Monoacylglycerols derived from butter oil by Penicillium roquefortii in suspension cultures

Structural isomers of monoacylglycerols (monoglycerides, MAGs) were identified and compared after degradation of butter oil by two strains of Penicillium roquefortii and a commercial lipase from P roquefortii (EC 3.1.1.3) at pH 7.0 and 10 °C. The conditions were selected as they were comparable with those used in the manufacture of blue mould‐ripened cheese. The commercial lipase was selected to compare with the fungal strains in terms of acyl migration. Results showed that the main isomers formed by lipolysis with the commercial lipase were sn‐2 MAGs (64 mol%), whilst spores and emerging mycelia of P roquefortii produced mainly sn‐1(3) MAGs (83–90 mol%). The work reported here may lead to further assessment of different MAG structural isomers as natural preservatives in foods and dairy products. © 2002 Society of Chemical Industry     

Impact of the addition of an enzyme pool on an activated sludge system treating dairy wastewater under fat shock loads

BACKGROUND: The application of lipase‐rich enzyme pools (such as the crude solid enzymatic preparation (SEP) obtained from Penicillium restrictum solid‐state fermentation of agro‐industrial wastes) to activated sludge systems may be an effective strategy for preventing various operational problems. The continuous addition of SEP to the treatment system can become cost‐prohibitive when in situ production and/or storage are factored in. The application of SEP to high‐fat wastewater treatment would only be justified as an emergency measure, such as a sudden increase in the fat content of the bioreactor influent. Therefore, the primary objective of this work was to investigate the efficiency of a crude SEP during fat shock loads, simulated through the periodic addition of dairy industry waste containing high fat concentrations to the feed stock of an activated sludge system, operated in continuous mode. RESULTS: The test bioreactor exhibited a higher average chemical oxygen demand (COD) removal efficiency than the control bioreactor (83% for control and 90% for test) and the fat accumulation in the biological flocs of the test bioreactor was 3.2 times lower than that in the control bioreactor. Turbidity was also lower in the effluent of the test bioreactor (123 and 66 FTU in control and test, respectively) and it had a shorter recovery time between shock loads, especially when the interval between loads was shorter than one month (biweekly and weekly shock loads). CONCLUSION: The addition of SEP during fat overloads in the reactor feed maintained efficient COD removal in the test bioreactor for 270 days without any operational problems. Copyright © 2008 Society of Chemical Industry     

Ecophysiology of ochratoxigenic Aspergillus ochraceus and Penicillium verrucosum isolates. Predictive models for fungal spoilage prevention - a review

Ochratoxin A (OTA) is a secondary metabolite produced by several species of Aspergillus and Penicillium; among them Aspergillus ochraceus and Penicillium verrucosum are two ochratoxigenic species capable of growing in different climates and thus contamination of food crops with OTA can occur worldwide. OTA can be found in a wide range of foods such as cereals, coffee, cocoa, spices, beer, wine, dried vine fruit, grapes and meat products. OTA is toxic to animals, it presents neurotoxic, immunotoxic and nephrotoxic effects. It has been implicated in a human kidney disorder known as Balkan Endemic Nephropathy. This review focuses on the ecophysiology of ochratoxin-producing Aspergillus ochraceus and Penicillium verrucosum, the effect of environmental factors on their germination, mycelial growth, and OTA production. Knowledge of environmental conditions required for sucessive stages of fungal development represent the first step towards preventing mycotoxin formation. Predictive models for different stages of fungal development are presented, which allow prediction of the time before spoilage as a function of the abiotic factors. Finally, the implications of these studies in management of barley, coffee and grapes are described. This can help to identify the critical control points in their production, storage and distribution processes.     

