• Title/Summary/Keyword: PKS

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Heterologous Expression of Hybrid Type II Polyketide Synthase System in Streptomyces Species

  • Kim, Chang-Young;Park, Hyun-Joo;Kim, Eung-Soo
    • Journal of Microbiology and Biotechnology
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    • v.13 no.5
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    • pp.819-822
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    • 2003
  • Polyketides are an extensive class of secondary metabolites with diverse molecular structures and biological activities. A plasmid-based minimal polyketide synthase (PKS) expression cassette was constructed using a subset of actinorhodin (act) biosynthetic genes (actI-orfl, actI-orf2, actI-orf3, actIII, actⅦ, and actIV) from Streptomyces coelicolor, which specify the construction of an orange-fluorescent anthraquinone product aloesaponarin II, a type II polyketide compound derived from one acetyl coenzyme A and 7 malonyl coenzyme A extender units. This system was designed as an indicator pathway in S. parvulus to generate a hybrid type II polyketide compound via gene-specific replacement. The act ${\beta}-ketoacyl$ synthase unit (actI-orfl and actI-orf2) in the expression cassette was specifically replaced with oxytetracycline ${\beta}-ketoacyl$ synthase otcY-orfl and otcY-orf2). This plasmid-based hybrid PKS cassette generated a novel orange-fluorescent compound structurally different from aloesaponarin II in both S. lividans and S. parvulus. In addition, several additional distinctive blue-fluorescent compounds were detected, when this hybrid PKS cassette was expressed in S. coelicolor B78 (actI-orf2 mutant), implying that the expression of plasmid-based hybrid PKS cassette in Streptomyces species should be an efficient way of generating hybrid type II polyketide compounds.

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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    • v.35 no.1
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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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A Study on the Characteristics of Torrefaction and Chlorine Release According to the Mild Pyrolysis Temperature Conditions of Biomass Fuels (WP·EFB·PKS) for Power Generation (발전용 바이오매스 연료(WP·EFB·PKS)의 열분해 온도 조건에 따른 반탄화 및 염소 방출 특성에 관한 연구)

  • KIM, JI-HUN;PARK, JAE-HEUN;CHOI, JAE-HYUN;JEON, CHUNG-HWAN
    • Transactions of the Korean hydrogen and new energy society
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    • v.28 no.6
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    • pp.683-690
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    • 2017
  • Wood pellet (WP), empty fruit bunch (EFB) and palm kernel shell (PKS) which are biomass fuels for power generation are selected to study the characteristics of torrefaction process. These biomass fuels are torrefied at $220^{\circ}C$, $250^{\circ}C$, and $280^{\circ}C$. The heating value of biomass fuels is increased depending on the torrefaction temperature. However, due to energy yield decline, it is not always desirable to torrefy biomass at higher temperature. Considering the mass yield and energy yield after torrefaction, the most proper temperature conditions for torrefaction of WP is $250-280^{\circ}C$ and for EFB, PKS are $220-250^{\circ}C$. Additionally, to investigate the phenomenons of chlorine release during torrefaction process, Ion Chromatography (IC) method was used. In the case of EFB and PKS torrefied at $300^{\circ}C$, the chlorine component has been reduced by 97.5% and 95.3% compared to the raw biomass, respectively. In conclusion, torrefied biomass can be used as alternative fuels in replacement of coals for both aspects of heating value and chlorine corrosion problems.

Draft Genome Analysis of Antimicrobial Streptomyces Isolated from Himalayan Lichen

  • Kim, Byeollee;Han, So-Ra;Lamichhane, Janardan;Park, Hyun;Oh, Tae-Jin
    • Journal of Microbiology and Biotechnology
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    • v.29 no.7
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    • pp.1144-1154
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    • 2019
  • There have been several studies regarding lichen-associated bacteria obtained from diverse environments. Our screening process identified 49 bacterial species in two lichens from the Himalayas: 17 species of Actinobacteria, 19 species of Firmicutes, and 13 species of Proteobacteria. We discovered five types of strong antimicrobial agent-producing bacteria. Although some strains exhibited weak antimicrobial activity, NP088, NP131, NP132, NP134, and NP160 exhibited strong antimicrobial activity against all multidrug-resistant strains. Polyketide synthase (PKS) fingerprinting revealed results for 69 of 148 strains; these had similar genes, such as fatty acid-related PKS, adenylation domain genes, PfaA, and PksD. Although the association between antimicrobial activity and the PKS fingerprinting results is poorly resolved, NP160 had six types of PKS fingerprinting genes, as well as strong antimicrobial activity. Therefore, we sequenced the draft genome of strain NP160, and predicted its secondary metabolism using antiSMASH version 4.2. NP160 had 46 clusters and was predicted to produce similar secondary metabolites with similarities of 5-100%. Although NP160 had 100% similarity with the alkylresorcinol biosynthetic gene cluster, our results showed low similarity with existing members of this biosynthetic gene cluster, and most have not yet been revealed. In conclusion, we expect that lichen-associated bacteria from the Himalayas can produce new secondary metabolites, and we found several secondary metabolite-related biosynthetic gene clusters to support this hypothesis.

