Life Sciences & Biotechnology
Title : | Engineering of PETase-silaffin fusion for diatom-mediated biodegradation of polyethylene terephthalate (PET) |
Area of research : | Life Sciences & Biotechnology |
Focus area : | Enzyme Engineering |
Principal Investigator : | Dr. Gayatri A Dave, Charotar University of science And Technology, Changa, Gujarat |
Timeline Start Year : | 2024 |
Timeline End Year : | 2027 |
Contact info : | gayatridave.bt@charusat.ac.in |
Details
Executive Summary : | since the discovery of IsPETase in 2016, researchers have been working on over-expressing PETase to improve structural and functional stability. However, to maintain bacteria in the natural environment, specific carbon sources need to be included on-site. Recently, photosynthetic microbes have gained attention as hosts for engineering PETase. Two previous studies have successfully expressed PETase in photosynthetic microalgae and seawater-containing mediums. This proposal aims to address the shortcomings of previous studies, which focused on intracellular expression and reduced catalytic stability. The research will focus on engineering surface active PETase in organic silaffin matrix using T. pseudonana, a marine-centric diatom, due to its availability of whole genome sequence data and ease in cultivation at environmental temperature. CRIsPR-based gene editing will be used for operational ease, targeting the delta-5 elongase gene for PETase knock-in. The engineered vector will be transformed into Thalassiosira pseudonana, and protein expression and validation will be confirmed through sDs-PAGE and Western blot analysis. The project aims to engineer PETase on the silaffin matrix of Thalassiosira pseudonana, which could be used for bioremediation of microplastics in water mesocosms. |
Co-PI: | Dr. Bragadish Dandapani Iyer, Charotar University of science & Technology, Changa,Gujarat |
Total Budget (INR): | 45,08,196 |
Organizations involved