• 제목/요약/키워드: aromatic polyketide

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Three New Non-reducing Polyketide Synthase Genes from the Lichen-Forming Fungus Usnea longissima

  • Wang, Yi;Wang, Juan;Cheong, Yong Hwa;Hur, Jae-Seoun
    • Mycobiology
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    • 제42권1호
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    • pp.34-40
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    • 2014
  • Usnea longissima has a long history of use as a traditional medicine. Several bioactive compounds, primarily belonging to the polyketide family, have been isolated from U. longissima. However, the genes for the biosynthesis of these compounds are yet to be identified. In the present study, three different types of non-reducing polyketide synthases (UlPKS2, UlPKS4, and UlPKS6) were identified from a cultured lichen-forming fungus of U. longissima. Phylogenetic analysis of product template domains showed that UlPKS2 and UlPKS4 belong to group IV, which includes the non-reducing polyketide synthases with an methyltransferase (MeT) domain that are involved in methylorcinol-based compound synthesis; UlPKS6 was found to belong to group I, which includes the non-reducing polyketide synthases that synthesize single aromatic ring polyketides, such as orsellinic acid. Reverse transcriptase-PCR analysis demonstrated that UlPKS2 and UlPKS4 were upregulated by sucrose; UlPKS6 was downregulated by asparagine, glycine, and alanine.

A New Antioxidant from the Marine Sponge-derived Fungus Aspergillus versicolor

  • Li, Jian Lin;Lee, Yoon-Mi;Hong, Jong-Ki;Bae, Kyung-Sook;Choi, Jae-Soo;Jung, Jee-H.
    • Natural Product Sciences
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    • 제17권1호
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    • pp.14-18
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    • 2011
  • A chemical investigation of the marine-derived fungi Aspergillus versicolor led to the isolation of a new aromatic polyketide (1), The structure was elucidated by spectroscopic analysis, and its radical-scavenging activity, reducing power, and inhibitory activity to lipid oxidation were investigated. Those activities of compound 1 were compared with standard antioxidants such as butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT), tertiarybutylhydroquinone (TBHQ), and ascorbic acid ($V_C$). Compound 1 showed antioxidant activity comparable to that of BHA, and siginificantly higher than that of BHT.

Genetic analysis of polyketide biosynthetic genes isolated from Streptomyces albus, a salinomycin producer.

  • JOO-WON SUH;KWON, HYOUNG-JIN;C.R. HUTCHINSON;HYUNG-JONG JIN;SOO-UN KIM;KYE-JOON LEE
    • Journal of Microbiology and Biotechnology
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    • 제5권2호
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    • pp.74-79
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    • 1995
  • Sequence analysis of a DNA region encompassing the site of hybridization to actl, the gene for type II minimal polyketide synthase (PKS) for actinorhodin biosynthesis, from Streptomyces ablus revealed three more complete open reading frames additional to the already found two genes, plausibly encoding ${\beta}-ketoacyl$ synthase/acyl transferase (KS/AT) and chain length determining factor (ClF). The open reading frames (ORFs) were named salA, salD, and salE, from the upstream. In the homology analysis of the deduced amino acid sequences, SalA resembles the Streptomyces glaucescens Tcml, decaketide cyclase, SalD resembles acyl carrier protein in type II PKS, and SalE resembles the Actlll ketoreductase, The whole 4.4 kb of DNA sequence obeys the same conservation pattern as other type II PKSs. Therefore, we suggest that the 4.4 kb DNA from Streptomyces albus encompasses genes encoding enzymes for polyketide biogenesis in the organism and its organization is type II. The exsitence of SaIA, an analogue of the aromatic cyclase, revealed a relatedness of the 4.4 kb DNA with the aromatic PKS.

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Building Triketide α-Pyrone-Producing Yeast Platform Using Heterologous Expression of Sporopollenin Biosynthetic Genes

  • Kim, Sung Soo
    • Journal of Microbiology and Biotechnology
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    • 제25권11호
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    • pp.1796-1800
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    • 2015
  • Sporopollenin is a poorly characterized mixed aliphatic and aromatic polymer with ester and ether linkages. Recent studies have reported that α-pyrone polyketide compounds generated by Arabidopsis thaliana, polyketide synthase A (PKSA) and tetraketide α-pyrone reductase 1 (TKPR1), are previously unknown sporopollenin precursors. Here, the yeast Saccharomyces cerevisiae was introduced to test potential sporopollenin biosynthetic pathways in vivo. A PKSA/TKPR1 dual expressor was generated and various chain-length alkyl α-pyrones were identified by GC-MS. The growth rate of the strain containing PKSA/TKPR1 appeared normal. These results indicate that PKSA/TKPR1-expressing yeast would be a starting platform to investigate in vivo sporopollenin metabolism.