Phosphagen Kinases of Parasites: Unexplored Chemotherapeutic Targets

  • Jarilla, Blanca R.;Agatsuma, Takeshi
    • Parasites, Hosts and Diseases
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    • v.48 no.4
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    • pp.281-284
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    • 2010
  • Due to the possible emergence of resistance and safety concerns on certain treatments, development of new drugs against parasites is essential for the effective control and subsequent eradication of parasitic infections. Several drug targets have been identified which are either genes or proteins essential for the parasite survival and distinct from the hosts. These include the phosphagen kinases (PKs) which are enzymes that playa key role in maintenance of homeostasis in cells exhibiting high or variable rates of energy turnover by catalizing the reversible transfer of a phosphate between ATP and naturally occurring guanidine compounds. PKs have been identified in a number of important human and animal parasites and were also shown to be significant in survival and adaptation to stress conditions. The potential of parasite PKs as novel chemotherapeutic targets remains to be explored.

The Growth and EPA Synthesis of Shewanella oneidensis MR-1 and Expectation of EPA Biosynthetic Pathway

  • Jeong, Young-Su;Song, Sang-Kyu;Lee, Su-Jin;Hur, Byung-Ki
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.11 no.2
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    • pp.127-133
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    • 2006
  • Shewanella oneidensis MR-1 has the ability to inhale certain metals and chemical compounds and exhale these materials in an altered state; as a result, this microorganism has been widely applied in bioremediation protocols. However, the relevant characteristics of cell growth and biosynthesis of PuFAs have yet to be thoroughly investigated. Therefore, in this study, we have attempted to characterize the growth and fatty acid profiles of S. oneidensis MR-1 under a variety of temperature conditions. The fastest growth of S. oneidensis MR-1 was observed at $30^{\circ}C$, with a specific growth rate and doubling time of $0.6885h^{-1}\;and\;1.007 h$. The maximum cell mass of this microorganism was elicited at a temperature of $4^{\circ}C$. The eicosapentaenoic acid (EPA) synthesis of S. oneidensis MR-1 was evaluated under these different culture temperatures. S. oneidensis MR-1 was found not to synthesize EPA at temperatures in excess of $30^{\circ}C$, but was shown to synthesize EPA at temperatures below $30^{\circ}C$. The EPA content was found to increase with decreases in temperature. We then evaluated the EPA biosynthetic pathway, using a phylogenetic tree predicted on 16s rRNA sequences, and the homology of ORFs between S. oneidensis MR-1 and Shewanella putrefaciens SCRC-2738, which is known to harbor a polyketide synthase (PKS)-like module. The phylogenetic tree revealed that MR-1 was very closely related to both Moritella sp., which is known to synthesize DHA via a PKS-like pathway, and S. putrefaciens, which has been reported to synthesize EPA via an identical pathway. The homology between the PKS-like module of S. putrefaciens SCRC-2738 and the entire genome of S. oneidensis MR-1 was also analyzed, in order to mine the genes associated with the PKS-like pathway in S. oneidensis MR-1. A putative PKS-like module for EPA biosynthesis was verified by this analysis, and was also corroborated by the experimental finding that S. oneidensis MR-1 was able to synthesize EPA without the expression of $dihomo-{\gamma}-linoleic$ acid (DGLA) and arachidonic acid (AA) formed during EPA synthesis via the FAS pathway.

Generation of Hybrid Polyketides through Combinatorial Biosynthesis of Polyketide Synthase (PKS) and Modification of Post-PKS Tailoring Steps

  • Yoon, Yeo-Joon
    • Proceedings of the PSK Conference
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    • 2003.10a
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    • pp.75-78
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    • 2003
  • Polyketides are a class of structurally diverse natural products which possess a wide range of biological activities. These compounds are used throughout medicine and agriculture as antimicrobials, immunosuppressants, antiparasitics, and anticancer agents. While structurally diverse, polyketides are assembled by a common mechanism of decarboxylative condensations of simple malonate derivatives by polyketide synthases (PKSs) in a manner very similar to fatty acid biosynthesis (Fig 1). (omitted)

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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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    • v.5 no.2
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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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Study of Oil Palm Biomass Resources (Part 5) - Torrefaction of Pellets Made from Oil Palm Biomass - (오일팜 바이오매스의 자원화 연구 V - 오일팜 바이오매스 펠릿의 반탄화 연구 -)

