Compositions and methods for enzymatic production of polyphenol aglycone compounds
AssigneeShreenika Pioneering Inc.
InventorsJose A. AMAYA, Jeffrey EHRHARDT, Jingyi LI, Drew Taylor WAGNER
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Abstract
The present disclosure provides compositions comprising a glycoside hydrolase and one or both of hesperidin and hesperetin, wherein an active site of the glycoside hydrolase comprises first, second, third, fourth, and fifth regions as described herein. The present disclosure also relates to methods for producing a polyphenol aglycone, comprising contacting a phenolic glycoside with a glycoside hydrolase described herein. Also provided are host cells and heterologous expression systems capable of expressing a heterologous glycoside hydrolase, and nucleic acids encoding the glycoside hydrolase.
Claims28
A composition comprising a glycoside hydrolase and one or both of hesperidin and hesperetin, wherein an active site of the glycoside hydrolase comprises:
- a first region comprising at least 80%, at least 90%, or 100% identity to HQEAII (SEQ ID NO:1);
- a second region comprising at least 80%, at least 90%, or 100% identity to DIHSLPGGLNGMGLGE (SEQ ID NO:2);
- a third region comprising at least 80%, at least 90%, or 100% identity to PINEPVDNRDITKFGTP (SEQ ID NO:3);
- a fourth region comprising at least 80%, at least 90%, or 100% identity to FRPVDYWAKHFAASTNIVFDVHNYYFAGRPT (SEQ ID NO:4);
- and
- a fifth region comprising at least 80%, at least 90%, or 100% identity to GSAYWTWKFFGNVPVDGEGTQGDYWNY (SEQ ID NO:5).
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5-7. (canceled)
9-12. (canceled)
A method of producing a polyphenol aglycone, comprising contacting a phenolic glycoside with a glycoside hydrolase, wherein an active site of the glycoside hydrolase comprises:
- a first region comprising at least 80%, at least 90%, or 100% identity to HQEAII (SEQ ID NO:1);
- a second region comprising at least 80%, at least 90%, or 100% identity to DIHSLPGGLNGMGLGE (SEQ ID NO:2);
- a third region comprising at least 80%, at least 90%, or 100% identity to PINEPVDNRDITKFGTP (SEQ ID NO:3);
- a fourth region comprising at least 80%, at least 90%, or 100% identity to FRPVDYWAKHFAASTNIVFDVHNYYFAGRPT (SEQ ID NO:4);
- and
- a fifth region comprising at least 80%, at least 90%, or 100% identity to GSAYWTWKFFGNVPVDGEGTQGDYWNY (SEQ ID NO:5);
- wherein the glycoside hydrolase hydrolyzes the phenolic glycoside to form the polyphenol aglycone.
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23-30. (canceled)
A host cell capable of expressing a heterologous glycoside hydrolase, wherein an active site of the glycoside hydrolase comprises:
- a first region comprising at least 80% identity to HQEAII (SEQ ID NO:1);
- a second region comprising at least 80% identity to DIHSLPGGLNGMGLGE (SEQ ID NO:2);
- a third region comprising at least 80% identity to PINEPVDNRDITKFGTP (SEQ ID NO:
- 3);
- a fourth region comprising at least 80% identity to FRPVDYWAKHFAASTNIVFDVHNYYFAGRPT (SEQ ID NO:4);
- and
- a fifth region comprising at least 80% identity to GSAYWTWKFFGNVPVDGEGTQGDYWNY (SEQ ID NO:5),
- wherein the glycoside hydrolase is capable of hydrolyzing hesperidin to form hesperetin.
35-38. (canceled)
A nucleic acid encoding a glycosidic hydrolase, wherein an active site of the glycoside hydrolase comprises:
- a first region comprising at least 80% or at least 90% identity to HQEAII (SEQ ID NO:
- 1);
- a second region comprising at least 80% or at least 90% identity to DIHSLPGGLNGMGLGE (SEQ ID NO:2);
- a third region comprising at least 80% or at least 90% identity to PINEPVDNRDITKFGTP (SEQ ID NO:3);
- a fourth region comprising at least 80% or at least 90% identity to FRPVDYWAKHFAASTNIVFDVHNYYFAGRPT (SEQ ID NO:4);
- and
- a fifth region comprising at least 80% or at least 90% identity to GSAYWTWKFFGNVPVDGEGTQGDYWNY (SEQ ID NO:5),
- wherein the glycoside hydrolase is capable of hydrolyzing hesperidin to form hesperetin, and wherein the nucleic acid is operably linked to a heterologous regulatory element.
41-46. (canceled)
Dependencies are inferred from the claim text.