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Proximate analysis of lignocellulosic biomass and its utilization for production, purification and characterization of ligninolytic enzymes by Aspergillus flavus

Khan Jehangir, Asad Muahammad Javaid, Mahmood Raja Tahir, Wattoo Feeroza Hamid, Zainab Tayyaba, Nazir Sidrah, Shah Muhammad Basir, Ahmed Dawood

Archives of Environmental Protection

2020

Vol. 46, no. 1

3--13

Ligninolytic enzymes are employed for the production of second-generation biofuel to minimize fuel crisis. Additionally, they play a crucial role in global carbon cycle and a variety of applications in food, agriculture, paper and textile industries. On a large scale production of ligninolytic enzymes, microorganisms can be cultured on lignocellulosic wastes. In the present study, proximate analysis including acid detergent lignin (ADL), acid detergent cellulose (ADC), acid detergent fiber (ADF) and acid insoluble ash (AIA) were performed for Platanus orientalis (chinar), Bauhinia variegata (orchid tree), Pinus roxburghii (chirpine), wheat straw and wheat husk. Platanus orientalis was selected as substrate because of higher lignin contents for the production of ligninolytic enzymes by Aspergillus flavus. Solid State Fermentation was used and Response Surface Methodology was employed for optimizing various parameters and enzymes production. Maximum production was achieved at temperature 32°C, fermentation period 120 hours, pH 4.5, inoculums size 3.5 mL, substrate mesh size 80 mm, substrate size 7 g. Maximum purification of laccase, manganese peroxidase (MnP) and lignin peroxidase (LiP) was achieved with 50%, 60% and 40% ammonium sulfate respectively and it was enhanced by gel filtration chromatography. Characterization of enzymes shows that Laccase has 35°C optimum temperature, 4.5 pH, 0.289 mM Km and 227.27 μM/ml Vmax. Manganese peroxidase has 30°C optimum temperature, 5.5 pH, 0.538 mM Km and 203.08 μM/ml Vmax. Lignin peroxidase has 30°C optimum temperature, 3 pH, 2 mM Km and 2000 uM/ml Vmax. Protein concentrations found in crude extracts and partially purified enzymes are respectively: laccase 1.78 and 0.71 mg/ml, MnP 1.59 and 0.68 mg/ml. LiP, 1.70 and 0.69 mg/ml.

First report on implementation of response surface methodology for the biodegradation of textile industrial effluents by Coniophora puteana IEBL-1

Mahmood Raja T., Asad Muhammad J., Asgher Muhammad, Zainab Tayyaba, Zafar Mudassar, Hadri Saqib H., Ali Imran, Zaman Nasib, Wattoo Feroza H.

Archives of Environmental Protection

2019

Vol. 45, no. 4

48--59

The current study was aimed to evaluate the industrial effluents biodegradation potential of an indigenous microorganism which reduced water pollution caused by these effluents. In the present study biodegradation of three textile industrial effluents was performed with locally isolated brown rot fungi named Coniophora puteana IEBL-1. Response Surface Methodology (RSM) was employed under Box Bhenken Design (BBD) for the optimization of physical and nutritional parameters for maximum biodegradation. Quality of treated effluents was checked by study of BOD, COD and analysis through HPLC. Three ligninolytic enzymes named lignin peroxidase, manganese peroxidase and laccase were also studied during the biodegradation process. The results showed that there was more than 85% biodegradation achieved for all three effluents with decrease in Biological Oxygen Demand (BOD) and Chemical Oxygen Demand (COD) below the recommended values for industrial effluent i.e. 80 mg/L for BOD and 220 mg/L for COD after optimization of nutritional parameters in the second stage. Analysis of samples through HPLC revealed the formation of less toxic diphenylamine, 3-methyldiphenylamine and N-methylaniline after treatment. The ligninolytic enzymes assays confirmed the role of lignin peroxidase (LiP), manganese peroxidase (MnP) and laccase in biodegradation process. Lignin peroxidase with higher activity has more contribution in biodegradation of effluents under study. It can be concluded through the results that Coniophora buteana IEBL-1 is a potential fungus for the treatment of industrial effluents.