The results of this study also demonstrate the generation of a Gram-positive operon that is functional in the synthesis of PDC and ADH proteins that actively convert pyruvate to ethanol. Our construction and expression of this G+ PET operon using the SvPDC and G. stearothermophilus ADH have demonstrated that high-level synthesis of active PDC and ADH no longer limits ethanol production in Grampositive biocatalysts. Optimization of the conversion of biomass to pyruvate and minimization of the pathways that compete for available pyruvate are now needed to ensure high yields of ethanol in recombinant Gram-positive bacteria.