Plastic pollution is a global environmental, social, and economic threat and renewable alternatives have been sought upon. In nature, poly-3-hydroxybutyrate (PHB) occurs naturally as carbon storage in various microbes and can be used as bioplastics. However, large-scale production of PHB nowadays requires economically competitive carbon sources and is not affordable nor sustainable in the long term. Acetate is a renewable and economical carbon source that can be derived from CO2 as by-product of industrial processes through energy-efficient chemical processes. Our research uses acetate as a sole carbon source to produce PHB with Vibrio natriegens. This bacterium is an emerging chassis of synthetic biology, due to its rapid growth, extremely high substrate uptake and exceptional protein expression capabilities. We used adaptive laboratory evolution and genome engineering to optimise the metabolic pathways in V. natriegens for acetate utilisation and PHB production. The growth rate of V. natriegens on acetate increased and a greater tolerance to higher levels of acetate was observed after 1000 consecutive generations. V. natriegens was able to produce PHB from acetate as a sole carbon source after upregulating the native phaBAC operon. Overall, V. natriegens shows promising results to be a candidate for microbial PHB production from acetate.