Removal of cadmium by Pleurotus sajor-caju basidiomycetes

The bioaccumulation of cadmium by the white rot fungus Pleurotus sajor-caju onto dry biomass was investigated using aqueous media with concentrations in the range of 0.125 mM–1.0 mM The highest cadmium uptake (between 88.9 and 91.8%) was observed with aerobic fungal biomass from the exponential growth phase. Up to 1.0 mM cadmium gradually inhibited mycelium development, but never blocked it completely. Freeze-dried, oven-dried and non-metabolizing live Pleurotus sajor-caju biomass types were tested for their capacity to adsorb the test ion Cd²⁺ within the pH range of 4.5 to 6.0. Freeze–dried biomass proved to be the most efficient biomass type for Cd²⁺ metal adsorption. Therefore, Pleurotus sajor-caju may be used for heavy metal removal and bioremediation.     

Decolourization of Municipal Effluent and Sludge by Pleurotus sajor-caju and Pleurotus ostreatus

Summary The Americana Municipal Treatment Station, S?o Paulo, Brazil, manages 400 l of effluent s⁻¹, from domestic and textile origin, which produces an average of 20 t of sludge per day. The decolourization of the effluent and sludge by three strains of Pleurotus (Pleurotus sajor-caju F2, F6 and Pleurotus ostreatus) was evaluated. The strains of P. sajor-caju F2 and F6 were able to decolourize the sludge, while P. ostreatus was less efficient. Detoxification was appraised with three bioassays comprising the cnidarian Hydra attenuata, the alga Selenastrum capricornutum and lettuce seeds. After exposure to fungi, effluent toxicity decreased but not that of its sludge. Strain P. sajor-caju F6 presented signs of toxicity shown by electron microscopy in the presence of the effluent. The three strains produced high amounts of manganese-peroxidase (Mn–P) and laccase in the presence of the sludge. Although P. ostreatus produced large amount of Mn–P and laccase enzymes, these enzymes did not result in decolourization of the sludge, suggesting that other factors are likely to be involved. Carbon content decreased only in the treatment with P. ostreatus.     

Degradation of tannins in spent coffee grounds by Pleurotus sajor-caju

Pleurotus sajor-caju PL27, a white rot fungus, degraded up to 87% of the tannins in spent coffee grounds as a solid substrate over 32 days. Degradation of tannins was enhanced if potato and dextrose were included. The potential nutritive value of the substrate as animal feed may be improved by this process.Yum-Shing Wong is with the Biology Department, Chinese University of Hong Kong, Shatin, NT, Hong Kong. Xun Wang is a visiting scientist from the Research Institute of Food and Fermentation Industry, People's Republic of China.     

Yield and size of Pleurotus ostreatus and Pleurotus sajor-caju as effected by delayed-release nutrient

Summary Pleurotus ostreatus (444) and P. sajorcaju (537) were grown on a pasteurized mixture of chopped wheat straw (70%) and milled corncobs (30%) nonsupplemented and supplemented with two levels of delayed-release nutrient. Yields for both lines increased 2.3 and 3.2 fold on substrate containing 16% and 32% (dry weight basis) delayed-release nutrient additions, respectively. For mushroom size, a differential response was observed for genotype and delayed-release nutrient. Line 444, a wild isolate from Missouri, produced larger mushrooms on substrate containing higher levels of delayed-release nutrient. Line 537, a commercial isolate used in Italy, produced smaller mushrooms with higher levels of nutrient. Mushrooms of line 537 were harvested 3 to 4 days earlier and line 444 was harvested 12 to 14 days earlier from supplemented substrate.Contribution No. 1575, Department of Plant Pathology, The Pennsylvania Agricultural Experiment Station. Authorized for publication April, 1986, as Journal Series Paper No. 7394     

凤尾菇和桃红平菇种间原生质体电融合获杂种菌株

以有自然标记的双核风尾菇(Pleurotus sajor-caju)和桃红平菇(P．Rhodophyllus)为出发菌株,以它们携带营养缺陷型标记的单孢菌株为直接亲本,采用BAEKON 2 000基因转移仪进行侧耳属种间原生质体电融合,成功地获得若干株融合体,其中有一株编号为F57的融台体已培育出子实体。比较了融合体F57与亲本的形态、生理、生化和遗传等性状,结果证实,融合体F57是一株新的种间杂种菌株。     

