Team:IIT Madras/Project

Project Sunscreen
The energy advantage provided to cells due to Proteorhodopsin(PR) expression increases the growth rate of PR expressing cells. This difference in growth rate has been hypothesized to be used for screening transformed cells. We propose a plasmid vector for screening, where the antibiotic resistance gene is replaced by Proteorhodopsin generator. The following is the modified protocol for screening:

• Clone the part of interest into the pSB1Pc cloning vector’s MCS.
• Transform bacterial cells (using standard protocol).
• Plate cells on Minimal media agar containing Retinal (10uM).
• Place the plate under green light (525 nm) and incubate at 37 C.
• Pick transformed colonies after 12 hours of incubation.

Project Artemis
Artemis is the Greek Goddess of Light, and is known as the “Protector of the Vulnerable”. Our team has designed an Expression vector based on the Carbon Stress Buster device, for improving yield of Recombinant protein in substrate limiting conditions. In times of carbohydrate stress, there would be low availability of energy for Protein production. This is a serious issue in Bioprocess industry. Recombinant Production drops with exhaustion of carbohydrates in the media. Our team has combined the effects of Carbon stress induced promoter PcstA, which is triggered in carbohydrate starvation, with the energy advantage provided by Proteorhodopsin. Thus, the recombinant protein needs to be cloned after the PcstA promoter. And voila, improved recombinant protein yield even when the carbohydrate levels in the media drops. In other words, more protein produced for lesser input of carbohydrates, and thereby more economical.

Hyaluronic Acid and Curdlan Synthesis
Hyaluronic Acid (HA) is an industrially important biopolymer, whose production has been shown to increase in situations where glycolysis is inhibited inside the cell. Inhibition of glycolysis redirects the cellular resources towards HA synthesis, but in turn decreases cell viability. Under conditions of glyolysis inhibition, our construct will help provide the energy required to maintain cell viability as close to normal as possible. This emerges as a novel method to optimize carbon source flux within the cell, by directing as much of it as possible towards the exopolysaccharide synthesis of HA.
Exopolysaccharide biosynthesis is a process in which the ATP demand is a significant proportion of total cellular ATP demand. One such exopolysaccharide of high industrial relevance is Curdlan or beta-1,3-glucan. Therefore our construct, which enhances the ATP production in the cell, is expected to supplement the ATP requirement during the synthesis of Curdlan and increase its rate of production for commercial purposes.