Enantioselective oligomerization of alpha-hydroxy carboxylic acids and alpha-amino acids

• US 20030143661A1
• Filed: 05/02/2002
• Published: 07/31/2003
• Est. Priority Date: 10/29/1999
• Status: Active Grant

• Current Assignee: Novus International, Inc. (Nippon Soda Company Limited)
• Original Assignee: Novus International, Inc. (Nippon Soda Company Limited)
• Inventors: Kapila, Shubhender, Lorbert, Stephen J., Schasteen, Charles S., Forciniti, Daniel, Nam, Paul K.S., Rajesh, Mathur P.
• Granted Patent: US 6,939,693 B2
• Time in Patent Office: Days
• US Class Current: 435/68.1
• CPC Class Codes: A23K 20/105 Aliphatic or alicyclic compounds
• CPC Class Codes: A23K 20/142 Amino acids; Derivatives thereof
• CPC Class Codes: A23K 20/147 Polymeric derivatives, e.g. peptides or proteins
• CPC Class Codes: A23K 20/189 Enzymes
• CPC Class Codes: A23K 50/00 Feeding-stuffs specially adapted for particular animals
• CPC Class Codes: A23K 50/10 for ruminants
• CPC Class Codes: A23K 50/80 for aquatic animals, e.g. fish, crustaceans or molluscs
• CPC Class Codes: C07K 2/00 Peptides of undefined number of amino acids; Derivatives thereof
• CPC Class Codes: C07K 7/06 having 5 to 11 amino acids
• CPC Class Codes: C12P 21/00 Preparation of peptides or proteins single cell protein C12N1/00

Abstract

An enzymatic synthesis and composition of oligomers and co-oligomers comprised of α-hydroxy carboxylic acids and α-amino acids or peptides is disclosed. In a preferred embodiment, a α-hydroxy carboxylic acid with a specific chiral configuration is linked by an amide linkage to a α-amino acid specific with a specific chiral configuration or linked by an amide linkage to a peptide made up of α-amino acid monomers having identical chiral configurations. Proteolytic enzymes catalyze oligomerization of the α-hydroxy carboxylic acid and α-amino acid. The degree and distribution of oligomerization varies upon the type and concentrations of different reaction mixtures utilized and upon the length of allowed reaction time. The resultant oligomers may be provided to animals such as ruminants as bioavailable amino acid supplements that are resistant to degradation in the rumen and other animals such as swine, poultry and aquatic animals.

Claims

1. A process for the preparation of an oligomer, the process comprising:

forming a reaction mixture comprising an enzyme, and an enantiomeric mixture of an α

-amino acid or a derivative thereof; and

forming an oligomer from the reaction mixture, said oligomer incorporating one of the members of the enantiomeric mixture of the α

-amino acid or derivative thereof in preference to the other member.

2. A process as set forth in claim 1, wherein the enantiomeric mixture is a racemic mixture.

3. A process as set forth in claim 1, wherein the oligomer comprises an α

-amino acid or a derivative thereof comprising an oligomer of two or more α

-amino acid monomers.

4. A process as set forth in claim 3, wherein the α

-amino acid monomers comprise L-α

-amino acid monomers.

5. A process as set forth in claim 1, wherein the reaction mixture further comprises an α

-hydroxy carboxylic acid or a derivative thereof and the formed oligomer comprises an oligomer of two or more α

-amino acid monomers bonded to a residue of the α

-hydroxy carboxylic acid or a derivative thereof by an amide or ester linkage.

6. A process as set forth in claim 5 wherein said α

-hydroxy carboxylic acid or a derivative thereof is present in said reaction mixture in an enantiomeric mixture.

7. A process as set forth in claim 6 wherein one α

-hydroxy carboxylic acid enantiomer of the enantiomeric mixture is bonded to the oligomer in preference to the other enantiomer of said enantiomeric mixture.

8. A process as set forth in claim 6 wherein the α

-hydroxy carboxylic acid comprises 2-hydroxy-4-(methylthio)butyric acid or a derivative thereof.

9. A process as set forth in claim 8, wherein the α

-amino acid monomers in the oligomer comprise L-α

-amino acid monomers.

10. A process as set forth in claim 1 wherein the α

-amino acid is selected from the group consisting of methionine, lysine, or a derivative thereof.

11. A process as set forth in claim 1 wherein said enantiomeric mixture of α

-amino acids comprises methionine and lysine.

