Abiotic and Biotic Degradation of Oxo-Biodegradable Plastic Bags by Pleurotus ostreatus

In this study, we evaluated the growth of Pleurotus ostreatus PLO6 using oxo-biodegradable plastics as a carbon and energy source. Oxo-biodegradable polymers contain pro-oxidants that accelerate their physical and biological degradation. These polymers were developed to decrease the accumulation of plastic waste in landfills. To study the degradation of the plastic polymers, oxo-biodegradable plastic bags were exposed to sunlight for up to 120 days, and fragments of these bags were used as substrates for P. ostreatus. We observed that physical treatment alone was not sufficient to initiate degradation. Instead, mechanical modifications and reduced titanium oxide (TiO₂) concentrations caused by sunlight exposure triggered microbial degradation. The low specificity of lignocellulolytic enzymes and presence of endomycotic nitrogen-fixing microorganisms were also contributing factors in this process.     

Degradation of Oxo-Biodegradable Plastic by Pleurotus ostreatus

Growing concerns regarding the impact of the accumulation of plastic waste over several decades on the environmental have led to the development of biodegradable plastic. These plastics can be degraded by microorganisms and absorbed by the environment and are therefore gaining public support as a possible alternative to petroleum-derived plastics. Among the developed biodegradable plastics, oxo-biodegradable polymers have been used to produce plastic bags. Exposure of this waste plastic to ultraviolet light (UV) or heat can lead to breakage of the polymer chains in the plastic, and the resulting compounds are easily degraded by microorganisms. However, few studies have characterized the microbial degradation of oxo-biodegradable plastics. In this study, we tested the capability of Pleurotus ostreatus to degrade oxo-biodegradable (D₂W) plastic without prior physical treatment, such as exposure to UV or thermal heating. After 45 d of incubation in substrate-containing plastic bags, the oxo-biodegradable plastic, which is commonly used in supermarkets, developed cracks and small holes in the plastic surface as a result of the formation of hydroxyl groups and carbon-oxygen bonds. These alterations may be due to laccase activity. Furthermore, we observed the degradation of the dye found in these bags as well as mushroom formation. Thus, P. ostreatus degrades oxo-biodegradable plastics and produces mushrooms using this plastic as substrate.     

Novel salt and alkali tolerant neotropical basidiomycetes for dye decolorisation in simulated textile effluent

The initial alkaline pH and the concentration of sodium chloride in the synthetic liquid medium were a key factor in the capability of twenty-five white-rot fungi strains to decolorise the dye Reactive Blue 19. Six strains decolorised 90% of the dye at pH 8.0, and only Peniophora cinerea decolorized 90% of the dye at pH 9.0. Fourteen strains were capable of decolorising the dye in saline medium (sodium chloride 10 g l⁻¹). P. ostreatus, P. cinerea and T. villosa were able to decolorize the dye both in medium with initial pH 8.0 or in saline medium. These three strains were selected and evaluated for simulated alkali-saline textile effluent decolorisation in different conditions: time of cultivation for effluent addition (0, 5, 7 and 9 days), initial pH (4.5 and 8.0) and agitation (0 and 120 rpm). P. ostreatus and P. cinerea decolorised the alkali-saline textile effluent by 93.0 and 25.4%, when the medium’s initial pH was 8.0 or 4.5, respectively, and the effluent was added in the 7th day of growth. T. villosa decolorized 40% when the effluent was added on the 9th day of cultivation at pH 4.5. Agitation increased the effluent decolorisation by T. villosa, but inhibition was observed for P. cinerea and P. ostreatus. The results showed that each fungus presented a specific behavior in relation to the best culture conditions for decolorisation of alkali-saline effluent containing reactive dyes. The strains of P. ostreatus, P. cinerea and T. villosa were considered as promising alternative for the biodegradation of this effluent, employing the strategy of effluent addition after a certain period of fungal growth.     

