猴头菌和金针菇漆酶对不同染料的降解

利用漆酶（laccase）处理染料废水是近年来研究的热点。本研究以猴头菌Hericium erinaceus和金针菇Flammulina filiformis的发酵液为试验材料,通过硫酸铵沉淀、离子交换层析和超滤等方法,对发酵液中的漆酶进行了初步的分离纯化,然后分别研究了两种初提纯漆酶及其与小分子介体组成的漆酶介体系统（laccase-mediator system,LMS）对12种常用染料的降解效果。结果表明,猴头菌的初提纯漆酶对甲基红、铬黑T、孔雀绿和活性蓝R都具有很好的降解效果,反应24h后,降解率分别为79.6%、66.8%、80.3%和64.6%,且表现出时间依赖性,而对其他染料的降解效果不明显。当反应体系中加入介体2,2′-连氮基-双-(3-乙基苯并二氢噻唑啉-6-磺酸)二铵盐[2,2′-azinobis-(3-ethylbenzothiazoline-6-sulphate),ABTS]后,可增加猴头菌漆酶对考马斯亮蓝、结晶紫和孔雀绿的降解率,分别提高到33.7%、45.6%和94.2%。金针菇初提纯漆酶对孔雀绿、活性艳橙K-7R和活性红KD-8B都具有一定的降解效果,降解率分别为37.8%、39.9%和49%,也表现出了明显的时间依赖性。当反应体系中也加入ABTS后,金针菇漆酶对活性红KM-8B的降解率明显增加,由9.7%增加到58.2%。以上研究为猴头菌和金针菇漆酶在染料废水处理领域的应用提供了科学依据。     

丛枝菌根真菌与高羊茅协同修复镉污染土壤

本研究采用温室盆栽试验,利用丛枝菌根（AM）真菌摩西管柄囊霉Funneliformis mosseae进行接种试验,研究了在Cd胁迫下（0、5、15和30mg/kg）接种AM真菌对高羊茅Festuca elata ‘Crossfire II’的生物量、防御酶活性、磷和镉（Cd）含量的影响。结果表明,随着Cd浓度的增加,高羊茅的菌根侵染率和菌根相对依赖性有所增加。接种AM真菌改善了磷从植株根系向地上部的转运,有助于植株在地上部积累更多的磷。此外,AM真菌和Cd胁迫对高羊茅植株抗氧化酶活性都有显著影响,在镉胁迫下,与未接种植株相比,接种AM真菌显著提高了植株的过氧化氢酶活性,而显著降低了植株的丙二醛含量。与未接种植株相比,接种摩西管柄囊霉显著提高了寄主植物对Cd的富集能力,有利于重金属在根部的积累,同时降低了地上部的Cd含量。本研究表明,高羊茅-丛枝菌根共生体在Cd污染土壤的修复中具有潜在应用价值。     

Lichenochemical Screening and Antioxidant Capacity of Four Tunisian Lichen Species

The phenolic composition and antioxidant capacity of four Tunisian lichen species, Cladonia rangiformis, Flavoparmelia caperata, Squamarina cartilaginea and Xanthoria parietina, were determined in order to provide a better understanding of their lichenochemical composition. Powdered material of F. caperata was the richest in total phenolic content (956.68 μg GAE g⁻¹ DW) and S. cartilaginea in proanthocyanidin content (77.31 μg CE g⁻¹ DW), while the acetone extract of X. parietina showed the highest flavonoid content (9.56 μg CE g⁻¹ DW). The antioxidant capacity of all lichen extracts and crude material was evaluated by DPPH^. scavenging, iron-chelating, and iron-reducing powers. Results showed that methanol extracts of S. cartilaginea had the highest DPPH^. antioxidant capacity (IC₅₀=0.9 μg mL⁻¹) and the highest iron-reducing power was attributed to the acetone extract of this species. All extracts of all species were further screened by Fourier-transform infrared spectroscopy (FT-IR) and nuclear resonance spectroscopy (NMR); results showed an abundance of phenols, aromatic compounds, and fatty acids. Overall, our results showed that the investigated species are a rich source of potentially bioactive compounds with valuable properties.     