12. A process as set forth in claim 1 wherein the enzyme comprises a protease.

13. A process as set forth in claim 12 wherein the enzyme comprises a protease selected from the group consisting of papain, bromelain, cathepsin s, cathepsin b, capthesin c, and substilisin.

14. A process as set forth in claim 1 wherein the reaction mixture comprises an aqueous solution.

15. A process as set forth in claim 1 wherein the reaction mixture comprises an aqueous phase and an organic solvent phase.

16. A process as set forth in claim 1 wherein the reaction mixture comprises an α

-hydroxy carboxylic acid derivative comprising a free acid, an acid halide, an amide, an ester or an anhydride.

17. A process as set forth in claim 1 wherein the reaction mixture comprises an α

-amino acid derivative comprising a free acid, an acid halide, an amide, an ester or an anhydride.

18. A composition comprising a residue of an α

-hydroxy carboxylic acid bonded to a peptide by an amide linkage, said peptide comprising two or more α

-amino acid residues, each of said α

-amino acids being independently selected from the group consisting of α

-amino acids.

19. A composition as set forth in claim 18 wherein more than 50% of the α

-amino acid residues in the peptide are of identical chirality.

20. A composition as set forth in claim 19 wherein essentially all of the α

-amino acid residues in the peptide are of identical chirality.

21. A composition comprising a residue of an α

-hydroxy carboxylic acid bonded to a peptide by an ester linkage, said peptide comprising two or more α

-amino acid residues, each of said α

-amino acids being independently selected from the group consisting of α

-amino acids.

22. A composition as set forth in claim 21 wherein more than 50% of the α

-amino acid residues in the peptide are of identical chirality.

23. A composition as set forth in claim 22 wherein essentially all of the α

-amino acid residues in the peptide are of identical chirality.

24. A composition comprising:

25. The composition of claim 24 wherein R¹ is CH₃SCH₂CH₂—

.

26. The composition of claim 25 wherein R² is H.

27. The composition of claim 24 wherein R² is H.

28. The composition of claim 24 wherein n is at least 4 and no more than 12 and each AA is methionine.

29. The composition of claim 24 wherein n is at least 3 and no more than 5 and each AA is selected from the group consisting of methionine and lysine.

30. The composition of claim 24 wherein n is at least 2 and no more than 10 and each AA is selected from the group consisting of methionine and lysine.

31. A process of providing a food ration to an animal comprising providing the composition of claim 24 to the animal wherein the method of administration is selected from the group consisting of oral administration, eye spray, placement in ear, placement in nasal cavity, bucchal administration, sublingual administration, rectal administration and injection.

32. A process as set forth in claim 31 wherein the composition is orally administered to the animals.

33. A process as set forth in claim 32 wherein the animal is selected from the group consisting of ruminants, swine, poultry, and aquatic animals.

34. A process as set forth in claim 33 wherein the ruminant is a dairy cow or beef cattle.

35. A process as set forth in claim 34 wherein the cow is a lactating dairy cow.

36. A process as set forth in claim 33 wherein the aquatic animal is a fish or crustacean.

37. A process as set forth in claim 36 wherein the fish is selected from the group consisting of freshwater and salt water fish.

38. A process as set forth in claim 37 wherein the freshwater and salt water fish are selected from the group consisting of carp, trout, catfish, bass, sea bass, cod, and salmon.

39. A process as set forth in claim 36 wherein the crustaceans are selected from the group consisting of shrimp, crabs, lobster, and freshwater shrimp.

40. A process for providing a food ration to an animal, the process comprising:

providing an oligomer or co-oligomer composition prepared from a mixture containing an enzyme, an α

-amino acid or a derivative thereof, and optionally, an α

-hydroxy carboxylic acid or a derivative thereof,

wherein the method of administration is selected from the group consisting of oral administration, eye spray, placement in ear, placement in nasal cavity, bucchal administration, sublingual administration, rectal administration and injection.

41. A process as set forth in claim 40 wherein the animal is selected from the group consisting of ruminants, swine, poultry and aquatic animals.

42. A process as set forth in claim 41 wherein the ruminant is a dairy cow or beef cattle.

43. A process as set forth in claim 42 wherein the cow is a lactating dairy cow.

44. A process as set forth in claim 41 wherein the aquatic animal is a fish or crustacean.

45. A process as set forth in claim 44 wherein the fish is selected from the group consisting of freshwater and salt water fish.