Biodegradation Kinetics of Naphthalene in Soil Medium Using Pleurotus Ostreatus in Batch Mode with Addition of Fibrous Biomass as a Nutrient

The efficiency and kinetics of naphthalene biodegradation in a soil medium using Pleurotus ostreatus (a type of white rot fungus) in batch mode with and without the addition of oil palm fiber (OPF) as a nutrient are evaluated in this study. Three batches are considered in the biodegradation study: (i) control—spiked soil; (ii) spiked soil with fungus; and (iii) spiked soil with both fungus and OPF. Biodegradation is conducted over a period of 22 days for which soil naphthalene concentrations are determined with respect to microwave extraction and high-performance liquid chromatography (HPLC) analysis. The results indicate that inoculation with Pleurotus ostreatus significantly enhances soil naphthalene biodegradation to 84%, which is further enhanced upon the addition of OPF to 98% with respect to the degradation rate. The high carbon content in OPF (>40%) affords it the capacity to be a viable nutrient supplement for Pleurotus ostreatus, thereby enhancing the potential of Pleurotus ostreatus in the biodegradation of polycylic aromatic hydrocarbons (PAHs), and indicating the potential of OPF as a nutrient for PAH biodegradation. A relationship between OPF mass and the biodegradation rate constant has been determined to be linear according to the following equation: k = 0.0429 × OPF + 0.1291.     

Lignocellulose Degradation during Solid-State Fermentation: Pleurotus ostreatus versus Phanerochaete chrysosporium

Lignocellulose degradation and activities related to lignin degradation were studied in the solid-state fermentation of cotton stalks by comparing two white rot fungi, Pleurotus ostreatus and Phanerochaete chrysosporium. P. chrysosporium grew vigorously, resulting in rapid, nonselective degradation of 55% of the organic components of the cotton stalks within 15 days. In contrast, P. ostreatus grew more slowly with obvious selectivity for lignin degradation and resulting in the degradation of only 20% of the organic matter after 30 days of incubation. The kinetics of ¹⁴C-lignin mineralization exhibited similar differences. In cultures of P. chrysosporium, mineralization ceased after 18 days, resulting in the release of 12% of the total radioactivity as ¹⁴CO₂. In P. ostreatus, on the other hand, 17% of the total radioactivity was released in a steady rate throughout a period of 60 days of incubation. Laccase activity was only detected in water extracts of the P. ostreatus fermentation. No lignin peroxidase activity was detected in either the water extract or liquid cultures of this fungus. 2-Keto-4-thiomethyl butyric acid cleavage to ethylene correlated to lignin degradation in both fungi. A study of fungal activity under solid-state conditions, in contrast to those done under defined liquid culture, may help to better understand the mechanisms involved in lignocellulose degradation.     

Kinetics and pathway of biodegradation of dibutyl phthalate by Pleurotus ostreatus

Dibutyl phthalate (DBP) is a plasticizer, whose presence in the environment as a pollutant has attained a great deal of attention due to its reported association with endocrine system disturbances on animals. Growth parameters, glucose uptake, percentage of removal efficiency (%E) of DBP, biodegradation constant of DBP (k) and half-life of DBP biodegradation (t_1/2) were evaluated for Pleurotus ostreatus grown on media containing glucose and different concentrations of DBP (0, 500 and 1000 mg l^?1). P. ostreatus degraded 99.6 % and 94 % of 500 and 1000 mg of DBP l^?1 after 312 h and 504 h, respectively. The k was 0.0155 h^?1 and 0.0043 h^?1 for 500 and 1000 mg of DBP l^?1, respectively. t_1/2 was 44.7 h and 161 h for 500 and 1000 mg of DBP l^?1, respectively. Intermediate compounds of biodegraded DBP were identified by GC-MS and a DBP biodegradation pathway was proposed using quantum chemical calculation. DBP might be metabolized to benzene and acetyl acetate, the first would be oxidated to muconic acid and the latter would enter into the Krebs cycle. P. ostreatus has the ability to degrade DBP and utilizes it as source of carbon and energy.     

