ABSTRACT
Two Indonesian fungi Aspergillus assiutensis BioMCC-f.T.7495 and Penicillium pedernalense BioMCC-f.T.5350 along with a Japanese fungus Hypomyces pseudocorticiicola FKI-9008 have been found to produce gentisyl alcohol (1), which inhibits Plasmodium falciparum dihydroorotate dehydrogenase (PfDHODH) with an IC50 value of 3.4 µM. Another Indonesian fungus, Penicillium citrinum BioMCC-f.T.6730, produced an analog of 1, homogentisic acid (4), which also inhibits PfDHODH with an IC50 value of 47.6 µM.
Subject(s)
Benzyl Alcohols/pharmacology , Enzyme Inhibitors/pharmacology , Fungi/chemistry , Homogentisic Acid/pharmacology , Oxidoreductases Acting on CH-CH Group Donors/antagonists & inhibitors , Plasmodium falciparum/enzymology , Antimalarials/chemistry , Antimalarials/isolation & purification , Antimalarials/pharmacology , Benzyl Alcohols/chemistry , Benzyl Alcohols/isolation & purification , Dihydroorotate Dehydrogenase , Enzyme Inhibitors/chemistry , Enzyme Inhibitors/isolation & purification , Fungi/classification , Homogentisic Acid/chemistry , Homogentisic Acid/isolation & purification , Inhibitory Concentration 50 , Molecular Structure , Plasmodium falciparum/drug effects , Protozoan Proteins/antagonists & inhibitorsABSTRACT
Pyomelanin mimics from homogentisic acid (HGA) and gentisic acid (GA) were biosynthesized by the oxidative enzyme T. versicolor laccase at physiological pH to obtain water soluble melanins. The pigments show brown-black color, broad band visible light absorption, a persistent paramagnetism and high antioxidant activity. The EPR approach shows that at least two different radical species are present in both cases, contributing to the paramagnetism of the samples. This achievement can also shed light on the composition of the ochronotic pigment in the Alkaptonuria disease. On the other hand, these soluble pyomelanin mimics, sharing physico-chemical properties with eumelanin, can represent a suitable alternative to replace the insoluble melanin pigment in biotechnological applications.
Subject(s)
Antioxidants/pharmacology , Gentisates/pharmacology , Homogentisic Acid/pharmacology , Antioxidants/chemistry , Antioxidants/isolation & purification , Antioxidants/metabolism , Biotechnology/methods , Fungal Proteins/metabolism , Gentisates/chemistry , Gentisates/isolation & purification , Gentisates/metabolism , Homogentisic Acid/chemistry , Homogentisic Acid/isolation & purification , Homogentisic Acid/metabolism , Laccase/metabolism , Melanins/chemistry , Polyporaceae/enzymologyABSTRACT
Chromatographic separation of fruits and flowers of the Thai medicinal plant, Miliusa velutina, resulted in the isolation of five new rare homogentisic acid derivatives, miliusanal (1) and miliusanones A-D (2-5), together with fifteen known secondary metabolites (6-20). Their structures were determined through the use of extensive spectroscopic data. The absolute configurations of homogentisic acid derivatives 2-7 were identified using NOESY data and a comparison of experimental and calculated ECD spectral data. Compounds 2, 3, 6, and 7 showed antimalarial activity with IC50 values in the range of 3.3-5.2⯵g/mL. Compound 6 also showed activity against Mycobacterium tuberculosis with an MIC value of 50⯵g/mL. Compounds 1-3, 6 and 7 exhibited cytotoxicity againt KB, MCF-7, NCI-H187 and Vero cell lines with IC50 values in the range of 5.8-40.4⯵g/mL. In addition, compounds 1, 2 and 6 showed moderate antibacterial activities against three Gram-positive bacteria (Bacillus cereus, Staphylococcus aureus, and Methicillin resistant S. aureus) with MICs in the range of 32-64⯵g/mL.
Subject(s)
Annonaceae/chemistry , Anti-Bacterial Agents/pharmacology , Antineoplastic Agents, Phytogenic/pharmacology , Homogentisic Acid/pharmacology , Animals , Anti-Bacterial Agents/isolation & purification , Antineoplastic Agents, Phytogenic/isolation & purification , Cell Line, Tumor , Chlorocebus aethiops , Flowers/chemistry , Fruit/chemistry , Gram-Positive Bacteria/drug effects , Homogentisic Acid/isolation & purification , Humans , Microbial Sensitivity Tests , Molecular Structure , Mycobacterium tuberculosis/drug effects , Phytochemicals/isolation & purification , Phytochemicals/pharmacology , Thailand , Vero CellsABSTRACT
As part of our search for new antimalarial drugs in South Pacific marine sponges, we have looked for inhibitors of Pfnek-1, a specific protein kinase of Plasmodium falciparum. On the basis of promising activity in a preliminary screening, the ethanolic crude extract of a new species of Pseudoceratina collected in Vanuatu was selected for further investigation. A bioassay-guided fractionation led to the isolation of a derivative of homogentisic acid [methyl (2,4-dibromo-3,6-dihydroxyphenyl)acetate, 4a] which inhibited Pfnek-1 with an IC(50) around 1.8 muM. This product was moderately active in vitro against a FcB1 P. falciparum strain (IC(50) = 12 muM). From the same sponge, we isolated three known compounds [11,19-dideoxyfistularin-3 (1), 11-deoxyfistularin-3 (2) and dibromo-verongiaquinol (3)] which were inactive against Pfnek-1. Synthesis and biological evaluation of some derivatives of 4a are reported.