  • Lee, Ji-Young;Kim, Chul-Hwan;Sung, Yong Joo;Nam, Hye-Gyeong;Park, Hyeong-Hun;Kwon, Sol;Park, Dong-Hun;Joo, Su-Yeon;Yim, Hyun-Tek;Lee, Min-Seok;Kim, Se-Bin
    • Journal of Korea Technical Association of The Pulp and Paper Industry
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    • v.48 no.2
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    • pp.34-45
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    • 2016
  • Global warming and climate change have been caused by combustion of fossil fuels. The greenhouse gases contributed to the rise of temperature between $0.6^{\circ}C$ and $0.9^{\circ}C$ over the past century. Presently, fossil fuels account for about 88% of the commercial energy sources used. In developing countries, fossil fuels are a very attractive energy source because they are available and relatively inexpensive. The environmental problems with fossil fuels have been aggravating stress from already existing factors including acid deposition, urban air pollution, and climate change. In order to control greenhouse gas emissions, particularly CO2, fossil fuels must be replaced by eco-friendly fuels such as biomass. The use of renewable energy sources is becoming increasingly necessary. The biomass resources are the most common form of renewable energy. The conversion of biomass into energy can be achieved in a number of ways. The most common form of converted biomass is pellet fuels as biofuels made from compressed organic matter or biomass. Pellets from lignocellulosic biomass has compared to conventional fuels with a relatively low bulk and energy density and a low degree of homogeneity. Thermal pretreatment technology like torrefaction is applied to improve fuel efficiency of lignocellulosic biomass, i.e., less moisture and oxygen in the product, preferrable grinding properties, storage properties, etc.. During torrefacton, lignocelluosic biomass such as palm kernell shell (PKS) and empty fruit bunch (EFB) was roasted under an oxygen-depleted enviroment at temperature between 200 and $300^{\circ}C$. Low degree of thermal treatment led to the removal of moisture and low molecular volatile matters with low O/C and H/C elemental ratios. The mechanical characteristics of torrefied biomass have also been altered to a brittle and partly hydrophobic materials. Unfortunately, it was much harder to form pellets from torrefied PKS and EFB due to thermal degradation of lignin as a natural binder during torrefaction compared to non-torrefied ones. For easy pelletization of biomass with torrefaction, pellets from PKS and EFB were manufactured before torrefaction, and thereafter they were torrefied at different temperature. Even after torrefaction of pellets from PKS and EFB, their appearance was well preserved with better fuel efficiency than non-torrefied ones. The physical properties of the torrefied pellets largely depended on the torrefaction condition such as reaction time and reaction temperature. Temperature over $250^{\circ}C$ during torrefaction gave a significant impact on the fuel properties of the pellets. In particular, torrefied EFB pellets displayed much faster development of the fuel properties than did torrefied PKS pellets. During torrefaction, extensive carbonization with the increase of fixed carbons, the behavior of thermal degradation of torrefied biomass became significantly different according to the increase of torrefaction temperature. In conclusion, pelletization of PKS and EFB before torrefaction made it much easier to proceed with torrefaction of pellets from PKS and EFB, leading to excellent eco-friendly fuels.

Cloning and Characterization of a Gene Cluster for the Production of Polyketide Macrolide Dihydrochalcomycin in Streptomyces sp. KCTC 0041BP

  • Jaishy Bharat Prasad;Lim Si-Kyu;Yoo Ick-Dong;Yoo Jin-Cheol;Sohng Jae-Kyung;Nam Doo-Hyun
    • Journal of Microbiology and Biotechnology
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    • v.16 no.5
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    • pp.764-770
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    • 2006
  • Dihydrochalcomycin (GERI-155), produced by Streptomyces sp. KCTC-0041BP isolated from Korean soil, is a 16-membered macrolide antibiotic consisting of two deoxysugar moieties at C-5 and C-20 positions of a branched lactone ring. The cloning and sequencing of a gene cluster for dihydrochalcomycin biosynthesis revealed a 63-kb nucleotide region containing 25 open reading frames (ORFs). The products of all of these 25 ORFs playa role in dihydrochalcomycin biosynthesis and self-resistance against the compounds synthesized. At the core of this cluster lies a 39.6-kb polyketide synthase (PKS) region encoding eight modules in five giant multifunctional protein-coding genes (gerSI-SV). The genes responsible for the biosynthesis of deoxysugar moieties, D-chalcose and D-mycinose, and their modification and attachment were found on either side of this PKS region. The involvement of this gene cluster in dihydrochalcomycin biosynthesis was confirmed by disruption of the dehydratase (DH) domain in module 3 of the PKS gene and by metabolite analysis.