2018 |
Cassels, B K; Saez-Briones, P Dark Classics in Chemical Neuroscience: Mescaline Artículo de revista Acs Chemical Neuroscience, 9 (10), pp. 2448-2458, 2018, ISSN: 1948-7193. Resumen | Enlaces | BibTeX | Etiquetas: analogs, biosynthesis, cancer, derivatives, diethylamide, hallucinogen, hallucinogenic life-threatening lysergic-acid mescaline, metabolism, pedro, peyote, pharmacology, phenethylamine, phenyl-aethylamine, properties, relationships, san serotonin stimulus structure-activity synthesis, wachuma @article{RN389, title = {Dark Classics in Chemical Neuroscience: Mescaline}, author = { B.K. Cassels and P. Saez-Briones}, url = {/brokenurl#<Go to ISI>://WOS:000447954300015}, doi = {10.1021/acschemneuro.8b00215}, issn = {1948-7193}, year = {2018}, date = {2018-01-01}, journal = {Acs Chemical Neuroscience}, volume = {9}, number = {10}, pages = {2448-2458}, abstract = {Archeological studies in the United States, Mexico, and Peru suggest that mescaline, as a cactus constituent, has been used for more than 6000 years. Although it is a widespread cactus alkaloid, it is present in high concentrations in few species, notably the North American peyote (Lophophora williamsii) and the South American wachuma (Trichocereus pachanoi, T. peruvianus, and T. bridgesii). Spanish 16th century chroniclers considered these cacti "diabolic", leading to their prohibition, but their use persisted to our days and has been spreading for the last 150 years. In the late 1800s, peyote attracted scientific attention; mescaline was isolated, and its role in the psychedelic effects of peyote tops or "mescal buttons" was demonstrated. Its structure was established by synthesis in 1929, and alternative routes were developed, providing larger amounts for pharmacological and biosynthetic research. Although its effects are attributed mainly to its action as a 5-HT2A serotonin receptor agonist, mescaline binds in a similar concentration range to 5-HT1A and alpha(2A) receptors. It is largely excreted unchanged in human urine, and its metabolic products are apparently unrelated to its psychedelic properties. Its low potency is probably responsible for its relative neglect by recreational substance users, as the successful search for structure-activity relationships in the hallucinogen field focused largely on finding more potent analogues. Renewed interest in the possible therapeutic applications of psychedelic drugs may hopefully lead to novel insights regarding the commonalities and differences between the actions of individual classic hallucinogens.}, keywords = {analogs, biosynthesis, cancer, derivatives, diethylamide, hallucinogen, hallucinogenic life-threatening lysergic-acid mescaline, metabolism, pedro, peyote, pharmacology, phenethylamine, phenyl-aethylamine, properties, relationships, san serotonin stimulus structure-activity synthesis, wachuma}, pubstate = {published}, tppubtype = {article} } Archeological studies in the United States, Mexico, and Peru suggest that mescaline, as a cactus constituent, has been used for more than 6000 years. Although it is a widespread cactus alkaloid, it is present in high concentrations in few species, notably the North American peyote (Lophophora williamsii) and the South American wachuma (Trichocereus pachanoi, T. peruvianus, and T. bridgesii). Spanish 16th century chroniclers considered these cacti "diabolic", leading to their prohibition, but their use persisted to our days and has been spreading for the last 150 years. In the late 1800s, peyote attracted scientific attention; mescaline was isolated, and its role in the psychedelic effects of peyote tops or "mescal buttons" was demonstrated. Its structure was established by synthesis in 1929, and alternative routes were developed, providing larger amounts for pharmacological and biosynthetic research. Although its effects are attributed mainly to its action as a 5-HT2A serotonin receptor agonist, mescaline binds in a similar concentration range to 5-HT1A and alpha(2A) receptors. It is largely excreted unchanged in human urine, and its metabolic products are apparently unrelated to its psychedelic properties. Its low potency is probably responsible for its relative neglect by recreational substance users, as the successful search for structure-activity relationships in the hallucinogen field focused largely on finding more potent analogues. Renewed interest in the possible therapeutic applications of psychedelic drugs may hopefully lead to novel insights regarding the commonalities and differences between the actions of individual classic hallucinogens. |