Fatty acid composition and lipid profiles as chemotaxonomic markers of phytopathogenic fungi Puccinia malvacearum and P. glechomatis

The analysis of the overall fatty acid pattern as well as their distribution in various lipid classes of phytopathogenic fungi Puccinia malvacearum and P. glechomatis are considered as chemotaxonomic biomarkers. Puccinia malvacearum on Alcea rosea and P. glechomatis on Glechoma hederacea collected from plants grown in various localities were analysed to determine their fatty acid composition. Both species synthesised significant amounts of saturated palmitic and stearic acids as well as 9,10-epoxy-octadecanoic acid, which rarely occurs in the nature. Both species synthesised hydroxy FAs including 9,10-dihydroxy octadecanoic acid and long-chain 2-hydroxy fatty acids.2-hydroxy 18:0 and 3-hydroxy 20:0 fatty acids were present only in P. malvacearum spores, and these may be the chemotaxonomic markers of the species. Ultra-high performance liquid chromatography mass spectrometry was performed for a comparative lipidomic analysis of P. malvacearum and P. glechomatis. The results revealed the complexity of molecular lipid species of these fungi. P. malvacearum and P. glechomatis lipids were characterised by the presence of a high number of triglyceride (TG) species. 9,10-epoxy octadecanoic fatty acid was found in TGs. Among the many types of oxidised TGs identified in P. glechomatis lipids, the most abundant species corresponds to TG(22:5+6O_17:0_18:2). P. malvacearum and P. glechomatis produced various ceramide species with different FAs from 14 to 24 chain-length. Unusual lipids like (O-acyl)-ω-hydroxy FA 18:0/18:0 in P. glechomatis and (O-acyl)-ω-hydroxy FA 18:0/20:0 and 18:0/22:0 in P. malvacearum were detected. The analysis of the polar lipid composition showed the presence of phosphatidylcholine and phosphatidylethanolamine as the main phospholipid classes of Puccinia spp. with the highest diversity of molecular species. Other phospholipids phosphatidic acid, phosphatidylglycerol phosphatidylserine and phosphatidylinositol were present in smaller amounts.The diversity of the neutral and polar lipid composition and fatty acid profile of P. malvacearum and P. glechomatis can be used in chemotaxonomic studies.     

Studies toward the comprehension of fungal-macroalgae interaction in cold marine regions from a biotechnological perspective

In marine ecosystems, macroalgae are the habitat for several microorganisms, fungi being among them. In the Antarctic benthic coastal ecosystem, macroalgae play a key role in organic matter cycling. In this study, 13 different macroalgae from Potter Cove and surrounding areas were sampled and 48 fungal isolates were obtained from six species, four Rhodophyta Ballia callitricha, Gigartina skottsbergii, Neuroglossum delesseriae and Palmaria decipiens, and two Phaeophyceae: Adenocystis utricularis and Ascoseira mirabilis. Fungal isolates mostly belonged to the Ascomycota phylum (Antarctomyces, Cadophora, Cladosporium, Penicillium, Phialocephala, and Pseudogymnoascus) and only one to the phylum Mucoromycota. Two of the isolates could not be identified to genus level, implying that Antarctica is a source of probable novel fungal taxa with enormous bioprospecting and biotechnological potential. 73% of the fungal isolates were moderate eurypsychrophilic (they grew at 5–25 °C), 12.5% were eurypsychrophilic and grew in the whole range, 12.5% of the isolates were narrow eurypsychrophilic (growth at 15–25 °C), and Mucoromycota AUe4 was classified as stenopsychrophilic as it grew at 5–15 °C. Organic extracts of seven macroalgae from which no fungal growth was obtained (three red algae Georgiella confluens, Gymnogongrus turquetii, Plocamium cartlagineum, and four brown algae Desmarestia anceps, D. Antarctica, Desmarestia menziesii, Himantothallus grandifolius) were tested against representative fungi of the genera isolated in this work. All extracts presented fungal inhibition, those from Plocamium cartilagineum and G. turquetii showed the best results, and for most of these macroalgae, this represents the first report of antifungal activity and constitute a promising source of compounds for future evaluation.     

Microbial transformation of curcumin by Rhizopus chinensis

Abstract

Curcumin (1) is a potent antioxidant and antitumor natural product. In spite of its efficacy and safety, its clinical use is hindered mainly by poor water solubility and bioavailability. Structural modification to introduce hydrophilic functions is a promising approach to resolve this problem. In the present study we first found that curcumin could be efficiently converted into glucosides by filamentous fungi including Rhizopus chinensis IFFI 03043, Absidia coerulea AS 3.3389 and Cunninghamella elegans AS 3.1207. Curcumin 4′-O-β-d-glucoside (2), together with hexahydrocurcumin (3), was isolated from a preparative-scale biotransformation with R. chinensis IFFI 03043 and characterized fully by NMR and MS. A time-course study revealed that curcumin could be efficiently converted into curcumin 4′-O-β-d-glucoside within 8 h when administered at 0.05 mmol L^?1 and the productivity was 57%. Additionally, the biotransformation products of curcumin by different fungal strains were analyzed by LC/MS. At least 15 metabolites were detected, and the predominant biotransformation reaction was glucosylation. This study provides a simple, efficient and less expensive approach for the preparation of curcumin glucosides. The introduction of the glucosyl function might be able to enhance the bioavailability of curcumin.     