Enhancement of biodegradability of disposable polyethylene in controlled biological soil

Plastics as polyethylene are widely used in packaging and other agricultural applications. They accumulate in the environment at a rate of 25 million tons per year. Thus, the development and use of degradable plastics was proposed as a solution for plastic waste problem. Because of the ever-increasing use of plastic films, nowadays, biodegradability has become a useful characteristic for plastics. Conversely, the introduction of biodegradable plastics has generated a need for methods to evaluate the biodegradation of these polymers in landfills and solid waste treatment systems such as composting or anaerobic digestion treatment plants. The purpose of this study was to investigate the biodegradation of disposable low-density polyethylene bags containing starch (12%), autoxidizable fatty acid ester and catalytic agents in soil. Structurally this work intended to evaluate the capacity of Phanerochaete chrysosporium (ATCC 34541) to enhance polyethylene film biodegradation in soil microcosms. Soil samples inoculated with P. chrysosporium were mixed with LDPE/starch blend films and biological changes of the films and soil were monitored for 6 months. The biodegradation of polyethylene starch blend film has been determined by the physical, chemical and biological properties of the samples such as pH, biomass, CO₂ formation, percentage elongation, relative viscosity and FTIR spectrum.     

Biodegradation of entomopathogenic hyphomycetes: Influence of clay coating on Beauveria bassiana blastospore survival in soil

The survival of naked and clay-coated Beauveria blastospores in soil was investigated with an experimental biodegradation method using a trap technique. At various times of incubation, traps were collected to study changes in biomass, inoculum potential, and infection potential of degraded blastospores, and to investigate the colonization of blastospore biomass by antagonistic soil-borne microorganisms. Infection potential was tested on larvae of Plutella maculipennis and Leptinotarsa decemlineata. Naked blastospores were inactivated after 3 weeks incubation, while clay-coated blastospores were still active after 2 months incubation in soil at 20°C. All investigations demonstrated the protective role of clay coating against biodegradation of fungal propagules by soil bacteria and protozoa. Consequently, clay coating of blastospores is proposed for the formulation of entomopathogenic hyphomycetes propagules. Moreover, antagonists implicated in lysis of blastospores, must be considered as an important part of the environmental response to a massive introduction of a fungus used for insect control.     

Effect of carbon source on the degradation of 2-naphthalenesulfonic acid polymers mixture by Pleurotus ostreatus in petrochemical wastewater

The ability of the white rot fungus Pleurotus ostreatus to biodegrade 2-naphthalenesulfonic acid polymers (2-NSAP) contained in a petrochemical wastewater was evaluated. The characterization of the wastewater and a LC–MS identification of polymers were performed. Biodegradation experiment was conducted in batches with and without the addition of a carbon source (glucose, corn starch, cellulose and lignin). It has been demonstrated that, in presence of a suitable carbon source, P. ostreatus is able to remove NSAP ranging from trimers to decamers. Interestingly, all the carbon sources utilized, despite the different velocities, allowed to degrade about 60–70% of the oligomers.Furthermore, respirometric tests showed that the fungal treatment was also able to significantly increase the bCOD/COD ratio, going from 9% (raw wastewater) up to 57%. This is clearly correlated with the enhancement in oligomers depolymerization confirming that the fungal action is not a mineralization of NSAP, but an increase of their biodegradability.     

Differentially Regulated, Vegetative-Mycelium-Specific Hydrophobins of the Edible Basidiomycete Pleurotus ostreatus

Three different hydrophobins (Vmh1, Vmh2, and Vmh3) were isolated from monokaryotic and dikaryotic vegetative cultures of the edible fungus Pleurotus ostreatus. Their corresponding genes have a number of introns different from those of other P. ostreatus hydrophobins previously described. Two genes (vmh1 and vmh2) were expressed only at the vegetative stage, whereas vmh3 expression was also found in the fruit bodies. Furthermore, the expression of the three hydrophobins varied significantly with culture time and nutritional conditions. The three genes were mapped in the genomic linkage map of P. ostreatus, and evidence is presented for the allelic nature of vmh2 and POH3 and for the different locations of the genes coding for the glycosylated hydrophobins Vmh3 and POH2. The glycosylated nature of Vmh3 and its expression during vegetative growth and in fruit bodies suggest that it should play a role in development similar to that proposed for SC3 in Schizophyllum commune.     