Rojas-Aedo, J F; Gil-Duran, C; Goity, A; Vaca, I; Levican, G; Larrondo, L F; Chavez, R The Developmental Regulator Pcz1 Affects the Production of Secondary Metabolites in the Filamentous Fungus Penicillium Roqueforti Artículo de revista Microbiological Research, 212 , pp. 67-74, 2018, ISSN: 0944-5013. Resumen | Enlaces | BibTeX | Etiquetas: biosynthesis, chrysogenum complex, cross-talk, discovery, expression, gene-cluster, laea, metabolism, mycophenolic-acid, pathway pcz1, penicillium protein, roqueforti, secondary subunit @article{RN382, title = {The Developmental Regulator Pcz1 Affects the Production of Secondary Metabolites in the Filamentous Fungus Penicillium Roqueforti}, author = { J.F. Rojas-Aedo and C. Gil-Duran and A. Goity and I. Vaca and G. Levican and L.F. Larrondo and R. Chavez}, url = {/brokenurl#<Go to ISI>://WOS:000438320100007}, doi = {10.1016/j.micres.2018.05.005}, issn = {0944-5013}, year = {2018}, date = {2018-01-01}, journal = {Microbiological Research}, volume = {212}, pages = {67-74}, abstract = {Penicillium roqueforti is used in the production of several kinds of ripened blue-veined cheeses. In addition, this fungus produces interesting secondary metabolites such as roquefortine C, andrastin A and mycophenolic acid. To date, there is scarce information concerning the regulation of the production of these secondary metabolites. Recently, the gene named pcz1 (Penicillium C6 zinc domain protein 1) was described in P. roqueforti, which encodes for a Zn(II)(2)Cys(6) protein that controls growth and developmental processes in this fungus. However, its effect on secondary metabolism is currently unknown. In this work, we have analyzed how the overexpression and down-regulation of pcz1 affect the production of roquefortine C, andrastin A and mycophenolic acid in P. roqueforti. The three metabolites were drastically reduced in the pcz1 down-regulated strains. However, when pcz1 was overexpressed, only mycophenolic acid was overproduced while, on the contrary, levels of roquefortine C and andrastin A were diminished. Importantly, these results match the expression pattern of keywords genes involved in the biosynthesis of these metabolites. Taken together, our results suggest that Pcz1 plays a keywords role in regulating secondary metabolism in the fungus Penicillium roqueforti.}, keywords = {biosynthesis, chrysogenum complex, cross-talk, discovery, expression, gene-cluster, laea, metabolism, mycophenolic-acid, pathway pcz1, penicillium protein, roqueforti, secondary subunit}, pubstate = {published}, tppubtype = {article} } Penicillium roqueforti is used in the production of several kinds of ripened blue-veined cheeses. In addition, this fungus produces interesting secondary metabolites such as roquefortine C, andrastin A and mycophenolic acid. To date, there is scarce information concerning the regulation of the production of these secondary metabolites. Recently, the gene named pcz1 (Penicillium C6 zinc domain protein 1) was described in P. roqueforti, which encodes for a Zn(II)(2)Cys(6) protein that controls growth and developmental processes in this fungus. However, its effect on secondary metabolism is currently unknown. In this work, we have analyzed how the overexpression and down-regulation of pcz1 affect the production of roquefortine C, andrastin A and mycophenolic acid in P. roqueforti. The three metabolites were drastically reduced in the pcz1 down-regulated strains. However, when pcz1 was overexpressed, only mycophenolic acid was overproduced while, on the contrary, levels of roquefortine C and andrastin A were diminished. Importantly, these results match the expression pattern of keywords genes involved in the biosynthesis of these metabolites. Taken together, our results suggest that Pcz1 plays a keywords role in regulating secondary metabolism in the fungus Penicillium roqueforti. |
2017 |
