Compositions and methods for producing podophyllotoxin derivatives
AssigneeThe Board of Trustees of the Leland Stanford Junior University
InventorsWarren Lau, Elizabeth Sattely
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Abstract
The present invention provides compositions and methods for biosynthetically producing podophyllotoxin intermediates and derivatives including enzymes and their equivalents involved in the biosynthetic production of podophyllotoxin intermediates and derivatives.
Claims22
An isolated nucleic acid molecule encoding an enzyme involved in synthesizing podophyllotoxin intermediates and derivatives and having at least 90% sequence identity to nucleic acid SEQ ID NO:1, and conservative variants thereof.
An isolated nucleic acid molecule encoding an enzyme involved in synthesizing podophyllotoxin intermediates and derivatives and having at least 90% sequence identity to nucleic acid SEQ ID NO:2, and conservative variants thereof.
An isolated nucleic acid molecule encoding an enzyme involved in synthesizing podophyllotoxin intermediates and derivatives and having at least 90% sequence identity to nucleic acid SEQ ID NO:3, and conservative variants thereof.
An isolated nucleic acid molecule encoding an enzyme involved in synthesizing podophyllotoxin intermediates and derivatives and having at least 90% sequence identity to nucleic acid SEQ ID NO:4, and conservative variants thereof.
An isolated nucleic acid molecule encoding an enzyme involved in synthesizing podophyllotoxin intermediates and derivatives and having at least 90% sequence identity to nucleic acid SEQ ID NO:5, and conservative variants thereof.
An isolated nucleic acid molecule encoding an enzyme involved in synthesizing podophyllotoxin intermediates and derivatives and having at least 90% sequence identity to nucleic acid SEQ ID NO:6, and conservative variants thereof. An isolated nucleic acid molecule encoding an enzyme involved in synthesizing podophyllotoxin intermediates and derivatives and having at least 90% sequence identity to nucleic acid SEQ ID NO:7, and conservative variants thereof.
An isolated nucleic acid molecule encoding an enzyme involved in synthesizing podophyllotoxin intermediates and derivatives and having at least 90% sequence identity to nucleic acid SEQ ID NO:8, and conservative variants thereof.
An isolated nucleic acid molecule encoding an enzyme involved in synthesizing podophyllotoxin intermediates and derivatives and having at least 90% sequence identity to nucleic acid SEQ ID NO:9, and conservative variants thereof.
An isolated nucleic acid molecule encoding an enzyme involved in synthesizing podophyllotoxin intermediates and derivatives and having at least 90% sequence identity to nucleic acid SEQ ID NO:10, and conservative variants thereof.
A host cell for producing 4′-desmethylepipodophyllotoxin, comprising nucleic acid SEQ ID NO:1 for converting pinoresinol to secoisolariciresinol;
- nucleic acid SEQ ID NO:2 for converting secoisolariciresinol to matairesinol;
- nucleic acid SEQ ID NO:3 for converting matairesinol to pluviatolide;
- nucleic acid SEQ ID NO:4 for converting pluviatolide to 5′-desmethoxy-yatein;
- nucleic acid SEQ ID NO:5 for converting 5′-desmethoxy-yatein into 5′-desmethyl-yatein;
- nucleic acid SEQ ID NO:6 for converting 5′-desmethyl-yatein into yatein;
- nucleic acid SEQ ID NO:7 for converting yatein into deoxypodophyllotoxin;
- nucleic acid SEQ ID NO:8 for converting deoxypodophyllotoxin into 4′-desmethyl-deoxypodophyllotoxin;
- and nucleic acid SEQ ID NO:9 for converting 4′-desmethyl-deoxypodophyllotoxin into 4′-desmethyl-epipodophyllotoxin.
A method for producing 4′-desmethyl-epipodophyllotoxin in a host cell, comprising culturing and transforming the host cell with nucleic acid SEQ ID NO:1 for converting pinoresinol to secoisolariciresinol;
- nucleic acid SEQ ID NO:2 for converting secoisolariciresinol to matairesinol;
- nucleic acid SEQ ID NO:3 for converting matairesinol to pluviatolide;
- nucleic acid SEQ ID NO:4 for converting pluviatolide to 5′-desmethoxy-yatein;
- nucleic acid SEQ ID NO:5 for converting 5′-desmethoxy-yatein into 5′-desmethyl-yatein;
- nucleic acid SEQ ID NO:6 for converting 5′-desmethyl-yatein into yatein;
- nucleic acid SEQ ID NO:7 for converting yatein into deoxypodophyllotoxin;
- nucleic acid SEQ ID NO:8 for converting deoxypodophyllotoxin into 4′-desmethyl-deoxypodophyllotoxin;
- and nucleic acid SEQ ID NO:9 for converting 4′-desmethyl-deoxypodophyllotoxin into 4′-desmethyl-epipodophyllotoxin;
- adding pinoresinol as starting material to culture medium;
- and recovering 4′-desmethylepipodophyllotoxin from culture medium.
Dependencies are inferred from the claim text.