METHODS FOR PRODUCING PHYCOTOXINS

• US 20120053344A1
• Filed: 09/23/2011
• Published: 03/01/2012
• Est. Priority Date: 03/24/2009
• Status: Active Grant

• Current Assignee: Proteus Corporation
• Original Assignee: Proteus Corporation
• Inventors: Lagos González, Marcelo Santiago
• Granted Patent: US 8,957,207 B2
• Time in Patent Office: Days
• US Class Current: 544/251
• CPC Class Codes: A61P 29/00 Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
• CPC Class Codes: C07D 473/00 Heterocyclic compounds containing purine ring systems
• CPC Class Codes: C07D 487/14 Ortho-condensed systems
• CPC Class Codes: C12N 1/12 Unicellular algae; Culture media therefor as new plants A01H13/00
• CPC Class Codes: C12P 17/182 Heterocyclic compounds containing nitrogen atoms as the only ring heteroatoms in the condensed system alloxazine or isoalloxazine, e.g. riboflavine C12P25/00

Abstract

Methods for producing phycotoxins from natural sources, wherein the phycotoxins have a definite compositional profile are described herein. In one embodiment, the phycotoxins are produced by cyanobacteria. In one embodiment, the phycotoxins are produced by continuously culturing cyanobacteria under strictly controlled conditions in order to produce a definite compositional profile. In another embodiment, organic nutrients are added to the culture that allows for higher concentrations of neosaxitoxin and saxitoxin or gonyaulatoxins 2 and 3 per weight of the algae. The phycotoxins are isolated primarily from the bacteria but can also be isolated from the culture medium. In one embodiment, the cyanobacteria produce only neosaxitoxin and saxitoxin in a ratio of about 6:1, 5:1, 4:1, or 3:1. In a preferred embodiment, the amount of saxitoxin is less than 20% by weight of the total amount of neosaxitoxin and saxitoxin produced. In another embodiment, the cyanobacteria produce only GTX2 and GTX 3.

Claims

1. A method for the continuous production of phycotoxins from cyanobacteria, the method comprising

inoculating a culture medium with the cyanobacteria, wherein the culture medium comprises one or more organic nutrients that stimulate production of saxitoxin, neosaxitoxin, gonyaulatoxin 2, gonyaulatoxin 3, and combinations thereof in the cyanobacteria;

and

harvesting the cyanobacteria to isolate the phycotoxins.

2. The method of claim 1, wherein the cyanobacteria is selected from the group consisting of Cylindrospermopsis raciborskii;

Anphanizomenon flos-aquae;

Aphanizomenon (APh) issatschenkoi (usaceb) Proskina-Lavrenco;

Aphanizomenon gracile (Lemm) Lemm;

Anabaena circinalis;

Lyngbya wollei; and

combinations thereof.

3. The method of claim 2, wherein the cyanobacteria is Aphanizomenon gracile (Lemm) Lemm.

4. The method of claim 1, wherein the organic nutrients are selected from the group consisting of arginine, methionine, allantoic acid, and combinations thereof.

5. The method of claim 4, wherein the micronutrients comprise arginine, methionine, and allantoic acid.

6. The method of claim 1, wherein the cyanobacteria produce only neosaxitoxin and saxitoxin.

7. The method of claim 6, wherein the ratio of neosaxitoxin to saxitoxin is approximately 6:

1.

8. The method of claim 6, wherein the ratio of neosaxitoxin to saxitoxin is approximately 5:

1.

9. The method of claim 6, wherein the ratio of neosaxitoxin to saxitoxin is approximately 4:

1.

10. The method of claim 6, wherein the ratio of neosaxitoxin to saxitoxin is approximately 3:

1.

11. The method of claim 1, wherein the cyanobacteria produce only gonyaulatoxin 2 and gonyaulatoxin 3.

12. Neosaxitoxin produced by the method of claim 1.

13. The neosaxitoxin of claim 12, wherein the structure is

14. Saxitoxin produced by the method of claim 1.

15. The saxitoxin of claim 14, wherein the structure is