Torrent, C; Gil-Duran, C; Rojas-Aedo, J F; Medina, E; Vaca, I; Castro, P; Garcia-Rico, R O; Cotoras, M; Mendoza, L; Levican, G; Chavez, R Role of Sfk1 Gene in the Filamentous Fungus Penicillium Roqueforti Artículo de revista Frontiers in Microbiology, 8 , 2017, ISSN: 1664-302x. Resumen | Enlaces | BibTeX | Etiquetas: alpha-subunit, biosynthesis, botrytis-cinerea, changes, chrysogenum, expression, four gene germination, growth kinase, metabolites, of pathways, penicillium phenotypic protein rna-mediated roqueforti, saccharomyces-cerevisiae, secondary signaling silencing, stress, suppressor @article{RN338, title = {Role of Sfk1 Gene in the Filamentous Fungus Penicillium Roqueforti}, author = { C. Torrent and C. Gil-Duran and J.F. Rojas-Aedo and E. Medina and I. Vaca and P. Castro and R.O. Garcia-Rico and M. Cotoras and L. Mendoza and G. Levican and R. Chavez}, url = {/brokenurl#<Go to ISI>://WOS:000417142700001}, doi = {10.3389/fmicb.2017.02424}, issn = {1664-302x}, year = {2017}, date = {2017-01-01}, journal = {Frontiers in Microbiology}, volume = {8}, abstract = {The sfk1 (suppressor of four kinase) gene has been mainly studied in Saccharomyces cerevisiae, where it was shown to be involved in growth and thermal stress resistance. This gene is widely conserved within the phylum Ascomycota. Despite this, to date sfk1 has not been studied in any filamentous fungus. Previously, we found that the orthologous of sfk1 was differentially expressed in a strain of Penicillium roqueforti with an altered phenotype. In this work, we have performed a functional characterization of this gene by using RNAi-silencing technology. The silencing of sfk1 in P. roqueforti resulted in decreased apical growth and the promotion of conidial germination, but interesting, it had no effect on conidiation. In addition, the attenuation of the sfk1 expression sensitized the fungus to osmotic stress, but not to thermal stress. RNA-mediated gene-silencing of sfk1 also affected cell wall integrity in the fungus. Finally, the silencing of sfk1 depleted the production of the main secondary metabolites of P. roqueforti, namely roquefortine C, andrastin A, and mycophenolic acid. To the best of our knowledge this is the first study of the sfk1 gene in filamentous fungi.}, keywords = {alpha-subunit, biosynthesis, botrytis-cinerea, changes, chrysogenum, expression, four gene germination, growth kinase, metabolites, of pathways, penicillium phenotypic protein rna-mediated roqueforti, saccharomyces-cerevisiae, secondary signaling silencing, stress, suppressor}, pubstate = {published}, tppubtype = {article} } The sfk1 (suppressor of four kinase) gene has been mainly studied in Saccharomyces cerevisiae, where it was shown to be involved in growth and thermal stress resistance. This gene is widely conserved within the phylum Ascomycota. Despite this, to date sfk1 has not been studied in any filamentous fungus. Previously, we found that the orthologous of sfk1 was differentially expressed in a strain of Penicillium roqueforti with an altered phenotype. In this work, we have performed a functional characterization of this gene by using RNAi-silencing technology. The silencing of sfk1 in P. roqueforti resulted in decreased apical growth and the promotion of conidial germination, but interesting, it had no effect on conidiation. In addition, the attenuation of the sfk1 expression sensitized the fungus to osmotic stress, but not to thermal stress. RNA-mediated gene-silencing of sfk1 also affected cell wall integrity in the fungus. Finally, the silencing of sfk1 depleted the production of the main secondary metabolites of P. roqueforti, namely roquefortine C, andrastin A, and mycophenolic acid. To the best of our knowledge this is the first study of the sfk1 gene in filamentous fungi. |
2016 |
