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PloS One 2011Penicillium dipodomyis is thought to be an exclusively asexual fungus associated with Kangaroo Rats, Dipodomys species, and is unique among Penicillium species in...
Penicillium dipodomyis is thought to be an exclusively asexual fungus associated with Kangaroo Rats, Dipodomys species, and is unique among Penicillium species in growing at 37°C but producing no known toxins. Lack of recombination within P. dipodomyis would result in limited adaptive flexibility but possibly enhance local adaptation and host selection via maintenance of favourable genotypes. Here, analysis of DNA sequence data from five protein-coding genes shows that recombination occurs within P. dipodomyis on a small spatial scale. Furthermore, detection of mating-type alleles supports outcrossing and a sexual cycle in P. dipodomyis. P. dipodomyis was a weaker competitor in in vitro assays with other Penicillium species found in association with Kanagaroo rats. Bayesian species level analysis suggests that the P. dipodomyis lineage diverged from closely related species also found in cheek pouches of Kangaroo Rats and their stored seeds about 11 million years ago, a similar divergence time as Dipodomys from its sister rodent taxa.
Topics: Animals; Dipodomys; Genetic Variation; Penicillium; Rats; Recombination, Genetic
PubMed: 21850241
DOI: 10.1371/journal.pone.0022883 -
Scientific Reports May 2020We present a Penicillium rubens strain with an industrial background in which the four highly expressed biosynthetic gene clusters (BGC) required to produce penicillin,...
We present a Penicillium rubens strain with an industrial background in which the four highly expressed biosynthetic gene clusters (BGC) required to produce penicillin, roquefortine, chrysogine and fungisporin were removed. This resulted in a minimal secondary metabolite background. Amino acid pools under steady-state growth conditions showed reduced levels of methionine and increased intracellular aromatic amino acids. Expression profiling of remaining BGC core genes and untargeted mass spectrometry did not identify products from uncharacterized BGCs. This platform strain was repurposed for expression of the recently identified polyketide calbistrin gene cluster and achieved high yields of decumbenone A, B and C. The penicillin BGC could be restored through in vivo assembly with eight DNA segments with short overlaps. Our study paves the way for fast combinatorial assembly and expression of biosynthetic pathways in a fungal strain with low endogenous secondary metabolite burden.
Topics: Biosynthetic Pathways; Gene Expression Regulation, Enzymologic; Gene Expression Regulation, Fungal; Genome, Fungal; Genomics; Metabolic Engineering; Multigene Family; Penicillium; Peptide Synthases; Secondary Metabolism; Transcriptome
PubMed: 32376967
DOI: 10.1038/s41598-020-64893-6 -
Microbial Biotechnology Feb 2022Fungal antifungal proteins (AFPs) have attracted attention as novel biofungicides. Their exploitation requires safe and cost-effective producing biofactories....
Fungal antifungal proteins (AFPs) have attracted attention as novel biofungicides. Their exploitation requires safe and cost-effective producing biofactories. Previously, Penicillium chrysogenum and Penicillium digitatum produced recombinant AFPs with the use of a P. chrysogenum-based expression system that consisted of the paf gene promoter, signal peptide (SP)-pro sequence and terminator. Here, the regulatory elements of the afpA gene encoding the highly produced PeAfpA from Penicillium expansum were developed as an expression system for AFP production through the FungalBraid platform. The afpA cassette was tested to produce PeAfpA and P. digitatum PdAfpB in P. chrysogenum and P. digitatum, and its efficiency was compared to that of the paf cassette. Recombinant PeAfpA production was only achieved using the afpA cassette, being P. chrysogenum a more efficient biofactory than P. digitatum. Conversely, P. chrysogenum only produced PdAfpB under the control of the paf cassette. In P. digitatum, both expression systems allowed PdAfpB production, with the paf cassette resulting in higher protein yields. Interestingly, these results did not correlate with the performance of both promoters in a luciferase reporter system. In conclusion, AFP production is a complex outcome that depends on the regulatory sequences driving afp expression, the fungal biofactory and the AFP sequence.
