We calculate gauge instanton corrections to a class of higher derivative string effective couplings introduced in [1]. We work in Type I string theory compactified on K3×T2 and realise gauge instantons in terms of D5-branes wrapping the internal space. In the field theory limit we reproduce the deformed ADHM action on a general Ω-background from which one can compute the non-perturbative gauge theory partition function using localisation. This is a non-perturbative extension of [1] and provides further evidence for our proposal of a string theory realisation of the Ω-background.