Coenzyme F is synthesized via a multi-step pathway:
* 7,8-didemethyl-8-hydroxy-5-deazariboflavin synthase (FbiC) produces Coenzyme FO (also written F0), itself a cofactor of DNA photolyase (antenna). This is the head portion of the molecule.
* 2-phospho-L-lactate transferase (FbiA) produces Coenzyme F-0, the portion containing the head, the diphosphate bridge, and ending with a carboxylic acid group.
* Coenzyme F420-0:L-glutamate ligase (one part of FbiB) puts a glutamate residue at the -COOH end, producing Coenzyme F-1.
* Coenzyme F420-1:gamma-L-glutamate ligase (other part of FbiB) puts a gamma-glutamate residue at the -COOH end, producing Coenzyme F-2, the final compound (in its oxidized form). Also responsible for adding additional units.
Oxidized F can be converted to reduced F-H by multiple enzymes such as Glucose-6-phosphate dehydrogenase (coenzyme-F420) (Fgd1).