46. A process as set forth in claim 45 wherein the freshwater and salt water fish are selected from the group consisting of carp, trout, catfish, bass, sea bass, cod, and salmon.

47. A process as set forth in claim 44 wherein the crustaceans are selected from the group consisting of shrimp, crabs, lobster, and freshwater shrimp.

48. An orally administered dietary supplement comprising a vitamin, a mineral or a nutrient coated with an oligomeric coating, said coating comprising a residue of an α

-hydroxy carboxylic acid bonded to a peptide by an amide linkage, said peptide comprising two or more α

-amino acid residues, each of said α

-amino acids being independently selected from the group consisting of α

-amino acids.

49. A process for providing an animal with a dietary supplement comprising a vitamin, mineral or nutrient, the process comprising:

coating said vitamin, mineral or nutrient with a composition to form a dietary supplement, said composition comprising a residue of an α

-hydroxy carboxylic acid bonded to a peptide by an amide linkage, said peptide comprising two or more α

-amino acid residues, each of said α

-amino acids being independently selected from the group consisting of α

-amino acids; and

orally administering the dietary supplement to the animal.

50. A process as set forth in claim 49 wherein the peptide comprises methionine.

51. A process as set forth in claim 49 wherein the animal is selected from the group consisting of ruminants, swine, poultry, and aquatic animals.

52. A process as set forth in claim 51 wherein the ruminant comprises a dairy cow or beef cattle.

53. A process as set forth in claim 52 wherein the ruminant comprises a lactating dairy cow.

54. A process as set forth in claim 51 wherein the aquatic animal comprises a fish or crustacean.

55. A process as set forth in claim 54 wherein the fish is selected from the group consisting of freshwater and salt water fish.

56. A process as set forth in claim 55 wherein the freshwater and salt water fish are selected from the group consisting of carp, trout, catfish, bass, sea bass, cod, and salmon.

57. A process as set forth in claim 54 wherein the crustaceans are selected from the group consisting of shrimp, crabs, lobster, and freshwater shrimp.

58. A process for providing an animal with a dietary supplement comprising a vitamin, mineral or nutrient, the process comprising:

preparing a mixture comprising an enzyme and at least one α

-amino acid, each α

-amino acid being present in the mixture as a free acid, acid halide, amide, ester or anhydride independently of the other,

forming an α

-amino acid oligomer in the mixture,

coating said vitamin, mineral or nutrient with the α

-amino acid oligomer to form an α

-amino acid oligomer coated dietary supplement, and

orally administering the dietary supplement to the animal.

59. A process as set forth in claim 58 wherein the animal comprises an aquatic animal selected from the group consisting of shrimp, carp, and trout.

60. A process as set forth in claim 59, wherein the α

-amino acid oligomeric coating comprises methionine.

61. A process for purifying an α

-hydroxy carboxylic acid enantiomer or derivative thereof from an enantiomeric mixture, the process comprising;

forming a reaction mixture comprising (i) an enantioselective enzyme, (ii) an enantiomeric mixture of an α

-hydroxy carboxylic acid or a derivative thereof, and (iii) an enantiomeric mixture of an α

-amino acid or a derivative thereof;

forming a reaction product from the reaction mixture, the reaction product comprising (i) an oligomer having a first α

-hydroxy carboxylic acid enantiomer of the enantiomeric mixture incorporated therein in preference to the second enantiomer, and (ii) unreacted second enantiomer; and

separating the oligomer and unreacted second enantiomer from the reaction product and each other.

62. A process as set forth in claim 61, wherein the enantioselective enzyme promotes an esterification reaction.

63. A process as set forth in claim 62, wherein the enantioselective enzyme is a lipase enzyme.

64. A process as set forth in claim 61, wherein the first enantiomer is recovered from the separated oligomer by hydrolyzing the oligomer with acid and separating the first α

-hydroxy carboxylic acid enantiomer or derivative thereof from other hydrolyzates.

65. A process as set forth in claim 64, wherein the first α

-hydroxy carboxylic acid enantiomer or derivative thereof is separated by subjection to enantioselective chromatography.

66. A process as set forth in claim 61, wherein the second unreacted α

-hydroxy carboxylic acid enantiomer or derivative thereof is recovered from the reaction mixture by rotary evaporation.

