• Title/Summary/Keyword: Secondary metabolite(s)

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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.

Evaluation of the Biological Activities of Marine Bacteria Collected from Jeju Island, Korea, and Isolation of Active Compounds from their Secondary Metabolites

  • Kim, Hyun-Soo;Zhang, Chao;Lee, Ji-Hyeok;Ko, Ju-Young;Kim, Eun-A;Kang, Nalae;Jeon, You-Jin
    • Fisheries and Aquatic Sciences
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    • v.17 no.2
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    • pp.215-222
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    • 2014
  • To explore marine microorganisms with medical potential, we isolated and identified marine bacteria from floats, marine algae, animals, and sponges collected from Jeju Island, Korea. We isolated and identified 21 different strains from the marine samples by 16S rRNA analysis, cultured them in marine broth, and extracted them with ethyl acetate (EtOAc) to collect secondary metabolite fractions. Next, we evaluated their anti-oxidative and anti-inflammatory effects. Among the 21 strains, the secondary metabolite fraction of Bacillus badius had both strong antioxidant and anti-inflammatory activity, and thus was selected for further experiments. An antioxidant compound detected from the secondary metabolite fraction of B. badius was purified by preparative centrifugal partition chromatography (n-hexane:EtOAc:methanol:water, 4:6:4:6, v/v), and identified as diolmycin A2. Additionally, diolmycin A2 strongly inhibited nitric oxide production. Thus, we successfully identified a significant bioactive compound from B. badius among the bacterial strains collected from Jeju Island.

Growth, secondary metabolite production and antioxidant enzyme response of Morinda citrifolia adventitious root as affected by auxin and cytokinin

  • Baque, Md. Abdullahil;Hahn, Eun-Joo;Paek, Kee-Yoeup
    • Plant Biotechnology Reports
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    • v.4 no.2
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    • pp.109-116
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    • 2010
  • Morinda citrifolia adventitious roots were cultured in shake flasks using Murashige and Skoog medium with different types and concentrations of auxin and cytokinin. Root (fresh weight and dry weight) accumulation was enhanced at 5 $mg\;l^{-1}$ indole butyric acid (IBA) and at 7 and 9 $mg\;l^{-1}$ naphthalene acetic acid (NAA). On the other hand, 9 $mg\;l^{-1}$ NAA decreased the anthraquinone, phenolic and flavonoid contents more severely than 9 $mg\;l^{-1}$ IBA. When adventitious roots were treated with kinetin (0.1, 0.3 and 0.5 $mg\;l^{-1}$) and thidiazuron (TDZ; 0.1, 0.3 and 0.5 $mg\;l^{-1}$) in combination with 5 $mg\;l^{-1}$ IBA, fresh weight and dry weight decreased but secondary metabolite content increased. The secondary metabolite content (including 1,1-diphenyl-2-picrylhydrazyl activity) increased more in TDZ-treated than in kinetin-treated roots. Antioxidative enzymes such as catalase (CAT) and guaiacol peroxidase (G-POD), which play important roles in plant defense, also increased. A strong decrease in ascorbate peroxidase activity resulted in a high accumulation of hydrogen peroxide. This indicates that adventitious roots can grow under stress conditions with induced CAT and G-POD activities and higher accumulations of secondary metabolites. These results suggest that 5 $mg\;l^{-1}$ IBA supplementation is useful for growth and secondary metabolite production in adventitious roots of M. citrifolia.

Correlation Between Sorangium cellulosum Subgroups and Their Potential for Secondary Metabolite Production

  • Lee, Chayul;An, Dongju;Lee, Hanbit;Cho, Kyungyun
    • Journal of Microbiology and Biotechnology
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    • v.23 no.3
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    • pp.297-303
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    • 2013
  • Phylogenetic analysis of the groEL1 and xynB1 gene sequences from Sorangium cellulosum strains isolated in Korea previously revealed the existence of at least 5 subgroups (A-E). In the present study, we used sequence analysis of polymerase chain reaction-amplified biosynthetic genes of strains from the 5 subgroups to indicate correlations between S. cellulosum subgroups and their secondary metabolic gene categories. We detected putative biosynthetic genes for disorazol, epothilone, ambruticin, and soraphen in group A, group C, group D, and group E strains, respectively. With the exception of KYC3204, culture extracts from group A, group B, and group C strains exhibited no noticeable antimicrobial inhibitory activities. By contrast, culture extracts from group D strains inhibited the growth of Candida albicans, whereas culture extracts from group E strains inhibited the growth of C. albicans and Staphylococcus aureus. High performance liquid chromatography analysis of the culture extracts from the strains of each subgroup revealed unique peak patterns. Our findings indicate the existence of at least 5 subgroups of S. cellulosum strains, each of which has the potential to produce a unique set of secondary metabolites.

Effects of Short Microwave Irradiation Time at the Seedlings Stage on the Growth and Secondary Metabolite Contents of Lettuce (Lactuca sativa L.) (유묘단계에서 단시간 마이크로웨이브 처리가 상추의 생육 및 이차대사산물 함량에 미치는 영향)

  • Yong Jae Lee;Su Yong Park;Ju Hyung Shin;Seung Yong Hahm;Gwang Ya Lee;Jong Seok Park
    • Journal of Bio-Environment Control
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    • v.32 no.3
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    • pp.217-225
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    • 2023
  • This experiment was conducted to investigate the effects of microwave irradiation on the growth and secondary metabolite contents of lettuce seedlings. Seedlings at three weeks after sowing were treated by a microwave oven for 0, 4, 8, and 12 seconds with 200 W. After cultivation in a close plant production system for 4 weeks, plant growth measurements and secondary metabolite analysis were performed. The results showed that the fresh and dry weights of the shoot and root, leaf area, leaf length, and the number of leaves were decreased as increasing the microwave treatment times. Chlorophyll a and b, total carotenoids were increased and total phenolics were decreased at the 12-second treatment compared to the 4-second treatment. Total flavonoid contents were decreased at the 8-second treatment compared to the control. These results suggest that the changes in the levels of secondary metabolites were caused by oxidative stress. Although there was no significant difference in secondary metabolite contents excluding total flavonoid contents on the microwave treatments compared to the control, the significant difference suggests that the microwave treatment of 200 W and 2.45 GHz may alter secondary metabolite contents of lettuce after 4 weeks.

