Circadian clocks are the internal time-keeping mechanisms for organisms to synchronize their cellular and physiological processes to the daily light/dark cycles. The molecular mechanisms underlying circadian clocks are remarkably similar in eukaryotes. Neurospora crassa, a filamentous fungus, is one of the best understood model organisms for circadian research. In recent years, accumulating data have revealed complex regulation in the Neurospora circadian clock at transcriptional, posttranscriptional, post-translational and epigenetic levels. Here we review the recent progress towards our understanding of the molecular mechanism of the Neurospora circadian oscillator. These advances have provided novel insights and furthered our understanding of the mechanism of eukaryotic circadian clocks.
Circadian Clocks ; Epigenomics ; Neurospora ; genetics ; metabolism ; physiology ; Neurospora crassa ; genetics ; metabolism ; physiology

Six kinds of elicitors were prepared respectively from Neurospora crassa, Monascus purpureus, Sporobolomyces roseus, Rhodotorula rubra, Nocardia sp. N89 and Actinoplanes sp. A05. When Penicillium sp. PT95 was incubated in Czapek's agar plates containing appropriate amounts of elicitors, both its sclerotia biomass and carotenoid content accumulated in sclerotia were enhanced significantly (P < 0.01). Among tested elicitors, the elicitors from the fungi N. crassa, M. purpureus, S.-roseus and R. rubra were more effective than those from the actinomycetes Nocardia sp. N89 and Actinoplanes sp. A05; the elicitor from M. purpureus gave the highest carotenoid yield of 599 micrograms/plate, 2.76 times higher than that of control. Every one of elicitors except that from M. purpureus could increase significantly the proportion of beta-carotene in total carotenoids (P < 0.01).
Actinomycetales ; physiology ; Biomass ; Carotenoids ; biosynthesis ; Neurospora crassa ; physiology ; Nocardia ; physiology ; Penicillium ; metabolism ; Rhodotorula ; physiology

Filamentous fungi are widely used for large-scale production of cellulases. Morphological characteristics of mycelia under submerged condition are closely correlated with cellulases productivity. In order to find out the critical genes involved in the mycelial morphology development and cellulases production in liquid fermentation, 95 Neurospora crassa morphological mutants (named as SZY1-95) were screened for cellulases production. Compared with the wild type, cellulases production in four mutants SZY32, SZY35, SZY39 and SZY43 were significantly decreased, whereas mutants SZY63, SZY69, SZY87 and SZY11 produced much more cellulases than that of the wild type strain. Meanwhile, endo-beta-1,4-glucanase activity, beta-glucosidase activity, viscosity of broth and dry weight of these mutants were measured. The mycelial morphology of the mutants was also studied by microscope. Particularly, pellets were formed in mutant SZY11 and SZY43, whose viscosities were 25% and 50% of the wild type strain, respectively. Mutant SZY87 appeared long hyphae, and the viscosity of its broth was at least 2 folds of the wild type strain. These results indicate that a single gene deletion could influence the mycelial morphology in liquid fermentation, and increased the cellulases production. The low-viscosity related genes identified in our study will be the potential candidates for genetic modification of filamentous fungi.
Cellulases ; biosynthesis ; Fermentation ; Gene Deletion ; Industrial Microbiology ; Neurospora crassa ; genetics ; metabolism

OBJECTIVETo identify the function of the gene encoding Neurospora crassa EAA33149.1 protein which has 46.85% similarity with Aspergillus niger phA gene.

METHODSThe bioinformatics analysis was conducted using the prediction algorithms SignalP v3.0, arginine and lysine propeptide cleavage sites in eukaryotic protein sequence prediction algorithms ProP 1.0 server, transmembrane domain prediction algorithms Tmpred and TMHMM v2.0, potential GP I-anchor sites prediction algorithm big-P I Predictor and the subcellular protein location prediction algorithms TargetP v1.01. According to the analysis results, the gene was cloned into Saccharomyces cerevisiae.

RESULTSThe signal peptide, the cleavage site and the secretion pathway were determined, and the expressed recombinant protein with 54,000 displayed phytase activity.

CONCLUSIONThe gene has been identified to encode N. crassa phyA.

Algorithms ; Amino Acid Sequence ; Computational Biology ; methods ; Fungal Proteins ; genetics ; metabolism ; Molecular Sequence Data ; Neurospora crassa ; genetics ; Phytochrome A ; genetics ; metabolism ; Recombinant Proteins ; genetics ; metabolism ; Saccharomyces cerevisiae ; genetics ; metabolism

The key and crucial step of metabolic engineering during quinic acid biosynthesize using shikimic acid pathway is high expression of quinate 5-dehydrogenase. The gene qa-3 which code quinate 5-dehydrogenase from Neurospora crassa doesn't express in Escherichia coli. By contrast with codon usage in Escherichia coli, there are 27 rare codons in qa-3, including eight AGG/AGA (Arg) and nine GGG (Gly). Two AGG are joined together (called box R) and some GGG codons are relative concentrate (called box G). Along with the secondary structure of mRNA analysed in computer, the free energy of mRNA changes a lot from -374.3 kJ/mol to least -80.5 kJ/mol when some bases in the end of qa-3 were transformed, and moreover, the change of free energy is quite small when only some bases in the box G and box R transformed. After the change of rare codon and optimization of some bases in the end, qa-3 was expression in E. coli and also the enzyme activity of quinate 5-dehydrogenase can be surveyed accurately. All the work above benefit the further research on producing quinic acid engineering bacterium.
Alcohol Oxidoreductases ; biosynthesis ; genetics ; Base Sequence ; Codon ; chemistry ; genetics ; Escherichia coli ; genetics ; metabolism ; Hydro-Lyases ; genetics ; Molecular Sequence Data ; Neurospora crassa ; enzymology ; genetics ; RNA, Messenger ; chemistry ; genetics ; Recombinant Proteins ; biosynthesis ; genetics ; Shikimic Acid ; metabolism ; Transformation, Bacterial