67. A process as set forth in claim 61, wherein the α

-hydroxy carboxylic acid comprises 2-hydroxy-4-(methylthio)butyric acid or a derivative thereof.

68. A process as set forth in claim 61 wherein the process further comprises recovering the first enantiomer from said separated oligomer by by hydrolyzing the oligomer with acid and separating the first α

-hydroxy carboxylic acid enantiomer or derivative thereof from other hydrolyzates; and

converting a fraction of the recovered first enantiomer to the stereochemical form of the second enantiomer thereby forming an enantiomeric mixture comprising the first and second enantiomers.

69. A process as set forth in claim 61 wherein the process further comprises recovering the unreacted second enantiomer from the reaction product;

and

converting a fraction of the separated unreacted second enantiomer to the stereochemical form of the first enantiomer thereby forming an enantiomeric mixture comprising the first and second enantiomers, at least a portion of the first enantiomer in the enantiomeric mixture being derived from separated unreacted second enantiomer.

70. A process as set forth in claim 69 wherein the enantiomeric mixture formed from the recovered second enantiomer is recycled for re-use in the formation of a reaction mixture.

71. A process as set forth in claim 69, wherein the enzyme is removed from the reaction mixture and recycled.

72. A process as set forth in claim 71, wherein the enzyme is removed from the reaction mixture by size exclusion chromatography.

73. A process as set forth in claim 69 wherein the second unreacted α

-amino acid enantiomers or derivatives thereof are separated from the reaction mixture by rotary evaporation.

74. A process for purifying an enantiomeric mixture of α

-amino acid or derivative thereof, the process comprising;

forming a reaction mixture comprising (i) an enzyme, (ii) an enantiomeric mixture of α

-amino acid or a derivative thereof, and (iii) an α

-hydroxy carboxylic acid or a derivative thereof;

forming a reaction product from the reaction mixture, the reaction product comprising (i) an oligomer incorporating a first member of the enantiomeric mixture of α

-amino acid or derivative thereof in preference to a second enantiomer of the enantiomeric mixture or derivative thereof, and (ii) unreacted second enantiomer; and

separating the oligomer and unreacted second enantiomer from the reaction product and each other.

75. A process as set forth in claim 74, wherein the first enantiomer is recovered from the separated oligomer by hydrolyzing the oligomer with acid and separating the α

-amino acid enantiomer or derivative thereof from other hydrolyzates.

76. A process as set forth in claim 75, wherein the α

-amino acid enantiomer or derivative thereof is separated by subjection to enantioselective chromatography.

77. A process as set forth in claim 74, wherein the unreacted α

-amino acid enantiomer or derivative thereof is recovered from the reaction mixture by rotary evaporation.

78. A process as set forth in claim 74, wherein the α

-amino acid is selected from the group consisting of methionine and lysine.

79. A process as set forth in claim 74, wherein the α

-hydroxy carboxylic acid comprises 2-hydroxy-4-(methylthio)butyric acid or a derivative thereof.

80. A process for purifying an α

-amino acid enantiomer or derivative thereof in an enantiomeric mixture, the process comprising;

forming a reaction mixture comprising (i) an enantioselective enzyme and (ii) an enantiomeric mixture of an α

-amino acid or derivative thereof;

forming a peptide reaction product mixture comprising (i) an oligomer or a co-oligomer having a first enantiomer of the enantiomeric mixture incorporated therein in preference to the second enantiomer of the enantiomeric mixture, and (ii) unreacted second enantiomer; and

separating the oligomer or co-oligomer and unreacted second enantiomer from the reaction product mixture and each other

81. A process as set forth in claim 80, wherein the first enantiomer is recovered from the separated oligomer or co-oligomer by hydrolyzing the oligomer or co-oligomer with acid and separating the α

-amino acid enantiomer or derivative thereof from other hydrolyzates.

82. A process as set forth in claim 81, wherein the first enantiomer or derivative thereof is separated by subjection to enantioselective chromatography.

83. A process as set forth in claim 80, wherein the unreacted α

-amino acid enantiomer or derivative thereof is recovered from the reaction mixture by rotary evaporation.

84. A process as set forth in claim 80 wherein the α

-hydroxy carboxylic acid derivative is a free acid, acid halide, amide, ester or anhydride.

85. A process as set forth in claim 84 wherein the α

-amino acid derivative is a free acid, acid halide, amide, ester or anhydride.