US6004787A
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Method of directing biosynthesis of specific polyketides
Assignee: Abbott Laboratories
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Filing: 1995-05-11
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Granted: 1999-12-21
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Expires: 2015-05-11
Abstract
Abstract A method to produce novel polyketide structures by designing and introducing specified changes in the DNA governing the synthesis of the polyketide is disclosed. The biosynthesis of specific polyketide analogs is accomplished by genetic manipulation of a polyketide-producing microorganism by isolating a polyketide biosynthetic gene-containing DNA sequence, identifying enzymatic activities associated within the DNA sequence, introducing one or more specified changes into the DNA sequence which codes for one of the enzymatic activities which results in an altered DNA sequence, introducing the altered DNA sequence into the polyketide-producing microorganism to replace the original sequence, growing a culture of the altered microorganism under conditions suitable for the formation of the specific polyketide analog, and isolating the specific polyketide analog from the culture. The method is most useful when the segment of the chromosome modified is involved in an enzymatic activity associated with polyketide biosynthesis, particularly for manipulating polyketide synthase genes from Saccarharopolyspora or Streptomyces.
Inventors
- 👤 Leonard Katz
- 👤 Stefano Donadio
- 👤 James B. McAlpine
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-11-08 23:53
CPC Classifications
C; C07; C07H; C07H17/00; C07H17/04; C07H17/08; C12; C12N; C12N15/00; C12N15/09; C12N15/11; C12N15/52; C12P; C12P19/00; C12P19/44; C12P19/60; C12P19/62
Claims Analysis
Total Claims: 43
Claim 1
Independent Claim
115 words
A method for directing the biosynthesis of specific macrolide polyketide analogs by genetic manipulation of a polyketide-producing microorganism, said method comprising the steps of: (1) isolating a macrolide polyketide biosynthetic gene-containing DNA sequence; (2) identifying enzymatic activities associated within said gene-containing DNA sequence; (3) introducing one or more specified changes into said gene-containing DNA sequence which codes for one of said enzymatic activities resulting in an altered DNA sequence; (4) introducing said altered DNA sequence into a polyketide-producing microorganism to replace the original sequence; (5) growing a culture of the altered microorganism under conditions suitable for the formation of the specific macrolide polyketide analog; and (6) isolating said specific macrolide polyketide analog from the culture.
Claim 2
Independent Claim
115 words
A method for directing the biosynthesis of specific macrolide polyketide analogs by genetic manipulation of a polyketide-producing microorganism, said method comprising the steps of: (1) isolating a macrolide polyketide biosynthetic gene-containing DNA sequence; (2) identifying enzymatic activities associated within said gene-containing DNA sequence; (3) introducing one or more specified changes into said gene-containing DNA sequence which codes for one of said enzymatic activities resulting in an altered DNA sequence; (4) introducing said altered DNA sequence into a polyketide-producing microorganism to replace the original sequence; (5) growing a culture of the altered microorganism under conditions suitable for the formation of the specific macrolide polyketide analog; and (6) isolating said specific macrolide polyketide analog from the culture.
Claim 3
Dependent Claim
15 words
An isolated polynucleotide segment that encodes a biologically active 6-deoxyerythronolide B synthase from Saccharopolyspora erythraea.
Claim 4
Dependent Claim
15 words
An isolated polynucleotide segment that encodes a biologically active 6-deoxyerythronolide B synthase from Saccharopolyspora erythraea.
Claim 5
Dependent Claim
24 words
The polynucleotide segment of claim 2 wherein said 6-deoxyerythronolide B synthase possesses one or more enzymatic activities associated with the production of 6-deoxyerythronolide B.
Claim 6
Dependent Claim
27 words
The polynucleotide segment of claim 3 wherein said enzymatic activities are selected from the group consisting of β-ketoreductase, dehydratase, enoylreductase, acyl carrier protein, β-ketoacyl ACP and acyltransferase.
Claim 7
Dependent Claim
19 words
The polynucleotide segment of claim 3 selected from the group consisting of SEQ ID NO:1 and SEQ ID NO:3.
Claim 8
Dependent Claim
25 words
The polynucleotide segment of claim 4 wherein said polynucleotide segment is from about nucleotide position 2250 to about nucleotide position 3626 of SEQ ID NO:1.
Claim 9
Dependent Claim
25 words
The polynucleotide segment of claim 4 wherein said polynucleotide segment is from about nucleotide position 3831 to about nucleotide position 4814 of SEQ ID NO:1.
Claim 10
Dependent Claim
25 words
The polynucleotide segment of claim 4 wherein said polynucleotide segment is from about nucleotide position 5574 to about nucleotide position 6125 of SEQ ID NO:1.
Claim 11
Dependent Claim
25 words
The polynucleotide segment of claim 4 wherein said polynucleotide segment is from about nucleotide position 6369 to about nucleotide position 6626 of SEQ ID NO:1.
Claim 12
Dependent Claim
25 words
The polynucleotide segment of claim 4 wherein said polynucleotide segment is from about nucleotide position 6675 to about nucleotide position 8057 of SEQ ID NO:1.
Claim 13
Dependent Claim
25 words
The polynucleotide segment of claim 4 wherein said polynucleotide segment is from about nucleotide position 8262 to about nucleotide position 9236 of SEQ ID NO:1.
Claim 14
Dependent Claim
25 words
The polynucleotide segment of claim 4 wherein said polynucleotide segment is from about nucleotide position 9906 to about nucleotide position 10454 of SEQ ID NO:1.
Claim 15
Dependent Claim
25 words
The polynucleotide segment of claim 4 wherein said polynucleotide segment is from about nucleotide position 10707 to about nucleotide position 10964 of SEQ ID NO:1.
