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71.
Alcohol dehydrogenase (ADH) is a key enzyme in the production and utilization of alcohols. Some also catalyze the formation
of carboxylate esters from alcohols and aldehydes. The ADH1 and ADH3 genes of Neurospora crassa FGSC2489 were cloned and expressed in recombinant Escherichia coli to investigate their alcohol dehydrogenation and carboxylate ester formation abilities. Homology analysis and sequence alignment
of amino acid sequence indicated that ADH1 and ADH3 of N. crassa contained a zinc-binding consensus sequence and a NAD+-binding motif and showed 54–75% identity with fungi ADHs. N. crassa ADH1 was expressed in E. coli to give a specific activity of 289 ± 9 mU/mg using ethanol and NAD+ as substrate and cofactor, respectively. Corresponding experiments on the expression and activity of ADH3 gave 4 mU/mg of specific activity. N. crassa
ADH1 preferred primary alcohols containing C3–C8 carbons to secondary alcohols such as 2-propanol and 2-butanol. N. crassa
ADH1 possessed 5.3 mU/mg of specific carboxylate ester-forming activity accumulating 0.4 mM of ethyl acetate in 18 h. Substrate
specificity of various linear alcohols and aldehydes indicated that short chain-length alcohols and aldehydes were good substrates
for carboxylate ester production. N. crassa
ADH1 was a primary alcohol dehydrogenase using cofactor NAD+ preferably and possessed carboxylate ester-forming activity with short chain alcohols and aldehydes. 相似文献
72.
We report structural features and distribution patterns of 26 different group I introns located at three distinct nucleotide
positions in nuclear small subunit ribosomal DNA (SSU-rDNA) of 10 Septoria and 4 other anamorphic species related to the teleomorphic genus Mycosphaerella. Secondary structure and sequence characteristics assigned the introns to the common IC1 and IE groups. Intron distribution
patterns and phylogenetic relationships strongly suggested that some horizontal transfer events have occurred among the closely
related fungal species sampled. To test this hypothesis, we used a comparative approach of intron- and rDNA-based phylogenies
through MP- and ML-based topology tests. Our results showed two statistically well-supported major incongruences between the
intron and the equivalent internal transcribed spacer (ITS) tree comparisons made. Such absence of a co-evolutive history
between group I introns and host sequences is discussed relatively to the intron structures, the mechanisms of intron movement,
and the biology of the Mycosphaerella pathogenic fungi.
Electronic Supplementary Material The online version of this article (doi:) contains supplementary material, which is available to authorized users.
Reviewing Editor: Debashish Bhattacharya 相似文献
73.
Fungal contamination is a major problem in cell culture, and the antifungal compounds currently in use can affect cultured
cells. Echinocandins are antifungal drugs that inhibit fungal cell wall synthesis by targeting an enzyme that has no counterpart
in mammalian cells. We evaluated whether the echinocandin caspofungin affected the growth or morphology of six murine cell
lines (a macrophage-like cell line (J774.16) and five hybridoma lines), or primary human endothelial cells. The antifungal
did not influence cellular characteristics at concentrations less than 512 μg/ml, while effectively reducing the incidence
of fungal contamination. Also, caspofungin did not affect the production of antibody by hybridoma cells, or alter the cytokine
production of J774.16 cells, although modest increases in IL-4 and IFN-γ occurred upon LPS stimulation. Hence, echinocandins
appear to be relatively non-toxic, and protect against fungal contamination in cell culture. 相似文献
74.
外科深部真菌感染的病原学分析 总被引:1,自引:0,他引:1
目的分析外科住院患者合并真菌感染状况。方法分离的真菌用API 20C AUX真菌鉴定条,用ATB FUNGUS-2进行药敏试验。结果真菌的种类分布中第1位是白色念珠菌占53%,第2位是热带念珠菌占20%,第3位是光滑念珠菌占14%,第4位是近平滑念珠菌占4%,第5位是其余念珠菌占2%;真菌在外科各区分布中,外科ICU占42%,移植外科占25%;真菌在各类标本分布中,痰占43%,尿液占13%,粪便占10%,引流液占9%,血液占7%;真菌对药物5-氟胞嘧啶、两性霉素B、氟康唑、伊曲康唑的药物敏感性分别是93%、98.9%、90.8%、59.8%。结论外科真菌感染集中在重症病区(ICU),分离的致病真菌主要是白色念珠菌、热带念珠菌、光滑念珠菌,真菌对两性霉素B敏感性最高。 相似文献
75.