Del-Cid, A; Gil-Duran, C; Vaca, I; Rojas-Aedo, J F; Garcia-Rico, R O; Levican, G; Chavez, R Identification and Functional Analysis of the Mycophenolic Acid Gene Cluster of Penicillium Roqueforti Artículo de revista Plos One, 11 (1), 2016, ISSN: 1932-6203. Resumen | Enlaces | BibTeX | Etiquetas: biosynthesis, cheese, chrysogenum, dehydrogenase, expression, fungi, immunosuppressant, imp liquid-chromatography, mycotoxins, strains @article{RN290, title = {Identification and Functional Analysis of the Mycophenolic Acid Gene Cluster of Penicillium Roqueforti}, author = { A. Del-Cid and C. Gil-Duran and I. Vaca and J.F. Rojas-Aedo and R.O. Garcia-Rico and G. Levican and R. Chavez}, url = {/brokenurl#<Go to ISI>://WOS:000367888100190}, doi = {10.1371/journal.pone.0147047}, issn = {1932-6203}, year = {2016}, date = {2016-01-01}, journal = {Plos One}, volume = {11}, number = {1}, abstract = {The filamentous fungus Penicillium roqueforti is widely known as the ripening agent of blue-veined cheeses. Additionally, this fungus is able to produce several secondary metabolites, including the meroterpenoid compound mycophenolic acid (MPA). Cheeses ripened with P. roqueforti are usually contaminated with MPA. On the other hand, MPA is a commercially valuable immunosuppressant. However, to date the molecular basis of the production of MPA by P. roqueforti is still unknown. Using a bioinformatic approach, we have identified a genomic region of approximately 24.4 kbp containing a seven-gene cluster that may be involved in the MPA biosynthesis in P. roqueforti. Gene silencing of each of these seven genes (named mpaA, mpaB, mpaC, mpaDE, mpaF, mpaG and mpaH) resulted in dramatic reductions in MPA production, confirming that all of these genes are involved in the biosynthesis of the compound. Interestingly, the mpaF gene, originally described in P. brevicompactum as a MPA self-resistance gene, also exerts the same function in P. roqueforti, suggesting that this gene has a dual function in MPA metabolism. The knowledge of the biosynthetic pathway of MPA in P. roqueforti will be important for the future control of MPA contamination in cheeses and the improvement of MPA production for commercial purposes.}, keywords = {biosynthesis, cheese, chrysogenum, dehydrogenase, expression, fungi, immunosuppressant, imp liquid-chromatography, mycotoxins, strains}, pubstate = {published}, tppubtype = {article} } The filamentous fungus Penicillium roqueforti is widely known as the ripening agent of blue-veined cheeses. Additionally, this fungus is able to produce several secondary metabolites, including the meroterpenoid compound mycophenolic acid (MPA). Cheeses ripened with P. roqueforti are usually contaminated with MPA. On the other hand, MPA is a commercially valuable immunosuppressant. However, to date the molecular basis of the production of MPA by P. roqueforti is still unknown. Using a bioinformatic approach, we have identified a genomic region of approximately 24.4 kbp containing a seven-gene cluster that may be involved in the MPA biosynthesis in P. roqueforti. Gene silencing of each of these seven genes (named mpaA, mpaB, mpaC, mpaDE, mpaF, mpaG and mpaH) resulted in dramatic reductions in MPA production, confirming that all of these genes are involved in the biosynthesis of the compound. Interestingly, the mpaF gene, originally described in P. brevicompactum as a MPA self-resistance gene, also exerts the same function in P. roqueforti, suggesting that this gene has a dual function in MPA metabolism. The knowledge of the biosynthetic pathway of MPA in P. roqueforti will be important for the future control of MPA contamination in cheeses and the improvement of MPA production for commercial purposes. |
2012 |
Wu, X B; Garcia-Estrada, C; Vaca, I; Martin, J F Motifs in the C-Terminal Region of the Penicillium Chrysogenum Acv Synthetase Are Essential for Valine Epimerization and Processivity of Tripeptide Formation Artículo de revista Biochimie, 94 (2), pp. 354-364, 2012, ISSN: 0300-9084. Resumen | Enlaces | BibTeX | Etiquetas: biosynthesis, biosynthetic cephalosporin cluster, enzyme, epimerase, functional genes, isopenicillin-n, l-valine non-ribosomal nonribosomal penicillin penicillin, peptide peptide-synthesis, repeats, syntethase, synthetase, synthetases, tandem @article{RN30d, title = {Motifs in the C-Terminal Region of the Penicillium Chrysogenum Acv Synthetase Are Essential for Valine Epimerization and Processivity of Tripeptide Formation}, author = { X.B. Wu and C. Garcia-Estrada and I. Vaca and J.F. Martin}, url = {/brokenurl#<Go to ISI>://WOS:000300270900010}, doi = {10.1016/j.biochi.2011.08.002}, issn = {0300-9084}, year = {2012}, date = {2012-01-01}, journal = {Biochimie}, volume = {94}, number = {2}, pages = {354-364}, publisher = {2011 Elsevier Masson SAS.