Rare sugars hold significant promise for application in a variety of industries, including food, pharma and agriculture. Unfortunately, their further exploration is hampered by a low availability and/or high price. To solve this problem, new enzymes will be developed that enable the large-scale production of the target sugars starting from sucrose as cheap and abundant substrate. The enzymes will be optimized through semi-rational engineering, i.e. site-saturation mutagenesis of so-called hotspots in the protein sequence or structure. The generated libraries will then be screened with a robotic platform available at the Core Facility “HTS for SynBio”. The selection of hotspots will be supported by state-of-the-art computational tools for modelling, docking and data analysis.