Characterization of aerobic bacterial and fungal microbiota on surfaces of historic Scottish monuments 总被引:1,自引:0,他引:1
Suihko ML Alakomi HL Gorbushina A Fortune I Marquardt J Saarela M 《Systematic and applied microbiology》2007,30(6):494-508
Twenty samples were taken from the inner or outer surfaces of stone monuments of six historic Scottish buildings and ruins. Biofilms developing on mineral substrates were analysed by in situ scanning electron microscopy and cultivation. Various methods were used to characterize the isolates including automated ribotyping, RAPD and sequencing of the 16S rRNA gene for bacteria, and stereomicroscopy and sequencing of the Internal Transcribed Spacers (ITS) for fungi. Most samples contained microbes between 10(5) and 10(7)cfug(-1) substrate. Actinobacteria belonging to the genus Streptomyces (17 samples/5 monuments) or Arthrobacter (12/3) and Pseudomonas (9/3) were frequently detected. Most streptomycetes were in terms of their 16S rRNA gene sequence most closely related to S. microflavus (10/3) or to the undescribed species S. "vulgaris" (8/3). Indoor and outdoor biofilms exhibited significant differences in their microbiota, as shown by both microscopy and isolation studies. Pigmented coccoid Arthrobacter species were typical for the outdoor samples, whereas Pseudomonas species were common in the indoor samples. Based on the low phylogenetic relationship to a known species (type strain), potential novel pigmented bacterial species belonging to the genera Arthrobacter, Brevundimonas, Cryseobacterium, Deinococcus and Dyadobacter were detected from the outdoor samples and to Pseudomonas from the indoor samples. Hyaline fungal species of Acremonium (10/4) mainly occurred in indoor samples, whereas pigmented species of Cladosporium (8/3), Penicillium (6/3) and Phialophora (6/2) were found outdoors. Using in situ microscopy diatom algae were also detected. 相似文献
76.
The Internal Transcribed Spacer (ITS) regions of ribosomal DNA are widely used as markers for phylogenetic analyses and environmental
sampling from a variety of organisms including fungi, plants, and animals. In theory, concerted evolution homogenizes multicopy
genes so that little or no variation exists within populations or individuals. However, contrary to theory, ITS variation
has been confirmed in populations and individuals from a diverse range of eukaryotes. The presence of intraspecific and intra-individual
variation in multicopy genes has important implications for ecological and phylogenetic studies, yet relatively little is
known about natural variation of these genes, particularly at the community level. In this study, we examined intraspecific
and intra-sporocarp ITS variation by DNA sequencing from sporocarps and pooled roots from 68 species of ectomycorrhizal fungi
collected at a single site in a Quercus woodland. We detected ITS variation in 27 species, roughly 40% of the taxa examined. Although intraspecific ITS variation
was generally low (0.16–2.85%, mean = 0.74%), it was widespread within this fungal community. We detected ITS variation in
both sporocarps and ectomycorrhizal roots, and variation was present within species of Ascomycota and Basidiomycota, two distantly
related lineages within the Fungi. We discuss the implications of such widespread ITS variability with special reference to
DNA-based environmental sampling from diverse fungal communities.
Electronic supplementary material The online version of this article (doi:) contains supplementary material, which is available to authorized users. 相似文献
77.
Background
Transposable Elements (TEs) are key components that shape the organization and evolution of genomes. Fungi have developed defense mechanisms against TE invasion such as RIP (Repeat-Induced Point mutation), MIP (Methylation Induced Premeiotically) and Quelling (RNA interference). RIP inactivates repeated sequences by promoting Cytosine to Thymine mutations, whereas MIP only methylates TEs at C residues. Both mechanisms require specific cytosine DNA Methyltransferases (RID1/Masc1) of the Dnmt1 superfamily.Results
We annotated TE sequences from 10 fungal genomes with different TE content (1-70%). We then used these TE sequences to carry out a genome-wide analysis of C to T mutations biases. Genomes from either Ascomycota or Basidiomycota that were massively invaded by TEs (Blumeria, Melampsora, Puccinia) were characterized by a low frequency of C to T mutation bias (10-20%), whereas other genomes displayed intermediate to high frequencies (25-75%). We identified several dinucleotide signatures at these C to T mutation sites (CpA, CpT, and CpG). Phylogenomic analysis of fungal Dnmt1 MTases revealed a previously unreported association between these dinucleotide signatures and the presence/absence of sub-classes of Dnmt1.Conclusions
We identified fungal genomes containing large numbers of TEs with many C to T mutations associated with species-specific dinucleotide signatures. This bias suggests that a basic defense mechanism against TE invasion similar to RIP is widespread in fungi, although the efficiency and specificity of this mechanism differs between species. Our analysis revealed that dinucleotide signatures are associated with the presence/absence of specific Dnmt1 subfamilies. In particular, an RID1-dependent RIP mechanism was found only in Ascomycota.Electronic supplementary material
The online version of this article (doi:10.1186/s12864-015-1347-1) contains supplementary material, which is available to authorized users. 相似文献78.
