Semi pilot scale production of Lipase by Solid State Fermentation (SSF) for biodiesel production

Nema, Ashutosh and Devarai, Santhosh Kumar (2019) Semi pilot scale production of Lipase by Solid State Fermentation (SSF) for biodiesel production. Masters thesis, Indian institute of technology Hyderabad.

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Abstract

Global depletion of crude oil resources has led to an extensive exploration of alternative sources to manage the increasing demand for fuels. This is becoming a greater challenge to the future of the energy sources. Biodiesel is found to be the promising alternative to the diminishing fossil fuels as it is low emission, renewable, biodegradable and non-toxic diesel fuel. Lipases are serine hydrolases that degrade triglycerides, an attribute that treasures wide applications in production of biodiesel, paper and pulp industries, pharmaceutical and other chemical industries, etc. The most sought-after application is in the high quality and economical production of biodiesel under mild reaction conditions and simplified product separation. For the said application, microbial lipases are ideal catalysts that could effectively catalyze esterification and transesterification reactions because of their specific ability to release fatty acids from 1,3 positions of acylglycerols. In the initial part of the work the chosen fungal strain was evaluated for lipase production using a mixture of agro-industrial substrates viz. rice husk, cotton seed cake and red gram husk in various combinations at flask level. Tri-substrate mixture (rice husk, cottonseed cake and red gram husk) combined in the ratio of 2:1:1 has shown the maximum lipase activity 28.19 U/gds at optimum cultivation conditions of temperature 40 °C, moisture content 75% (v/w), pH 6.0 and initial spore concentration of 5.4 million spores per mL. Further studies were performed for scale up of lipase from flask level to lab scale using tray fermenter. Lipase activity was found to be 24.38 U/gds and 21.62 U/gds for 100 g and 1000 g substrate respectively. A novel solid substrate Prosopis juliflora (PJ) has been studied for the production of lipase using Aspergillus niger MTCC 872. Simplex centroid mixture design (SCMD) was implemented to optimize the mixture composition of the tri-substrate consisting of Prosopis juliflora (PJ), red gram husk (RGH) and cottonseed cake (CSC). Mixture taken in the ratio of 6.67:1.66:1.66 for PJ:RGH:CSC has shown highest lipase activity of 212.2 U/gds at 30°C, 7 pH and 70% initial moisture content (v/w). Sequential optimization of physical parameters was done using the central composite face-centered design (CCFD). The optimum mixture composition has shown the highest lipase activity of 269.87 U/gds at 35°C, 7 pH and 75% initial moisture content (v/w). Large scale production using 1kg substrate was carried out in tray bioreactor and the highest lipase activity of 197.057 U/gds is obtained.

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