Research Projects

The booming energy demands of today and the urgent need to mitigate climate change and reduce reliance on fossil fuels have encouraged the development of sustainable biofuel production technologies. One such chemical is Isoprene, a high-value, volatile natural hydrocarbon, used to produce synthetic rubber and drop in fuel by dimerization process [1-3]. Isoprene can be industrially produced by the thermal cracking of crude oil at high temperatures which proves to be an energy-intensive, non-eco-friendly process [4,5]. Plants naturally produce Isoprene, however extraction from the plant system is tedious [4,6]. Metabolic engineering of microorganisms has emerged as an alternative innovative approach for sustainable bio-isoprene production. So far, mainly worked on genetic modification of E. coli and cyanobacterium for bio-isoprene accumulation by modulating the expression of key enzymes in the MVA and MEP pathways, aim for scalable yet eco-friendly alternatives for isoprene production [7]. Genetic modification of E. coli studies demonstrated the presence of unwanted byproducts hindered the production in large scale and yield reduction [8]. Many researchers also studying cyanobacteria as a host due to its high photosynthetic efficiency and ability to grow in extreme environments [9,10]. In our lab, research focuses on the fastest-growing cyanobacteria Synechococcus elongatus UTEX 2973, known for its relatively high isoprene yield among cyanobacteria. However, its production levels still fall short compared to those reported in E. coli and other host system [11]. To address such limitations, Yeast can be emerged as an innovative yet promising host for isoprene biosynthesis due to its advantages, such as ease of manipulation, high sugar catabolic rate, and tolerance to diverse industrial conditions [12]. Various large-scale metabolites production and high-cell-density fermentation (HCDF) of recombinant Pichia pastoris has attracted significant attention recently [13,14]. To date, no study has demonstrated bio-isoprene production from the P. pastoris host system. In this study, we propose to explore the biosynthesis of Isoprene by targeted metabolic engineering in the P. pastoris expression system. We aim to enhance isoprene production by upregulating key bottleneck enzymes that play a role in precursor synthesis, as well as downregulating the gene that deviates the substrate utilization in the MVA pathway, thus reducing the metabolic burden and conserving substrate availability for isoprene synthesis. Furthermore, we intend to perform large-scale fermentation of the modified yeast strain under optimized conditions to improvise isoprene yield. The isoprene chemical dimerization will be studied and characterized to develop it into jet fuel-grade fuel. This approach seeks to transform P. pastoris into a high-performance bio-factory for Isoprene, offering a renewable, scalable, and sustainable pathway for producing high-performance jet fuels.