Claim 16
Dependent Claim
25 words
The polynucleotide segment of claim 4 wherein said polynucleotide segment is from about nucleotide position 103 to about nucleotide position 1482 of SEQ ID NO:3.
Claim 17
Dependent Claim
25 words
The polynucleotide segment of claim 4 wherein said polynucleotide segment is from about nucleotide position 1693 to about nucleotide position 2670 of SEQ ID NO:3.
Claim 18
Dependent Claim
25 words
The polynucleotide segment of claim 4 wherein said polynucleotide segment is from about nucleotide position 3406 to about nucleotide position 3921 of SEQ ID NO:3.
Claim 19
Dependent Claim
25 words
The polynucleotide segment of claim 4 wherein said polynucleotide segment is from about nucleotide position 4171 to about nucleotide position 4428 of SEQ ID NO:3.
Claim 20
Dependent Claim
25 words
The polynucleotide segment of claim 4 wherein said polynucleotide segment is from about nucleotide position 4477 to about nucleotide position 5847 of SEQ ID NO:3.
Claim 21
Dependent Claim
25 words
The polynucleotide segment of claim 4 wherein said polynucleotide segment is from about nucleotide position 6055 to about nucleotide position 7026 of SEQ ID NO:3.
Claim 22
Dependent Claim
25 words
The polynucleotide segment of claim 4 wherein said polynucleotide segment is from about nucleotide position 7165 to about nucleotide position 7638 of SEQ ID NO:3.
Claim 23
Dependent Claim
25 words
The polynucleotide segment of claim 4 wherein said polynucleotide segment is from about nucleotide position 9433 to about nucleotide position 9984 of SEQ ID NO:3.
Claim 24
Dependent Claim
25 words
The polynucleotide segment of claim 4 wherein said polynucleotide segment is from about nucleotide position 10225 to about nucleotide position 10482 of SEQ ID NO:3.
Claim 25
Dependent Claim
25 words
The polynucleotide segment of claim 4 wherein said polynucleotide segment is from about nucleotide position 10831 to about nucleotide position 12174 of SEQ ID NO:3.
Claim 26
Dependent Claim
25 words
The polynucleotide segment of claim 4 wherein said polynucleotide segment is from about nucleotide position 12379 to about nucleotide position 13350 of SEQ ID NO:3.
Claim 27
Dependent Claim
25 words
The polynucleotide segment of claim 4 wherein said polynucleotide segment is from about nucleotide position 14062 to about nucleotide position 14610 of SEQ ID NO:3.
Claim 28
Dependent Claim
25 words
The polynucleotide segment of claim 4 wherein said polynucleotide segment is from about nucleotide position 14857 to about nucleotide position 15114 of SEQ ID NO:3.
Claim 29
Dependent Claim
25 words
The polynucleotide segment of claim 4 wherein said polynucleotide segment is from about nucleotide position 15172 to about nucleotide position 16569 of SEQ ID NO:3.
Claim 30
Dependent Claim
25 words
The polynucleotide segment of claim 4 wherein said polynucleotide segment is from about nucleotide position 16768 to about nucleotide position 17721 of SEQ ID NO:3.
Claim 31
Dependent Claim
25 words
The polynucleotide segment of claim 4 wherein said polynucleotide segment is from about nucleotide position 18379 to about nucleotide position 18921 of SEQ ID NO:3.
Claim 32
Dependent Claim
25 words
The polynucleotide segment of claim 4 wherein said polynucleotide segment is from about nucleotide position 19141 to about nucleotide position 19398 of SEQ ID NO:3.
Claim 33
Dependent Claim
17 words
A vector comprising a polynucleotide segment that encodes a biologically active 6-deoxyerythronolide B synthase from Saccharopolyspora erythraea.
Claim 34
Dependent Claim
17 words
A vector comprising a polynucleotide segment that encodes a biologically active 6-deoxyerythronolide B synthase from Saccharopolyspora erythraea.
Claim 35
Dependent Claim
23 words
The vector of claim 31 wherein said 6-deoxyerythronolide B synthase possesses one or more enzymatic activities associated with the production of 6-deoxyerythronolide B.
Claim 36
Dependent Claim
26 words
The vector of claim 32 wherein said enzymatic activities are selected from the group consisting of β-ketoreductase, dehydratase, enoylreductase, acyl carrier protein, β-ketoacyl ACP and acyltransferase.
Claim 37
Dependent Claim
9 words
A host cell comprising the vector of claim 31.
Claim 38
Dependent Claim
9 words
A host cell comprising the vector of claim 32.
Claim 39
Dependent Claim
9 words
A host cell comprising the vector of claim 33.
Claim 40
Dependent Claim
14 words
An isolated polypeptide having the biological activity of 6-deoxyerythronolide B synthase from Saccharopolyspora erythraea.
Claim 41
Dependent Claim
14 words
An isolated polypeptide having the biological activity of 6-deoxyerythronolide B synthase from Saccharopolyspora erythraea.
Claim 42
Dependent Claim
23 words
The polypeptide of claim 37 wherein said 6-deoxyerythronolide B synthase possesses one or more enzymatic activities associated with the production of 6-deoxyerythronolide B.
Claim 43
Dependent Claim
26 words
The polypeptide of claim 38 wherein said enzymatic activities are selected from the group consisting of β-ketoreductase, dehydratase, enoylreductase, acyl carrier protein, β-ketoacyl ACP and acyltransferase.
References Cited (Backward Citations)
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US4874748A
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US5110728A
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USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 2002-12-20 | 2003-06-21 | 2003-12-21 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 2006-12-20 | 2007-06-21 | 2007-12-21 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 2010-12-20 | 2011-06-21 | 2011-12-21 | $7700.00 | Window Closed / Lapsed |