Research Projects

Our recent patented and published work has established Thermobifida fusca, as an efficient thermophilic bacterial-bacterial synthetic microbial consortium (SynCONS) partner for cellulose valorisation (Joshi et al., 2023, Masakapalli Lab-Patent No 411230). In this study, T. fusca played a critical role in the saccharification of cellulose through the secretion of thermostable cellulases and showed remarkable robustness under controlled aerophilic conditions in bioreactor. Further comparative analysis of publicly available T.fusca genomes highlight its potential as a host for thermostable carbohydrate-active enzymes (CAZymes) along with the metabolic versatility to produce valuable platform chemicals (Pyruvate, acetate, succinate etc). These features render T.fusca as a promising candidate for consolidated bioprocessing (CBP), where renewable lignocellulosic biomass deconstruction and conversion can occur in a single organism. In spite of the abilities, T. fusca remains underexploited as a microbial chassis, primarily due to a lack of systems-level understanding of its metabolic pathways and physiological responses under industrially relevant conditions. Building upon our findings in the last few years, the proposed research is aimed at establishing T. fusca as a microbial platform for the dual production of thermostable cellulolytic-hemicellulolytic enzymes (Cellulases and Xylanases) and valuable platform chemicals through the integration of 13C-Fluxomics with other multi-omics approaches and rational scalable bioprocess optimization. This hypothesis will be tested through a series of interrelated objectives aimed at deciphering the metabolic and biocatalytic potential of T. fusca and achieve TRL-3 to TRL-6. The specific scientific objectives of the project are: 1. To map and quantify key metabolic pathways of Thermobifida fusca including the bifido shunt and transport systems involved in sugar assimilation and product formation using an integrated omics approach comprising of genomics, transcriptomics, metabolomics and secretome profiling. 2. To identify metabolic flux bottlenecks under different substrate conditions (glucose and xylose) using 13C-metabolic flux analysis (MFA), thereby providing a quantitative framework for understanding pathway dynamics and energy/redox balancing. 3. Metabolic Flux and Omics data guided optimisation of bioprocessing conditions and bioreactor scaleup of T.fusca cell factories for enhanced co-production of CAZymes and platform chemicals (e.g., acetate, pyruvate, lactate, and succinate) The significance of this research lies in both its fundamental and translational potential. Scientifically, it will provide unprecedented systems-level insights into the metabolism of a thermophilic, non-model actinobacterium. Technologically, the project aims to establish a low-input, high-yield microbial platform capable of converting lignocellulosic biomass directly into industrial enzymes and valuable chemical intermediates. This aligns closely with national and global mandates (e.g., BioE3) aimed at reducing reliance on fossil carbon and enhancing the efficiency of biomass-to-product pathways. In summary, this project represents a first-of-its-kind effort to integrate systems biology and scalable bioprocess optimisations for developing T. fusca into a thermophilic microbial chassis tailored for CAZymes (Cellulases and Xylanases) production and carbon valorisation. Success in this endeavour will advance both the fundamental biology of thermophiles and the applied science of sustainable biomanufacturing.