Patent EP0215594B1 successfully refreshed from Google Patents.
EP0215594B1
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Heterologous polypeptide expressed in filamentous fungi, processes for their preparation, and vectors for their preparation
Assignee: Danisco US Inc
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Filing: 1986-08-27
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Granted: 1995-01-11
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Expires: 2006-08-27
Abstract
Abstract Novel vectors are disclosed for expressing and secreting heterologous polypeptides from filamentous fungi. Such vectors are used in novel processes to express and secrete such heterologous polypeptides. The vectors used for transforming a filamentous fungus to express and secrete a heterologous polypeptide include a DNA sequence encoding a heterologous polypeptide and a DNA sequence encoding a signal sequence which is functional in a secretory system in a given filamentous fungus and which is operably linked to the sequence encoding the heterologous polypeptide. Such signal sequences may be the signal sequence normally associated with the heterologous polypeptides or may be derived from other sources. The vector may also contain DNA sequences encoding a promoter sequence which is functionally recognized by the filamentous fungus and which is operably linked to the DNA sequence encoding the signal sequence. Preferably functional polyadenylation sequences are operably linked to the 3' terminus of the DNA sequence encoding the heterologous polypeptides. Each of the above described vectors are used in novel processes to transform a filamentous fungus wherein the DNA sequences encoding the signal sequence and heterologous polypeptide are expressed. The thus synthesized polypeptide is thereafter secreted from the filamentous fungus.
Inventors
- 👤 Randy M. Berka
- 👤 Daniel Cullen
- 👤 Gregory L. Gray
- 👤 Kirk J. Hayenga
- 👤 Virgil B. Lawlis
Bibliographic Details
Application Number
—
Jurisdiction
EP
Data Source
google_patents
Last Synchronized
2026-11-09 01:50
CPC Classifications
C; C12; C12N; C12N9/00; C12N9/14; C12N9/24; C12N9/2402; C12N9/2405; C12N9/2408; C12N9/2411; C12N9/2428; C12N15/00; C12N15/09; C12N15/63; C12N15/79; C12N15/80; C12N9/48; C12N9/50; C12N9/58; C12N9/64; C12N9/6421; C12N9/6478; C12N9/6481; C07; C07K; C07K2319/00; C07K2319/01; C07K2319/02; C07K2319/50; C07K2319/61
Claims Analysis
Total Claims: 30
Claim 1
Independent Claim
67 words
A process for making a heterologous polypeptide comprising: transforming a filamentous fungus from the subdivision Eumycotina with a vector containing DNA sequences capable of expressing a heterologous polypeptide and of causing secretion of the heterologous polypeptide from the filamentous fungus, said filamentous fungus being selected from members of the subdivision Eumycotina that are capable of being propagated in filamentous form, and expressing and secreting said heterologous polypeptide.
Claim 2
Dependent Claim
67 words
A process for making a heterologous polypeptide comprising: transforming a filamentous fungus from the subdivision Eumycotina with a vector containing DNA sequences capable of expressing a heterologous polypeptide and of causing secretion of the heterologous polypeptide from the filamentous fungus, said filamentous fungus being selected from members of the subdivision Eumycotina that are capable of being propagated in filamentous form, and expressing and secreting said heterologous polypeptide.
Claim 3
Dependent Claim
48 words
A process as claimed in claim 1 wherein said vector comprises a DNA sequence encoding said heterologous polypeptide, a DNA sequence encoding a signal sequence and a promoter sequence operably linked to said DNA encoding the signal sequence, said promoter sequence being functionally recognized by said filamentous fungus.
Claim 4
Dependent Claim
17 words
A process as claimed in claim 2 wherein the signal sequence is native to the heterologous polypeptide.
Claim 5
Dependent Claim
30 words
A process as claimed in claim 3 wherein the native signal sequence and the heterologous sequence are DNA sequences encoding bovine preprochymosin, Mucor meihei preprocarboxyl protease or Aspergillus niger preproglucoamylase.
Claim 6
Dependent Claim
17 words
A process as claimed in claim 2 wherein the signal sequence is foreign to the heterologous polypeptide.
Claim 7
Dependent Claim
31 words
A process as claimed in claim 5 wherein the foreign signal sequence is a DNA sequence encoding the signal sequence of bovine preprochymosin Aspergillus niger glucoamylase or Mucor meihei carboxyl protease.
Claim 8
Dependent Claim
20 words
A process as claimed in claim 2 wherein said promoter sequence is from an endogenous gene from a filamentous fungus.
