RESUMO
The cyclipostins are a group of hormone-sensitive lipase inhibitors produced by a Streptomyces species. Having verticillate spore chains this strain exhibits significant differences to the known species of the former genus Streptoverticillium. Taxonomic studies and fermentation results are presented.
Assuntos
Inibidores Enzimáticos/metabolismo , Esterol Esterase/antagonistas & inibidores , Streptomyces/metabolismo , Biomassa , Metabolismo dos Carboidratos , Meios de Cultura , Fermentação , Microscopia Eletrônica de Varredura , Dados de Sequência Molecular , Esporos Bacterianos/fisiologia , Esporos Bacterianos/ultraestrutura , Streptomyces/química , Streptomyces/classificação , Streptomyces/fisiologiaRESUMO
A strain that produces new lipopeptide antibiotics is a new species of the genus Actinoplanes for which we propose the name Actinoplanes friuliensis (type strain: HAG 010964). The strain is an actinoplanete actinomycete having cell wall II composition and forming sporangia. Comparisons with Actinoplanes spp. which have similarities with our isolate, including fatty acid analysis, showed that the isolate belongs to a new species. Taxonomic studies and fermentation are presented.
Assuntos
Actinomycetales/classificação , Actinomycetales/metabolismo , Antibacterianos/farmacologia , Inibidores da Síntese de Proteínas/farmacologia , Actinomycetales/fisiologia , Antibacterianos/metabolismo , Avaliação Pré-Clínica de Medicamentos/métodos , Ácidos Graxos/metabolismo , Fermentação , Testes de Sensibilidade Microbiana , Peptidoglicano/biossíntese , Peptidoglicano/efeitos dos fármacos , Inibidores da Síntese de Proteínas/metabolismoRESUMO
The actagardine-producing strain Actinoplanes liguriae ATCC 31048, forms an additional lantibiotic when it is cultured on mannitol and soya meal. The new compound, Ala(0)-actagardine (1), has been isolated by solid-phase extraction followed by a two-step chromatographic separation. The molecular formula of 1 is C84H129N21O25S4. Its chemical structure was determined by 2D-NMR analysis and was further confirmed by an amino acid analysis, Edman degradation, and partial synthesis from actagardine. 1 exhibits a slightly higher biological activity than the parent compound actagardine. The synthetic analogs Lys(0)-actagardine (2) and Ile(0)-actagardine (3) demonstrate also antibacterial activities and emphasize the importance of the N-terminus for further derivatization.
Assuntos
Actinomycetales/metabolismo , Antibacterianos/química , Antibacterianos/farmacologia , Peptídeos , Sequência de Aminoácidos , Antibacterianos/isolamento & purificação , Bacteriocinas , Fermentação , Espectroscopia de Ressonância Magnética , Testes de Sensibilidade Microbiana , Dados de Sequência Molecular , Relação Estrutura-AtividadeRESUMO
The novel peptide feglymycin has been isolated from cultures of Streptomyces sp. DSM 11171 by solid phase extraction, size exclusion chromatography and repeated reversed-phase chromatography. The molecular weight was found to be 1900.90 g/mol and the molecular formula is C95H97Nl3O30. Feglymycin contains 13 amino acids of which four are 3-hydroxyphenylglycine and five are 3,5-dihydroxyphenylglycine residues. The structure of the linear peptide has been determined by 1H and 13C NMR spectroscopy. The sequence was confirmed by the observed mass spectroscopic fragmentation pattern. As well as having weak antibacterial activity, feglymycin inhibits the replication of the human immunodeficiency virus (HIV) in vitro.
Assuntos
Antivirais/isolamento & purificação , HIV/efeitos dos fármacos , Peptídeos/isolamento & purificação , Proteínas/isolamento & purificação , Streptomyces/química , Replicação Viral/efeitos dos fármacos , Antivirais/química , Antivirais/farmacologia , Cromatografia em Gel , Escherichia coli/efeitos dos fármacos , Fermentação , Cromatografia Gasosa-Espectrometria de Massas , Células Gigantes/efeitos dos fármacos , HIV/fisiologia , Espectroscopia de Ressonância Magnética , Testes de Sensibilidade Microbiana , Peso Molecular , Peptídeos/química , Peptídeos/farmacologia , Proteínas/química , Proteínas/farmacologia , Pseudomonas aeruginosa/efeitos dos fármacos , Staphylococcus aureus/efeitos dos fármacos , Staphylococcus epidermidis/efeitos dos fármacos , Streptococcus pyogenes/efeitos dos fármacos , Streptomyces/metabolismoRESUMO
New antifungal antibiotics, designated as 3874 H1 and H3, were discovered in the fermentation broth of the strain Streptomyces sp. HAG 003874. The compounds were obtained as yellow powders after sequential purification by chromatography on MCI Gel CHP20P, Fractogel HW-40 and ODS reversed phase chromatography. On the basis of the results of spectroscopic analysis, it was found that 3874 H1, C58H86N2O18, MW 1098, belongs to the p-aminoacetophenone containing family of heptaene antibiotics, while 3874 H3, C57H87NO18, MW 1073, is a non-aromatic heptaene. In addition to these, a minor component, 3874 H2, C59H88N2O18, MW 1112, a N-methyl derivative of 3874 H1 has been detected. The structures were elucidated through mass spectral analyses and 1-D and 2-D homonuclear and heteronuclear NMR data. The outstanding physico-chemical feature of 3874 H3 is its improved solubility. The new heptaenes are potent antifungal compounds with broad activity spectra, encompassing dermatophytes, yeasts and filamentous fungi.