Reaction Mechanism of Vanadium Haloperoxidase and Marine Natural Products (Vanadium Haloperoxidase의 구조와 작용 메커니즘과 해양천연물질)

  • Han, Jae-Hong
    • Korean Journal of Crystallography
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    • v.16 no.2
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    • pp.66-74
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    • 2005
  • Marine natural products with various bioactivities are featured with similar structure to the common secondary metabolites and generally modified by halogenides, such as chloride, bromide, and iodide ions. Vanadium haloperoxidase is a key enzyme for the production of marine natural products and a metalloenzyme which requires a cofactor of vanadate. This review will cover isolation of vanadium haloperoxidase and the protein structures, as well as reaction mechanism of the metalloenzyme. Finally, reactivity of vanadium haloperoxidase and the biosynthesis of the secondary metabolites of indole, terpenoids, and acetogenins will be described.

Functional Expression of Proteomics-guided AfsR2-dependent Genes in Avermectin-producing Streptomyces avermitilis (Avermectin을 생산하는 Streptomyces avermitilis에서의 Proteomics-guided AfsR2-dependent 유전자의 발현)

  • Kim Myung-Gun;Park Hyun-Joo;Im Jong-Hyuk;Kim Eung-Soo
    • Microbiology and Biotechnology Letters
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    • v.34 no.3
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    • pp.211-215
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    • 2006
  • AfsR2 is a global regulatory protein involved in the stimulation of secondary metabolite biosynthesis in various Streptomyces species including avermectin-producing S. avermitilis. Among several AfsR2-dependent genes identified from the comparative proteomics, the polyribonucleotide nucleotidyltransferase (PNP) and the glyceraldehyde-3-phosphate dehydrogenase (GPD) genes were previously proposed to regulate the actinorhodin production in S. lividans upon afsR2 over-expression positively and negatively, respectively. To show the biological significance of the PNP and GPD genes in the S. avermitilis strains, these two genes were functionally expressed in both the wild-type and the avermectin-overproducing mutant strains. The PNP gene expression stimulated secondary metabolite production in the wild-type S. avermitilis ATCC31267, but not in the avermectin-overproducing S. avermitilis ATCC31780. Interestingly, the GDP gene expression stimulated secondary metabolite production by 4-fold in the wild-type S. avermitilis ATCC31267 and by 2.5-fold in the avermectin-overproducing S. avermitilis ATCC31780, respectively. These results suggest that the biological significance of the afsR2-dependent PNP and GPD gene expressions on antibiotic biosynthetic regulation could be significantly different depending on Streptomyces species.

Overexpression of Shinorhizobium meliloti Hemoprotein in Streptomyces lividans to Enhance Secondary Metabolite Production

  • Kim, Yoon-Jung;Sa, Soon-Ok;Chang, Yong-Keun;Hong, Soon-Kwang;Hong, Young-Soo
    • Journal of Microbiology and Biotechnology
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    • v.17 no.12
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    • pp.2066-2070
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    • 2007
  • It was found that Shinorhizobium meliloti hemoprotein (SM) was more effective than Vitreoscilla hemoglobin (Vhb) in promoting secondary metabolites production when overexpressed in Streptomyces lividans TK24. The transformant with sm (sm-transformant) produced 2.7-times and 3-times larger amounts of actinorhodin than the vhb-transformant in solid culture and flask culture, respectively. In both solid and flask cultures, a larger amount of undecylprodigiocin was produced by the sm-transformant. It is considered that the overexpression of SM especially has activated the pentose phosphate pathway through oxidative stress, as evidenced by an increased NADPH production observed, and that it has promoted secondary metabolites biosynthesis.

Metabolomics-Based Chemotaxonomic Classification of Streptomyces spp. and Its Correlation with Antibacterial Activity

  • Lee, Mee Youn;Kim, Hyang Yeon;Lee, Sarah;Kim, Jeong-Gu;Suh, Joo-Won;Lee, Choong Hwan
    • Journal of Microbiology and Biotechnology
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    • v.25 no.8
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    • pp.1265-1274
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    • 2015
  • Secondary metabolite-based chemotaxonomic classification of Streptomyces (8 species, 14 strains) was performed using ultraperformance liquid chromatography-quadrupole-time-offlight-mass spectrometry with multivariate statistical analysis. Most strains were generally well separated by grouping under each species. In particular, S. rimosus was discriminated from the remaining sevens pecies (S. coelicolor, S. griseus, S. indigoferus, S. peucetius, S. rubrolavendulae, S. scabiei, and S. virginiae) in partial least squares discriminant analysis, and oxytetracycline and rimocidin were identified as S. rimosus-specific metabolites. S. rimosus also showed high antibacterial activity against Xanthomonas oryzae pv. oryzae, the pathogen responsible for rice bacterial blight. This study demonstrated that metabolite-based chemotaxonomic classification is an effective tool for distinguishing Streptomyces spp. and for determining their species-specific metabolites.