Subject(s)
Antimalarials/pharmacology , Homogentisic Acid/analogs & derivatives , Homogentisic Acid/pharmacology , Plasmodium falciparum/drug effects , Porifera/chemistry , Protein Kinase Inhibitors/pharmacology , Animals , Anti-Bacterial Agents/isolation & purification , Anti-Bacterial Agents/pharmacology , Antimalarials/isolation & purification , Enzyme Assays , Homogentisic Acid/isolation & purification , Protein Kinase Inhibitors/isolation & purification , VanuatuABSTRACT
The new homogentisic acid derivatives miliusol (1b) and miliusolide (2) from Miliusa balansae were isolated and structurally determined by spectroscopic means. The relative configurations of the new 1b and its known acetate 1a were established. Furthermore, the symmetric ether bis(2-hydroxyphenyl)methyl ether 3, which was isolated for the first time from a natural source, the known flavonoids pachypodol and chrysosplenol C, and sodium benzoate were identified.
Subject(s)
Annonaceae/chemistry , Benzofurans/isolation & purification , Homogentisic Acid/isolation & purification , Plants, Medicinal/chemistry , Quercetin/analogs & derivatives , Benzofurans/chemistry , Flavonoids/chemistry , Flavonoids/isolation & purification , Homogentisic Acid/analogs & derivatives , Homogentisic Acid/chemistry , Molecular Structure , Nuclear Magnetic Resonance, Biomolecular , Quercetin/chemistry , Quercetin/isolation & purification , Stereoisomerism , VietnamABSTRACT
The lly locus confers fluorescence, haemolysis, brown pigmentation and an increased resistance to light in Legionella pneumophila. In this study, we correlated the pigment production of two lly-positive L. pneumophila isolates and a recombinant lly-positive Escherichia coli strain with the presence of homogentisic acid (HGA) in the culture supernatant. The detection of HGA by high performance liquid chromatography and the data analysis of the deduced amino acid sequence of the lly gene indicate that the lly locus codes for a p-hydroxyphenylpyruvate dioxygenase (HPPD). This enzyme catalyses the transformation of p-hydroxyphenylpyruvate into HGA, which subsequently oxidises and polymerises into a melanin-like pigment. One open reading frame (ORF 1) in the lly region exhibited homologies with genes of Synechocystis sp., Petroselium crispum and Streptomyces mycarofaciens that code for methyltransferases. By screening a genomic library of L. pneumophila (serogroup 1) strain Corby with a monoclonal antibody against the legiolysin (lly), we identified two recombinant E. coli clones that did not produce the brown pigment and showed no haemolysis and fluorescence. DNA sequencing revealed that both clones contained 874 nucleotides of the N-terminal part of the lly gene. The recombinant strains expressed truncated legiolysin proteins of 39.5 and 35.7 kDa and did not produce HGA. Considering the highly conserved structure of legiolysin-like HPPD genes from other organisms, we suggest that the C-terminus of the legiolysin may be essential for the enzymatic activity that conferred pigmentation via HGA polymerisation, haemolysis and fluorescence.
Subject(s)
4-Hydroxyphenylpyruvate Dioxygenase/metabolism , Bacterial Proteins/physiology , Legionella pneumophila/metabolism , Bacterial Proteins/biosynthesis , Bacterial Proteins/genetics , Blotting, Western , Chromatography, High Pressure Liquid , Escherichia coli/genetics , Homogentisic Acid/isolation & purification , Homogentisic Acid/metabolism , Legionella pneumophila/genetics , Molecular Sequence Data , Open Reading Frames , Phenylpyruvic Acids/metabolism , Pigments, Biological/isolation & purification , Pigments, Biological/metabolism , Transformation, BacterialABSTRACT
Analysis of organic acids in strawberry-tree (Arbutus unedo) honey showed the presence of an unknown acid as the main constituent. This compound was isolated and identified as homogentisic acid (2, 5-dihydroxyphenylacetic acid) by MS and NMR techniques. Its average content in honey was 378 +/- 92 mg/kg. Analysis of nectar confirmed the floral origin of the compound found in honey. Since this acid was not detected in any of the different monofloral honeys, it could be used as a marker of strawberry-tree (A. unedo) honey.