Biodegradation of aflatoxin B1 in contaminated rice straw by Pleurotus ostreatus MTCC 142 and Pleurotus ostreatus GHBBF10 in the presence of metal salts and surfactants

Aflatoxin B1 (AFB1) is a highly toxic fungal metabolite having carcinogenic, mutagenic and teratogenic effects on human and animal health. Accidental feeding of aflatoxin-contaminated rice straw may be detrimental for ruminant livestock and can lead to transmission of this toxin or its metabolites into the milk of dairy cattle. White-rot basidiomycetous fungus Pleurotus ostreatus produces ligninolytic enzymes like laccase and manganese peroxidase (MnP). These extracellular enzymes have been reported to degrade many environmentally hazardous compounds. The present study examines the ability of P. ostreatus strains to degrade AFB1 in rice straw in the presence of metal salts and surfactants. Laccase and MnP activities were determined spectrophotometrically. The efficiency of AFB1 degradation was evaluated by high performance liquid chromatography. Highest degradation was recorded for both P. ostreatus MTCC 142 (89.14 %) and P. ostreatus GHBBF10 (91.76 %) at 0.5 µg mL^?1 initial concentration of AFB1. Enhanced degradation was noted for P. ostreatus MTCC 142 in the presence of Cu²⁺ and Triton X-100, at toxin concentration of 5 µg mL^?1. P. ostreatus GHBBF10 showed highest degradation in the presence of Zn²⁺ and Tween 80. Liquid chromatography-mass spectrometric analysis revealed the formation of hydrated, decarbonylated and O-dealkylated products. The present findings suggested that supplementation of AFB1-contaminated rice straw by certain metal salts and surfactants can improve the enzymatic degradation of this mycotoxin by P. ostreatus strains.     

Bioremediation of oil-polluted soil with an association including the fungus Pleurotus ostreatus and soil microflora

The possibility of application of the Pleurotus ostreatus D1-soil microflora to bioremediation of oil-polluted soils was studied. The fungus degraded mainly the aromatic fractions, whereas soil microflora intensely degraded paraffin and naphthene oil fractions. Introduction of the fungus Pleurotus ostreatus D1 to soil induces degradation of a wider range of oil hydrocarbons. It is reasonable to further investigate the discovered phenomenon in order to improve procedures of remediation of oil-polluted soils.     

Biodegradation of Degradable Plastic Polyethylene by Phanerochaete and Streptomyces Species

The ability of lignin-degrading microorganisms to attack degradable plastics was investigated in pure shake flask culture studies. The degradable plastic used in this study was produced commercially by using the Archer-Daniels-Midland POLYCLEAN masterbatch and contained pro-oxidant and 6% starch. The known lignin-degrading bacteria Streptomyces viridosporus T7A, S. badius 252, and S. setonii 75Vi2 and fungus Phanerochaete chrysosporium were used. Pro-oxidant activity was accelerated by placing a sheet of plastic into a drying oven at 70°C under atmospheric pressure and air for 0, 4, 8, 12, 16, or 20 days. The effect of 2-, 4-, and 8-week longwave UV irradiation at 365 nm on plastic biodegradability was also investigated. For shake flask cultures, plastics were chemically disinfected and incubated-shaken at 125 rpm at 37°C in 0.6% yeast extract medium (pH 7.1) for Streptomyces spp. and at 30°C for the fungus in 3% malt extract medium (pH 4.5) for 4 weeks along with an uninoculated control for each treatment. Weight loss data were inconclusive because of cell mass accumulation. For almost every 70°C heat-treated film, the Streptomyces spp. demonstrated a further reduction in percent elongation and polyethylene molecular weight average when compared with the corresponding uninoculated control. Significant (P < 0.05) reductions were demonstrated for the 4- and 8-day heat-treated films by all three bacteria. Heat-treated films incubated with P. chrysosporium consistently demonstrated higher percent elongation and molecular weight average than the corresponding uninoculated controls, but were lower than the corresponding zero controls (heat-treated films without 4-week incubation). The 2- and 4-week UV-treated films showed the greatest biodegradation by all three bacteria. Virtually no degradation by the fungus was observed. To our knowledge, this is the first report demonstrating bacterial degradation of these oxidized polyethylenes in pure culture.     

