatty acid administration in rats induced in vivo lipid peroxidation and resulted in increased urinary excretion of a number of lipophilic aldehydes and related carbonyl compounds. These secondary lipid peroxiation products, measured as 2, 4-dinitrophenylhydrazine deritives, were detected and identified by the newly developed HPLC method. The identified urinary lipophilic nonpolar aldehydes and related carbonyl compounds were butanal, butan-2-one, pentan-2-one, hexanal, hex-2-enal, hepta-2, 4-dienal, hept-2-enal, octanal, and oct-2-enal. Lipophilic polar aldehydes such as 4-hydroxyhex-2-enal and 4-hydroxyoct-2-enal were also identified. A polyunsaturated fatty acid diet containing n-3 fatty acids generally caused high levels of urinary excretion of lipophilic aldehydes and related carbonyl compounds in rats than a normal diet. Significantly increased secondary lipid peroxidation products were hexanal, hepta-2, 4-dienal, octanal, 4-hydroxyhex-2-exal, 4-hydroxyoct-2-enal, and a number of unidentified compunds.

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Structure and Bioactivity of Boholamide A from a Tidal Mudflat Actinomycete (갯벌 방선균 유래 Boholamide A의 구조 및 생리활성에 대한 연구)

  • Seo, Jeongwon;Moon, Kyuho
    • Korean Journal of Pharmacognosy
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    • v.52 no.4
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    • pp.203-207
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    • 2021
  • LC/MS approach targeting secondary metabolites of bacterial strain resulted in the discovery of boholamide A (1), from the culture of marine actinomycete strain which was isolated from a tidal mudflat in Muan, Republic of Korea. Boholamide A (1), a cyclodepsipeptide with HDMN, APD, glycine, and valine was structurally determined by using 1D/2D NMR spectroscopy, mass spectrometry and UV spectroscopy. Boholamide A (1) showed the inhibitory activity against Bacillus subtilis, with IC50 value of 0.08 mM.

Streptomyces Cytochrome P450 Enzymes and Their Roles in the Biosynthesis of Macrolide Therapeutic Agents

  • Cho, Myung-A;Han, Songhee;Lim, Young-Ran;Kim, Vitchan;Kim, Harim;Kim, Donghak
    • Biomolecules & Therapeutics
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    • v.27 no.2
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    • pp.127-133
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    • 2019
  • The study of the genus Streptomyces is of particular interest because it produces a wide array of clinically important bioactive molecules. The genomic sequencing of many Streptomyces species has revealed unusually large numbers of cytochrome P450 genes, which are involved in the biosynthesis of secondary metabolites. Many macrolide biosynthetic pathways are catalyzed by a series of enzymes in gene clusters including polyketide and non-ribosomal peptide synthesis. In general, Streptomyces P450 enzymes accelerate the final, post-polyketide synthesis steps to enhance the structural architecture of macrolide chemistry. In this review, we discuss the major Streptomyces P450 enzymes research focused on the biosynthetic processing of macrolide therapeutic agents, with an emphasis on their biochemical mechanisms and structural insights.

Screening of Myxobacteria Carrying Tubulysin Biosynthetic Genes

  • Hyun, Hyesook;Choi, Juo;Kang, Daun;Kim, Yungpil;Lee, Pilgoo;Chung, Gregory J.Y.;Cho, Kyungyun
    • Microbiology and Biotechnology Letters
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    • v.49 no.1
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    • pp.32-38
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    • 2021
  • Tubulysins are a group of secondary metabolites produced by myxobacteria that inhibit the function of the eukayotic cytoskeleton. We developed a pair of PCR primers that specifically amplified tubulysin biosynthetic genes. Using these primers, eight out of the eighty-one strains of myxobacteria belonging to the Cystobacteraceae family that harbored putative tubulysin biosynthetic genes were screened through PCR analysis. The selected strains included two Archangium gephyra, two Stigmatella sp., two Vitiosangium cumulatum, and two unidentified myxobacteria. LC-MS analysis of the culture extracts from the selected strains revealed that A. gephyra KYC4066 produced putative tubulysin A and B.

Effects of Increased NADPH Concentration by Metabolic Engineering of the Pentose Phosphate Pathway on Antibiotic Production and Sporulation in Streptomyces lividans TK24

  • Jin, Xue-Mei;Chang, Yong-Keun;Lee, Jae Hag;Hong, Soon-Kwang
    • Journal of Microbiology and Biotechnology
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    • v.27 no.10
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    • pp.1867-1876
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    • 2017
  • Most of the biosynthetic pathways for secondary metabolites are influenced by carbon metabolism and supply of cytosolic NADPH. We engineered carbon distribution to the pentose phosphate pathway (PPP) and redesigned the host to produce high levels of NADPH and primary intermediates from the PPP. The main enzymes producing NADPH in the PPP, glucose 6-phosphate dehydrogenase (encoded by zwf1 and zwf2) and 6-phosphogluconate dehydrogenase (encoded by zwf3), were overexpressed with opc encoding a positive allosteric effector essential for Zwf activity in various combinations in Streptomyces lividans TK24. Most S. lividans transformants showed better cell growth and higher concentration of cytosolic NADPH than those of the control, and S. lividans TK24/pWHM3-Z23O2 containing zwf2+zwf3+opc2 showed the highest NADPH concentration but poor sporulation in R2YE medium. S. lividans TK24/pWHM3-Z23O2 in minimal medium showed the maximum growth (6.2 mg/ml) at day 4. Thereafter, a gradual decrease of biomass and a sharp increase of cytosolic NADPH and sedoheptulose 7-phosphate between days 2 and 4 and between days 1 and 3, respectively, were observed. Moreover, S. lividans TK24/pWHM3-Z23O2 produced 0.9 times less actinorhodin but 1.8 times more undecylprodigiosin than the control. These results suggested that the increased NADPH concentration and various intermediates from the PPP specifically triggered undecylprodigiosin biosynthesis that required many precursors and NADPH-dependent reduction reaction. This study is the first report on bespoke metabolic engineering of PPP routes especially suitable for producing secondary metabolites that need diverse primary precursors and NADPH, which is useful information for metabolic engineering in Streptomyces.

