Hops represent an important natural source of bioactive polyphenols, particularly proanthocyanidins, which can contribute to prevention of several civilization diseases, owing to their antioxidant and radical scavenging activity. We have developed a high-throughput ultra-high-performance liquid chromatography time-of-flight mass spectrometry profiling method, which can be used for monitoring of bioactive proanthocyanidins in hops. The method was applied for analysis of hops of four Czech varieties (Saaz, Sladek, Preminat and Agnus) from the 2011 crop (9 localities, 11 samples) and the 2012 crop (24 localities, 40 samples). Hop samples were extracted by acetone and the analytes were separated on the Acquity UPLC BEH Shield RP18 column. Partial validation of the method revealed a satisfactory intra-day repeatability of the method for retention times (relative standard deviation within 1.39%) as well as areas under the peaks (within 9.89%). Experimental data were evaluated using principal component analysis and cluster analysis. Significant amounts of di-, tri- and tetramer proanthocyanidins consisting of (epi)catechin and (epi)gallocatechin were found in the hop samples. The dependence of the proantocyanidin composition on both the variety and the growing locality was observed. Specifically, the traditional Saaz variety contained more frequently oligomers formed by (epi)catechin units only, whereas the varieties Premiant and Agnus produced oligomers consisting of (epi)catechin as well as (epi)gallocatechin units. The relative abundance of proanthocyanidins in studied hop varieties from the two crops, 2011 and 2012, did correspond to each other. In the further perspective, the method may also be used for prediction of qualitative marks or authenticity verification of hops.
- Klíčová slova
- Catechin, Cluster analysis, Flavan-3-ol, Hops, PCA, Proanthocyanidins, Profiling, TOF MS, UHPLC,
- MeSH
- aceton chemie MeSH
- analýza hlavních komponent MeSH
- Humulus chemie MeSH
- katechin analogy a deriváty izolace a purifikace MeSH
- proantokyanidiny analýza chemie MeSH
- reprodukovatelnost výsledků MeSH
- rozpouštědla chemie MeSH
- rychlé screeningové testy MeSH
- shluková analýza MeSH
- spektrometrie hmotnostní - ionizace laserem za účasti matrice MeSH
- vysokoúčinná kapalinová chromatografie metody MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Geografické názvy
- Česká republika MeSH
- Názvy látek
- aceton MeSH
- gallocatechol MeSH Prohlížeč
- katechin MeSH
- proantokyanidiny MeSH
- rozpouštědla MeSH
AIMS: Production of minor asukamycin congeners and its new derivatives by combination of targeted genetic manipulations with specific precursor feeding in the producer of asukamycin, Streptomyces nodosus ssp. asukaensis. METHODS AND RESULTS: Structural variations of manumycins lie only in the diverse initiation of the 'upper' polyketide chain. Inactivation of the gene involved in the biosynthesis of cyclohexanecarboxylic acid (CHC) turned off the production of asukamycin in the mutant strain and allowed an increased production of other manumycins with the branched end of the upper chain. The ratio of produced metabolites was further affected by specific precursor feeding. Precursor-directed biosynthesis of a new asukamycin analogue (asukamycin I, 28%) with linear initiation of the upper chain was achieved by feeding norleucine to the mutant strain. Another asukamycin analogue with the unbranched upper chain (asukamycin H, 14%) was formed by the CHC-deficient strain expressing a heterologous gene putatively involved in the formation of the n-butyryl-CoA starter unit of manumycin A. CONCLUSIONS: Combination of the described techniques proved to be an efficient tool for the biosynthesis of minor or novel manumycins. SIGNIFICANCE AND IMPACT OF THE STUDY: Production of two novel asukamycin derivatives, asukamycins H and I, was achieved. Variations appeared in the upper polyketide chain, the major determinant of enzyme-inhibitory features of manumycins, affecting their cancerostatic or anti-inflammatory features.
