Compositions and methods comprising the use of
An isolated and/or purified α-glucanotransferase from Exiguobacterium acetylicum , recombinantly engineered variants thereof, active fragments thereof, synthetic nucleic acids encoding the α-glucanotransferase and variants thereof, host cells comprising the synthetic nucleic acids, and compositions comprising the α-glucanotransferase are provided. Methods of using the compositions include the manufacture of oligosaccharides.
1. A composition comprising at least:
(i) an isolated GH70 subfamily 4 α-glucanotransferase comprising an amino acid sequence that is at least 95% identical to amino acids 33-736 of SEQ ID NO:7 or amino acids 22-726 of SEQ ID NO:11, and
(ii) a glucooligosaccharide with α-(1→4) and α-(1→6) glycosidic linkages.
2. The composition of claim 1 , wherein the α-glucanotransferase comprises an amino acid sequence that is at least 98% identical to amino acids 33-736 of SEQ ID NO:7 or amino acids 22-726 of SEQ ID NO:11.
3. The composition of claim 1 , wherein the α-glucanotransferase comprises an amino acid sequence that is at least 99% identical to amino acids 33-736 of SEQ ID NO:7 or amino acids 22-726 of SEQ ID NO:11.
4. The composition of claim 1 , wherein the α-glucanotransferase comprises an amino acid sequence that is at least 99.5% identical to amino acids 33-736 of SEQ ID NO:7 or amino acids 22-726 of SEQ ID NO:11.
5. The composition of claim 1 , wherein the α-glucanotransferase comprises amino acids 33-736 of SEQ ID NO:7 or amino acids 22-726 of SEQ ID NO:11.
6. The composition of claim 1 , wherein the composition is in a lyophilized powder form, an encapsulated form, a coated form, a granulated form, or a liquid formulation.
7. The composition of claim 1 , further comprising a diluent.
8. The composition of claim 1 , wherein the α-glucanotransferase comprises said amino acid sequence that is at least 95% identical to amino acids 33-736 of SEQ ID NO:7.
9. The composition of claim 1 , wherein the α-glucanotransferase comprises said amino acid sequence that is at least 95% identical to amino acids 22-726 of SEQ ID NO:11.
10. A method of producing saccharide products, said method comprising contacting the α-glucanotransferase according to claim 1 with a substrate comprising a maltooligosaccharide or maltodextrin under conditions of pH 3-10 and 30-70° C. to produce at least one glucooligosaccharide with α-(1→4) and α-(1→6) glycosidic linkages.
11. The method of claim 10 , wherein the substrate comprises malto-tetraose, malto-pentaose, malto-hexaose, or malto-heptaose.
12. The method of claim 10 , wherein the substrate is comprised in a beverage or food, wherein said glucooligosaccharide with α-(1→4) and α-(1→6) glycosidic linkages is produced in the beverage or food in situ.
13. The method of claim 12 , wherein the substrate is comprised in said beverage.
14. The method of claim 12 , wherein the substrate is comprised in said food.
15. The method of claim 10 , wherein the α-glucanotransferase comprises said amino acid sequence that is at least 95% identical to amino acids 33-736 of SEQ ID NO:7.
16. The method of claim 10 , wherein the α-glucanotransferase comprises said amino acid sequence that is at least 95% identical to amino acids 22-726 of SEQ ID NO:11.