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Применить Всего найдено 16. Отображено 16.
30-03-2017 дата публикации

Methods and Compositions for Synthesizing Improved Silk Fibers

Номер: US20170088675A1
Автор: Daniel M. Widmaier, David N. Breslauer, Joshua Kittleson, Brendan Turner, Lindsay Wray, WIDMAIER DANIEL M, BRESLAUER DAVID N, KITTLESON JOSHUA, TURNER BRENDAN, WRAY LINDSAY, Widmaier Daniel M., Breslauer David N., Kittleson Joshua, Turner Brendan, Wray Lindsay
Принадлежит:

The present disclosure provides methods and compositions for directed to synthetic block copolymer proteins, expression constructs for their secretion, recombinant microorganisms for their production, and synthetic fibers (including advantageously, microfibers) comprising these proteins that recapitulate many properties of natural silk. The recombinant microorganisms can be used for the commercial production of silk-like fibers. 1. A fiber spinning solution , comprisinga polypeptide, the polypeptide comprising from 2 to 8 concatenated repeats of SEQ ID NO: 1396 or circularly permuted variants thereof, the polypeptide further comprising a FLAG tag; anda spinning solvent, wherein said polypeptide is dissolved in said spinning solvent.2. The fiber spinning solution of claim 1 , wherein said spinning solvent comprises formic acid.3. The fiber spinning solution of claim 1 , wherein said spinning solvent is formic acid.4. The fiber spinning solution of claim 1 , wherein said fiber spinning solution comprises from 20-30% of said polypeptide by weight.5. The fiber spinning solution of claim 1 , wherein the FLAG tag is on the C-terminal end of the polypeptide.6. The fiber spinning solution of claim 1 , wherein said FLAG tag is a 3×FLAG tag.7. The fiber spinning solution of claim 1 , wherein said FLAG tag is encoded by a polynucleotide comprising SEQ ID NO: 1409.8. The fiber spinning solution of claim 1 , wherein the polypeptide comprises a property selected from the group consisting of an alanine composition from 12 to 40% claim 1 , a glycine composition from 25 to 50% claim 1 , a proline composition from 9 to 20% claim 1 , a β-turn composition from 15 to 37% claim 1 , a GPG amino acid motif content from 18 to 55% claim 1 , and a poly-alanine amino acid motif content from 9 to 35%.9. The fiber spinning solution of claim 1 , wherein the polypeptide comprises exactly 3 concatenated repeats of SEQ ID NO: 1396.10. A method of making a silk fiber claim 1 , comprising:{'claim-ref ...

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Номер записи: 1
04-08-2016 дата публикации

Method and Compositions for Synthesizing Improved Silk Fibers

Номер: US20160222174A1
Автор: Daniel M. Widmaier, David N. Breslauer, Joshua Kittleson, Brendan Turner, Lindsay Wray, WIDMAIER DANIEL M, BRESLAUER DAVID N, KITTLESON JOSHUA, TURNER BRENDAN, WRAY LINDSAY, Widmaier Daniel M., Breslauer David N., Kittleson Joshua, Turner Brendan, Wray Lindsay
Принадлежит:

The present disclosure provides methods and compositions for directed to synthetic block copolymer proteins, expression constructs for their secretion, recombinant microorganisms for their production, and synthetic fibers (including advantageously, microfibers) comprising these proteins that recapitulate many properties of natural silk. The recombinant microorganisms can be used for the commercial production of silk-like fibers. 1. A proteinaceous block co-polymer , comprising:a polypeptide, the polypeptide comprising from 2 to 8 concatenated repeats of SEQ ID NO: 1396 or circularly permuted variants thereof.2. The proteinaceous block co-polymer of claim 1 , wherein the polypeptide comprises a property selected from the group consisting of an alanine composition from 12 to 40% claim 1 , a glycine composition from 25 to 50% claim 1 , a proline composition from 9 to 20% claim 1 , a β-turn composition from 15 to 37% claim 1 , a GPG amino acid motif content from 18 to 55% claim 1 , and a poly-alanine amino acid motif content from 9 to 35%.3. The proteinaceous block co-polymer of claim 1 , consisting of: 3 concatenated repeats of SEQ ID NO: 1396.4. A fiber comprising the block co-polymer of .5. The fiber of claim 4 , comprising a circular cross-section and a diameter from 4.48 to 12.7 μm.6. The fiber of claim 4 , wherein the fiber has a yield stress from 24 to 172 MPa.7. The fiber of claim 4 , wherein the fiber has a maximum stress from 54 to 310 MPa.8. The fiber of claim 4 , wherein the fiber has a breaking strain from 2 to 200%.9. The fiber of claim 4 , wherein the fiber has an initial modulus from 1617 to 5820 MPa.10. The fiber of claim 4 , wherein the fiber has a toughness value from 0.5 MJ/mto 59.2 MJ/m.11. An expression construct encoding the proteinaceous block co-polymer of .12. The expression construct of claim 11 , further comprising a secretion signal operatively linked to the coding sequence for the block copolymer polypeptide.13. The expression construct of ...