Topics: Antifungal Agents; Fungal Proteins; Penicillium; Penicillium chrysogenum; alpha-Fetoproteins
PubMed: 35084102
DOI: 10.1111/1751-7915.14006 -
Sensors (Basel, Switzerland) Mar 2019In this study, the PEN3 electronic nose was used to detect and recognize fresh and moldy apples inoculated with and , taking Golden Delicious apples as the model...
In this study, the PEN3 electronic nose was used to detect and recognize fresh and moldy apples inoculated with and , taking Golden Delicious apples as the model subject. Firstly, the apples were divided into two groups: individual apple inoculated only with/without different molds (Group A) and mixed apples of inoculated apples with fresh apples (Group B). Then, the characteristic gas sensors of the PEN3 electronic nose that were most closely correlated with the flavor information of the moldy apples were optimized and determined to simplify the analysis process and improve the accuracy of the results. Four pattern recognition methods, including linear discriminant analysis (LDA), backpropagation neural network (BPNN), support vector machines (SVM), and radial basis function neural network (RBFNN), were applied to analyze the data obtained from the characteristic sensors, aiming at establishing the prediction model of the flavor information and fresh/moldy apples. The results showed that only the gas sensors of W1S, W2S, W5S, W1W, and W2W in the PEN3 electronic nose exhibited a strong signal response to the flavor information, indicating most were closely correlated with the characteristic flavor of apples and thus the data obtained from these characteristic sensors were used for modeling. The results of the four pattern recognition methods showed that BPNN had the best prediction performance for the training and testing sets for both Groups A and B, with prediction accuracies of 96.3% and 90.0% (Group A), 77.7% and 72.0% (Group B), respectively. Therefore, we demonstrate that the PEN3 electronic nose not only effectively detects and recognizes fresh and moldy apples, but also can distinguish apples inoculated with different molds.
Topics: Aspergillus niger; Discriminant Analysis; Electronic Nose; Fruit; Gases; Malus; Neural Networks, Computer; Pattern Recognition, Automated; Penicillium; Support Vector Machine
PubMed: 30934812
DOI: 10.3390/s19071526 -
Journal of Applied Microbiology Jul 2015Different entrapment matrices were screened to immobilize two strains of Penicillium roqueforti (AG101 and LG109) for more effective production of mycophenolic acid...
AIMS
Different entrapment matrices were screened to immobilize two strains of Penicillium roqueforti (AG101 and LG109) for more effective production of mycophenolic acid (MPA). Further improvement in the MPA productivity from immobilization of spores and mycelia was adopted by UV and gamma irradiation.
METHODS AND RESULTS
Penicillium roqueforti strains were immobilized in different entrapping carriers and used for MPA production in shake flask cultures. Maximum MPA production was achieved on using an alginate concentration of 3·0% (w/v) and a mycelial fresh weight of 10% (w/v). MPA produced by alginate-immobilized spores and mycelia was almost double in comparison to the free system. The MPA-producing ability of immobilized AG101 and LG109 strain was significantly enhanced by mutagenesis through irradiation by UV (254 nm) for 120 and 90 min, respectively and gamma rays at 0·75 KGy. The feasibility of MPA production in a semi-continuous form by immobilized cells as affected by irradiation was adopted.
CONCLUSIONS
MPA production by immobilized spores and mycelia was more intensified by UV and gamma irradiation. Moreover, the immobilized cell culture was superior to free-cell culture.
SIGNIFICANCE AND IMPACT OF THE STUDY
These findings indicate the future possibility to reduce the cost of producing fermentation-based drugs.
Topics: Alginates; Cells, Immobilized; Fermentation; Gamma Rays; Glucuronic Acid; Hexuronic Acids; Immunosuppressive Agents; Industrial Microbiology; Mutagenesis; Mycelium; Mycophenolic Acid; Penicillium
PubMed: 25892607
DOI: 10.1111/jam.12828 -
Journal of Applied Microbiology Jul 2013In this study, a gene that encodes a carboxylesterase (carb) in Penicillium expansum GF was cloned, sequenced and overexpressed by Penicillium griseoroseum PG63, and the...