Claim 9
Dependent Claim
23 words
A process as claimed in claim 7 wherein said promoter sequence is from a filamentous fungus other than that transformed with said vector.
Claim 10
Dependent Claim
26 words
A process as claimed in claim 7 wherein said promoter comprises the promoter sequence of Aspergillus niger glucoamylase, Humicola grisea glucoamylase or Mucor meihei carboxyl protease.
Claim 11
Dependent Claim
26 words
A process as claimed in claim 2 further comprising a functional transcription termination and polyadenylation sequence operably linked to the DNA sequence encoding the heterologous polypeptide.
Claim 12
Dependent Claim
24 words
A process as claimed in claim 10 wherein the polyadenylation sequence comprises the polyadenylation sequence of Aspergillus niger glucoamylase or Mucor meihei carboxyl protease.
Claim 13
Dependent Claim
21 words
A process as claimed in claim 2 further comprising a DNA sequence encoding a selection characteristic expressible in the filamentous fungus.
Claim 14
Dependent Claim
29 words
A process as claimed in claim 12 wherein the selection characteristic is a DNA sequence encoding Neurospora crassa pyr4, Aspergillus nidulans acetamidase, Aspergillus nidulans argB or Aspergillus nidulans trpC.
Claim 15
Dependent Claim
24 words
A process as claimed in claim 2 further comprising a DNA sequence capable of increasing the transformation efficiency of the vector into Aspergillus nidulans.
Claim 16
Dependent Claim
18 words
A process as claimed in claim 14 wherein the DNA sequence for increasing fungal transformation efficiency comprises ANS-1.
Claim 17
Dependent Claim
15 words
A process as claimed in claim 2 wherein the secreted heterologous polypeptide is an enzyme.
Claim 18
Dependent Claim
26 words
A process as claimed in claim 17 wherein the enzyme is bovine chymosin, bovine prochymosin, Aspergillus niger glucoamylase, Humicola grisea glucoamylase or Mucor meihei carboxyl protease.
Claim 19
Dependent Claim
16 words
A process as claimed in claim 2 wherein the secreted heterologous polypeptide is a mammalian polypeptide.
Claim 20
Dependent Claim
25 words
A process as claimed in claim 18 wherein said signal sequence comprises the signal sequence of bovine preprochymosin and said mammalian polypeptide comprises bovine prochymosin.
Claim 21
Dependent Claim
25 words
A process as claimed in claim 18 wherein said signal sequence comprises the signal sequence of Aspergillus glucoamylase and said mammalian polypeptide comprises bovine prochymosin.
Claim 22
Dependent Claim
33 words
A process as claimed in claim 2 wherein said promoter and signal sequence comprises a DNA sequence encoding the promoter and signal sequence from Aspergillus glucoamylase and said heterologous polypeptide comprises bovine prochymosin.
Claim 23
Dependent Claim
33 words
A process as claimed in claim 2 wherein said promoter comprises the promoter from Aspergillus glucoamylase, said signal sequence comprises the signal sequence from bovine preprochymosin and said heterologous polypeptide comprises bovine prochymosin.
Claim 24
Dependent Claim
20 words
A process as claimed in claim 2 wherein the secreted heterologous polypeptide comprises a polypeptide from a second filamentous fungus.
Claim 25
Dependent Claim
28 words
A process as claimed in claim 23 wherein said filamentous fungus is an Aspergillus species and said second filamentous fungus is a Humicola species or a Mucor species.
Claim 26
Dependent Claim
26 words
A process as claimed in claim 23 wherein said filamentous fungus is a first Aspergillus species and said second filamentous fungus is a second Aspergillus species.
Claim 27
Dependent Claim
15 words
A process as claimed in claim 2 wherein the secreted heterologous polypeptide is biochemically active.
Claim 28
Dependent Claim
35 words
A process as claimed in claim 2 wherein said filamentous fungus from the subdivision Eumycotina is selected from members of the group of genera consisting of Aspergillus, Trichoderma, Neurospora, Podospora, Endothia Mucor, Cochiobolus and Pyricularia.
Claim 29
Dependent Claim
29 words
A process as claimed in claim 2 wherein said filamentous fungus from the subdivision Eumycotina is selected from members of the group of genera consisting of Aspergillus and Trichoderma.
Claim 30
Dependent Claim
25 words
A process as claimed in claim 2 wherein said filamentous fungus is selected from the group consisting of Aspergillus nidulans, Aspergillus awamori and Trichoderma reesei.
References Cited (Backward Citations)
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