10123152Milk-free production of recombinant Osteopontin to enable supplementation in baby, elderly, and sports nutritionClosedCollaborative R&DInnovate UKOsteopontin (OPN) is a protein found at high levels in human milk and in many tissues including bone, skin, and kidneys. OPN is interesting for nutritional and medical uses as it plays a diverse and critical role: it's involved in immune system development, immune cell attraction, wound healing, bone formation, modulation of bone and tissue mineralisation, and prevention of calcification. Several food manufacturers add OPN to infant formulas and baby foods to better match human milk composition following studies establishing benefits for immune system development; recent research also indicates benefits for elderly and sports nutrition. **Problem:** OPN for human consumption can only be produced by purifying OPN from cow's milk, where it is present in low concentrations of 18 milligram per litre - 8 times less than in human milk. This means OPN production is extremely expensive and unsustainable: costs £3,000/kg and needs 40,000 L of water for a single gram! This makes it difficult for scientists to research its health benefits and for food companies to add it in their products. **Expertise:** Better Dairy is a London-based SME specialising in a method of food production called precision fermentation (PF) to make dairy proteins without animals, reducing water use and greenhouse gas emissions by over 80%. The Biorenewables Development Centre is a research organisation associated with the University of York, which helps companies optimise production processes. In PF, microorganisms like yeast are genetically programmed to make compounds usually made via other sources like cows, hence the compound is 'animal-free' . When the microorganisms are fermented, they produce the desired compound - just like when yeast is fermented, it produces alcohol to make beer. PF has been used for decades to make commonly eaten products like vitamins and natural flavours, and medicines like insulin for diabetic people. **Innovation:** In this project, we will produce OPN from yeast with post-translational modifications (PTMs) for the first time, using PF. OPN with PTMs is challenging to produce in this way because PTMs are dependent on a lot of different genetic pathways which have to be encoded in the microorganism. We will make new genetic modifications and optimise the fermentation conditions to achieve a high yield of OPN production at a small scale, showing that it is feasible to make OPN production sustainable and affordable. This will enable food companies to add OPN in their products and scientists to research more health benefits.