AIMS
In this study, a gene that encodes a carboxylesterase (carb) in Penicillium expansum GF was cloned, sequenced and overexpressed by Penicillium griseoroseum PG63, and the enzyme was characterized.
METHODS AND RESULTS
The recombinant strain, P. griseoroseum T55, obtained upon transformation using the plasmid pAN-52-1-carb, showed integration of the carb gene into at least two heterologous sites of the genome by Southern blotting. Furthermore, the recombinant strain T55 exhibited almost a fourfold increase in carboxylesterase activity compared with PG63 strain when both were cultured without inducers. Based on the secondary structure and multiple sequence alignments with carboxylesterases, cholinesterase and lipase, a three-dimensional model was obtained. The α/β barrel topology, that is typical of esterases and lipases, was indicated for the CARB protein with Ser(213)-Glu(341)-His(456) as the putative catalytic triad. CARB preferentially hydrolysed acyl chains with eight carbon atoms, and its activity was optimal at a pH of 7·0 and a temperature of 25°C. CARB exhibited stability in alkaline pH, high activity under mesophilic conditions and stability in organic solvents.
CONCLUSION
The CARB protein is potentially useful in bioremediation, food and chemical/pharmaceutical industries.
SIGNIFICANCE AND IMPACT OF THE STUDY
This study is the first to report the development of a recombinant strain superproducing a Penicillium sp. carboxylesterase.
Topics: Amino Acid Sequence; Base Sequence; Carboxylesterase; Cloning, Molecular; Molecular Sequence Data; Penicillium; Sequence Alignment
PubMed: 23581645
DOI: 10.1111/jam.12215 -
Marine Drugs Dec 2022Five new compounds, including two cyclopiane diterpenes conidiogenones J and K (-), a steroid andrastin H (), an alkaloid...
Five new compounds, including two cyclopiane diterpenes conidiogenones J and K (-), a steroid andrastin H (), an alkaloid ()-4-(5-acetoxy--hydroxy-3-methylpent-2-enamido) butanoate (), and an aliphatic acid ()-5-acetoxy-3-methylpent-2-enoic acid (), together with ten known compounds (- and -) were isolated from the EtOAc extract of the fermentation broth of the -derived fungus HLLG-13. Their structures were elucidated by 1D, 2D NMR, and HR-ESI-MS spectral analyses. The absolute configurations of , , and were determined by quantum chemical electronic circular dichroism (ECD) calculations, and the absolute configuration of was determined for the first time. Compound was a new natural product, and its NMR data were reported for the first time. Compounds and - exhibited antibacterial activities against and , with MIC values ranging from 6.25 to 25 μg/ mL. Compounds - and - showed significant growth inhibition activities against newly hatched Hubner larvae, with IC values ranging from 50 to 200 μg/mL.
Topics: Animals; Penicillium; Diterpenes; Magnetic Resonance Spectroscopy; Circular Dichroism; Molecular Structure
PubMed: 36662195
DOI: 10.3390/md21010022 -
Current Microbiology Jun 2021Soil-occupant fungi produce a variety of mycotoxins as secondary metabolites, one of which is mycophenolic acid (MPA), an antibiotic and immunosuppressive agent. MPA is...
Soil-occupant fungi produce a variety of mycotoxins as secondary metabolites, one of which is mycophenolic acid (MPA), an antibiotic and immunosuppressive agent. MPA is mainly produced by several species of Penicillium, especially Penicillium brevicompactum. Here, we present the first report of MPA production by a local strain belonging to Penicillium glabrum species. We screened ascomycete cultures isolated from moldy food and fruits, as well as soils, collected from different parts of Iran. MPA production of one hundred and forty Penicillium isolates was analyzed using HPLC. Three MPA producer isolates were identified, among which the most producer was subjected to further characterization, based on morphological and microscopic analysis, as well as molecular approach (ITS, rDNA and beta-tubulin gene sequences). The results revealed that the best MPA producer belongs to P. glabrum IBRC-M 30518, and can produce 1079 mg/L MPA in Czapek-Dox medium.