Manganese peroxidase isoenzymes produced by Pleurotus ostreatus grown on wood sawdust

The white rot basidiomycete Pleurotus ostreatus produces two manganese peroxidase (MnP) isoenzymes when grown in solid stationary conditions on poplar sawdust, whereas a lower production of these same enzymes is observed on fir sawdust. Addition of Mn(2+) to poplar culture resulted in a threefold increase of MnP activity; the same addition to fir culture was able to increase tenfold the MnP production. The two MnP isoenzymes (MnP2 and MnP3) were purified from P. ostreatus poplar culture. The isoenzymes differ in their pI values, molecular masses, and N-terminal sequences. MnP3 has the same N-terminal sequence as that of a P. ostreatus MnP previously reported. Both isoenzymes exhibit Mn(2+)-dependent and Mn(2+)-independent peroxidase activities when tested on phenolic substrates. The gene coding for the new isoenzyme MnP2 was cloned and sequenced and the promoter region analyzed. Furthermore, the chromosomal localization of all known P. ostreatus genes was determined.     

The cultivation and nutritional value of Pleurotus sajor-caju

Summary A method for the cultivation of Pleurotus sajorcaju on a relatively large scale using cotton waste as substrate has been developed, and the mushroom so obtained has higher protein content than and comparable carbohydrate content to Agaricus bisporus, Volvariella volvacea, Lentinus edodes and Pleurotus ostreatus. The crude fats, ash, energy value, vitamin and mineral contents are lower and yet the differences are not great. The biological efficiency from cotton waste compost is lower than that from straw compost, however, the former has the advantage of giving rather even yield over successive flushes. This mushroom has a high potential to be produced economically on a large scale in Hong Kong.     

Molecular characterization of beta-tubulin gene from Pleurotus sajor-caju.

A beta-tubulin gene (TUB1) from the basidiomycete Pleurotus sajor-caju was sequenced. TUB1 encodes a 446-amino-acid protein. The coding region is interrupted by 9 introns, all of which had a 5'-GTRNGT... YAG-3' sequence at the boundaries. Locations of the introns in TUB1 were common between the beta-tubulin genes of other basidiomycetes, but not with animals, ascomycetes, or plants. This suggests that the introns were inserted independently into the beta-tubulin gene after these divisions had diverged.     

Purification and Characterization of Extracellular Laccase Secreted by Pleurotus sajor-caju MTCC 141

The effect of lignin containing natural substrates corn-cob, coir-dust, saw-dust, wheat straw and bagasse particles on the extracellular secretion of laccase in the liquid culture growth medium of Pleurotus sajor-caju MTCC 141 has been studied. The culture conditions for maximum secretion of laccase by Pleurotus sajor-caju MTCC 141 have been optimized. Homogeneous preparation of laccase from the culture filtrate of the fungus has been achieved using ammonium sulphate precipitation, anion exchange chromatography on DEAE and gel filtration chromatography on Sephadex G-100. The purified enzyme preparation gave a single protein band in SDS-PAGE analysis indicating a molecular weight of 90 kD. The enzymatic characteristics Km, kcat, pH and temperature optima of the purified laccase have been determined using 2, 6-dimethoxyphenol as the substrate and have been found to be 35μmol/L, 0.30 min-1, 4.5 and 37℃ respectively. The Km values for the other substrate like catechol, m-cresol, pyrogallol and syringaldazine have also been determined which were found to be 216 μmol/L, 380 μmol/L, 370 μmol/L and 260 μmol/L respectively.     

Veratryl alcohol oxidases from the lignin-degrading basidiomycete Pleurotus sajor-caju.

