Hydroxytyrosol (HT) is a highly potent antioxidant originating in nature as a second metabolite of plants, most abundantly in olives (Olea europaea). In the last decade, numerous research studies showed the health benefits of antioxidants in general and those of HT in particular. As olive oil is a prime constituent of the health-promoting Mediterranean diet, HT has obtained recognition for its attributes, supported by a recent health claim of the European Food Safety Authority. HT is already used as a food supplement and in cosmetic products, but it has the potential to be used as a food additive and drug, based on its anticarcinogenic, anti-inflammatory, antiapoptotic and neuroprotective activity. Nevertheless, there is a large gap between the potential of HT and its current availability in the market due to its high price tag. In this review, the challenges of producing HT using biotechnological methods are described with an emphasis on the substrate source, the biocatalyst and the process parameters, in order to narrow the gap towards an efficient bio-based industrial process.