Methods for producing alpha-cyclodextrins
AssigneeBeren Therapeutics P.B.C.
InventorsEli GROBAN, Zhihao HU, Steven PFEIFFER, Benjamin RIZKIN, Gabor BENKOVICS
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Abstract
Provided herein are methods for the enzymatic production of alpha-cyclodextrin from sucrose. The methods involve contacting sucrose with one or more enzymes to convert sucrose to amylose, followed by contacting the amylose with one or more enzymes to convert the amylose to alpha-cyclodextrin. The methods produce higher yields of alpha-cyclodextrin relative to beta-cyclodextrin, gamma-cyclodextrin, or both.
Claims30
A method of producing a composition comprising cyclodextrin, the method comprising:
- (a) contacting sucrose with an enzyme, or an enzyme mixture, capable of converting sucrose to amylose under conditions that permit the conversion of the sucrose to amylose, thereby producing amylose;
- and
- (b) contacting the amylose produced in (a) with an enzyme capable of converting amylose to cyclodextrin under conditions that permit the conversion of the amylose to cyclodextrin, thereby producing the composition comprising cyclodextrin,
- wherein the enzyme capable of converting amylose to cyclodextrin in (b) is a variant enzyme capable of producing a greater amount and/or concentration of alpha-cyclodextrin than beta-cyclodextrin, gamma-cyclodextrin, or both, relative to a wild-type enzyme capable of converting amylose to cyclodextrin,
- wherein the composition comprising cyclodextrin comprises alpha-cyclodextrin in an amount and/or concentration greater than beta-cyclodextrin, gamma-cyclodextrin, or both.
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An enzyme comprising or consisting of an amino acid sequence of any one of SEQ ID NOs:
- 1-42, or an enzyme comprising or consisting of an amino acid sequence having at least about 70% sequence identity, preferably at least about 90% sequence identity, to the amino acid sequence of any one of SEQ ID NOs:
- 1-42.
Dependencies are inferred from the claim text.