Computational metabolic engineering of a cell-free enzymatic biosystem for psilocin production from L-tryptophan derived from ODE kinetic modeling, molecular docking, and parametric optimization. Seven testable hypotheses: (1) Methionine stoichiometric threshold (2:1 Met:Trp), (2) PsiH rate-limiting enzyme identification, (3) MAT product inhibition in SAM cycling, (4) pH optimization as multi-enzyme compromise, (5) Temperature-dependent enzyme stability, (6) NADPH regeneration sufficiency, (7) System scalability predictions. Generated via LiteFold Platform by Rosalind Scientific Agent.