Cultivation of oyster mushrooms (<Emphasis Type="Italic">Pleurotus</Emphasis> spp.) on various lignocellulosic wastes

Cultivation of speciality mushrooms on lignocellulosic wastes represents one of the most economically and cost-effective organic recycling processes. Three species of Pleurotus, namely P. columbinus, P. sajor-caju and P. ostreatus were experimentally evaluated on untreated organic wastes including chopped office papers, cardboard, sawdust and plant fibres. Production studies were carried out in polyethylene bags of about 1 kg wet weight with 5% spawning rates of substrate fresh weight in a custom-made growth room especially designed for spawn run and cropping. The conversion percentage from dry substrate weight to fresh mushroom weight (biological efficiency) was determined. The highest biological efficiency was noted with P. columbinuson cardboard (134.5%) and paper (100.8%), whereas P. ostreatus produced maximum yield on cardboard (117.5%) followed by paper (112.4%). The overall yield of P. sajor-cajuwas comparatively low (range 47–78.4%). The average number of sporophore flushings ranged between 5 and 6 times. The findings that P. columbinus and P. ostreatus are superior to P. sajor-caju are consistent with previous reports elsewhere. Further evaluation of P. columbinus alone on different bagging systems containing partially pasteurized office papers as a growing substrate revealed that polyethylene bags resulted in 109.4% biological efficiency in contrast to pottery (86%), plastic trays (72%) or polyester net (56%). The above findings reveal an opportunity for commercial implication of oyster mushroom especially P. columbinus for utilization of different feasible and cheap recyclable residues.     

大蒜浸出液对平菇细菌性褐斑病病原菌托拉斯假单胞杆菌的抑菌作用

【背景】由托拉斯假单胞杆菌(Pseudomonas tolaasii)引起的平菇细菌性褐斑病在国内外大面积发生,导致产量降低,并有潜在的安全风险,寻找安全有效的抑菌剂对产业发展具有重要意义。【目的】通过5种不同溶剂提取得到大蒜浸出液,测定其对平菇细菌性褐斑病病原菌托拉斯假单胞杆菌的抑制作用,同时检测其对平菇菌丝生长的作用。【方法】利用抑菌圈法测定5种不同的大蒜浸出液对托拉斯假单胞杆菌的抑菌作用,利用平板扩散法筛选能促进平菇菌丝生长的药剂及适宜的浓度。【结果】5种大蒜浸出液原液对托拉斯假单胞杆菌均有显著的抑菌活性,其中大蒜山杏壳木醋液浸出原液抑菌效果最强。不同浓度的大蒜浸出液抑菌作用比较发现,浓度为10%的大蒜山杏壳木醋液浸出液具有较好的抑菌效果,其抑菌效果与0.33 mg/mL的链霉素相当,并对平菇菌丝生长有显著的促进作用,菌丝生长速度显著大于对照,并且菌丝浓密,边缘整齐。【结论】本研究为大蒜与山杏壳木醋液复配药剂防治平菇细菌性褐斑病奠定了实验基础。     

Kinetics of wheat straw solid-state fermentation with Trametes versicolor and Pleurotus ostreatus — lignin and polysaccharide alteration and production of related enzymatic activities