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Номер записи: 2
20-02-2020 дата публикации

Methods and Compositions for Synthesizing Improved Silk Fibers

Номер: US20200055996A1
Автор: Breslauer David N., Kittleson Joshua, Turner Brendan, Widmaier Daniel M., Wray Lindsay
Принадлежит:

The present disclosure provides methods and compositions for directed to synthetic block copolymer proteins, expression constructs for their secretion, recombinant microorganisms for their production, and synthetic fibers (including advantageously, microfibers) comprising these proteins that recapitulate many properties of natural silk. The recombinant microorganisms can be used for the commercial production of silk-like fibers. 1. An expression construct encoding a proteinaceous block co-polymer , comprising a polypeptide , the polypeptide comprising from 2 to 8 concatenated repeats of SEQ ID NO: 1396 or circularly permuted variants thereof.2. The expression construct of claim 1 , further comprising a secretion signal operatively linked to the coding sequence for the block co-polymer polypeptide.3. The expression construct of claim 2 , wherein the secretion signal comprises an alpha mating factor leader and pro sequence.4. The expression construct of claim 1 , wherein the polypeptide comprises a property selected from the group consisting of an alanine composition from 12 to 40% claim 1 , a glycine composition from 25 to 50% claim 1 , a proline composition from 9 to 20% claim 1 , a β-turn composition from 15 to 37% claim 1 , a GPG amino acid motif content from 18 to 55% claim 1 , and a poly-alanine amino acid motif content from 9 to 35%.5. The expression construct of claim 1 , the polypeptide consisting of 3 concatenated repeats of SEQ ID NO: 1396.6. A host cell claim 1 , comprising: the expression construct of .7. The host cell of claim 6 , wherein the cell is selected from the group consisting of: a yeast cell claim 6 , a fungal cell claim 6 , and a gram positive bacteria cell.8Trichoderma.. The host cell of claim 7 , wherein the cell is from the genus9. A method of producing a proteinaceous block co-polymer comprising: a polypeptide comprising from 2 to 8 concatenated repeats of SEQ ID NO: 1396 or circularly permuted variants thereof claim 7 , the method ...

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Номер записи: 3
04-11-2021 дата публикации

Elastomeric Proteins

Номер: US20210340194A1
Автор: Daniel M. WIDMAIER, David N. Breslauer, Joshua KITTLESON, Michael Lee
Принадлежит: Bolt Threads Inc

This invention relates to elastomeric protein and elastomeric protein production. In particular, the invention is directed to elastomeric protein sequences, including methods and compositions for production of elastomeric protein sequences, such as expression constructs, and host cells, and including compositions generated from the elastomeric protein sequences.

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Номер записи: 4
27-09-2018 дата публикации

Methods and Compositions for Synthesizing Improved Silk Fibers

Номер: US20180273693A1
Автор: Brendan TURNER, Daniel M. WIDMAIER, David N. Breslauer, Joshua KITTLESON, Lindsay WRAY
Принадлежит: Bolt Threads Inc

The present disclosure provides methods and compositions for directed to synthetic block copolymer proteins, expression constructs for their secretion, recombinant microorganisms for their production, and synthetic fibers (including advantageously, microfibers) comprising these proteins that recapitulate many properties of natural silk. The recombinant microorganisms can be used for the commercial production of silk-like fibers.