}, abstract = {The first step in the penicillin biosynthetic pathway is the non-ribosomal condensation of L-alpha-aminoadipic acid, L-cysteine and L-valine into the tripeptide delta-(L-alpha-aminoadipyl)-L-cysteinyl-D-valine (ACV). This reaction is catalysed by the multienzyme ACV synthetase (ACVS), which is encoded in the filamentous fungus Penicillium chrysogenum by the pcbAB gene. This enzyme contains at least ten catalytic domains. The precise role of the C-terminal domain of this multidomain NRPS still remains obscure. The C-terminal region of ACVS bears the epimerase and the thioesterase domains and may be involved in the epimerization of LLL-ACV to LLD-ACV and in the hydrolysis of the thioester bond. In this work, the conserved motifs (3371)EGHGRE(3376) (located in the putative epimerase domain) and (3629)GWSFG(3633) (located in the thioesterase domain) were changed by site-directed-mutagenesis to LGFGLL and GWAFG, respectively. In addition, the whole thioesterase domain (230 amino acids) and the different parts of this domain were deleted. The activity of these mutant enzymes was assessed in vivo by two different procedures: i) through the quantification of bisACV produced by the fungus and ii) by quantifying the benzylpenicillin production using tailored strains of P. chrysogenum, which lack the pcbAB gene, as host strains. All indicated mutant enzymes showed lower or null activity than the control strain confirming that E3371, H3373, R3375 and E3376 belong to the epimerase active centre. Different fragments included in the C-terminal region of ACVS control thioester hydrolysis. Overexpression of the sequence encoding the ACVS integrated thioesterase domain as a separate (stand-alone) transcriptional unit complemented mutants lacking the integrated thioesterase domain, although with low ACV releasing activity, suggesting that the stand-alone thio-esterease interacts with the other ACVS domains.}, keywords = {biosynthesis, biosynthetic cephalosporin cluster, enzyme, epimerase, functional genes, isopenicillin-n, l-valine non-ribosomal nonribosomal penicillin penicillin, peptide peptide-synthesis, repeats, syntethase, synthetase, synthetases, tandem}, pubstate = {published}, tppubtype = {article} } The first step in the penicillin biosynthetic pathway is the non-ribosomal condensation of L-alpha-aminoadipic acid, L-cysteine and L-valine into the tripeptide delta-(L-alpha-aminoadipyl)-L-cysteinyl-D-valine (ACV). This reaction is catalysed by the multienzyme ACV synthetase (ACVS), which is encoded in the filamentous fungus Penicillium chrysogenum by the pcbAB gene. This enzyme contains at least ten catalytic domains. The precise role of the C-terminal domain of this multidomain NRPS still remains obscure. The C-terminal region of ACVS bears the epimerase and the thioesterase domains and may be involved in the epimerization of LLL-ACV to LLD-ACV and in the hydrolysis of the thioester bond. In this work, the conserved motifs (3371)EGHGRE(3376) (located in the putative epimerase domain) and (3629)GWSFG(3633) (located in the thioesterase domain) were changed by site-directed-mutagenesis to LGFGLL and GWAFG, respectively. In addition, the whole thioesterase domain (230 amino acids) and the different parts of this domain were deleted. The activity of these mutant enzymes was assessed in vivo by two different procedures: i) through the quantification of bisACV produced by the fungus and ii) by quantifying the benzylpenicillin production using tailored strains of P. chrysogenum, which lack the pcbAB gene, as host strains. All indicated mutant enzymes showed lower or null activity than the control strain confirming that E3371, H3373, R3375 and E3376 belong to the epimerase active centre. Different fragments included in the C-terminal region of ACVS control thioester hydrolysis. Overexpression of the sequence encoding the ACVS integrated thioesterase domain as a separate (stand-alone) transcriptional unit complemented mutants lacking the integrated thioesterase domain, although with low ACV releasing activity, suggesting that the stand-alone thio-esterease interacts with the other ACVS domains. |
2011 |
