Penicillium aurantiogriseum Dierckx 1901: Producer of Diketopiperazine Alkaloids (Roquefortine and 3,12-Dihydroroquefortine), Isolated from Permafrost

Secondary metabolites of three strains of Penicillium aurantiogriseumisolated from permafrost sediments were identified. It was found that these fungi synthesized the diketopiperazine alkaloids roquefortine and 3,12-dihydroroquefortine. The strain VKM FW-766 synthesized alkaloids in the course of certain growth-related processes. When the strain was grown on a mineral medium, the time courses of the roquefortine and 3,12-dihydroroquefortine concentrations were characterized by biphasic curves.     

The biosynthesis of low-molecular-weight nitrogen-containing secondary metabolite-alkaloids by the resident strains of Penicillium chrysogenum and Penicillium expansum isolated on the board of the Mir space station

The analysis of the absorption spectra of the low-molecular-weight nitrogen-containing secondary metabolites--alkaloids--of 4 Penicillium chrysogenum strains and 6 Penicillium expansum strains isolated on board the Mir space station showed that all these strains synthesize metabolites of alkaloid origin (roquefortine, 3,12-dihydroroquefortine, meleagrin, viridicatin, viridicatol, isorugulosuvin, rugulosuvin B, N-acetyl-tryptamine, and a "yellow metabolite" containing the benzoquinone chromophore).     

Isolation of Penicillium DIERCKX 1901--a producer of diketopiperazine alkaloids rokefortin and 3,12-dihydrokefortin--from permafrost

Secondary metabolites of the three strains of Penicillium aurantiogriseum, isolated from permafrost sediments, were identified. It was found that these fungi synthesized diketopiperazine alkaloids roquefortine and 3,12-dihydroroquefortine. The strain VKM FW-766 synthesized alkaloids in in the course of certain growth-related processes. When the strain was grown on a mineral medium, time courses of roquefortine and 3,12-dihydroroquefortine concentrations were characterized by biphasic curves.     

Study of conditions of production of roquefortine and other metabolites of Penicillin roqueforti.

Experiments to determine optimum yields of roquefortine, isofumigaclavine A, and PR toxin, metabolites from Penicillum roqueforti Thom, were performed. Four strains, isolated from blue cheese, and five liquid media were evaluated, although not all permutations were studied. Sucrose (15%)-yeast extract (2%) was the medium chosen for time-course studies at 25 and 15 degrees C using one favorable strain. At 25 degrees C, maximum estimated yields of roquefortine were about 100 mg/liter in the mycelium by 16 days, and no subsequent degradation of this alkaloid was observed. On the other hand, production of PR toxin in the medium peaked at 770 mg/liter at 21 days. At 15 degrees C, yields of roquefortine and PR toxin after 49 days were 60 to 70% of the maximum yields obtained at 25 degrees C. However, about three times more isofumigaclavine A (up to 11 mg/liter) was formed in the mycelium at 15 degrees C than at 25 degrees C. All four strains of P. roqueforti procedure both roquefortine and PR toxin on the sucrose-yeast extract medium at 25 degrees C; isofumigaclavine A was detected in all but one strain grown on this medium.     

Formation of alkaloids from Penicillium species fungi during growth on wheat kernels

The ability to produce alkaloids has been studied in 13 strains belonging to 10 species of the genus Penicillium. Most of these strains produce identical ranges of alkaloids when grown on wheat grain and synthetic Abe medium. They are roquefortine, 3,12-dihydroroquefortine, and glandicolines A and B in strain P. chrysogenum VKM F-1987; fumigaclavines A and B, festuclavine, and pyroclasine in P. commune VKM F-308, F-3491, and KBP4; agroclavine 1 and epoxyagroclavine 1 in P. fellutanum VKM F-1073; fellutanine A in P. fellutanum F-3020; roquefortine, 3,12-dihydroroquefortine, meleagrin, and glandicolines A and B in P. glandicola VKM F-743; aurantioclavine in P. nalgiovense VKM F-229; isofumigaclavines A and B, festuclavine, roquefortine, and 3,12-dihydroroquefortine in P. roquefortii VKM F-2389; roquefortine, 3,12-dihydroroquefortine, and meleagrin in P. vitale VKM F-3624; roquefortine and oxaline in P. vulpinum VKM F-256; and alpha-cyclopiazonic acid and rugulovasine B in P. viridicatum C-47. No alkaloids were found in P. rugulosum VKM F-352 grown on wheat grain. A simple method is proposed for isolating alkaloids from affected grains.     

