US2017051025A1PendingUtilityA1

Silk Proteins

51
Assignee: COMMW SCIENT IND RES ORGPriority: Oct 5, 2005Filed: Jun 29, 2016Published: Feb 23, 2017
Est. expiryOct 5, 2025(expired)· nominal 20-yr term from priority
Inventors:Tara D. SutherlandVictoria S. HaritosHolly TruemanAlagacone SriskanthaSarah WeismanPeter M. Campbell
A61P 43/00A61P 17/02C12P 21/02C12N 15/70C07K 2319/73C07K 2319/00C07K 14/43572C07K 14/43563C12N 15/8257C12N 15/63C07K 19/00C07K 14/435A61P 17/00A61K 47/42
51
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Claims

Abstract

The present invention provides silk proteins, as well as nucleic acids encoding these proteins. The present invention also provides recombinant cells and/or organisms which synthesize silk proteins. Silk proteins of the invention can be used for a variety of purposes such as in the manufacture of personal care products, plastics, textiles, and biomedical products.

Claims

exact text as granted — not AI-modified
1 - 45 . (canceled) 
     
     
         46 . A silk polypeptide, wherein at least a portion of the polypeptide has a coiled coil structure, and wherein the polypeptide comprises an amino acid sequence which is at least 40% identical to any one or more of SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:22, SEQ ID NO:23, SEQ ID NO:40, SEQ ID NO:41, SEQ ID NO:56, SEQ ID NO:57; SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:24, SEQ ID NO:25, SEQ ID NO:42, SEQ ID NO:43, SEQ ID NO:58, SEQ ID NO:59, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:26, SEQ ID NO:27, SEQ ID NO:44, SEQ ID NO:45, SEQ ID NO:60, SEQ ID NO:61, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:28, SEQ ID NO:29, SEQ ID NO:46, SEQ ID NO:47, SEQ ID NO:62, or SEQ ID NO:63, and
 wherein the silk polypeptide is fused to at least one other polypeptide; or   wherein the silk polypeptide is present in a silk fiber or copolymer and crosslinked to a surface of interest.   
     
     
         47 . The silk polypeptide of  claim 46 , wherein
 i) the portion of the polypeptide that has a coiled coil structure comprises at least 10 copies of the heptad sequence abcdefg, and wherein at least 25% of the amino acids at positions a and d are alanine residues, or   ii) the portion of the polypeptide that has a coiled coil structure comprises at least 10 copies of the heptad sequence abcdefg, and at least 25% of the amino acids at positions a, d and e are alanine residues.   
     
     
         48 . The silk polypeptide of  claim 46  which is fused to at least one other polypeptide. 
     
     
         49 . The silk polypeptide of  claim 46 , wherein the silk polypeptide is present in a silk fiber or copolymer and crosslinked to a surface of interest. 
     
     
         50 . A product comprising at least one silk polypeptide of  claim 46 . 
     
     
         51 . The product of  claim 50 , wherein the product is selected from the group consisting of: a personal care product, textiles, plastics, and biomedical products. 
     
     
         52 . A vector comprising: a polynucleotide which encodes a silk polypeptide, wherein at least a portion of the polypeptide has a coiled coil structure, and wherein the silk polypeptide comprises an amino acid sequence which is at least 40% identical to at least any one or more of SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:22, SEQ ID NO:23, SEQ ID NO:40, SEQ ID NO:41, SEQ ID NO:56, SEQ ID NO:57; SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:24, SEQ ID NO:25, SEQ ID NO:42, SEQ ID NO:43, SEQ ID NO:58, SEQ ID NO:59, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:26, SEQ ID NO:27, SEQ ID NO:44, SEQ ID NO:45, SEQ ID NO:60, SEQ ID NO:61, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:28, SEQ ID NO:29, SEQ ID NO:46, SEQ ID NO:47, SEQ ID NO:62, or SEQ ID NO:63; and
 a heterologous promoter operably linked to the polynucleotide.   
     
     
         53 . A composition comprising the vector of  claim 52 , and one or more acceptable carriers. 
     