Batch cultures of Penicillium camembertii on glucose and amino acids—model for growth and ammonium production

BACKGROUND: Three groups of amino acids were previously characterized based on their ability to be assimilated as carbon source by Penicillium camembertii. To describe the diauxic growth recorded on glucose and amino acids from the second group, such as arginine, an unstructured model was previously developed, based on the sequential consumption of both carbon substrates, glucose, followed after its exhaustion, by arginine. The model was modified to describe also the behaviour recorded during growth on other amino acids. RESULTS: The growth model involved the carbon substrate consumption (Verlhust model) and the biomass on carbon substrate yield. Glucose was therefore considered during P. camembertii growth on nitrogen source amino acids (lysine—first group); and amino acid consumption was considered during growth on carbon source amino acids (glutamate—third group), with glucose being dissimilated only for energy supply. The excess nitrogen from amino acids was released as ammonium; the linking of this production to growth was found to increase with the ability of the amino acid to be assimilated as carbon source by P. camembertii. CONCLUSION: The various metabolic behaviours recorded during P. camembertii growth on amino acids, in the presence of a primary carbon source such as glucose, were proved to be satisfactorily described by the model, showing the robustness of the model. Copyright © 2007 Society of Chemical Industry     

Critical temperature for production of MAG by esterification of different FA with glycerol using <Emphasis Type="Italic">Penicillium camembertii</Emphasis> lipase

Production of MAG by a lipase-catalyzed reaction is known to be effective at low temperature. This phenomenon can be explained by assuming that synthesized MAG are excluded from the reaction system because MAG, which have low m.p., are solidified at low temperatures. Consequently, MAG are efficiently accumulated and do not serve as the precursor of DAG. If this hypothesis is correct, the critical temperature for MAG production, defined as the highest temperature at which DAG synthesis is repressed, should depend on the m.p. of the MAG. Esterification of FFA with glycerol using Candida rugosa, Rhizopus oryzae, and Penicillium camembertii lipases produced MAG efficiently at low temperatures. However, Candida lipase showed very low esterification activity at high temperatures (>20°C), and Rhizopus lipase produced not only MAG but also DAG even at low temperatures. Meanwhile, P. camembertii lipase catalyzed synthesis of MAG only from FFA and glycerol at low temperatures, although the enzyme catalyzed synthesis of DAG from MAG in addition to synthesis of MAG at high temperatures. We thus studied the effect of temperature on esterification of C₁₀−C₁₈ FFA with glycerol using Penicillium lipase as a catalyst and determined the critical temperatures for production of MAG. The critical temperature for production of each MAG showed a linear correlation with m.p. of the MAG, which supported the hypothesis. In addition, because the m.p. of MAG are estimated from that of the constituent FA, the optimal temperature for production of MAG can be predicted from the m.p. of the FFA used as a substrate.     

Activity and stability ofPenicillium cyclopium lipase in surfactant and detergent solutions

Activity and stability of an alkaline lipase fromPenicillium cyclopium var.album (PG 37) were studied in surfactant and detergent solutions. Three anionic surfactants [Na salts of C₁₂SO₄ ^? (sodium dodecyl sulfate), C₁₂ØSO₃/^? (linear alkyl benzene sulfonate), and C₁₁COO^? (laurate)] and four homologous series of nonionic surfactants of C_12–15 polyoxethylenated fatty alcohols (AEO₃, AEO₅, AEO₇, and AEO₉) were evaluated. At a concentration range of 3.2–40 μM, sodium dodecyl sulfate and laurate stimulated the activity of PG 37 lipase. At concentrations greater than 5.6 μM, linear alkylbenzene sulfonate inhibited PG 37 lipase activity. Nonionic surfactants, AEO₅ and AEO₇, in the concentration range of 0.25–20 mM, enhanced and stabilized the activity of PG 37 lipase. The presence of PG 37 lipase in detergent formulaton improved detergency ～20%. The mechanism of inhibition of the lipolytic activity of PG 37 lipase is proposed to be partly due to the formation of inactive (BR)_n-E complex between the hydrophobic moiety of the surfactants and the surface of the lipase. Conversely, formation of a soluble (RB)_n-E complex between the hydrophilic group of the surfactant and lipase may account for the increased lipolytic activity of PG 37 lipase.     

