Enzymatic Synthesis of a Vitamin B-9 Derivative for More Cost-Efficient Biochemical Assays
Dr. Jeffrey Pruet
Arts and Sciences
Noah Moriarty (0000-0002-6022-9769)
Vitamin B-9, and its polyglutamated derivative, is a key component of biochemical assays aimed at testing novel therapeutic agents. Therefore, there is a need to produce this essential molecule in an efficient manner. Due to inherent difficulties in organic synthesis of a Vitamin B-9 derivative, in addition to the significantly high commercial price of this compound, a new synthetic route has been designed based on biocatalysis. This design utilized the enzyme folyl polyglutamate synthase (FPGS) for an enzymatic polyglutamation reaction on the commercially available 5-methyltetrahydrofolate (5MeTHF) to make polyglutamated Vitamin B-9. Production of the FPGS enzyme involved protein expression from a designed plasmid originating in b. licheniformis, followed by cell lysis and protein purification. The isolated FPGS enzyme was then tested for its ability to add additional glutamates to 5MeTHF through an ATP-promoted process. The viability of this biochemically produced Vitamin B9 derivative is being assessed using a fluorescence-based assay.
Moriarty, Noah; Burkhart, Grace; Bennett, Zachary; Markovich, Sam; and Pruet, Jeffrey, "Enzymatic Synthesis of a Vitamin B-9 Derivative for More Cost-Efficient Biochemical Assays" (2021). Symposium on Undergraduate Research and Creative Expression (SOURCE). 952.