947 resultados para YEAST CYTOCHROME-C


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Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

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Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

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Two ascomycetous yeast species, Candida flosculorum sp. nov. and Candida floris sp. nov., were isolated from tropical flowers and their associated insects. C. flosculorum was isolated from flower bracts of Heliconia velloziana and Heliconia episcopalis (Heliconiaceae) collected from two Atlantic rain forest sites in Brazil. C. floris was isolated from flowers of lpomoea sp. (Convolvulaceae) growing on the banks of the river Paraguai in the pantanal ecosystem in Brazil and from an adult of the stingless bee Trigona sp. and a flower of Merremia quinquefolia (Convolvulaceae) in Costa Rica. C. flosculorum belongs to the Metschnikowiaceae clade and C. floris belongs to the Starmerella clade. The type strain of C. flosculorum is UFMG-JL13(T) (=CBS 10566(T) =NRRL Y-48258(T)) and the type strain of C. floris is UWO(PS) 00-226.2(T) (=CBS 10593(T) =NRRL Y-48255(T)).

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Strains belonging to two novel yeast species, Candida bromeliacearum and Candida ubatubensis, were isolated from the bromeliad tank of Canistropsis seidelii (Bromeliaceae) in a sandy coastal plain (restinga) ecosystem site in an Atlantic rainforest of south-eastern Brazil. These species were genetically distinct from all other currently accepted ascomycetous yeasts, based on sequence divergence in the D1/D2 domains of the large-subunit rDNA and in the small-subunit rDNA. The species occupy basal positions in the Metschnikowiaceae clacle. The type strains are Candida bromeliacearum UNESP 00-103(T) (=CBS 10002(T) = NRRL Y-27811(T)) and Candida ubatubensis UNESP 01-247R(T) (=CBS 10003(T) = NRRL Y-27812(T)).

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The novel yeast species Candida leandrae is described based on eight isolates from decaying fruits of Leandra reversa Cogn. (Melastomataceae) in an Atlantic rainforest site in Brazil, one from a Convolvulaceae flower in Costa Rica and one from a drosophilid in Hawai'i. The strains differed in their colony morphology, one being butyrous and smooth and the other being filamentous and rugose. Sequences of the D1/D2 domains of the large-subunit rRNA gene from both morphotypes were identical. C. leandrae belongs to the Kodamaea clade and is closely related to Candida restingae. The two species can be separated on the basis of growth at 37degreesC and the assimilation of melezitose, negative in the novel species. The type culture of C. leandrae is strain UNESP 00-64R(T) (= CBS 9735(T) = NRRL Y-27757(T)).

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The yeasts are microorganisms with great potential for biotechnological applications in diverse areas. The biological control of phytopathogens by yeasts has showed satisfactory results under laboratory conditions, and it has already produced commercial formulations. With this as focus, this work aims to perform in vitro and in vivo evaluations of the action of a Torulaspora globosa yeast strain (1S112), isolated from sugarcane rhizosphere, against the phytopathogenic mold Colletotrichum sublineolum, the causative agent of anthracnose in sorghum. In vitro experiments included the antagonism test in Petri dishes with morphological hyphal evaluation; yeast killer activity; siderophore, volatile compound and hydrolytic enzyme production. In vivo experiments were conducted in greenhouse conditions with a sorghum variety susceptible to C. sublineolum by evaluating the anthracnose disease for 6 weeks. The results indicated that the yeast strain significantly controlled the fungal growth, either in vitro or in vivo. The strain of T. globosa exhibited killer activity against two sensitive strains, which is a novel capacity for this species. The yeast did not produce siderophores, volatile compounds or hydrolytic enzymes, although it has reduced the mycelial growth, resulting in hyphal deformities but not cell death. The yeast controlled the anthracnose disease in sorghum, either inoculated before or after the fungal spores, suggesting that the competition for space and nutrients to dominate the mold and killer toxin production, altering the hyphal morphology, are mechanisms utilized by the yeast in the biocontrol.

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Insecticide resistance in laboratory selected Drosophila strains has been associated with upregulation of a range of different cytochrome P450s, however in recent field isolates of D. melanogaster resistance to DDT and other compounds is conferred by one P450 gene, Cyp6g1. Using microarray analysis of all Drosophila P450 genes, here we show that different P450 genes such as Cyp12d1 and Cyp6a8 can also be selected using DDT in the laboratory. We also show, however, that a homolog of Cyp6g1 is over-expressed in a field resistant strain of D. simulans. In order to determine why Cyp6g1 is so widely selected in the field we examine the pattern of cross-resistance of both resistant strains and transgenic flies over-expressing Cyp6g1 alone. We show that all three DDT selected P450s can confer resistance to the neonicotinoid imidacloprid but that Cyp6a8 confers no cross-resistance to malathion. Transgenic flies over-expressing Cyp6g1 also show cross-resistance to other neonicotinoids such as acetamiprid and nitenpyram. We suggest that the broad level of cross-resistance shown by Cyp6g1 may have facilitated its selection as a resistance gene in natural Drosophila populations. (C) 2003 Elsevier B.V. Ltd. All rights reserved.