Secretome analysis of novel IgE-binding proteins from Penicillium citrinum

The Penicillium genus of fungi is a frequently reported cause of allergic reactions. However, only a limited number of allergens have been reported. In Penicillium spp., many allergens show higher IgE-binding activity in culture filtrate extracts than in cellular extracts. In order to investigate the IgE-reactive profile of mold-sensitized patients, secreted IgE-reactive proteins from Penicillium citrinum were identified by 2-DE, serum immunoblotting, and nanoLC-MS/MS. Among the IgE-reactive spots, one known allergen, Pen c 13, and four novel allergens were identified. The cDNAs coding for Pen c 32 and Pen c 30 were cloned using designed primers based on nanoLC-MS/MS analysis. The amino acid sequences of Pen c 32 and Pen c 30 were, respectively, found to have extensive similarity with those of pectate lyases and catalases from various fungi. Native Pen c 30 was shown to have catalase activity and to bind to serum IgE from 48% of mold-allergic patients and induced immediate type skin reactions in a sensitized patient. Here, we present a proteome approach which resulted in the identification of four novel secreted allergens. These novel allergens might be useful in allergy diagnosis and in the treatment of mold-allergic disorders.     

Chemical response of Picea glehnii seed-epiphytic Penicillium species to Pythium vexans under in vitro competitive conditions for mycelial growth

The potential protection of Picea glehnii seedlings from damping-off by seed-epiphytic Penicillium species was investigated. We studied the chemical response of seed-epiphytic Penicillium species (Pen. cyaneum, Pen. damascenum, and Pen. implicatum) to Pythium vexans, a damping-off fungus, in vitro. Penicillium species were cultured singly or cocultured with Pyt. vexans for 14 or 18 d, and mycelial growth, pH of culture filtrate, antifungal activity of the culture filtrate against Pyt. vexans, and the amount of antifungal compound produced by each Penicillium species, were examined. The filtrate of both the single culture of Penicillium and the coculture of Penicillium and Pyt. vexans showed antifungal activity against Pyt. vexans. In a coculture with Pyt. vexans, Pen. cyaneum produced an antifungal compound (patulin) as in the single culture. Pen. damascenum cocultured with Pyt. vexans produced an antifungal compound (citrinin), as it did in the single culture and in larger amounts on day 10. Pen. implicatum produced two antifungal compounds, frequentin and palitantin, and the ratio of frequentin (with higher antifungal activity than palitantin) to palitantin was higher in the coculture with Pyt. vexans than in the single culture. Our results indicate that these Penicillium species have the ability to produce antifungal compounds and to keep antifungal activity under competitive condition with Pyt. vexans. The chemical response of these Penicillium species to Pyt. vexans may contribute to protect P. glehnii seedlings from damage by Pyt. vexans.     

Purification,characterisation and application of α‐l‐rhamnosidase from Penicillium citrinum MTCC‐8897

An α‐l ‐rhamnosidase secreted by Penicillium citrinum MTCC‐8897 has been purified to homogeneity from the culture filtrate of the fungal strain using ammonium sulphate precipitation and cation‐exchange chromatography on carboxymethyl cellulose. The sodium dodecyl sulphate/polyacrylamide gel electrophoresis analysis of the purified enzyme gave a single protein band corresponding to the molecular mass 51.0 kDa. The native polyacrylamide gel electrophoresis also gave a single protein band confirming the enzyme purity. The K_m and V_max values of the enzyme for p‐nitrophenyl α‐l ‐rhamnopyranoside were 0.36 mm and 22.54 μmole min^?1 mg^?1, respectively, and k_cat value was 17.1 s^?1 giving k_cat/K_m value of 4.75 × 10⁴ m ^?1 s^?1. The pH and temperature optima of the enzyme were 7.0 and 60 °C, respectively. The purified enzyme liberated l ‐rhamnose from naringin, rutin, hesperidin and wine, indicating that it has biotechnological application potential for the preparation of l ‐rhamnose and other pharmaceutically important compounds from natural glycosides containing terminal α‐l ‐rhamnose and also in the enhancement of wine aroma.