Background
Maternal or uniparental inheritance (UPI) of mitochondria is generally observed in sexual eukaryotes, however, the underlying mechanisms are diverse and largely unknown. Recently, based on the use of mutants blocked in autophagy, it has been demonstrated that autophagy is required for strict maternal inheritance in the nematode Caenorhabditis elegans. Uniparental mitochondrial DNA (mtDNA) inheritance has been well documented for numerous fungal species, and in particular, has been shown to be genetically governed by the mating-type loci in the isogamous species Cryptococcus neoformans, Phycomyces blakesleeanus and Ustilago maydis. Previously, we have shown that the a2 mating-type locus gene lga2 is decisive for UPI during sexual development of U. maydis. In axenic culture, conditional overexpression of lga2 triggers efficient loss of mtDNA as well as mitophagy. To assess a functional relationship, we have investigated UPI in U. maydis Δatg11 mutants, which are blocked in mitophagy.Results
This study has revealed that Δatg11 mutants are not affected in pathogenic development and this has allowed us to analyse UPI under comparable developmental conditions between mating-compatible wild-type and mutant strain combinations. Explicitly, we have examined two independent strain combinations that gave rise to different efficiencies of UPI. We demonstrate that in both cases UPI is atg11-independent, providing evidence that mitophagy is not critical for UPI in U. maydis, even under conditions of strict UPI.Conclusions
Until now, analysis of a role of mitophagy in UPI has not been reported for microbial species. Our study suggests that selective autophagy does not contribute to UPI in U. maydis, but is rather a consequence of selective mtDNA elimination in response to mitochondrial damage.Electronic supplementary material
The online version of this article (doi:10.1186/s12866-015-0358-z) contains supplementary material, which is available to authorized users. 相似文献79.
M.A. ShirakawaK. Loh V.M. JohnM.E.S. Silva C.C. Gaylarde 《International biodeterioration & biodegradation》2011,65(5):669-674
Four different architectural acrylic paint formulations were tested by exposure to weathering for 7 years in the urban site of São Paulo and the coastal site of Ubatuba, South-East Brazil. Surface discolorations and detachment of coatings were assessed and the components of the biofilms were identified by standard microbiological methods. The painted surfaces of the mortar panels were much more discolored in Ubatuba, where major components of the biofilms were the cyanobacteria Gloeocapsa and Scytonema. In two of the four paint films, a pink coloration on the surface at this coastal site, caused mainly by red-pigmented Gloeocapsa, produced high discoloration ratings, but low degradation (as measured by detachment). Biofilms in São Paulo contained the same range of phototrophs, but in lesser quantity. However, fungal numbers, as determined by plating, were higher. Detachment ratings in this urban site were only slightly lower than in Ubatuba. The matt paint performed worst of the four, with silk and semi-gloss finishes giving lowest biodeterioration ratings. The matt elastomeric paint performed well at both sites, apart from becoming almost 100% covered by the pink biofilm in Ubatuba. Unpainted mortar panels became intensely discolored with a black biofilm, showing that all the paints had achieved one of their objectives, that of surface protection of the substrate. The value of PVC (pigment volume content) as an indicator of coatings biosusceptibility, is questioned. 相似文献
80.
Two new sorbicillinoids, 1 and 2 , together with a novel benzofuranone derivative named phialofurone ( 3 ), were isolated from a deep‐sea sediment‐derived fungus, Phialocephala sp. Their structures were established on the basis of spectroscopic data. All compounds displayed cytotoxic effects against P388 (IC50 values of 11.5±1.4, 0.1±0.1, and 0.2±0.01 μM , resp.) and K562 (IC50 values of 22.9±0.8, 4.8±0.3 and 22.4±0.9 μM , resp.) cell lines. 相似文献