Patent US6258559B1 successfully refreshed from Google Patents.
US6258559B1
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Method for producing proteins in transformed Pichia
Assignee: Zymogenetics Inc
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Filing: 2000-03-21
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Granted: 2001-07-10
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Expires: 2020-03-21
Abstract
Abstract Methylotrophic yeasts are useful hosts for the production of commercially valuable recombinant proteins. However, the development of large-scale cultures of recombinant methylotrophic yeasts has been hindered by the formation of precipitation in culture media. A new soluble minimal medium overcomes this problem. Moreover, new feeding schemes provide cultures of high biomass, which produce biologically active recombinant protein.
Inventors
- 👤 Bruce L. Zamost
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-11-09 22:02
CPC Classifications
C; C12; C12P; C12P21/00; C12P21/02; C12N; C12N1/00; C12N1/14; C12N1/16
Claims Analysis
Total Claims: 44
Claim 1
Independent Claim
72 words
A method for producing a peptide or polypeptide by a recombinant Pichia methanolica host, comprising the step of incubating the recombinant Pichia methanolica host in a medium to produce a Pichia methanolica culture, wherein the cultured Pichia methanolica express the peptide or polypeptide under the control of an alcohol-inducible promoter, wherein the medium comprises sugar but is not supplemented with alcohol, and wherein the incubated Pichia methanolica produce the peptide or polypeptide.
Claim 2
Independent Claim
72 words
A method for producing a peptide or polypeptide by a recombinant Pichia methanolica host, comprising the step of incubating the recombinant Pichia methanolica host in a medium to produce a Pichia methanolica culture, wherein the cultured Pichia methanolica express the peptide or polypeptide under the control of an alcohol-inducible promoter, wherein the medium comprises sugar but is not supplemented with alcohol, and wherein the incubated Pichia methanolica produce the peptide or polypeptide.
Claim 3
Dependent Claim
13 words
The method of claim 1 , wherein the medium is a minimal medium.
Claim 4
Dependent Claim
24 words
The method of claim 2 , wherein the minimal medium consists essentially of water, sugar, inorganic ammonia, potassium, phosphate, iron, biotin, and citric acid.
Claim 5
Dependent Claim
13 words
The method of claim 1 , wherein the medium is a rich medium.
Claim 6
Dependent Claim
20 words
The method of claim 1 , wherein the sugar is selected from the group consisting of glucose, fructose, and mannose.
Claim 7
Dependent Claim
11 words
The method of claim 5 , wherein the sugar is fructose.
Claim 8
Dependent Claim
11 words
The method of claim 5 , wherein the sugar is mannose.
Claim 9
Dependent Claim
11 words
The method of claim 5 , wherein the sugar is glucose.
Claim 10
Dependent Claim
13 words
The method of claim 8 , wherein the medium is a minimal medium.
Claim 11
Dependent Claim
14 words
The method of claim 8 , wherein the alcohol-inducible promoter is a methanol-inducible promoter.
Claim 12
Dependent Claim
13 words
The method of claim 10 , wherein the medium is a minimal medium.
Claim 13
Dependent Claim
27 words
The method of claim 11 , further comprising the step of isolating the peptide or polypeptide either from the culture medium, or from the cultured Pichia host.
Claim 14
Dependent Claim
27 words
The method of claim 8 , further comprising the step of isolating the peptide or polypeptide either from the culture medium, or from the cultured Pichia host.
Claim 15
Dependent Claim
16 words
The method of claim 10 , wherein the methanol-inducible promoter is a Pichia methanolica AUG1 promoter.
Claim 16
Dependent Claim
27 words
The method of claim 10 , further comprising the step of isolating the peptide or polypeptide either from the culture medium, or from the cultured Pichia host.
Claim 17
Dependent Claim
14 words
The method of claim 1 , wherein the alcohol-inducible promoter is a methanol-inducible promoter.
Claim 18
Dependent Claim
13 words
The method of claim 16 , wherein the medium is a minimal medium.
Claim 19
Dependent Claim
16 words
The method of claim 16 , wherein the methanol-inducible promoter is a Pichia methanolica AUG1 promoter.
Claim 20
Dependent Claim
27 words
The method of claim 18 , further comprising the step of isolating the peptide or polypeptide either from the culture medium, or from the cultured Pichia host.
Claim 21
Dependent Claim
13 words
The method of claim 1 , wherein incubation is performed by batch fermentation.
Claim 22
Dependent Claim
13 words
The method of claim 1 , wherein incubation is performed by fed-batch fermentation.
Claim 23
Dependent Claim
13 words
The method of claim 21 , wherein the medium is a minimal medium.
Claim 24
Dependent Claim
13 words
The method of claim 1 , wherein incubation is performed by continuous fermentation.
