Method for extracting a macrolide from biomatter

1. A method for obtaining a macrolide comprising the step of extracting macrolide-containing biomatter that is whole fermentation broth with a hydrophobic extraction solvent to obtain a solution of the macrolide in the hydrophobic extraction solvent, wherein the pH of the macrolide-containing biomatter being extracted is between 8 and about 13 and recovering said macrolide from said solution.

2. The method of claim 1 wherein the hydrophobic extraction solvent is selected from the group consisting of n-butanol, iso-butanol, C 2 -C 6 linear and branched esters of acetic acid or formic acid, C 3 -C 6 linear or branched aliphatic ketones, halogenated methanes, halogenated ethanes, and aromatic hydrocarbons that are liquid at 25° C. and that have a boiling point at atmospheric pressure less than about 150° C.

3. The method of claim 2 wherein the hydrophobic extraction solvent is selected from the group consisting of n-butanol, iso-butanol, n-butyl acetate, iso-butyl acetate, t-butyl acetate, ethyl acetate, propyl acetate, ethyl formate, butyl methyl ketone, methyl iso-butyl ketone, dichloromethane, chloroform, tetrachloromethane, dichloroethane, and toluene.

4. The method of claim 3 wherein the hydrophobic extraction solvent is ethyl acetate, iso-butyl acetate, or a mixture of these.

5. The method of claim 4 wherein the hydrophobic extraction solvent is iso-butyl acetate.

6. The method of claim 1 wherein the extraction is at a temperature between about 2° C. and about 70° C.

7. The method of claim 6 wherein the extraction is at a temperature between about 15° C. and about 35° C.

8. The method of claim 1 wherein the pH of the biomatter extracted is between 8 and about 10.

9. The method of claim 1 wherein the macrolide-containing biomatter is obtained from a microorganism selected from Streptomyces tsukubaensis, Streptomyces hygroscopicus , and Streptomyces lividans.

10. The method of claim 1 wherein the macrolide is tacrolimus.

11. The method of claim 10 wherein the pH of the biomatter is between 8 and about 10.

12. The method of claim 11 wherein the hydrophobic extraction solvent is iso-butyl acetate.

13. The method of claim 1 further comprising the steps of, after extraction;

separating the solution containing the macrolide from the extracted macrolide-containing biomatter to obtain a separated macrolide-containing solution,

concentrating the separated macrolide-containing solution to form a concentrated macrolide-containing solution,

loading the concentrated macrolide-containing solution onto a silica gel column and eluting with an eluent to obtain at least one macrolide-containing fraction that is a macrolide-containing solution, and

isolating the macrolide.

14. The method of claim 1 wherein the pH of the biomatter is between about 10 and about 13.

15. The method of claim 1 , wherein macrolide is obtained from a macrolide-containing biomatter and the yield of the macrolide is about 83 % to about 94 %.

16. The method of claim 1 comprising:

a) extracting macrolide-containing biomatter that is whole fermentation broth obtained by fermentation of a microorganism selected from the group consisting of Streptomyces tsukubaensis, Streptomyces hygroscopicus , and Streptomyces lividans with a hydrophobic extraction solvent selected form the group consisting of n-butanol, iso-butanol, n-butyl acetate, iso-butyl acetate, t-butyl acetate, ethyl acetate, propyl acetate, ethyl formate, butyl methyl ketone, dichloromethane, chloroform, tetrachloromethane, dichloroethane and toluene to obtain a solution of the macrolide in hydrophobic extraction solvent at a temperature between about 2° C. and about 70° C., wherein the macrolide is tacrolimus.

17. The method of claim 16 , further comprising the steps of:

b) isolating the macrolide-containing solution from the extracted macrolide-containing biomatter,

c) concentrating the macrolide-containing solution,

d) purifying the concentrated macrolide-containing solution on a silica gel chromatography column.

18. The method of claim 16 wherein the pH of the whole fermentation broth is between 8 and about 10.

19. The method of claim 16 wherein the hydrophobic extraction solvent is iso-butyl acetate.

20. The method of claim 16 wherein the extraction is at a temperature between about 15° C. and about 30° C.

21. A method of obtaining a macrolide from a macrolide-containing biomatter comprising the steps of:

a) extracting macrolide-containing biomatter that is whole fermentation broth obtained by fermentation of a microorganism selected from the group consisting of Streptomyces tsukubaensis, Streptomyces hygroscopicus , and Streptomyces lividans with iso-butyl acetate wherein the pH of the macrolide-containing biomatter is between 8 and about 13 to obtain a solution of a macrolide in iso-butyl acetate at a temperature between about 2° C. and about 70° C.,

b) isolating the macrolide-containing solution from the extracted macrolide-containing biomatter,

c) concentrating the macrolide-containing solution,

d) purifying the concentrated macrolide-containing solution on a silica gel chromatography column to obtain at least one macrolide-containing fraction that is a macrolide-containing solution,

e) concentrating at least one macrolide-containing fraction, wherein the macrolide is tacrolimus, and

(f) recovering said tacrolimus from step (e).

22. The method of claim 21 wherein the pH of the macrolide-containing biomatter is between 8 and about 10.

23. The method of claim 21 wherein the temperature of extraction is between about 15° C. and about 35° C.

24. The method of claim 21 , wherein macrolide is obtained from a macrolide-containing biomatter and the yield of the macrolide is about 83% to about 94%.