Metabolic Engineering and Enzyme Engineering for Microbial Limonene Production

Borkar, Shreyash Pramod (2023) Metabolic Engineering and Enzyme Engineering for Microbial Limonene Production. Masters thesis, Indian Institute of Science Education and Research Kolkata.

Text (MS dissertation of Shreyash Pramod Borkar (18MS024)) Thesis_18MS024.pdf - Submitted Version Restricted to Repository staff only Download (3MB)

Abstract

Limonene is a compound widely used in various industries, including food, pharmaceuticals, flavouring, cleaning and solvents. It is a major component of citrus essential oils and is responsible for their characteristic aroma and flavour. However, the traditional methods of limonene production from citrus fruits suffer from several limitations, such as seasonal availability, quality fluctuations, and food-grade issues. With the increasing effects of climate change, these limitations are expected to accelerate further, highlighting the need for sustainable and reliable alternative methods of limonene production. This thesis presents a solution to the challenges of traditional limonene production by focusing on microbial production in E. coli through metabolic and enzyme engineering. The study employs several strategies to achieve the objective, including improving the availability of limonene precursors, exploring orthogonal limonene precursors, enhancing the catalytic activity and substrate specificity of limonene synthase, and developing a high-throughput workflow for limonene synthase screening via enzyme engineering. Moreover, a part of the study explores the possibility of switching the enantiospecificity of the product produced by limonene synthases to expand the potential applications of microbial limonene production. Follow-up projects built on this study would provide a sustainable, stable alternative to the current limonene production methods and meet the growing global needs while mitigating environmental impact. By developing new strategies to enhance the catalytic activity and product specificity of limonene synthases, this study could also significantly impact other terpene synthases used in various industries. Overall, this thesis presents a comprehensive and innovative approach to limonene production that has the potential to revolutionise the industry and provide a sustainable and reliable source of this valuable compound for various applications.

Actions (login required)

View Item