DNA encoding triol polyketide synthase

• US 5,849,541 A
• Filed: 08/23/1996
• Issued: 12/15/1998
• Est. Priority Date: 11/02/1993
• Status: Expired due to Fees

• Current Assignee: Merck Sharp & Dohme Corporation (Merck & Co., Inc.)
• Original Assignee: Merck & Co., Inc.
• Inventors: Reeves, Christopher D., Hendrickson, Lee E., Rambosek, John, Conder, Michael J., Vinci, Victor A., McAda, Phyllis C., Davis, Charles Ray
• Primary Examiner(s): Wax, Robert A.
• Assistant Examiner(s): Slobodyansky, Elizabeth
• Application Number:
  US08/637,640
• Time in Patent Office: 844 Days
• Field of Search: 435/69.2, 435/91.1, 435/125, 435/254.3, 435/254.5, 435/254.6, 435/320.1, 435/183, 536/23.2, 536/24.32
• US Class Current: 435/91.1
• CPC Class Codes: C12N 9/88 Lyases (4.)
• CPC Class Codes: C12P 17/06 containing a six-membered hetero ring, e.g. fluorescein

Abstract

DNA encoding triol polyketide synthase (TPKS) from Aspergillus terreus has been isolated, purified and sequenced. Expression vectors comprising said DNA, cells transformed with the expression vectors, and processes employing the transformed cells are provided.

Claims

1. A purified and isolated DNA molecule encoding a triol polyketide synthase of Aspergillus terreus said DNA molecule having a nucleotide sequence set forth in SEQ ID NO:

1.

2. An expression vector comprising a DNA molecule of claim 1.

3. A host cell transformed with an expression vector of claim 2.

4. The expression vector of claim 2 which is pTPKS100 (ATCC 69416).

5. A host cell transformed with the expression vector of claim 4.

6. A process for producing HMG-CoA reductase inhibitors, comprising:

(a) transforming a cell with a DNA molecule of claim 1;

(b) cultivating the transformed cell under conditions that permit the expression of the DNA molecule; and

(c) recovering the HMG-CoA reductase inhibitor.

7. The process of claim 6 wherein the HMG-CoA reductase inhibitors are selected from the group consisting of lovastatin, triol and compactin.

8. The process of claim 6 wherein said transformed cell is selected from the group consisting of cells of Aspergillus terreus, Monascus ruber, Penicillum citrinum, Penicillum brevicompactum, Hypomyces chrysospermus, Paecilomyces viridis, Paecilomyces sp. M2016, Eupenicillium sp. MM603, Trichoderma longibrachiatum M6735 and Trichoderma pseudokoningii M6828.

9. A method of isolating DNA encoding polyketide synthase, comprising:

(a) hybridizing a DNA of claim 1 to a sample containing DNA encoding polyketide synthase to form a complex; and

(b) purifying the complex.

10. The method of claim 9 wherein the sample is derived from a microorganism, the microorganism being selected from the group consisting of Aspergillus terreus, Monascus ruber, Penicillum citrinum, Penicillum brevicompactum, Hypomyces chrysopermus, Paecilomyces viridis, Paecilomyces sp. M2016, Eupenicillium sp. MM603, Trichoderma longibrachiatum. M6735 and Trichoderma pseudokoningii M6828.