Claim 25
Independent Claim
96 words
A method for producing a peptide or polypeptide by a recombinant Pichia methanolica host, comprising the step of incubating the recombinant Pichia methanolica host in a medium to produce a Pichia methanolica culture, wherein the cultured Pichia methanolica express the peptide or polypeptide under the control of a methanol-inducible promoter, wherein the medium comprises sugar but is not supplemented with alcohol, wherein the sugar is selected from the group consisting of glucose, fructose, and mannose, wherein the incubated Pichia methanolica produce the peptide or polypeptide, and wherein the incubation step is performed by fed batch fermentation.
Claim 26
Independent Claim
96 words
A method for producing a peptide or polypeptide by a recombinant Pichia methanolica host, comprising the step of incubating the recombinant Pichia methanolica host in a medium to produce a Pichia methanolica culture, wherein the cultured Pichia methanolica express the peptide or polypeptide under the control of a methanol-inducible promoter, wherein the medium comprises sugar but is not supplemented with alcohol, wherein the sugar is selected from the group consisting of glucose, fructose, and mannose, wherein the incubated Pichia methanolica produce the peptide or polypeptide, and wherein the incubation step is performed by fed batch fermentation.
Claim 27
Dependent Claim
13 words
The method of claim 24 , wherein the medium is a minimal medium.
Claim 28
Dependent Claim
24 words
The method of claim 25 , wherein the minimal medium consists essentially of water, sugar, inorganic ammonia, potassium, phosphate, iron, biotin, and citric acid.
Claim 29
Dependent Claim
27 words
The method of claim 25 , further comprising the step of isolating the peptide or polypeptide either from the culture medium, or from the cultured Pichia host.
Claim 30
Dependent Claim
16 words
The method of claim 24 , wherein the methanol-inducible promoter is a Pichia methanolica AUG1 promoter.
Claim 31
Dependent Claim
13 words
The method of claim 28 , wherein the medium is a minimal medium.
Claim 32
Dependent Claim
27 words
The method of claim 29 , further comprising the step of isolating the peptide or polypeptide either from the culture medium, or from the cultured Pichia host.
Claim 33
Dependent Claim
27 words
The method of claim 28 , further comprising the step of isolating the peptide or polypeptide either from the culture medium, or from the cultured Pichia host.
Claim 34
Dependent Claim
27 words
The method of claim 24 , further comprising the step of isolating the peptide or polypeptide either from the culture medium, or from the cultured Pichia host.
Claim 35
Independent Claim
82 words
A method for producing a peptide or polypeptide by a recombinant Pichia methanolica host, comprising the step of incubating the recombinant Pichia methanolica host in a medium to produce a Pichia methanolica culture, wherein the cultured Pichia methanolica express the peptide or polypeptide under the control of a methanol-inducible promoter, wherein the medium comprises glucose but is not supplemented with alcohol, wherein the incubated Pichia methanolica produce the peptide or polypeptide, and wherein the incubation step is performed by fed batch fermentation.
Claim 36
Independent Claim
82 words
A method for producing a peptide or polypeptide by a recombinant Pichia methanolica host, comprising the step of incubating the recombinant Pichia methanolica host in a medium to produce a Pichia methanolica culture, wherein the cultured Pichia methanolica express the peptide or polypeptide under the control of a methanol-inducible promoter, wherein the medium comprises glucose but is not supplemented with alcohol, wherein the incubated Pichia methanolica produce the peptide or polypeptide, and wherein the incubation step is performed by fed batch fermentation.
Claim 37
Dependent Claim
13 words
The method of claim 33 , wherein the medium is a minimal medium.
Claim 38
Dependent Claim
24 words
The method of claim 34 , wherein the minimal medium consists essentially of water, sugar, inorganic ammonia, potassium, phosphate, iron, biotin, and citric acid.
Claim 39
Dependent Claim
27 words
The method of claim 34 , further comprising the step of isolating the peptide or polypeptide either from the culture medium, or from the cultured Pichia host.
Claim 40
Dependent Claim
16 words
The method of claim 33 , wherein the methanol-inducible promoter is a Pichia methanolica AUG1 promoter.
Claim 41
Dependent Claim
13 words
The method of claim 37 , wherein the medium is a minimal medium.
Claim 42
Dependent Claim
27 words
The method of claim 38 , further comprising the step of isolating the peptide or polypeptide either from the culture medium, or from the cultured Pichia host.
Claim 43
Dependent Claim
27 words
The method of claim 37 , further comprising the step of isolating the peptide or polypeptide either from the culture medium, or from the cultured Pichia host.
Claim 44
Dependent Claim
27 words
The method of claim 33 , further comprising the step of isolating the peptide or polypeptide either from the culture medium, or from the cultured Pichia host.
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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) | 2004-07-09 | 2005-01-08 | 2005-07-10 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 2008-07-09 | 2009-01-08 | 2009-07-10 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 2012-07-09 | 2013-01-08 | 2013-07-10 | $7700.00 | Window Closed / Lapsed |