A two-component sensor kinase (GacS) mediated signal transduction pathway involved in production of antifungal compounds in Pseudomonas chlororaphis O6.

  • Kang, Beom-Ryong;Lee, Jung-Hoon;Kim, Hyun-Jung;Cho, Baik-Ho;Kim, Young-Cheol
    • Proceedings of the Korean Society of Plant Pathology Conference
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    • 2003.10a
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    • pp.99.1-100
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    • 2003
  • E. intermedium Blocontrol activity of a P. chlororaphis rhizobacteium O6, depends to the synthesis of extracellular secondary metabolites and exoenzymes, thought to antagonize the pathogenicity of a variety of phytopathogenic fungi. The production of secondary metabolites and exoenzymes in O6, depends essentially on the GacS-mediated signal transduction pathway, which activates largely unknown signal transduction pathway. To exploit the GacS-mediated signal transdcution pathway involved in activation of ph genes that are necessary for biosynthesis of phenazine from P. chlororaphis O6, we cloned and sequenced the phz operon, rpoS gene encoding stationary specific sigma factor, ppx gene encoding polyphosphatase, and lon gene encoding ion protease. Expression of each gene in wild type and GacS mutant were analyzed by RT-PCR. Transcripts from rpoS, phzI enconing acylhomoserine lactone (AHL) synthase, and ph structural genes in the GacS mutant were reduced in each of these growth phases compared to the wild type. The GacS or Lon mutant was found to be deficient in the production of phenzines, exoenzymes, and the acylhomoserine lactone. These mutants were not complemented by ph operon and addition of exogenous AHL. These results indicate that the GacS global regulatory systems controls phenazine production at multiple levels. Future research will focus to identifying the GacS-mediated regulatory cascade involving in production of phenazine in P. chlororaphis.

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Effects of Growth Regulators on Adventitious Root Growth and Eleutherosides and Chlorogenic Acid Accumulation in Air Lift Bioreactor Cultures of Eleutherococcus koreanum (생물반응기 배양에서 생장조절제에 따른 섬오갈피 부정근 증식 및 Eleutheroside와 Chlorogenic Acid 생산)

  • Ahn Jin-Kwon;Park So-Young;Lee Wi-Young;Lee Jeong-Ju
    • Journal of Plant Biotechnology
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    • v.32 no.1
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    • pp.57-61
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    • 2005
  • The influence of different growth regulators on biomass of adventitious roots and secondary metabolites accumulation in bioreactor cultures of Eleutherococcus koreanum was studied. The maximum growth of adventitious roots was observed in the presence of $2.0\;\cal{mg/L}$ IBA (6.7 g DW/L). However the productivity of eleutheroside E was not significantly different among IBA levels. High level of thidiazuron (TDZ) efficiently increased both root growth and secondary metabolites production. Especially when $0.1\;\cal{mg/L}$ TDZ was combined with $3.0\;\cal{mg/L}$ IBA, the highest eleutheroside E accumulation ($3,327{\mu}g/g $DW) was observed. When the same TDZ level was treated in combination with $5.0\;\cal{mg/L}$ IBA, both chlorogrnic acid and eleutheroside B were accumulated to maximum level. In contrast, zeatin remarkably suppressed the accumulation of both eleutheroside B and chlorogenic acid.

Saponarin content and biosynthesis-related gene expression in young barley (Hordeum vulgare L.) seedlings

  • Lee, HanGyeol;Woo, So-Yeun;Ra, Ji-Eun;Lee, Kwang-Sik;Seo, Woo Duck;Lee, Jeong Hwan
    • Journal of Plant Biotechnology
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    • v.46 no.4
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    • pp.247-254
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    • 2019
  • Flavonoids are widely distributed secondary metabolites in plants that have a variety biological functions, as well as beneficial biological and pharmacological activities. In barley (Hordeum vulgare L.), for example, high levels of saponarin accumulate during primary leaf development. However, the effect of saponarin biosynthetic pathway genes on the accumulation of saponarin in barley is poorly understood. Accordingly, the aim of the present study was to examine the saponarin contents and expression levels of saponarin biosynthetic pathway genes [chalcone synthase (CHS), chalcone isomerase (CHI), and UDP-Glc:isovitexin 7-O-glucosyltransferase (OGT)] during early seedling developmental and under several abiotic stress conditions. Interestingly, the upregulation of HvCHS, HvCHI, and HvOGT during early development was associated with saponarin accumulation during later stages. In addition, exposure to abiotic stress conditions (e.g., light/dark transition, drought, and low or high temperature) significantly affected the expression of HvCHS and HvCHI but failed to affect either HvOGT expression or saponarin accumulation. These findings suggested that the expression of HvOGT, which encodes an enzyme that catalyzes the final step of saponarin biosynthesis, is required for saponarin accumulation. Taken together, the results of the present study provide a basis for metabolic engineering in barley plants, especially in regards to enhancing the contents of useful secondary metabolites, such as saponarin.