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Bioethanol Production from Lignocellulosic Biomass using Immobilized Cellulolytic Enzymes

Abstract : The overall objective of the study was to produce bioethanol from lignocellulosic biomass by using free and immobilized xylanase, cellulase and β-1, 3-glucanase. Specifically, this study was focused on the isolation of Trichoderma citrinoviride strain AUKAR04 and it produces xylanase (55,000 IU/gds), Cellulase (385 IU/gds) and β-1, 3-glucanase (695 IU/gds) in solid state fermentation. Then the free enzymes were biochemically characterized such as effect of pH, temperature and metal ion concentration and kinetics parameters. Then the enzymes were subjected to two types of immobilization using carrier-free co-immobilization (combi-CLEAs) method and immobilized on bifunctionalized magnetic nanoparticles (ISN-CLEAs) with higher thermal stability, extended reusability and good storage stability. Liquid ammonia pretreatment removed 40% lignin from the biomass and retained 95% of glucan, 65% of xylan and 41% of arabinan in sugarcane bagasse (SCB). SCB was enzymatically hydrolyzed and converted to 87% glucose from cellulose and 74% of xylose, 64% of arabinose from the hemicelluloses which is remarkably higher than the activity of the free enzymes. Chemical and structural analysis of SCB was done by ATR-FTIR, TGA and XRD. FTIR result showed a successful pretreatment of the SCB raw material. It showed that hemicelluloses and cellulose are partially depolymerized by the action of xylanase, cellulase and β-1,3-glucanase in ISN-CLEAs. TGA studies showed that the thermal stability of the ammonia pretreated and enzymatically treated samples have improved remarkably. XRD results showed that the crystallinity index of the ISN-CLEAs treated SCB increased to 61.3±1% when compared to the ammonia-treated SCB. Mono-culture fermentation using Saccharomyces cerevisiae LGP2Y1 utilized SCB hydrolysate containing 103.8 g/L of glucose and produced 42 g/L ethanol in 36 h of fermentation. The overall metabolic yield achieved was about 79% of theoretical yield. Co-culture fermentation using Saccharomyces cerevisiae LGP2Y1 and Candida utilis ATCC 22023 utilized SCB hydrolysate containing 107.6 g/L of glucose and 41.5 g/L xylose and produced 65 g/L ethanol in 42 h of fermentation. The overall metabolic yield in co-culture fermentation achieved was about 88 % of the theoretical yield.
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Submitted on : Thursday, November 15, 2018 - 9:03:06 AM
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  • HAL Id : tel-01923141, version 1




Karthik Periyasamy. Bioethanol Production from Lignocellulosic Biomass using Immobilized Cellulolytic Enzymes. Other. Anna University (Chennai, Inde), 2018. English. ⟨NNT : 2018GREAI024⟩. ⟨tel-01923141⟩



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