The basidiomycete Pleurotus sajor-caju mineralizes ring-14C-labelled lignin (dehydrogenative polymer) when grown in mycological broth. Under these conditions, two veratryl alcohol oxidase (VAO) enzymes were found in the culture medium. They oxidized a number of aromatic alcohols to aldehydes and reduced O2 to H2O2. The enzymes were purified by ion-exchange and gel-permeation chromatography. The final step of purification on Mono Q resolved the activity into two peaks (VAO I and VAO II). Both enzymes had the same Mr, approx. 71,000, but their isoelectric points differed slightly, 3.8 for VAO I and 4.0 for VAO II. Their amino acid compositions were similar except for aspartic acid/asparagine and glycine. Both enzymes are glycoproteins and contain flavin prosthetic groups. Their pH optima were around 5, and kinetic constants and specificities were similar. 4-Methoxybenzyl alcohol was oxidized the most rapidly, followed by veratryl alcohol. Not all aromatic alcohols were oxidized, neither were non-aromatic alcohols. Cinnamyl alcohol was oxidized at the gamma position. The VAO enzymes thus represent a significantly different route for veratryl alcohol oxidation from that catalysed by the previously found lignin peroxidases from Phanerochaete chrysosporium. The role of the oxidases in biodegradation might be to produce H2O2 during oxidation of lignin fragments.     

Effect of nutrient nitrogen and manganese on manganese peroxidase and laccase production by Pleurotus sajor-caju

Pleurotus sajor-caju, strain Pl-27, produces manganese-dependent peroxidase (MnP) and laccase, but not lignin peroxidase, when grown on a defined medium with glucose as sole carbon source. MnP activity was detected in fungal cultures supplemented with both high (26 mM-N) and low (2.6 mM-N) nutrient nitrogen although higher specific activity values were recorded under the latter conditions. Conversely, laccase production was not influenced by nutrient nitrogen levels under the growth conditions adopted. Both the titre and time of appearence of MnP were also affected by the concentration of Mn in the culture medium with highest enzyme levels recorded in cultures supplemented with 15 ppm Mn. Two MnP and five laccase isoforms were identified by FPLC and gel electrophoresis.     

Studies on cultivation,biological efficiency and chemical analysis of Pleurotus sajor-caju (FR.) SINGER on different bio-wastes

Three different kinds of biomass, namely Populus deltoides, Eupatorium adenophorum and sericulture waste were used individually for the cultivation of Pleurotus sajor-caju, alone and mixed with paddy straw. P. sajor-caju, when used alone, exhibited a very good colonizing ability on these substrates, except in sericulture waste. The biological efficiency of P. deltoides and E. adenophorum when used as pure substrate was 75 and 77%, respectively, but it increased to 102% when P. sajor-caju was cultivated in a mixture with paddy straw in a ratio of 1:2. Experiments examining the growth on sericulture waste in both pure and mixed substrate are encouraging. From the analysis of substrate before and after the cultivation of P. sajor-caju it was noted that subsstrates were enriched in their protein content as a result of growth of this mushroom. The percentage of degradation of cellulose, hemicellulose and lignin showed that P. sajor-caju is capable of utilizing all three major components. The fruit bodies of P. sajor-caju were analyzed for crude protein content, crude fat and carbohydrate content. The energy values in the fruit bodies of P. sajor-caju and different organic wastes were found to vary from 282 to 309 kcal/100 g and from 319 to 467 kcal/100 g, respectively. It was found, however, that the energy recovery from organic wastes by fruit bodies was very low, i.e. 4.19-8.73 kcal/100g of dry substrate.     

Biological efficiency and nutritive value of Pleurotus sajor-caju cultivated on different agro-wastes

The oyster mushroom, Pleurotus sajor-caju (Fr.) Singer, was cultivated on a number of agro-residues and their mixtures. Biological efficiency, defined as the percentage conversion of substrate into fruit bodies on a dry weight basis, was found to be maximum on paddy straw supplemented with cotton seeds (12·82 g/100 g substrate). Biochemical changes effected in the substrates as a result of mushroom growth, in terms of nitrogen content and degradation of cellulose, hemicellulose and lignin components, were monitored. The proximate compositions of fruit bodies of the mushroom in terms of protein, total carbohydrates, fat, fibre and ash were determined. The crude protein and total carbohydrate contents varied from 26·3% to 36·7% and 41·2% to 47·1%, respectively, on various residues. The variation in the contents of crude fat, crude fibre and ash ranged from 1·7% to 2·0%, 11·7% to 17·0% and 6·1%, respectively, on different residues. The energy value of the fruit bodies was found to be between 275 and 300 kCal/100 g, whereas the energy recovery of the substrate in the fruit bodies was from 5% to 10%.     