The Ecology of Pneumocystis: Perspectives,Personal Recollections,and Future Research Opportunities

I am honored to receive the second Lifetime Achievement Award by International Workshops on Opportunistic Protists and to give this lecture. My research involves Pneumocystis, an opportunistic pulmonary fungus that is a major cause of pneumonia (“PcP”) in the immunocompromised host. I decided to focus on Pneumocystis ecology here because it has not attracted much interest. Pneumocystis infection is acquired by inhalation, and the cyst stage appears to be the infective form. Several fungal lung infections, such as coccidiomycosis, are not communicable, but occur by inhaling < 5 μm spores from environmental sources (buildings, parks), and can be affected by environmental factors. PcP risk factors include environmental constituents (temperature, humidity, SO₂, CO) and outdoor activities (camping). Clusters of PcP have occurred, but no environmental source has been found. Pneumocystis is communicable and outbreaks of PcP, especially in renal transplant patients, are an ongoing problem. Recent evidence suggests that most viable Pneumocystis organisms detected in the air are confined to a patient's room. Further efforts are needed to define the risk of Pneumocystis transmission in health care facilities; to develop more robust preventive measures; and to characterize the effects of climatological and air pollutant factors on Pneumocystis transmission in animal models similar to those used for respiratory viruses.     

Detection of a 3′, 5′-cyclic-AMP phosphodiesterase activity in the lichenized fungus Evernia prunastri

Abstract

A 3′, 5′-cyclic-AMP phosphodiesterase (PDE) was detected and measured in the lichen Evernia prunastri. The percentage of hydrolysis of tritiated 3′, 5′-cyclic-adenosine monophosphate ([³H]-cAMP) and 3′, 5′-cyclic-guanosine monophosphate ([³H]-cGMP) by the PDE enzyme into tritiated 5′-adenosine-monophospahte ([³H]-AMP) and tritiated 5′-guanosine-monophospahte ([³H]-GMP) was measured by treating the PDE products with a 5′-nucleotidase enzyme present in snake venom. The lysate fraction (L) (plasma membranes and cell walls) and the supernatant (S) (soluble fraction of the cells) were tested. In both fractions, competition of unlabelled cAMP, but not unlabelled cGMP, was revealed. Specific competitive PDE inhibitors such as IBMX inhibited enzymatic activity. Although it is thought that in this species cAMP is regulated by red/far red light through PDE activity, this is the first report that seems to suggest the presence of a PDE activity specific for cAMP in lichenized fungi. However, this work is at a preliminary stage and despite the high levels of enzymatic activity with cAMP found in both fractions, data are still insufficient to state the absolute specificity for this nucleotide.     

Notes on the Laboulbeniales (Ascomycota) parasitic on Diptera from Portugal and other countries

Eighteen species of Stigmatomyces are reported for the first time from continental Portugal and/or from the Azores. These are Stigmatomyces asteiae W. Rossi et Cesari, Stigmatomyces athyroglossae W. Rossi et Cesari, Stigmatomycescanzonerii W. Rossi et Cesari, Stigmatomyces ceratophorus Whisler, Stigmatomyces constrictus Thaxt., Stigmatomyces crassicollis Thaxt., Stigmatomyces divergatus Thaxt., Stigmatomyces discocerinae Thaxt., Stigmatomyces ensinae Thaxt., Stigmatomyces cf. ephydrae L. Mercier et R. A. Poiss., Stigmatomycesgeomyzae W. Rossi et Cesari, Stigmatomyces limnophorae Thaxt., Stigmatomyces majewskii H. L. Dainat, Manier et Balazuc, Stigmatomyces papuanus Thaxt., Stigmatomycesplatensis Speg., Stigmatomyces ptylomyiae Thaxt. and Stigmatomyces purpureus Thaxt., Stigmatomyces rugosus Thaxt. New records of these species are also reported from Australia, Canada, Canary Islands (Spain), Costa Rica, Cuba, Ecuador, Finland, Germany, Great Britain, Greece, Kenya, Hungary, Israel, Kyrgyzstan, Lebanon, Morocco, Saudi Arabia, Sierra Leone, Taiwan, Thailand, Turkey, Uganda, United Arab Emirates, USA, Yemen and Zimbabwe. The new records increase, sometimes considerably, information about distribution of these parasites. Two synonymies are also established: Stigmatomyces autriquei Balazuc = Stigmatomyces ensinae Thaxt.; Stigmatomyces psilopae Thaxt. var. camarguensis H. L. Dainat et J. Dainat = S. rugosus Thaxt.