Assuntos
Antibacterianos/isolamento & purificação , Antifúngicos/isolamento & purificação , Macrolídeos , Polienos/isolamento & purificação , Streptomyces/química , Antibacterianos/química , Antibacterianos/farmacologia , Antifúngicos/química , Antifúngicos/farmacologia , Contagem de Colônia Microbiana , Fungos/efeitos dos fármacos , Espectroscopia de Ressonância Magnética/métodos , Espectrometria de Massas , Testes de Sensibilidade Microbiana , Polienos/química , Polienos/farmacologia , Streptomyces/classificaçãoRESUMO
An extracellular esterase from the actinomycetes Amycolatopsis orientalis was found by screening. It is capable of splitting the isomeric mixture (K/J) of (I, Scheme 1) into 7-amino-3-methoxymethyl-3-cephem-4-carboxylic acid, pivalic acid, and acetaldehyde with a high yield. The purified enzyme of 55.4 Kd by SDS-PAGE shows an N-terminal sequence of VRTCADLVRTYDLPGAVTH. The isoelectric point is 8.9 +/- 0.1. It can be immobilized with good yield to VA-Epoxy Biosynth. Besides the above-mentioned reaction, the esterase cleaves many other esters such as methyl-2-chloropropionic acid.
Assuntos
Cefalosporinas/síntese química , Esterases/química , Esterases/metabolismo , Ácidos Pentanoicos , Pró-Fármacos , Actinobacteria/enzimologia , Sequência de Aminoácidos , Cefalosporinas/metabolismo , Esterases/isolamento & purificação , Ésteres , Fermentação , Cinética , Dados de Sequência Molecular , Fragmentos de Peptídeos/química , Especificidade por SubstratoRESUMO
Different proteases from the culture fluids of recombinant Streptomyces lividans strains were isolated. Several individual proteases were separated and characterized. A chymotrypsin-chylike activity (CLA) was identified that specifically degrades a fusion protein between the alpha-amylase inhibitor from S. tendae (Tendamistat, HOE467) and proinsulin from the monkey Macaca fascicularis. The effective chemical inhibition of the degrading enzyme is demonstrated.
Assuntos
Peptídeo Hidrolases/metabolismo , Streptomyces/enzimologia , Sequência de Aminoácidos , Animais , Proteínas de Bactérias/metabolismo , Cromatografia por Troca Iônica , Quimotripsina/antagonistas & inibidores , Quimotripsina/metabolismo , Metais/farmacologia , Dados de Sequência Molecular , Peptídeo Hidrolases/isolamento & purificação , Proteínas Recombinantes de Fusão/metabolismo , Streptomyces/genética , Streptomyces/crescimento & desenvolvimento , Fatores de TempoRESUMO
A flow cytometry (FCM) system was chosen to analyze and sort microbiological samples, e.g., bacteria, bacterial spores, yeasts, and fungal spores, without major changes in the commercially available state. The system was further improved by addition of a stepping motor-driven scanning table that accepts standard petri dishes or microtiter plates. The electronics of the sorting system were changed to enable the sorter to deliver only one particle at a time, working in a "handshake" mode with the scanning table. Appropriate parameters, depending on the biological material and including all fluorescent stains that do not impair growth and productivity of cells were chosen to sort distinct bioparticles under aseptic conditions and to clone colonies or cultures out of them. A mutagenized sample of spores entering the germination cycle can be followed and thus provide a means to pick only viable growing cells despite the killing effect of the mutagen. One example of a typical strain improvement is illustrated. From a spore suspension of Rhizopus arrhizus, a subpopulation of morphologically different spores comprising about 5-10% of the whole population was cloned. From approximately 8,000 clones, 10 were isolated that produced approximately five- to six-fold the amount of fungal lipase activity, compared to the original strain or to reisolated clones from the mean population of clones.