Biotransformation of banana waste into protein by pleurotus sajor-caju

Pseudostems and leaves from banana waste were used for biotransformation into protein by using P. sajor-caju, an oyster mushroom. Treatment of formalin (500 ppm) + carbendazim (12.5 ppm) of these substrates was found to favour relatively high percentage biological efficiency (BE) of P. sajor-caju.. Steam sterilization also exhibited comparable yield performance by P. sajor-caju. Fruiting bodies harvested from all the treatments had relatively higher protein contents. The spent substrate (steam sterilized) was found to be suitable as an ideal animal feed because of its rich nutritive composition.     

Incorporation of caffeine into the macromicete fungus Pleurotus sajor-caju growing on coffee pulp

TWhen the chemical composition of secondary metabolites from the Pleurotus sajor-caju growing on coffee pulp were study, it was found that the fungus has the faculty of incorporating caffeine inside its fructiferous body. Component of the substrate (around 1.3% on dry basis) did not show a structural change over the alkaloid; this constitutes an unexpected outcome for a species belonging to realm of the fungi.     

Pleurotus sajor-caju HSP100 complements a thermotolerance defect inhsp104 mutantSaccharomyces cerevisiae

A putativeHsp100 gene was cloned from the fungusPleurotus sajor-caju. mRNA expression studies demonstrated that this gene (designatedPsHsp100) is highly induced by high temperature, induced less strongly by exposure to ethanol, and not induced by drought or salinity. Heat shock induction is detectable at 37‡C and reaches a maximum level at 42‡C.PsHsp100 mRNA levels sharply increased within 15 min of exposure to high temperature, and reached a maximum expression level at 2 h that was maintained for several hours. These results indicate that PsHsp100 could work at an early step in thermotolerance. To examine its function,PsHsp100 was transformed into a temperature-sensitivehsp104 deletion mutantSaccharomyces cerevisiae strain to test the hypothesis that PsHSP100 is an protein that functions in thermotolerance. Overexpression of PsHSP100 complemented the thermotolerance defect of thehsp104 mutant yeast, allowing them survive even at 50‡C for 4 h. These results indicate that PsHSP100 protein is functional as an HSP100 in yeast and could play an important role in thermotolerance inP. sajor-caju. An erratum to this article is available at .     

Extracellular enzyme production during anamorphic growth in the edible mushroom,Pleurotus sajor-caju

Cultivation of mushrooms on lignocellulosic wastes represents a cost-effective organic recycling process. Pleurotus sajor-caju grown on cotton-waste produced relatively low levels of three components of the cellulase complex namely cellobiohydrolase (EC 3.2.1.91), CMCase (EC 3.2.1.4) and -glucosidase (EC 3.2.1.21) with specific activity values of 10.0, 71.4 and 21.6U (mg protein)–1 respectively after 15days. Higher specific activity was registered in alkali-treated cotton with 15.6, 83.4 and 56.1U (mg protein)–1 respectively after 20days. Lower levels were noted on rubber-tree sawdust substrate with specific activity values of 0.28, 0.62 and 0.75U (mg protein)–1 for the respective enzymes after 28–35days growth. The maximum production of xylanase (EC 3.2.1.8) of 0.63U (mg protein)–1 occurred after 20days while a relatively higher level of the phenoloxidase enzyme, laccase (EC 1.14.18.1) of 27.4U (mg protein)–1 (maximum) was found after 35days. Laccase, the activity of which is associated with morphogenesis, increased with mycelial growth, peaked at maximum growth and thereafter decreased rapidly. This could prove important commercially in timing the end of spawn-run in preparation for initiation of fruiting.