Characterization and industrial application of oxidative/hydrolytic enzymes of Pleurotus florida (Mont.) Sing. and Calocybe indica (Pur. & Chan.)

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dc.contributor.advisorSharma, Shivani
dc.contributor.authorManmeet Kaur
dc.date.accessioned2023-12-12T16:22:20Z
dc.date.available2023-12-12T16:22:20Z
dc.date.issued2023
dc.description.abstractPleurotus florida and Calocybe indica are the edible mushrooms that have been widely accepted due to their ability to grow on a variety of substrates and possess the potential to produce oxidative and hydrolytic enzymes. The present work involved the study and characterization of oxidative/hydrolytic enzymes of P. florida and C. indica for their potential applications in alcohol fermentation, dye decolorization, biobleaching and mushroom production. The oxidative and hydrolytic enzymes extracted from P. florida and C. indica were estimated at different mycelial growth stages for enzyme production. The intracellular and extracellular enzyme activity of P. florida and C. indica increased with time and maximum activity of lignocellulolytic enzymes was found to be on 14th day of incubation. In both mushrooms, ligninolytic enzyme activity increased during substrate colonisation but quickly decreased during fruiting body development. On the other hand, P. florida and C. indica showed relatively modest hydrolase activity during substrate colonisation. The activity of hydrolytic enzymes increased dramatically during primordial formation and peaked at the fruiting body stage. The purification of laccase, lignin peroxidase, manganese peroxidase and endoxylanase from the fruiting body of P. florida resulted in maximum purification fold of 21.49, 17.73, 16.81 and 12.78 with yield of 24.98%, 20.60%, 19.53% and 14.86%, respectively. However, the purification of laccase, lignin peroxidase, manganese peroxidase and endoxylanase from the fruiting body of C. indica resulted in maximum yield of 21.36%, 18.92%, 17.37% and 16.62% with purification fold of 25.6, 25.12, 20.88 and 19.97, respectively. The SDS-PAGE of the purified enzyme laccase and lignin peroxidase isolated from P. florida showed a single prominent band at 66 kDa and 55 kDa respectively. The SDS-PAGE of the purified enzyme laccase and lignin peroxidase isolated from C. indica showed a single prominent band at 64 kDa and 47 kDa, respectively. FTIR spectra of the purified enzymes indicated a secondary structure that reflected the amide I and amide II bands, respectively. The pretreatment of paddy straw and wheat straw with P. florida, C. indica and their ligninolytic enzyme resulted in decrease in lignin and hemicellulose content, respectively during the incubation period of 30 days. However, the decrease in cellulose content occurred during the pretreatment of substrates with fungus while relative increase in cellulose content during the treatment with ligninolytic enzymes was observed over the incubation period of 30 days. Under optimized conditions, saccharification of biologically pretreated paddy and wheat straw resulted in release of 0.415 and 0.389 g/gds reducing sugars, respectively. The fermentation of biologically pretreated and commercial cellulase saccharified paddy and wheat straw hydrolysate resulted in 0.129 and 0.119 g/g ethanol, respectively. The crude enzyme extract of P. florida and C. indica were able to degrade RBBR (25.74 %, 22.06%) and Amido Black (19.76%, 17.58%) dyes maximally after 96 hours of incubation at 30°C and pH 7.0. The paddy and wheat straw was treated with a ligninolytic crude enzyme, which stimulated faster mushroom growth and fructification. The present study thus revealed that expression of biosynthetic potential of P. florida and C. indica is highly dependent on the method of fungi cultivation. These ligninolytic enzymes showed a unique profile in terms of versatility, greenness, pollutant removal and efficiency in lignin degradation for the exploitation and valorization of agro-wastes.
dc.identifier.citationManmeet Kaur (2023). Characterization and industrial application of oxidative/hydrolytic enzymes of Pleurotus florida (Mont.) Sing. and Calocybe indica (Pur. & Chan.) (Unpublished Ph.D. Dissertation). Punjab Agricultural University, Ludhiana, Punjab, India.
dc.identifier.urihttps://krishikosh.egranth.ac.in/handle/1/5810202750
dc.keywordsBioethanol
dc.keywordsCalocybe indica
dc.keywordsCharacterization
dc.keywordsEnzyme purification
dc.keywordsHydrolytic enzymes
dc.keywordsOxidative enzymes
dc.keywordsPleurotus florida
dc.keywordsSaccharification
dc.language.isoEnglish
dc.pages200
dc.publisherPunjab Agricultural University
dc.research.problemCharacterization and industrial application of oxidative/hydrolytic enzymes of Pleurotus florida (Mont.) Sing. and Calocybe indica (Pur. & Chan.)
dc.subMicrobiology
dc.themeCharacterization and industrial application of oxidative/hydrolytic enzymes of Pleurotus florida (Mont.) Sing. and Calocybe indica (Pur. & Chan.)
dc.these.typePh.D
dc.titleCharacterization and industrial application of oxidative/hydrolytic enzymes of Pleurotus florida (Mont.) Sing. and Calocybe indica (Pur. & Chan.)
dc.typeThesis
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