     
         54 . A recombinant host cell comprising a polynucleotide which encodes a silk polypeptide, wherein at least a portion of the polypeptide has a coiled coil structure, and wherein the silk polypeptide comprises an amino acid sequence which is at least 40% identical to at least any one or more of SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:22, SEQ ID NO:23, SEQ ID NO:40, SEQ ID NO:41, SEQ ID NO:56, SEQ ID NO:57; SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:24, SEQ ID NO:25, SEQ ID NO:42, SEQ ID NO:43, SEQ ID NO:58, SEQ ID NO:59, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:26, SEQ ID NO:27, SEQ ID NO:44, SEQ ID NO:45, SEQ ID NO:60, SEQ ID NO:61, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:28, SEQ ID NO:29, SEQ ID NO:46, SEQ ID NO:47, SEQ ID NO:62, or SEQ ID NO:63; and
 wherein   a) the polynucleotide is operably linked to a heterologous promoter, and/or   b) the recombinant host cell is a bacterial, yeast or plant cell.   
     
     
         55 . The recombinant host cell of  claim 54 , wherein the silk polypeptide comprises an amino acid sequence which is at least 50% identical to at least any one or more of SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:22, SEQ ID NO:23, SEQ ID NO:40, SEQ ID NO:41, SEQ ID NO:56, SEQ ID NO:57; SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:24, SEQ ID NO:25, SEQ ID NO:42, SEQ ID NO:43, SEQ ID NO:58, SEQ ID NO:59, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:26, SEQ ID NO:27, SEQ ID NO:44, SEQ ID NO:45, SEQ ID NO:60, SEQ ID NO:61, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:28, SEQ ID NO:29, SEQ ID NO:46, SEQ ID NO:47, SEQ ID NO:62, or SEQ ID NO:63. 
     
     
         56 . The recombinant host cell of  claim 54 , wherein the silk polypeptide comprises an amino acid sequence which is at least 70% identical to at least any one or more of SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:22, SEQ ID NO:23, SEQ ID NO:40, SEQ ID NO:41, SEQ ID NO:56, SEQ ID NO:57; SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:24, SEQ ID NO:25, SEQ ID NO:42, SEQ ID NO:43, SEQ ID NO:58, SEQ ID NO:59, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:26, SEQ ID NO:27, SEQ ID NO:44, SEQ ID NO:45, SEQ ID NO:60, SEQ ID NO:61, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:28, SEQ ID NO:29, SEQ ID NO:46, SEQ ID NO:47, SEQ ID NO:62, or SEQ ID NO:63. 
     
     
         57 . The recombinant host cell of  claim 54 , wherein the silk polypeptide comprises an amino acid sequence which is at least 80% identical to at least any one or more of SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:22, SEQ ID NO:23, SEQ ID NO:40, SEQ ID NO:41, SEQ ID NO:56, SEQ ID NO:57; SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:24, SEQ ID NO:25, SEQ ID NO:42, SEQ ID NO:43, SEQ ID NO:58, SEQ ID NO:59, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:26, SEQ ID NO:27, SEQ ID NO:44, SEQ ID NO:45, SEQ ID NO:60, SEQ ID NO:61, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:28, SEQ ID NO:29, SEQ ID NO:46, SEQ ID NO:47, SEQ ID NO:62, or SEQ ID NO:63. 
     
     
         58 . The recombinant host cell of  claim 54 , wherein the polynucleotide comprises a nucleic acid sequence which is at least 40% identical to any one or more of SEQ ID NO:11, SEQ ID NO:12, SEQ ID NO:31, SEQ ID NO:32, SEQ ID NO:48, SEQ ID NO:49, SEQ ID NO:64, SEQ ID NO:65, SEQ ID NO:13, SEQ ID NO:14, SEQ ID NO:33, SEQ ID NO:34, SEQ ID NO:50, SEQ ID NO:51, SEQ ID NO:66, SEQ ID NO:67, SEQ ID NO:15, SEQ ID NO:16, SEQ ID NO:35, SEQ ID NO:36, SEQ ID NO:52, SEQ ID NO:53, SEQ ID NO:68, SEQ ID NO:69, SEQ ID NO:17, SEQ ID NO:18, SEQ ID NO:37, SEQ ID NO:38, SEQ ID NO:54, SEQ ID NO:55, SEQ ID NO:70, SEQ ID NO:71, or SEQ ID NO:76. 
     