Topics: Iran; Mycophenolic Acid; Penicillium
PubMed: 34019120
DOI: 10.1007/s00284-021-02509-6 -
Molecular Microbiology Dec 2021Numerous transcription factors (TFs) in ascomycete fungi play crucial roles in cellular processes; however, how most of them function is poorly understood. Here, we...
Numerous transcription factors (TFs) in ascomycete fungi play crucial roles in cellular processes; however, how most of them function is poorly understood. Here, we identified and characterized a novel TF, CxrC (POX01387), acting downstream of the key TF CxrA, which is essential for plant-biomass-degrading-enzyme (PBDE) production in Penicillium oxalicum. Deletion of cxrC in P. oxalicum significantly affected the production of PBDEs, as well as mycelial growth and conidiospore production. CxrA directly repressed the expression of cxrC after about 12 hr following switch to Avicel culture. CxrC bound the promoters of major PBDE genes and genes involved in conidiospore development. CxrC was found to bind the TSSGTYR core sequence (S: C and G; Y: T and C; R: G and A) of the important cellulase genes cbh1 and eg1. Both N- and C-terminal peptides of CxrC and the CxrC phosphorylation were found to mediate its homodimerization. The conserved motif LPSVRSLLTP (65-74) in CxrC was found to be required for regulating cellulase production. This study reveals novel mechanisms of TF-mediated regulation of the expression of PBDE genes and genes involved in cellular processes in an ascomycete fungus.
Topics: Amino Acid Motifs; Cellulase; Fungal Proteins; Gene Expression Regulation, Fungal; Penicillium; Promoter Regions, Genetic; Spores, Fungal; Transcription Factors
PubMed: 34797006
DOI: 10.1111/mmi.14843 -
PloS One 2020Various marine fungi have been shown to produce interesting, bioactive compounds, but scaling up the production of these compounds can be challenging, particularly...
Various marine fungi have been shown to produce interesting, bioactive compounds, but scaling up the production of these compounds can be challenging, particularly because little is generally known about how the producing organisms grow. Here we assessed the suitability of using 100-well BioScreen plates or 96-well plates incubated in a robot hotel to cultivate eight filamentous marine fungi, six sporulating and two non-sporulating, to obtain data on growth and substrate (glucose, xylose, galactose or glycerol) utilisation in a high throughput manner. All eight fungi grew in both cultivation systems, but growth was more variable and with more noise in the data in the Cytomat plate hotel than in the BioScreen. Specific growth rates between 0.01 (no added substrate) and 0.07 h-1 were measured for strains growing in the BioScreen and between 0.01 and 0.27 h-1 for strains in the plate hotel. Three strains, Dendryphiella salina LF304, Penicillium chrysogenum KF657 and Penicillium pinophilum LF458, consistently had higher specific growth rates on glucose and xylose in the plate hotel than in the BioScreen, but otherwise results were similar in the two systems. However, because of the noise in data from the plate hotel, the data obtained from it could only be used to distinguish between substrates which did or did not support growth, whereas data from BioScreen also provided information on substrate preference. Glucose was the preferred substrate for all strains, followed by xylose and galactose. Five strains also grew on glycerol. Therefore it was important to minimise the amount of glycerol introduced with the inoculum to avoid misinterpreting the results for growth on poor substrates. We concluded that both systems could provide physiological data with filamentous fungi, provided sufficient replicates are included in the measurements.
Topics: Ascomycota; Culture Media; Glucose; Glycerol; Penicillium; Seawater; Xylose
PubMed: 32764772
DOI: 10.1371/journal.pone.0236822