Development of an Oxygen and NAD+ Regenerating System for in the Cell-Free Synthesis of b-Lactam Antibiotics
Authors: Chas Smith, Aria Rose Neiman, Yingyi Huang
Faculty Supervisor: George Gassner
Department: Chemistry & Biochemistry
b-lactam antibiotics are the first-line treatment for common bacterial infections, but current industrial methods used for their production are both inefficient and environmentally harmful. Moreover, pharmaceutical companies, driven by more profitable exploits are producing these antibiotics at a rate that fails to meet annual demands. This has resulted globally in recurrent antibiotic shortages that negatively impact vulnerable populations unable to access treatment. To help offset this problem, our laboratory is developing a modular green-chemical approach that joins solvent tolerant bacterial and fungal enzymes in a cell-free pathway for the synthesis of b-lactam antibiotics using alcohol, oxide, aldehyde, or styrene as starting materials. Sustainable operation of this pathway requires the regeneration of NAD+ and oxygen, which are reduced to yield NADH and hydrogen peroxide as products. This poster presents the development of sensitive stopped-flow luminescence assay for the detection of hydrogen peroxide and its use in establishing the conditions needed to optimize the enzymatic regeneration of NAD+ and the electrochemical regeneration of oxygen.