Acetylation, through the post-transcriptional modification of histones, is a well-established regulator of gene transcription. More recent research has also identified an important role for acetylation in the regulation of non-histone proteins, both inside and outside the nucleus. As a fast (and reversible) post-translational process, acetylation allows cells to rapidly alter the function of existing proteins, making it ideally suited to biological programmes that require an immediate response to changing conditions. Using metabolic programmes as an example, the present chapter looks at how reversible acetylation can be used to regulate important enzymes in an ever-changing cellular environment.