Optimizing a Synthetic Microbial Consortium using lux and agr Quorum Sensing System in Gram Negative and Gram Positive Bacteria

Chatterjee, Debdeep (2022) Optimizing a Synthetic Microbial Consortium using lux and agr Quorum Sensing System in Gram Negative and Gram Positive Bacteria. Masters thesis, Indian Institute of Science Education and Research Kolkata.

Text (MS dissertation of Debdeep Chatterjee (17MS003)) 17MS003_Thesis_file.pdf - Submitted Version Restricted to Repository staff only Download (4MB)

Abstract

For scaling up the production of biofuels, a stable and tunable microbial consortia population would be helpful. However, developing microbial co-cultures can be challenging. This project aimed to optimize a synthetic microbial consortium using the quorum sensing technique. Gram-negative (E. coli) bacteria have been used to design this microbial consortium that will be controlled by the Quorum-sensing system (QS) and eventually produce cellulases. Two separate QS systems have been used to engineer this microbial consortium, one is the agr QS system discovered initially from a gram-positive strain S. aureus, and the other is the lux QS system initially discovered from gram-negative strain Vibrio fischeri to control the luminescence. The lux and agr QS were established, and their expression was regulated using a library of promoters in E. coli. The cloning and pathway assembly was based on an adapted modular assembly method involving different assembly levels through the golden gate assembly method. This establishment of these QS systems will further allow us to study the synergistic expression of cellulase.

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