Mutualisms require protection from non-reciprocating exploiters. Pseudomyrmex workers that engage in an obligate defensive mutualism with Acacia hosts feed exclusively on the sucrose-free extrafloral nectar (EFN) that is secreted by their hosts, a behaviour linking ant energy supply directly to host performance and thus favouring reciprocating behaviour. We tested the hypothesis that Acacia hosts manipulate this digestive specialisation of their ant mutualists. Invertase (sucrose hydrolytic) activity in the ant midguts was inhibited by chitinase, a dominant EFN protein. The inhibition occurred quickly in cell-free gut liquids and in native gels and thus likely results from an enzyme–enzyme interaction. Once a freshly eclosed worker ingests EFN as the first diet available, her invertase becomes inhibited and she, thus, continues feeding on host-derived EFN. Partner manipulation acts at the phenotypic level and means that one partner actively controls the phenotype of the other partner to enhance its dependency on host-derived rewards.