Moenomycin biosynthesis-related compositions and methods of use thereof

• US 9,115,358 B2
• Filed: 08/13/2007
• Issued: 08/25/2015
• Est. Priority Date: 08/11/2006
• Status: Active Grant

• Current Assignee: President and Fellows of Harvard College (Harvard Corporation)
• Original Assignee: President and Fellows of Harvard College (Harvard Corporation)
• Inventors: Walker, Suzanne, Omelyanovich Ostash, Bohdan
• Primary Examiner(s): Joike, Michele K
• Application Number:
  US12/377,117
• Publication Number:
  US 20100279980A1
• Time in Patent Office: 2,934 Days
• Field of Search: None
• US Class Current: 1/1
• CPC Class Codes: A61K 31/7028 Compounds having saccharide radicals attached to non-saccharide compounds by glycosidic linkages
• CPC Class Codes: C12N 15/52 Genes encoding for enzymes or proenzymes
• CPC Class Codes: C12P 19/44 Preparation of O-glycosides, e.g. glucosides polysaccharides and not substituted disaccharides C12P19/04, C12P19/12

Abstract

The methods and compositions described herein relate to the identification, isolation, and characterization of genes which encode proteins useful for the biosynthesis of transglycosylase inhibitors such as moes. The methods and compositions also relate to the production of such proteins, and their use in the synthesis of moes, the expression of moes, and the production of modified moes.

Claims

1. An isolated recombinant cell expressing one or more polypeptides that comprise a moenomycin biosynthesis-related polypeptide selected from the group consisting of:

MoeA4 (SEQ ID NO;

     29), MoeB4 (SEQ ID NO;

     30), MoeC4 (SEQ ID NO;

     28), MoeB5 (SEQ ID NO;

     27), Moe A5 (SEQ ID NO;

     26), MoeD5 (SEQ ID NO;

     47), MoeJ5 (SEQ ID NO;

     48), MoeE5 (SEQ ID NO;

     42), MoeF5 (SEQ ID NO;

     36), MoeH5 (SEQ ID NO;

     37), MoeK5 (SEQ ID NO;

     38), MoeM5 (SEQ ID NO;

     39), MoeN5 (SEQ ID NO;

     43), MoeO5 (SEQ ID NO;

     44), MoeX5 (SEQ ID NO;

     46), MoeP5 (SEQ ID NO;

     45), MoeR5 (SEQ ID NO;

     40), MoeS5 (SEQ ID NO;

     41), MoeGT1 (SEQ ID NO;

     31), MoeGT2 (SEQ ID NO;

     32), MoeGT3 (SEQ ID NO;

     33), MoeGT4 (SEQ ID NO;

     34), MoeGT5 (SEQ ID NO;

     35), and variants thereof having at least about 95% sequence identity to the corresponding natural moenomycin biosynthesis-related polypeptide, wherein the cell is selected from the group consisting of;

Streptomyces lividans TK24, E. coli, a mammalian cell, a yeast cell, and an insect cell.

2. The isolated recombinant cell of claim 1, wherein the cell is Streptomyces lividans TK24.

3. The isolated recombinant cell of claim 1, wherein the cell is E. coli.

4. The isolated recombinant cell of claim 1, wherein the cell is a mammalian cell.

5. The isolated recombinant cell of claim 1, wherein the cell is a yeast cell.

6. The isolated recombinant cell of claim 1, wherein the cell is an insect cell.

7. The isolated recombinant cell of claim 1, wherein the cell expresses two or more polypeptides that comprise the moenomycin biosynthesis-related polypeptide and variants thereof.

8. The isolated recombinant cell of claim 1, wherein the cell expresses three or more polypeptides that comprise the moenomycin biosynthesis-related polypeptide and variants thereof.

9. The isolated recombinant cell of claim 1, wherein the cell expresses four or more polypeptides that comprise the moenomycin biosynthesis-related polypeptide and variants thereof.

10. The isolated recombinant cell of claim 1, wherein the cell expresses five or more polypeptides that comprise the moenomycin biosynthesis-related polypeptide and variants thereof.

11. The isolated recombinant cell of claim 1, wherein the moenomycin biosynthesis-related polypeptide variants have at least about 98% sequence identity to the corresponding natural moenomycin biosynthesis-related polypeptide.

12. The isolated recombinant cell of claim 1, wherein the moenomycin biosynthesis-related polypeptide variants have at least about 99% sequence identity to the corresponding natural moenomycin biosynthesis-related polypeptide.