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A colorimetric method has been developed and optimized to measure L-malic acid in samples of fruit juices and wine. This method is based on oxidation of the analyte, catalyzed by malate dehydrogenase (MDH) from dry baker's yeast, and in combination with the reduction of a tetrazolium salt (MTT: 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide). In the present study, the method exhibited sensitivity in the range of 500-4000 mu M of L-malic acid in the reaction cuvette, with the lower detection limit of 6.7-10(-2) g/L, the upper limit of 53.6.10(-2) g/L and a maximum standard deviation of only 2.5 % for the analyzed samples. The MDH activity from baker's yeast was also optimized, the enzyme showed a high stability at pH=8.0-9.0 and the activity was maintained completely at temperatures up to 40 degrees C for 1 hour. The results show that the colorimetric method using enzymatic preparations from dry baker's yeast is a simple and low-cost method with possibility of wide application.

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Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

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Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

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This work investigated the effects of increasing temperature from 30 degrees C to 47 degrees C on the physiological and genetic characteristics of Saccharomyces cerevisiae strain 63M after continuous fermentation with cell recycling in a system of five reactors in series. Steady state was attained at 30 degrees C, and then the temperature of the system was raised so it ranged from 35 degrees C in the last reactor to 43 degrees C in the first reactor or feeding reactor with a 2 degrees C difference between reactors. After 15 days at steady state, the temperature was raised from 37 degrees C to 45 degrees C for 25 days at steady state, then from 39 degrees C to 47 degrees C for 20 days at steady state. Starter strain 63M was a hybrid strain constructed to have a MAT a/alpha, LYS/lys, URA/ura genotype. This hybrid yeast showed vigorous growth on plates at 40 degrees C, weak growth at 41 degrees C, positive assimilation of melibiose, positive fermentation of galactose, raffinose and sucrose. of 156 isolates obtained from this system at the end of the fermentation process, only 17.3% showed the same characteristics as starter strain 63M. Alterations in mating type reaction and in utilization of raffinose, melibiose, and sucrose were identified. Only 1.9% of the isolates lost the ability to grow at 40 degrees C. Isolates showing requirements for lysine and uracil were also obtained. In addition, cell survival was observed at 39-47 degrees C, but no isolates showing growth above 41 degrees C were obtained.

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Flotation is a process of cell separation based on the affinity of cells to air bubbles. In the present work, flotability and hydrophobicity were determined using cells from different yeasts (Hansenulla polymorpha, Saccharomyces cerevisiae, Candida albicans), which were propagated in different media and at different temperatures. Alterations to the supernatant of the cells were also carried out before the flotation assays. The results described here indicate that supernatants of the yeast cells can play a more important role on flotation than cell-wall hydrophobicity. For example, wall-hydrophobicity of strain FLT-01 of S. cerevisiae was high but flotation did not occur when their washed cells were resuspended in water. Additions of neopeptone to cultures of S. cerevisiae and H. polymorpha repressed flotation and increased the volume of foam. An additional task of the present work was to show that the relationship between cell-wall hydrophobicity and flotation performance was dependent on the method used for the measurement of hydrophobicity. Based on the assay procedure, two types of hydrophobicity were distinguished: (a) the apparent hydrophobicity for cells suspended in the medium and expressed by the degree of cell affinity to the organic solvent in the two-phase system supernatant/hexane; (b) the standard hydrophobicity, which was determined for cells suspended in a standard solution (acetate buffer, in the present work) within the acetate buffer/hexane system. Flotation of cells of S. cerevisiae and C albicans were best related to the degree of apparent hydrophobicity (varying with the supernatant composition at the cell/medium interface) rather than to the degree of standard hydrophobicity (varying with the alterations in the wall components, since the liquid phase was constant in the assay). However, depending on the yeast unpredictable results can be obtained. For example, cells of H. polymorpha exhibited good flotation associated to a high degree of standard hydrophobicity while having a lower degree of apparent hydrophobicity. Concerning growth temperature, flotation of cells of C albicans was strongly repressed when the temperature was raised from 30 to 38 degreesC while a similar effect was not observed in cultures of S. cerevisiae and H. polymorpha. It is difficult to understand and predict flotation of yeast cells but simple modifications made to the supernatant of cultures can activate or repress flotation. (C) 2003 Elsevier B.V. B.V. All rights reserved.