- MeSH
- acylkoenzym A metabolismus MeSH
- aminokyseliny metabolismus MeSH
- antibakteriální látky biosyntéza MeSH
- genetické inženýrství MeSH
- inzerční mutageneze MeSH
- kultivační média MeSH
- kyseliny cyklohexankarboxylové metabolismus MeSH
- mutace MeSH
- polyeny metabolismus MeSH
- polynenasycené alkamidy metabolismus MeSH
- Streptomyces genetika metabolismus MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- acylkoenzym A MeSH
- aminokyseliny MeSH
- antibakteriální látky MeSH
- asukamycin MeSH Prohlížeč
- butyryl-coenzyme A MeSH Prohlížeč
- cyclohexanecarboxylic acid MeSH Prohlížeč
- kultivační média MeSH
- kyseliny cyklohexankarboxylové MeSH
- manumycin MeSH Prohlížeč
- polyeny MeSH
- polynenasycené alkamidy MeSH
Geosmithia fungi are little known symbionts of bark beetles. Secondary metabolites of lilac colored species G. lavendula and other nine Geosmithia species were investigated in order to elucidate their possible role in the interactions of the fungi with environment. Hydroxylated anthraquinones (yellow, orange, and red pigments), were found to be the most abundant compounds produced into the medium during the submerged cultivation. Three main compounds were identified as 1,3,6,8-tetrahydroxyanthraquinone (1), rhodolamprometrin (1-acetyl-2,4,5,7-tetrahydroxyanthraquinone; 2), and 1-acetyl-2,4,5,7,8-pentahydroxyanthraquinone (3). Compounds 2 and 3 (representing the majority of produced metabolites) inhibited the growth of G+-bacteria Staphylococcus aureus and Bacillus subtilis with minimum inhibitory concentration of 64-512 microg/mL. Anti-inflammatory activity detected as inhibition of cyclooxygenase-2 was found only for compound 3 at 1 and 10 microg/mL. Compound 2 interfered with the morphology, compound 3 with cell-cycle dynamics of adherent mammalian cell lines.
- MeSH
- anthrachinony chemie metabolismus farmakologie MeSH
- antibakteriální látky chemie metabolismus farmakologie MeSH
- antiflogistika nesteroidní chemie metabolismus farmakologie MeSH
- Bacillus subtilis účinky léků MeSH
- biologické pigmenty chemie metabolismus farmakologie MeSH
- biotechnologie metody MeSH
- buněčný cyklus účinky léků MeSH
- Ficus parazitologie MeSH
- HeLa buňky MeSH
- hydroxylace MeSH
- Hypocreales růst a vývoj metabolismus MeSH
- inhibitory cyklooxygenasy chemie metabolismus farmakologie MeSH
- lidé MeSH
- nosatcovití mikrobiologie fyziologie MeSH
- spory hub růst a vývoj metabolismus MeSH
- Staphylococcus aureus účinky léků MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- 1-acetyl-2,4,5,7-tetrahydroxyanthraquinone MeSH Prohlížeč
- 1-acetyl-2,4,5,7,8-pentahydroxyanthraquinone MeSH Prohlížeč
- 1,3,6,8-tetrahydroxyanthraquinone MeSH Prohlížeč
- anthrachinony MeSH
- antibakteriální látky MeSH
- antiflogistika nesteroidní MeSH
- biologické pigmenty MeSH
- inhibitory cyklooxygenasy MeSH
A cosmid bearing an insert of 38 217 bp covering the gene cluster and its flanking regions of type strain Streptomyces lincolnensis ATCC 25466 was sequenced. Two relatively extensive sequence changes and several hundred point mutations were identified if compared with the previously published sequence of the lincomycin (Lin) industrial strain S. lincolnensis 78-11. Analysis of the cluster-flanking regions revealed its localization within the genome of the ATCC 25466 strain. The cluster-bearing cosmid was integrated into the chromosome of Lin non-producing strains S. coelicolor CH 999 and S. coelicolor M 145. The modified strains heterologously produced Lin but the level dropped to approximately 1-3% of the production in the ATCC 25466 strain.
