Recombinant chondroitinase ABC I and uses thereof
The invention relates to chondroitinase ABC I and uses thereof. In particular, the invention relates to recombinant and modified chondroitinase ABC I, their production and their uses. The chondroitinase ABC I enzymes of the invention are useful for a variety of purposes, including degrading and analyzing polysaccharides such as glycosaminoglycans (GAGs). These GAGs can include chondroitin sulfate, dermatan sulfate, unsulfated chondroitin and hyaluronan. The chondroitinase ABC I enzymes can also be used in therapeutic methods such as promoting nerve regeneration, promoting stroke recovery, treating spinal cord injury, treating epithelial disease, treating infections and treating cancer.
1. An composition comprising an isolated chondroitinase ABC I comprising the amino acid sequence of SEQ ID NO: 2.
2. The composition of claim 1 , wherein the isolated chondroitinase ABC I is in a substantially purified recombinant form.
3. The composition of claim 1 , wherein the isolated chondroitinase ABC I selectively degrades chondroitin sulfate, dermatan sulfate, chondroitin 6-sulfate or chondroitin 4-sulfate.
4. The composition of claim 1 further comprising a divalent ion.
5. The composition of claim 4 , wherein the divalent ion is not zinc.
6. The composition of claim 4 , wherein the divalent ion is calcium ion, manganese ion, copper ion, iron ion, barium ion, magnesium ion or a lanthanide.
7. The composition of claim 6 , wherein the divalent ion is terbium or lutetium.
8. The composition of claim 6 , wherein the divalent ion is calcium ion in the form of CaCl 2 .
9. The composition of claim 8 , wherein the CaCl 2 is present in the composition at a concentration of 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 mM.
10. The composition of claim 8 , wherein the CaCl 2 is present at a concentration of at least 5 mM.
11. The composition of claim 10 , wherein the CaCl 2 is present at a concentration of at least 10 mM.
12. The composition of claim 4 , further comprising a pharmaceutically or physiologically acceptable carrier.
13. The composition of claim 1 further comprising an enzyme stabilizer.
14. The composition of claim 13 , wherein the enzyme stabilizer is a protease inhibitor.
15. The composition of claim 14 , wherein the protease inhibitor is AEBSF, bestatin, E64 protease inhibitor, pepstatin A or phosphoramidon.
16. The composition of claim 13 , wherein the enzyme stabilizer is a water mimic.
17. The composition of claim 16 , wherein the water mimic is glycerol or dextran.
18. The composition of claim 13 , further comprising a pharmaceutically or a physiologically acceptable carrier.
19. The composition of claim 1 further comprising a pharmaceutically acceptable carrier or a physiologically acceptable carrier.
20. The composition of claim 11 , wherein the CaCl 2 is present at a concentration of 10 mM.