Method of preparing low-iron lactoferrin
View Patent ↗The present invention relates to methods for producing low-iron Lf having improved antimicrobial activity and to a low-iron Lf having improved antimicrobial activity.
1. A method for manufacturing low-iron lactoferrin (Lf) having improved antimicrobial activity from an aqueous Lf preparation, which comprises combining the aqueous Lf preparation with an alcohol solvent and a suitable acid such that the pH is between 2.5 and about 4.5 to release the iron from the Lf, and removing the released iron, alcohol solvent and acid.
2. The method according to claim 1 wherein the suitable acid is selected from any one or more of citric acid, tartaric acid, oxalic acid, nitrilotriacetic acid, or EDTA.
3. The method according to claim 1 , wherein the suitable acid is citric acid.
4. The method according to claim 1 , wherein the suitable acid can be formed from a citric acid salt and one or more organic or inorganic acid.
5. The method according to claim 1 , wherein the suitable acid can be formed from a citric acid salt and an acid selected from HCl, H 2 SO 4 , or acetic acid.
6. The method according to claim 1 , wherein the suitable acid is formed from sodium citrate or potassium citrate and one or more organic or inorganic acid.
7. The method according to claim 1 , wherein the alcohol solvent is selected from methanol, ethanol, or propanol.
8. The method according to claim 1 , wherein the released iron, alcohol solvent and acid are removed using ultrafiltration (UF) and/or diafiltration (DF).
9. The method according to claim 1 wherein the low-iron Lf produced contains less than about 14 mg/100 g iron or about 10% iron saturation.
10. A method for manufacturing low-iron Lf and with higher antimicrobial activity than normal commercial Lf, the method comprising:
a) mixing an aqueous Lf preparation with an alcohol solvent and sufficient acid solution such that the pH is lowered to between about 2.5 to about 4.5 to release the iron from the Lf in the aqueous Lf preparation;
b) removing the released iron, acid and alcohol by UF and/or DF to achieve a processing pH>about 5.5 and a conductivity <about 2 mS in the product;
c) further processing the low-iron Lf product produced in (b) to produce a liquid or dried low-iron Lf product.
11. The method according to claim 10 wherein the low-iron Lf produced contains less than about 14 mg/100 g iron or about 10% iron saturation.
12. The method according to claim 10 wherein the low-iron Lf product in step c) is freeze-dried or spray-dried.
13. The method according to claim 10 wherein the acid is selected from citric acid, tartaric acid, oxalic acid, nitrilotriacetic acid, or EDTA.
14. The method according to claim 10 wherein the pH in step a) is lowered to between about 3.5 and about 4.5.
15. The method according to claim 12 wherein the aqueous Lf preparation in step a) is treated with acid solution for a period from about 2 hours to about 3 days to release the iron.
16. The method according to claim 10 wherein the concentration of the acid solution used in step a) is between about 5 to about 20%.
17. The method according to claim 10 wherein the alcohol solvent is added prior the addition of the acid.
18. The method according to claim 10 wherein the alcohol solvent is selected from any one or more of methanol, ethanol, propanol or similar alcohol.
19. The method according to claim 10 wherein the alcohol solvent is added in an amount of between about 0.2 and about 2.5% of the Lf preparation.
20. The method according to claim 10 wherein the processing pH is between about 3.5 and about 4.5.
21. The method according to claim 10 wherein the temperature in step a) is between about 2 and about 30° C.
22. The method according to claim 10 wherein the temperature in step b) is between about 5 and about 10° C.
23. The method according to claim 10 wherein the Lf in the aqueous Lf preparation is sourced from colostrum, skim milk or whey of bovine, human or other mammals.
24. The method according to claim 10 wherein the Lf in the aqueous Lf preparation is isolated by chromatograph, ion-exchanger and molecular-sieve means.
25. The method according to claim 10 wherein the Lf in the aqueous Lf preparation can be an undried extract or solution, or a dried powder.
26. The method according to claim 10 wherein the concentration of Lf in the aqueous Lf preparation is between about 0.01 and about 35% wt/vol.