Cloning and Characterization of Actinorhodin Biosynthetic Gene Clusters from Streptomyces lividans TK24

  • Park, Kie-In
    • Animal cells and systems
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    • 제6권4호
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    • pp.305-309
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    • 2002
  • Actinorhodin antibiotics produced by Streptomyces lividans TK24 are blue pigments with a weak antibiotic activity, derived from one acetyl-CoA and 15 malonyl-CoA units via a typical ployketide pathway. In an attempt to clone polyketide biosynthetic genes of S. lividans TK24, hybridizing fragments in the genomic DNA of S. lividans TK24 were detected by use of acn and act III polyketide synthase gene probes. Since typical aromatic polyketide bio-synthetic gene clusters are roughly 22-34 Kb long, we constructed in E. coli XL-Blue MR using the Streptomyces-E. coli bifunctional shuttle cosmid vector (pojn46). Then, about 5,000 individual E. coii colonies were thor-oughly screened with acrl-ORFI and actIII probes. From these cosmid libra-ries, 12 positive clones were identified. Restriction analysis and southern hybridization showed two polyketide biosynthetic gene clusters in this organism. These cosmid clones can be transformed into Streptomyces parvulus 12434 for expression test that identify product of actinorhodin biosynthetic genes by heterologous expression. Thus, heterologous expres-sion of a derivative compound of a actinorhodin biosynthetic intermediate was obtained in pKE2430. Expression of these compounds by the trans-formants was detected by photodiode array HPLC analysis of crude extracts.

Nucleotide sequence analysis of a second set of the polyketide synthase .betha.-ketoacyl synthase and chain length factor genes from the salinomycin-producing streptomyces albus

  • Hyun, Chang-Gu;Park, Kwan-Hyung;C.Richard Hutchinson;Suh, Joo-Won
    • Journal of Microbiology
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    • 제35권1호
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    • pp.40-46
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    • 1997
  • The pWHM220 cosmid with a 24-kb insert cloned from Streptomyces albus ATCC 21838 induces the biosynthesis of a polysther antibiotic similar to salinomycin in Streptomyces invidans. We have analyzed this region by DNA sequencing as well as Southern blot hybridization with type I and type II polyketide synthase (PKS) probes. Surprisingly, we found another set of type II SKS genes only 10-kb from the original PKS genes, salABCDE. The DNA sequence revealed two complete open reading frames (ORFs) named salB2 and salC2, and one partial ORF that does not resemble any known DNA or deduced protein sequence. The salC2 should code for chain length determining factor while the deduced amino acid sequence encoded by salB2 exhibits high similarity to .betha.-ketoacyl synthase from different PKS gene clusters. The highest identity was found for .betha.-keetoacyl synthases from S. argillaceus (MtmP. 59.1% identity), the mithramycin producer and from S. venezuelae ISP5230 (JadA, 52.3% identity), the jadomycin producer. The SalC2 protein clearly resembles its counterparts in order aromatic PKS gene clusters that are believed to influence the length of the polyketide chain. The highest identities observed were to that of S. argillaceus (MtmK, 62.3%) and S. venezuelae ISP 5230 (JadB, 55.1%) proteins, Moreover, the deduced amino acid sequences of the salB2 and salC2 products were 29.0% identical.

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Streptomyces Peucetius에서의 ${\varepsilon}$-rhodomycinone 추출 및 이종균주에서의 rhodomycin D 생산 연구 (NDP-sugar production and glycosylation of ${\varepsilon}$-rhodomycinone in Streptomyces venezuelae)

  • 박성희;차민호;김은정;윤여준;송재경;이희찬;류광경;김병기
    • KSBB Journal
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    • 제23권1호
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    • pp.44-47
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    • 2008
  • Streptomyces peucetius가 생산하는 anthracycline 계열의 doxorubicin은 치료목적으로 사용되는 중요한 항암제 중 하나이다. Doxorubicin은 rhodomycin D에서부터 몇 단계의 생합성 과정을 더 거쳐 생산되는데, 생물학적 활성을 갖기 위해서는 deoxy-sugar의 전이가 반드시 일어나야 한다. 본 논문에서는 이종균주인 Streptomyces venezuelae에 11개의 유전자를 형질 전환하여 TDP-L-daunosamine를 생산하고 이것을 ${\varepsilon}$-rhodomycinone에 전이하여 rhodomycin D를 생산하는 연구를 수행하였다. S. peucetius 유래의 7개 유전자 dnmU, T, J, V, Z, Q, S.를 당 합성 및 전이를 위해 plasmid 형태로 전이하였으며, S. venezuelae의 desIII, IV와 doxorubicin 내성 유전자인 drrA, B는 chromosomal DNA에 삽입하였다. Aglycone 기질인 ${\varepsilon}$-rhodomycinone을 확보하기 위하여 6L의 고체 배지에 S. peucetius를 배양하여 유기용매로 추출하고 preparative HPLC로 분리 정제하였다. 결과적으로 이종균주인 S. venezuelae에서 ${\varepsilon}$-rhodomycinone에 당 전이가 일어난 생산물을 확인함으로써 deoxy-sugar의 생합성 및 전이에 필요한 최소한의 유전적 정보를 확인할 수 있었다. 또한, 유사서열 단백질 모델링을 통하여, 최초로 당 전이 반응에 필수적인 도움효소 DnrQ의 구조를 예측하였다.