Summary The kinetics of straw solid-state fermentation (SSF) with Trametes versicolor and Pleurotus ostreatus was investigated to characterize the delignification processes by these white-rot fungi. Two successive phases could be defined during straw transformation, characterized by changes in respiratory activity, changes in lignin and polysaccharide content and composition, increase in in-vitro digestibility, and enzymatic activities produced by the fungi. Lignin composition was analysed after CuO alkaline degradation, and decreases in syringyl/guaiacyl and syringyl/p-hydroxyphenyl ratios and cinnamic acid content were observed during the fungal treatment. An increase in the phenolic acid yield, revealing fungal degradation of side-chains in lignin, was produced by P. ostreatus. The highest xylanase level was produced by P. ostreatus, and exocellulase activity was nearly absent from straw treated with this fungus. Lactase activity was found in straw treated with both fungi, but lignin peroxidase was only detected during the initial phase of straw transformation with T. versicolor. High levels of H₂O₂-producing aryl-alcohol oxidase occurred throughout the straw SSF with P. ostreatus. Offprint requests to: A. T. Martínez     

Biodegradation and metabolite transformation of pyrene by basidiomycetes fungal isolate Armillaria sp. F022

Armillaria sp. F022 is a white-rot fungus isolated from a tropical rain forest in Indonesia that is capable of utilizing pyrene as a source of carbon and energy. Enzymes production during the degradation process by Armillaria sp. F022 was certainly related to the increase in biomass. In the first week after incubation, the growth rate rapidly increased, but enzyme production decreased. After 7 days of incubation, rapid growth was observed, whereas, the enzymes were produced only after a good amount of biomass was generated. About 63 % of pyrene underwent biodegradation when incubated with this fungus in a liquid medium on a rotary shaker (120 rpm, 25 °C) for 30 days; during this period, pyrene was transformed to five stable metabolic products. These metabolites were extracted in ethyl acetate, isolated by column chromatography, and then identified using thin layer chromatography (TLC) and gas chromatography–mass spectrometry (GC–MS). 1-Hydroxypyrene was directly identified by GC–MS, while 4-phenanthroic acid, 1-hydroxy-2-naphthoic acid, phthalic acid, and protocatechuic acid were identified to be present in their derivatized forms (methylated forms and silylated forms). Protocatechuic acid was the end product of pyrene degradation by Armillaria sp. F022. Dynamic profiles of two key enzymes, namely laccase and 1,2-dioxygenase, were revealed during the degradation process, and the results indicated the presence of a complicated mechanism in the regulation of pyrene-degrading enzymes. In conclusion, Armillaria sp. F022 is a white-rot fungus with potential for application in the degradation of polycyclic aromatic hydrocarbons such as pyrene in the environment.     

Atrazine,desethylatrazine (DEA) and desisopropylatrazine (DIA) degradation by Pleurotus ostreatus INCQS 40310

Abstract

Atrazine is the most common herbicide applied in crops of economic relevance, such as sugar cane, soybean, and corn. Atrazine and its derivatives desethylatrazine (DEA) and desisopropylatrazine (DIA) are toxic to the environment, affecting animal and human health. Thus, this study aimed to evaluate the degradation of atrazine and its derivatives by the fungus Pleurotus ostreatus INCQS 40310, as well as the potential of the enzymes involved in this process. P. ostreatus INCQS 40310 was able to degrade atrazine (82%), DEA (71%), and DIA (56%) over 22?days of fungal cultivation. Proteomic analysis indicated the participation of hydrolases and peroxidases during the degradation process. Additionally, resting cells of the fungus were tested to verify the action of intracellular enzymes in the degradation process, suggesting the participation of cytochrome P450 enzymatic complex. Resting cells experiments promoted the degradation of 50% of atrazine, 36% of DIA, 30% of DEA. So far, this is the first work evaluating the biodegradation of DEA and DIA by fungus.