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Номер записи: 5
04-10-2018 дата публикации

COMPOSITIONS AND METHODS FOR PRODUCING HIGH SECRETED YIELDS OF RECOMBINANT PROTEINS

Номер: US20180282380A1
Автор: Breslauer David N., Hill Rena, Kittleson Joshua, Pesce Carlos Gustavo, Stevens Thomas, Widmaier Daniel M.
Принадлежит:

The present disclosure relates to methods for producing recombinant proteins, as well as compositions used in and produced by such methods. Specifically, the present disclosure relates to methods for producing high secreted yields of recombinant proteins, and the compositions provided herein include expression constructs, recombinant vectors, and recombinant host cells that comprise polynucleotide sequences encoding proteins operably linked to recombinant secretion signals that comprise the leader peptide of the α-mating factor (αMF) of and a non-αMF signal peptide. 1. An expression construct comprising a polynucleotide sequence that encodes a protein operably linked to a recombinant secretion signal , wherein said recombinant secretion signal comprises a leader peptide and a signal peptide , wherein said leader peptide is native pro-αMF(sc) or a functional variant that has an at least 80% amino acid sequence identity to SEQ ID NO: 1 , and wherein said signal peptide does not comprise pre-αMF(sc).2. The expression construct of claim 1 , wherein said functional variant is native pro-αMF(sc) comprising one or two substituted amino acids.3. The expression construct of claim 1 , wherein said functional variant is SEQ ID NO: 2.4. The expression construct of claim 1 , wherein said functional variant has an at least 85% claim 1 , at least 90% claim 1 , at least 95% claim 1 , or at least 99% amino acid sequence identity to SEQ ID NO: 1.5. The expression construct of claim 1 , wherein said signal peptide is selected from Table 1 or is a functional variant that has an at least 80% amino acid sequence identity to a signal peptide selected from Table 1.6. (canceled)7. The expression construct of claim 1 , wherein said recombinant secretion signal is selected from Table 2 or is a functional variant that has an at least 80% amino acid sequence identity to a recombinant secretion signal selected from Table 2.8. (canceled)9. The expression construct of claim 1 , wherein said ...

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Номер записи: 6
18-10-2018 дата публикации

Methods and Compositions for Synthesizing Improved Silk Fibers

Номер: US20180298151A1
Автор: Breslauer David N., Kittleson Joshua, Turner Brendan, Widmaier Daniel M., Wray Lindsay
Принадлежит:

The present disclosure provides methods and compositions for directed to synthetic block copolymer proteins, expression constructs for their secretion, recombinant microorganisms for their production, and synthetic fibers (including advantageously, microfibers) comprising these proteins that recapitulate many properties of natural silk. The recombinant microorganisms can be used for the commercial production of silk-like fibers. 1. A fiber spinning solution , comprisinga polypeptide, the polypeptide comprising from 2 to 8 concatenated repeats of a recombinant silk polypeptide or circularly permuted variants thereof, the polypeptide further comprising a FLAG tag; anda spinning solvent, wherein said polypeptide is dissolved in said spinning solvent.2. The fiber spinning solution of claim 1 , wherein said spinning solvent comprises formic acid.3. The fiber spinning solution of claim 1 , wherein said spinning solvent is formic acid.4. The fiber spinning solution of claim 1 , wherein said fiber spinning solution comprises from 20-30% of said polypeptide by weight.5. The fiber spinning solution of claim 1 , wherein the FLAG tag is on the C-terminal end of the polypeptide.6. The fiber spinning solution of claim 1 , wherein said FLAG tag is a 3×FLAG tag.7. The fiber spinning solution of claim 1 , wherein said FLAG tag is encoded by a polynucleotide comprising SEQ ID NO: 1409.8. The fiber spinning solution of claim 1 , wherein the polypeptide comprises a property selected from the group consisting of an alanine composition from 12 to 40% claim 1 , a glycine composition from 25 to 50% claim 1 , a proline composition from 9 to 20% claim 1 , a β-turn composition from 15 to 37% claim 1 , a GPG amino acid motif content from 18 to 55% claim 1 , and a poly-alanine amino acid motif content from 9 to 35%.9. The fiber spinning solution of claim 1 , wherein said concatenated repeats comprise SEQ ID NO: 1396.10. A method of making a silk fiber claim 1 , comprising:{'claim-ref': {'@idref ...

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Номер записи: 7
31-10-2019 дата публикации

Elastomeric Proteins

Номер: US20190330287A1
Автор: Breslauer David N., Kittleson Joshua, Lee Michael, Widmaier Daniel M.
Принадлежит:

This invention relates to elastomeric protein and elastomeric protein production. In particular, the invention is directed to elastomeric protein sequences, including methods and compositions for production of elastomeric protein sequences, such as expression constructs, and host cells, and including compositions generated from the elastomeric protein sequences.