Vaca, I; Casqueiro, J; Ullan, R V; Rumbero, A; Chavez, R; Martin, J F Journal of Antibiotics, 64 (6), pp. 447-451, 2011, ISSN: 0021-8820. Resumen | Enlaces | BibTeX | Etiquetas: biosynthesis, cephalosporin-c, chrysogenum, conversion, hplc, isopenicillin liquid-chromatography n, natural-products, penicillium-chrysogenum, purification, stability, storage, system temperature @article{RN36h, title = {A Preparative Method for the Purification of Isopenicillin N from Genetically Blocked Acremonium Chrysogenum Strain Td189: Studies on the Degradation Kinetics and Storage Conditions}, author = { I. Vaca and J. Casqueiro and R.V. Ullan and A. Rumbero and R. Chavez and J.F. Martin}, url = {/brokenurl#<Go to ISI>://WOS:000292090800006}, doi = {10.1038/ja.2011.30}, issn = {0021-8820}, year = {2011}, date = {2011-01-01}, journal = {Journal of Antibiotics}, volume = {64}, number = {6}, pages = {447-451}, abstract = {A protocol for preparative isopenicillin N (IPN) purification, a highly interesting and hitherto unavailable intermediate of the penicillin and cephalosporin biosynthetic pathway due to its high unstability, is described. Culture broths of Acremonium chrysogenum TD189, a strain blocked in cephalosporin biosynthesis that accumulates this metabolite, were treated with acetone and filtered though charcoal and a hydrophobic resin in a single step as tandem columns. The cleared broth was then lyophilized and passed though a Sephadex G-25 column. The last step was the purification to homogeneity of IPN in a semipreparative HPLC equipment and, optionally, a desalting step by Sephadex G-10 column. Once purified, a complete analysis of the stability of the compound and the conditions for its long-term storage was carried out. Our results suggest a first-order model for IPN decomposition for all the pH and temperature analyzed. IPN is more stable at neutral pH, and once lyophilized, can be stored under vacuum and -75 degrees C with a half-life of 770 days. The Journal of Antibiotics (2011); published online 27 April 2011}, keywords = {biosynthesis, cephalosporin-c, chrysogenum, conversion, hplc, isopenicillin liquid-chromatography n, natural-products, penicillium-chrysogenum, purification, stability, storage, system temperature}, pubstate = {published}, tppubtype = {article} } A protocol for preparative isopenicillin N (IPN) purification, a highly interesting and hitherto unavailable intermediate of the penicillin and cephalosporin biosynthetic pathway due to its high unstability, is described. Culture broths of Acremonium chrysogenum TD189, a strain blocked in cephalosporin biosynthesis that accumulates this metabolite, were treated with acetone and filtered though charcoal and a hydrophobic resin in a single step as tandem columns. The cleared broth was then lyophilized and passed though a Sephadex G-25 column. The last step was the purification to homogeneity of IPN in a semipreparative HPLC equipment and, optionally, a desalting step by Sephadex G-10 column. Once purified, a complete analysis of the stability of the compound and the conditions for its long-term storage was carried out. Our results suggest a first-order model for IPN decomposition for all the pH and temperature analyzed. IPN is more stable at neutral pH, and once lyophilized, can be stored under vacuum and -75 degrees C with a half-life of 770 days. The Journal of Antibiotics (2011); published online 27 April 2011 |
2018 |
Dark Classics in Chemical Neuroscience: Mescaline Artículo de revista Acs Chemical Neuroscience, 9 (10), pp. 2448-2458, 2018, ISSN: 1948-7193. |
The Developmental Regulator Pcz1 Affects the Production of Secondary Metabolites in the Filamentous Fungus Penicillium Roqueforti Artículo de revista Microbiological Research, 212 , pp. 67-74, 2018, ISSN: 0944-5013. |
2017 |
Role of Sfk1 Gene in the Filamentous Fungus Penicillium Roqueforti Artículo de revista Frontiers in Microbiology, 8 , 2017, ISSN: 1664-302x. |
2016 |
Identification and Functional Analysis of the Mycophenolic Acid Gene Cluster of Penicillium Roqueforti Artículo de revista Plos One, 11 (1), 2016, ISSN: 1932-6203. |
2012 |
Motifs in the C-Terminal Region of the Penicillium Chrysogenum Acv Synthetase Are Essential for Valine Epimerization and Processivity of Tripeptide Formation Artículo de revista Biochimie, 94 (2), pp. 354-364, 2012, ISSN: 0300-9084. |
2011 |
Journal of Antibiotics, 64 (6), pp. 447-451, 2011, ISSN: 0021-8820. |