ME: Metabolic Engineering of Bacillus for Enhanced Product Yield

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The objective of the proposed research is the metabolic engineering of Bacillus subtilis for reduced acid formation and enhanced product yield. Preliminary work in both E. coli and B. subtilis showed that the addition of citrate allowed enhancements in growth and product formation. Acid formation was also substantially lowered. These improvements were tied to the level of pyruvate in the cell. Direct deletion of the pyk gene, NMR experiments, and flux modeling confirmed these results. The specific objectives of this research include: (1) improving the performance of B. subtilis which shows reduced growth with the pyk mutant, and (2) utilization of these results for the improvement of toxin production in Bacillus thuringiensis.