Production of Alkaloids by Fungi of the Genus PenicilliumGrown on Wheat Grain

The ability to produce alkaloids has been studied in 13 strains belonging to ten species of the genus Penicillium. Most of these strains produce identical ranges of alkaloids when grown on wheat grain and synthetic Abe's medium. These are roquefortine, 3,12-dihydroroquefortine, and glandicolines A and B in strain P. chrysogenum VKM F-1987; fumigaclavines A and B, festuclavine, and pyroclavine in P. commune VKM F-308, F-3491, and KBP4; agroclavine 1 and epoxyagroclavine 1 in P. fellutanum VKM F-1073; fellutanine A in P. fellutanum F-3020; roquefortine, 3,12-dihydroroquefortine, meleagrin, and glandicolines A and B in P. glandicola VKM F-743; aurantioclavine in P. nalgiovense VKM F-229; isofumigaclavines A and B, festuclavine, roquefortine, and 3,12-dihydroroquefortine in P. roquefortii VKM F-2389; roquefortine, 3,12-dihydroroquefortine, and meleagrin in P. vitale VKM F-3624; roquefortine and oxaline in P. vulpinum VKM F-256; and -cyclopiazonic acid and rugulovasine B in P. viridicatum C-47. No alkaloids were found in P. rugulosum VKM F-352 grown on wheat grain. A simple method is proposed for isolating alkaloids from affected grain.     

Minor Alkaloids of the Fungus Penicillium roquefortiiThom 1906

New isomers of clavine alkaloids with distinctively low chromatographic mobilities were isolated from the collection and mutant strains of Penicillium roquefortiiThom 1906, in addition to alkaloids roquefortine, 3,12-dihydroroquefortine, isofumigaclavines A and B, festuclavine, and chanoclavine-I, which are characteristic of this species. It was demonstrated that the collection strain produces isomers of agroclavine and epoxyagroclavine, whereas the mutant strain synthesizes isomers of fumigaclavines A and B, festuclavine, and chanoclavine.     

Mycotoxins from the fungi Penicillium vulpinum (Cooke & Massee) Seifert & Samson

Mycotoxins produced by seven strains of Penicillium vulpinum (formerly Penicillium claviforme) isolated from different sources were studied. The strains were characterized by specific profiles of secondary metabolites and produced mycotoxins of different structural types. In addition to toxins already known for this fungal species (patulin, roquefortine, 3,12-dihydroroquefortine, oxalin, viridicatin, cyclopenin, and alpha-cyclopiazonic acid), the strains studied also produced indolyl-3-acetic acid, griseofulvin, meleagrin, and cyclopeptin.     

Mycotoxin production profiles ofPenicillium vulpinum (Cooke & Massee) Seifert & Samson strains

Mycotoxins produced by seven strains ofPenicillium vulpinum (formerlyPenicillium claviforme) isolated from different sources were studied. The strains were characterized by specific profiles of secondary metabolites and produced mycotoxins of different structural types. In addition to toxins already known for this fungal species (patulin, roquefortine, 3,12-dihydroroquefortine, oxalin, viridicatin, cyclopenin, and α-cyclopiazonic acid), the strains studied also produced indolyl-3-acetic acid, griseofulvin, meleagrin, and cyclopeptin.     

The Biosynthesis of Low-Molecular-Weight Nitrogen-Containing Secondary Metabolites—Alkaloids—by the Resident Strains of Penicillium chrysogenum and Penicillium expansum Isolated on Board the Mir Space Station

The analysis of the absorption spectra of the low-molecular-weight nitrogen-containing secondary metabolites—alkaloids—of four Penicillium chrysogenum strains and six P. expansum strains isolated on board the Mir space station showed that all these strains synthesize metabolites of alkaloid origin (roquefortine, 3,12-dihydroroquefortine, meleagrin, viridicatin, viridicatol, isorugulosuvin, rugulosuvin B, N acetyltryptamine, and a yellow metabolite containing the benzoquinone chromophore).     

Diketopiperazine alkaloids from the fungus Penicillium piscarium Westling

Fungi of the species Penicillium piscarium produced diketopiperazine alkaloids (isorugulosuvine, puberuline, verrucosine, prolyltryptophanyldiketopiperazine, 12,13-dehydroprolyltryptophanyldiketopiperazine, fellutanine A, phenylalanylphenylalanyldiketopiperazine, as well as roquefortine and 3,12-dihydroroquefortine whose precursors are tryptophan, phenylalanine, leucine, proline, and histidine.     

Minor alkaloids from the fungus Penicillium roquefortii Thom 1906

New spatial of clavine alkaloids, distinguished by low chromatographical mobility, have been isolated from the collection and mutant strain of Penicillium roquefortii, in addition to alkaloids roquefortine, 3,12-dihydroroquefortine, isofumigaclavines A and B, festuclavine, and chanoclavine-I, characteristic of this fungal species. In has been shown that the collection strain produces isomers of agroclavine and epoxyagroclavine, and the mutant strain produces isomers of fumigaclavines A and B, festuclavine, and chanoclavine.     

Diketopiperazine Alkaloids from the fungusPenicillium piscarium westling

Fungi of the speciesPenicillium piscarium produced diketopiperazine alkaloids (isorugulosuvine, puberuline, verrucosine, prolyltryptophanyldiketopiperazine, 12,13-dehydroprolyltryptophanyldiketopiperazine, fellutanine A, phenylalanylphenylalanyldiketopiperazine, as well as roquefortine and 3,12-dihydroroquefortine, whose precursors are tryptophan, phenylalanine, leucine, proline, and histidine.     