- MeSH
- antibakteriální látky biosyntéza chemie MeSH
- bakteriální proteiny genetika metabolismus MeSH
- biotechnologie MeSH
- bodová mutace MeSH
- genová knihovna MeSH
- kosmidy MeSH
- linkomycin biosyntéza chemie MeSH
- multigenová rodina * MeSH
- sekvenční analýza DNA * MeSH
- Streptomyces coelicolor genetika metabolismus MeSH
- Streptomyces genetika růst a vývoj metabolismus MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- antibakteriální látky MeSH
- bakteriální proteiny MeSH
- linkomycin MeSH
We studied genetic variability of 100 isolates of Claviceps purpurea by using randomly amplified polymorphic DNA (RAPD), an EcoRI restriction site polymorphism in the 5.8S ribosomal DNA (rDNA), the alkaloids produced, and conidial morphology. We identified three groups: (i) group G1 from fields and open meadows (57 isolates), (ii) group G2 from shady or wet habitats (41 isolates), and (iii) group G3 from Spartina anglica from salt marshes (2 isolates). The sclerotia of G1 isolates contained ergotamines and ergotoxines; G2 isolates produced ergosine and ergocristine along with small amounts of ergocryptine; and G3 isolates produced ergocristine and ergocryptine. The conidia of G1 isolates were 5 to 8 microm long, the conidia of G2 isolates were 7 to 10 microm long, and the conidia of G3 isolates were 10 to 12 microm long. Sclerotia of the G2 and G3 isolates floated on water. In the 5.8S rDNA analysis, an EcoRI site was found in G1 and G3 isolates but not in G2 isolates. The host preferences of the groups were not absolute, and there were host genera that were common to both G1 and G2; the presence of members of different groups in the same locality was rare. Without the use of RAPD or rDNA polymorphism, it was not possible to distinguish the three groups solely on the basis of phenotype, host, or habitat. In general, populations of C. purpurea are not host specialized, as previously assumed, but they are habitat specialized, and collecting strategies and toxin risk assessments should be changed to reflect this paradigm shift.
- MeSH
- alkaloidy biosyntéza MeSH
- Claviceps genetika izolace a purifikace metabolismus MeSH
- DNA primery genetika MeSH
- fenotyp MeSH
- fungální RNA genetika MeSH
- genetická variace MeSH
- polymorfismus délky restrikčních fragmentů MeSH
- RNA ribozomální 5.8S genetika MeSH
- rostliny mikrobiologie MeSH
- sekvence nukleotidů MeSH
- technika náhodné amplifikace polymorfní DNA MeSH
- životní prostředí MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- alkaloidy MeSH
- DNA primery MeSH
- fungální RNA MeSH
- RNA ribozomální 5.8S MeSH
The enantioselective behaviour of some underivatized 2-arylpropionic acids (profens) and flobufen by HPLC using a terguride-based chiral stationary phase was tested. X-ray analysis of crystals of the chiral selector and its complexes with naproxen allowed a deeper insight into the enantiodiscriminative process. The column stability and reproducibility, and the potential of the packing for semipreparative scale separations were also determined. A method for determining flobufen enantiomers and metabolites in plasma samples is described.
- MeSH
- antiflogistika nesteroidní chemie izolace a purifikace MeSH
- butyráty chemie izolace a purifikace MeSH
- ergoliny chemie izolace a purifikace MeSH
- molekulární konformace MeSH
- molekulární modely MeSH
- molekulární struktura MeSH
- propionáty chemie izolace a purifikace MeSH
- stereoizomerie MeSH
- vysokoúčinná kapalinová chromatografie metody MeSH
- vztahy mezi strukturou a aktivitou MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- antiflogistika nesteroidní MeSH
- butyráty MeSH
- ergoliny MeSH
- flobufen MeSH Prohlížeč
- propionáty MeSH
