An extracellular lipase with 23,666.66 U/ml/min activity was produced by Aspergillus tamarii JGIF06 under submerged fermentation in mineral salt medium containing coconut oil (2.5 % v/v), tryptone (2 % w/v) and ammonium chloride (2 % w/v), with initial pH of 5 ± 0.2, incubated at 25 °C for 7 days on a rotary shaker at 120 rpm. A 7.9-fold increase in lipase-specific activity was recorded after purification by DEAE Sepharose ion exchange and Sephadex G200 column chromatography. The apparent molecular mass of this enzyme was revealed as 50 kDa by sodium dodecyl sulphate polyacrylamide gel electrophoresis. The optimal lipase activity was recorded at pH 4 and 37 °C. The enzyme revealed broad specificity towards different vegetable oils. The K m and V max of the lipase on olive oil was found to be 330.4 mg and 53,690 U/ml/min, respectively. The lipase activity was stable in the presence of surfactants such as cetrimonium bromide, sodium dodecyl sulphate and Tween 80, and metal ions and reagents such as Ca2+, Ba2+ and 2-mercaptoethanol. However, the activity was greatly reduced in the presence of organic solvents such as chloroform. The stain removal potential of the crude lipase was determined on polycotton fabric pieces stained with peanut oil. Lipase added to cold water alone significantly enhanced the removal of stain by 152 %. The addition of lipase also improved the stain removal efficiency of a commercially available detergent in the presence of either cold (25 ± 2 °C) or hot (65 ± 2 °C) water. The current findings suggest the potentiality of this enzyme for energy-efficient biocatalytic application.

Benzo[a]pyrene is considered as a priority pollutant because of its carcinogenic, teratogenic and mutagenic effects. The highly recalcitrant nature of Benzo[a]pyrene poses a major problem for its degradation. White-rot fungi such as Pleurotus ostreatus can degrade Benzo[a]pyrene by enzymes like laccase and manganese peroxidase. The present investigation was carried out to determine the extent of Benzo[a]pyrene degradation by the PO-3, a native isolate of P. ostreatus, in the presence of heavy metals and ligninolytic enzyme mediators. Modified mineral salt medium was supplemented with 5 mM concentration of different heavy metal salts and ethylenediaminetetraacetic acid. Vanillin and 2,2′-azinobis-(3-ethylbenzothiazoline-6-sulfonate) (1 and 5 mM) were used to study the effect of mediators. Results indicated that P. ostreatus PO-3 degraded 71.2 % of Benzo[a]pyrene in the presence of copper ions. Moderate degradation was observed in the presence of zinc and manganese. Both biomass formation and degradation were severely affected in the presence of all other heavy metal salts used in the study. Copper at 15 mM concentration supported the best degradation (74.2 %), beyond which the degradation progressively reduced. Among the mediators, 1 mM 2,2′-azinobis-(3-ethylbenzothiazoline-6-sulfonate) supported 78.7 % degradation and 83.6 % degradation was observed under the influence of 5 mM vanillin. Thus, metal ion like copper is essential for better biodegradation of Benzo[a]pyrene. Compared to synthetic laccase mediator like 2,2′-azinobis-(3-ethylbenzothiazoline-6-sulfonate), natural mediator such as vanillin may play a significant role in the degradation of aromatic compounds by white-rot fungi.

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 Cu(2+) and Triton X-100, at toxin concentration of 5 µg mL(-1). P. ostreatus GHBBF10 showed highest degradation in the presence of Zn(2+) 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.

A laccase produced by Pleurotus ostreatus MTCC 142 under solid-state fermentation using co-substrates of paddy straw and corn husk (1.5:1.5, g w/w) showed an activity of 2.54 U gds−1. Laccase activity was determined spectrophotometrically using 0.5 mM 2,2′- azino-bis (3-ethylbenzthiazoline-6-sulphonic acid) (ABTS). Supplementation with fructose and potassium nitrate resulted in maximum enzyme production at initial pH 5.8 ± 0.2 and initial moisture content of 70%. A carbon: nitrogen ratio of 0.5:0.1 yielded highest laccase activity in the presence of surfactant Tween 20 (0.05%, w/v). Incorporation of vanillin (5 mM) and copper sulphate (10 mM) facilitated enhanced synthesis of laccase. A 4.8-fold increase in enzyme activity was recorded after optimization of nutritional parameters. The apparent molecular mass of this enzyme was revealed as 43 kDa by sodium dodecyl sulphate–polyacrylamide gel electrophoresis. The laccase showed optimal activity at pH 3 and 35 °C with 82.8% residual activity after 1 h of incubation. The Km and Vmax values on ABTS were found to be 0.52 mM and 9.33 U gds−1, respectively. The enzyme activity was enhanced by Cu2+ and remained unaffected with Ba2+, Mn2+, Pb2+, Mg2+, Ca2+ and Fe3+. However, pre-incubation of the enzyme with reagents like sodium azide, sodium lauryl sulphate and 2-mercaptoethanol demonstrated an inhibition of its activity. Addition of crude laccase to Congo red dye solution resulted in 36.84% decolourization after 20 h of incubation at 35 ± 2 °C. This study discusses the production and characterization of a laccase from P. ostreatus strain with potential for azo dye decolourization.

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The present investigation highlights the process parameters influencing the submerged fermentation of chitinase by Bacillus pumilus JUBCH08, purification and characterization of the enzyme and determination of antagonistic activity of the bacterium against Fusarium oxysporum. Medium supplemented with 0.5 % chitin and peptone, at initial pH 8.0, when incubated at 35 °C for 72 h favored highest chitinase production. The enzyme was purified 25.1-fold to homogeneity. The chitinase was found to be thermostable and alkali-tolerant with maximum activity at pH 8.0 and 70 °C for 1 h. The molecular weight of chitinase was found to be 64 kDa by sodium dodecyl sulfate polyacrylamide gel electrophoresis. Mg2+, Co2+, Ca2+ and Mn2+ improved the chitinase activity. The K m and V max values of the enzyme were 0.13 mg/ml and 38.23 U/ml, respectively. When subjected to dual plate assay, the bacterium showed 45 % antagonism against F. oxysporum. Thus, it could be inferred that cultural conditions strongly affected the chitinase production by B. pumilus JUBCH08. The enzyme being thermostable and best functional under alkaline conditions could be useful for the feed industry and related biotechnological applications. Inhibition of F. oxysporum by the culture through lytic mechanism indicates its potentiality as a biocontrol agent.

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