     
         59 . The recombinant host cell of  claim 54 , wherein the polynucleotide comprises a nucleic acid sequence which is at least 50% identical to any one or more of SEQ ID NO:11, SEQ ID NO:12, SEQ ID NO:31, SEQ ID NO:32, SEQ ID NO:48, SEQ ID NO:49, SEQ ID NO:64, SEQ ID NO:65, SEQ ID NO:13, SEQ ID NO:14, SEQ ID NO:33, SEQ ID NO:34, SEQ ID NO:50, SEQ ID NO:51, SEQ ID NO:66, SEQ ID NO:67, SEQ ID NO:15, SEQ ID NO:16, SEQ ID NO:35, SEQ ID NO:36, SEQ ID NO:52, SEQ ID NO:53, SEQ ID NO:68, SEQ ID NO:69, SEQ ID NO:17, SEQ ID NO:18, SEQ ID NO:37, SEQ ID NO:38, SEQ ID NO:54, SEQ ID NO:55, SEQ ID NO:70, SEQ ID NO:71, or SEQ ID NO:76. 
     
     
         60 . The recombinant host cell of  claim 54 , wherein the polynucleotide comprises a nucleic acid sequence which is at least 70% identical to any one or more of SEQ ID NO:11, SEQ ID NO:12, SEQ ID NO:31, SEQ ID NO:32, SEQ ID NO:48, SEQ ID NO:49, SEQ ID NO:64, SEQ ID NO:65, SEQ ID NO:13, SEQ ID NO:14, SEQ ID NO:33, SEQ ID NO:34, SEQ ID NO:50, SEQ ID NO:51, SEQ ID NO:66, SEQ ID NO:67, SEQ ID NO:15, SEQ ID NO:16, SEQ ID NO:35, SEQ ID NO:36, SEQ ID NO:52, SEQ ID NO:53, SEQ ID NO:68, SEQ ID NO:69, SEQ ID NO:17, SEQ ID NO:18, SEQ ID NO:37, SEQ ID NO:38, SEQ ID NO:54, SEQ ID NO:55, SEQ ID NO:70, SEQ ID NO:71, or SEQ ID NO:76. 
     
     
         61 . The recombinant host cell of  claim 54 , wherein the polynucleotide comprises a nucleic acid sequence which is at least 80% identical to any one or more of SEQ ID NO:11, SEQ ID NO:12, SEQ ID NO:31, SEQ ID NO:32, SEQ ID NO:48, SEQ ID NO:49, SEQ ID NO:64, SEQ ID NO:65, SEQ ID NO:13, SEQ ID NO:14, SEQ ID NO:33, SEQ ID NO:34, SEQ ID NO:50, SEQ ID NO:51, SEQ ID NO:66, SEQ ID NO:67, SEQ ID NO:15, SEQ ID NO:16, SEQ ID NO:35, SEQ ID NO:36, SEQ ID NO:52, SEQ ID NO:53, SEQ ID NO:68, SEQ ID NO:69, SEQ ID NO:17, SEQ ID NO:18, SEQ ID NO:37, SEQ ID NO:38, SEQ ID NO:54, SEQ ID NO:55, SEQ ID NO:70, SEQ ID NO:71, or SEQ ID NO:76. 
     
     
         62 . The recombinant host cell of  claim 54 , wherein the portion of the silk polypeptide that has a coiled coil structure comprises at least 10 copies of the heptad sequence abcdefg, and wherein at least 25% of the amino acids at positions a and d are alanine residues. 
     
     
         63 . The recombinant host cell of  claim 54 , wherein the portion of the silk polypeptide that has a coiled coil structure comprises at least 18 copies of the heptad sequence abcdefg, and wherein at least 25% of the amino acids at positions a and d are alanine residues. 
     
     
         64 . The recombinant host cell of  claim 54  which is a bacterial cell. 
     
     
         65 . A process for preparing a silk polypeptide comprising cultivating the recombinant host cell of  claim 54 , under conditions which allow expression of the polynucleotide encoding the polypeptide, and recovering the expressed polypeptide.

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