Delaware State University

Awards Issued Prior to January 20, 2025, Were Funded Under Previous Administrations and May Not Reflect the Priorities and Policies of the Current Administration. the Use of Lignocellulosic Biomass to Produce Biofuels and Other Chemicals Has Been Impeded by the Resistance of Plant Cell Walls to Enzymatic Deconstruction, Primarily Due to the Highly Heterogenic Polymer, Lignin. Various Pretreatment Strategies Have Been Developed to Reduce This Recalcitrance. While Encouraging Results Have Been Achieved, the Methods Suffer From TECHNO[1]ECONOMIC Challenges. Advances in Molecular Genetics Have Offered Tremendous Opportunities for Producing Enzymes in Planta That Can Modify Cell Wall Composition to Overcome Biomass Recalcitrance. However, None of These Recombinant Enzymes Have Been Applied in the Biofuel Industry on a Commercial Scale Thus Far. This Approach Also Has Its Own Set of Drawbacks Including the Metabolic Load Associated With Plants Producing Large Quantities of Enzymes, Hence Requiring More Fertilizer Inputs, and the Risk of Undesirable Effects on Normal Plant Development. Our Recent Study Revealed That Heterologous Expression of a Bacterial Lignin-degrading Peroxidase Depolymerizes Lignin and Reduces Biomass Recalcitrance in Transgenic Tobacco (nicotiana Benthamiana). This Project Will Explore the Potential of Highly Efficient Bacterial and Fungal Lignin-degrading Peroxidases to Improve Biomass Digestibility. Specific Objectives Include: 1) Generating Expression Cassettes of a Fungal Pleurotus Eryngii (VPL2) and Bacterial Amycolatopsis SP. 75IV2 DYP2 Ligninases, a Construct Containing Both Genes for Co-expression, and a Chimeric VPL2::DYP2 Construct Obtained by Swapping N- and C-terminal Domains of the Two Proteins; 2) Developing and Characterizing Transgenic Switchgrass Lines Expressing One or Two Ligninases Using Molecular and Biochemical Tools; 3) Determining Lignin Modification, Saccharification, and Fermentation Efficiency of Bioengineered Biomass; 4) Providing Training Opportunities for Undergraduate Students and Junior Level Scientists. We Intend to Develop a Strategy That Will Address the Long-standing Challenge of Biomass Recalcitrance. the Platform Established Using Switchgrass Will Serve as a Model to Engineer Other Bioenergy Grasses and Trees. the Project Will Also Provide Unique Training Opportunities for Underrepresented Minority Students in Biotechnology and Biofuels and Prepare Them for the Scientific Workforce. It Will Also Enhance Faculty Skills to Develop Grant Proposals to Seek Funding From Federal Agencies and Private Foundations. Furthermore, It Will Strengthen Collaboration Between Delaware State University With Institutions Equipped With Advanced Research Facilities.