Dynamically Regulated Toehold Switch Genetic Circuits For Tailoring And Controlling Hyaluronic Acid Molecular Weights In Lactococcus Lactis
Date11th Dec 2023
Time03:30 PM
Venue BT Seminar Hall
PAST EVENT
Details
Synthetic Biology has allowed us to design engineering-driven approaches for programming of cellular functions that can build transformative strategies in advancing metabolic engineering. In this study we report the construction, characterization and employment of a library of synthetic Lactococcus specific Toehold Switch genetic circuits. These switches display dynamic ranges comparable to transcription-factor-dependent systems and provide the added advantages of negligible leakage. Furthermore, they are employed as Boolean logic function which can combine inputs of different tunable promoters and dynamically provide a single tunable output. We have utilized these Switches with diverse ON/OFF ratios for solving some of the challenges faced in the Metabolic Engineering of Hyaluronic Acid (HA) in Lactococcus lactis. Multiple studies have already detailed achieving high titres of HA in genetically engineered hosts. However, the recombinant production of controlled molecular weight HA at high titres, still remains a challenge. Our earlier studies have reported that HA molecular weight strongly correlates with the glycolytic flux and ratio of HA-precursors in the pathways. The current strategy involves AND-gate genetic logic circuits consisting of Toehold Switches under inducible and dynamically regulated promoters. These circuits can have the capabilities to sense the internal state of the bacteria and autonomously provide the information for the production and control of HA of different molecular weights. The incorporation of these SynBio tools has enabled us to enhance the HA molecular weight by over 40% for the control strain. The benefit of this design can be coupled with established bioprocess strategies to achieve an array of HA molecular weights of commercial significance.
Speakers
Mr. Sandeep Kumar Panda (BT16D032)
BIOTECHNOLOGY