αgalactosidases and methods of using them
View Patent ↗A thermostable alpha-glycosidase derived from various Thermococcus, alcaliphilus AEDII12RA is disclosed. The enzymes are produced from native or recombinant host cells and can be utilized in the food processing industry.
1. A method for hydrolyzing α-glycosidic bonds capable of being hydrolyzed by a polypeptide having an α-galactosidase activity comprising:
contacting a compound having the α-glycosidic bond with an effective amount of a polypeptide having at least a 95% amino acid identity to an amino acid sequence set forth in SEQ ID NO:4, having at least a 95% amino acid identity to an amino acid sequence as set forth in amino acids 1 to 364 of SEQ ID NO:4, or enzymatically active fragments thereof, wherein the polypeptide has α-galactosidase activity.
2. The method according to claim 1 wherein the polypeptide has the amino acid sequence as set forth in SEQ ID NO: 4 or an amino acid sequence as set forth in amino acids 1 to 364 of SEQ ID NO:4.
3. The method according to claim 1 wherein the polypeptide is recombinantly produced or is a synthetic enzyme.
4. The method according to claim 1 wherein the compound having the α-glycosidic bond is raffinose.
5. The method according to claim 4 wherein the raffinose is in raw beet sugar.
6. The method according to claim 1 wherein the compound comprises raffinose, stachyose, verbascose, or a combination thereof.
7. The method according to claim 6 wherein the compound is contained in a member of the lentil or bean family, or both.
8. The method according to claim 1 wherein the contacting is at a temperature of about 85° C.
9. The method according to claim 1 wherein the contacting is at a pH of about 9.5.
10. The method according to claim 1 wherein the contacting is at a temperature of about 85° C. and a pH of about 9.5.
11. The method according to claim 1 wherein the α-glycosidic bond is an α-1,6 galactosyl bond or an α-1,6 galactosidic bond.
12. The method according to claim 1 wherein the polypeptide comprises a sequence having at least 50 amino acids identical to a contiguous region of amino acids 1 to 364 of SEQ ID NO:4 or SEO ID NO:4.
13. The method according to claim 1 , wherein the polypeptide has at least 97% amino acid identity to SEQ ID NO: 4.
14. The method of claim 1 , wherein the compound is a food.
15. The method of claim 1 , wherein the compound having the α-glycosidic bond comprises a saccharide and the polypeptide is contacted with the saccharide under conditions which facilitate the hydrolysis of the saccharides, thereby catalyzing the hydrolysis of saccharides.
16. The method of claim 15 , wherein the saccharide is a polysaccharide or an oligosaccharide.
17. The method of claim 16 , wherein the polysaccharide or oligosaccharide is found in a legume.
18. The method of claim 16 , wherein the polysaccharide or oligosaccharide is raffinose, stachyose, verbascose, or a combination thereof.
19. A method for hydrolyzing α-glycosidic bonds in a compound comprising raffinose, stachyose, or verbascose comprising contacting the compound with an effective amount of a polypeptide having: an amino acid sequence having at least a 95% amino acid identity to an amino acid sequence set forth in SEQ ID NO: 4; an amino acid sequence having at least a 95% amino acid identity to an amino acid sequence as set forth in amino acids 1 to 364 of SEQ ID NO: 4; a sequence set forth in SEQ ID NO: 4; or enzymatically active fragments thereof, wherein the polypeptide has α-galactosidase activity.
20. The method of claim 19 , wherein the polypeptide has at least 97% amino acid identity to SEQ ID NO: 4.
21. The method of claim 19 wherein the contacting is at a temperature of about 85° C.
22. The method of claim 19 wherein the contacting is at a pH of about 9.5.
23. The method of claim 19 wherein the contacting is at a temperature of about 85° C. and a pH of about 9.5.
24. The method of claim 19 , wherein the polysaccharide or oligosaccharide is found in a legume.