Spray-dried, high-purity, neutral human milk oligosaccharides (HMOs) from microbial fermentation
The present application discloses a simple process for the purification of neutral human milk oligosaccharides (HMOs) produced by microbial fermentation. The process uses a combination of cationic ion exchanger treatment, an anionic ion exchanger treatment, and a nanofiltration and/or electrodialysis step, which allows efficient purification of large quantities of neutral HMOs at high purity. Contrary to the purification currently used in fermentative production of neutral HMOs, the presented process allows the provision of HMOs without the need of a chromatographic separation. The so purified HMOs may be obtained in solid form by spray drying, as crystalline material or as sterile filtered concentrate. The provided HMOs are free of proteins and recombinant material originating from the used recombinant microbial strains and thus very well-suited for use in food, medical food and feed (e.g. pet food) applications.
1. A spray-dried neutral human milk oligosaccharide (HMO) product comprising a neutral HMO of interest and one or more contaminants, wherein said spray-dried neutral HMO product is obtained from a microbial fermentation broth, wherein the microorganism is a recombinant microorganism engineered to produce the neutral HMO of interest, wherein the purity of the neutral HMO of interest is greater than or equal to 80 percent, and wherein the one or more contaminants comprise 3-fucosyllactose, difucosyllactose and lactose, and wherein said spray-dried neutral HMO product is free of recombinant DNA material, and wherein the neutral HMO of interest is 2′-fucosyllactose.
2. The spray-dried neutral HMO product according to claim 1 , wherein the spray-dried neutral HMO product is free of DNA derived from the recombinant microorganism.
3. The spray-dried neutral HMO product according to claim 1 , wherein the spray-dried neutral HMO product is free of proteins derived from the recombinant microorganism.
4. The spray-dried neutral HMO product according to claim 1 , wherein the spray-dried neutral HMO product is free of proteins.
5. The spray-dried neutral HMO product according to claim 1 , wherein the spray-dried neutral HMO product has a conductivity of less than 1 mSi/cm at a 300 g/l solution.
6. The spray-dried neutral HMO product according to claim 1 , wherein said spray-dried neutral HMO product is obtainable by filtration of a purified solution comprising the HMO produced from fermentation using a recombinant microorganism, through a 3 kDa filter, followed by spray-drying the HMO product.
7. The spray-dried neutral HMO product according to claim 1 , wherein the spray-dried neutral HMO product is obtained by purifying the neutral HMO from a fermentation broth obtained by microbial fermentation, the fermentation broth comprising the neutral HMO, biomass, medium components and contaminants, wherein the purity of the neutral HMO in the fermentation broth is <80 percent, wherein the fermentation broth is applied to the following purification steps:
i) separating the microbial biomass from the fermentation broth;
ii) subjecting the separated fermentation broth obtained in step i) to a cation exchanger treatment or to an anion exchanger treatment to obtain a solution containing the neutral HMO;
iii) subjecting the solution obtained in step ii) to the cation exchanger treatment or anion exchanger treatment not used in step ii);
iv) subjecting the solution obtained from step iii) to nanofiltration or electrodialysis of reverse osmosis or partial vacuum evaporation to obtain a purified solution of the neutral HMO,
wherein the neutral HMO in the purified solution obtained after step iv) has a purity of ≥80%.
8. A method for treating a gastrointestinal disorder comprising administering to a patient in need thereof the spray-dried neutral HMO product of claim 1 .