Taxonomic position and nitrogen-containing secondary metabolites of the fungus Penicillium vitale Pidoplichko Et Bilai Apud Bilai

The type strain Penicillium vitale Pidoplichko et Bilai apud Bilai 1961 VKM F-3624 was found to considerably differ from a sibling species P. janthinellium (syn. P. simplicissimum) in some physiological and morphological features (growth rates at different temperatures, the size of philiades, and the shape of conidia), as well as in the pattern of the nitrogen-containing secondary metabolites produced (roquefortine, 3,12-dihydroroquefortine, meleagrin, aurantioclavine, indole-3-acetic acid, and N-acetyltryptamine). The data obtained suggest that P. vitale represent an independent species.     

Taxonomic position and nitrogen-containing secondary metabolites of the fungusPenicillium vitale pidoplichko et bilai apud bilai

The type strainPenicillium vitale Pidoplichko et Bilai apud Bilai 1961 VKM F-3624 was found to considerably differ from a sibling speciesP. janthinellium (syn.P. simplicissimum) in some physiological and morphological features (growth rates at different temperatures, the size of philiades, and the shape of conidia), as well as in the pattern of the nitrogen-containing secondary metabolites produced (roquefortine, 3,12-dihydroroquefortine, meleagrin, aurantioclavine, indole-3-acetic acid, andN-acetyltryptamine). The data obtained suggest thatP. vitale represents an independent species.     

Silencing of a second dimethylallyltryptophan synthase of <Emphasis Type="Italic">Penicillium roqueforti</Emphasis> reveals a novel clavine alkaloid gene cluster

Penicillium roqueforti produces several prenylated indole alkaloids, including roquefortine C and clavine alkaloids. The first step in the biosynthesis of roquefortine C is the prenylation of tryptophan-derived dipeptides by a dimethylallyltryptophan synthase, specific for roquefortine biosynthesis (roquefortine prenyltransferase). A second dimethylallyltryptophan synthase, DmaW2, different from the roquefortine prenyltransferase, has been studied in this article. Silencing the gene encoding this second dimethylallyltryptophan synthase, dmaW2, proved that inactivation of this gene does not prevent the production of roquefortine C, but suppresses the formation of other indole alkaloids. Mass spectrometry studies have identified these compounds as isofumigaclavine A, the pathway final product and prenylated intermediates. The silencing does not affect the production of mycophenolic acid and andrastin A. A bioinformatic study of the genome of P. roqueforti revealed that DmaW2 (renamed IfgA) is a prenyltransferase involved in isofumigaclavine A biosynthesis encoded by a gene located in a six genes cluster (cluster A). A second three genes cluster (cluster B) encodes the so-called yellow enzyme and enzymes for the late steps for the conversion of festuclavine to isofumigaclavine A. The yellow enzyme contains a tyrosine-181 at its active center, as occurs in Neosartorya fumigata, but in contrast to the Clavicipitaceae fungi. A complete isofumigaclavines A and B biosynthetic pathway is proposed based on the finding of these studies on the biosynthesis of clavine alkaloids.     

Factors contributing to roquefortine yield variability during cultivation of penicillium roquefortii

Variability in the roquefortine yield was shown to be associated with its consumption by the mycelium during isolation of the end product, which depended on temperature, time of culture liquid storage, and biomass concentration. This was also related to the presence in chloroform of chlorocarbonic acid ethyl ester that reacted with roquefortine.     

Factors Contributing to Roquefortine Yield Variability during Cultivation of Penicillium roquefortii

Variability in the roquefortine yield was shown to be associated with its consumption by the mycelium during isolation of the end product, which depended on temperature, time of culture liquid storage, and biomass concentration. This was also related to the presence in chloroform of chlorocarbonic acid ethyl ester that reacted with roquefortine.     

Optimization of the medium and cultivation conditions of Penicillium roquefortii f39 producing the diketopiperazine alkaloid roquefortine

We optimized the medium for cultivation of Penicillium roquefortii f39, a producer of roquefortine. In this medium, the roquefortine yield increased 1.5-2-fold. The increase in roquefortine content was associated with high biomass yield, but not with an increase in biosynthetic activity of the mycelium. Direct correlation was found between extracellular roquefortine concentration and amount of the inoculum. The introduction of sucrose into the growth medium allowed us to increase the concentration of roquefortine during fermentation to 90 mg/l.     

Optimization of the Medium and Cultivation Conditions of Penicillium roquefortii f39 Producing the Diketopiperazine Alkaloid Roquefortine

We optimized the medium for cultivation of Penicillium roquefortii f39, a producer of roquefortine. In this medium, the roquefortine yield increased 1.5–2-fold. An increase in roquefortine content was associated with high biomass yield, but not with an increase in biosynthetic activity of the mycelium. Direct correlation was found between extracellular roquefortine concentration and the amount of the inoculum. The use of sucrose in the inoculum medium allowed us to increase the concentration of roquefortine during fermentation to 90 mg/l.