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Номер записи: 8
26-03-2015 дата публикации

Methods and compositions for synthesizing improved silk fibers

Номер: WO2015042164A2
Автор: Brendan TURNER, Daniel M. WIDMAIER, David N. Breslauer, Josh KITTLESON, Lindsay WRAY
Принадлежит: Refactored Materials, Inc.

The present disclosure provides methods and compositions for directed to synthetic block copolymer proteins, expression constructs for their secretion, recombinant microorganisms for their production, and synthetic fibers (including advantageously, microfibers) comprising these proteins that recapitulate many properties of natural silk. The recombinant microorganisms can be used for the commercial production of silk-like fibers.

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Номер записи: 9
31-07-2018 дата публикации

Methods and compositions for synthesizing improved silk fibers

Номер: US10035886B2
Автор: Brendan TURNER, Daniel M. WIDMAIER, David N. Breslauer, Joshua KITTLESON, Lindsay WRAY
Принадлежит: Bolt Threads Inc

The present disclosure provides methods and compositions for directed to synthetic block copolymer proteins, expression constructs for their secretion, recombinant microorganisms for their production, and synthetic fibers (including advantageously, microfibers) comprising these proteins that recapitulate many properties of natural silk. The recombinant microorganisms can be used for the commercial production of silk-like fibers.

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Номер записи: 10
20-11-2019 дата публикации

Elastomeric proteins

Номер: EP3568408A2
Автор: Daniel M. WIDMAIER, David N. Breslauer, Joshua KITTLESON, Michael Lee
Принадлежит: Bolt Threads Inc

This invention relates to elastomeric protein and elastomeric protein production. In particular, the invention is directed to elastomeric protein sequences, including methods and compositions for production of elastomeric protein sequences, such as expression constructs, and host cells, and including compositions generated from the elastomeric protein sequences.

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Номер записи: 11
27-07-2016 дата публикации

Methods and compositions for synthesizing improved silk fibers

Номер: EP3046585A2
Автор: Brendan TURNER, Daniel M. WIDMAIER, David N. Breslauer, Josh KITTLESON, Lindsay WRAY
Принадлежит: Bolt Threads Inc

The present disclosure provides methods and compositions for directed to synthetic block copolymer proteins, expression constructs for their secretion, recombinant microorganisms for their production, and synthetic fibers (including advantageously, microfibers) comprising these proteins that recapitulate many properties of natural silk. The recombinant microorganisms can be used for the commercial production of silk-like fibers.

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Номер записи: 12
08-11-2023 дата публикации

エラストマータンパク質

Номер: JP2023162366A
Автор: Kittleson Joshua, Lee Michael, M Widmaier Daniel, N Breslauer David, ジョシュア キトルソン, ダニエル エム． ウィドマイアー, デイビッド エヌ． ブレスロウアー, マイケル リー
Принадлежит: Bolt Threads Inc

【課題】細胞外部分からエラストマータンパク質を回収するための方法を含む、組換えエラストマータンパク質の発現および精製のための改善された方法を提供する。【解決手段】組換えレジリンタンパク質を含む組成物を作製するための方法は、発酵物中で組換え宿主細胞の集団を培養する工程であって、組換え宿主細胞が、分泌型レジリンコード配列を含むベクターを含み、かつ組換え宿主細胞が、分泌型レジリンコード配列によってコードされた組換えレジリンタンパク質を分泌する、工程；および発酵物から組換えレジリンタンパク質を精製する工程を含む。【選択図】図３

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Номер записи: 13
02-01-2024 дата публикации

Elastomeric proteins

Номер: US11858971B2
Автор: Daniel M. WIDMAIER, David N. Breslauer, Joshua KITTLESON, Michael Lee
Принадлежит: Bolt Threads Inc

This invention relates to elastomeric protein and elastomeric protein production. In particular, the invention is directed to elastomeric protein sequences, including methods and compositions for production of elastomeric protein sequences, such as expression constructs, and host cells, and including compositions generated from the elastomeric protein sequences.

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Номер записи: 14
09-05-2024 дата публикации

Compositions and Methods for Producing High Secreted Yields of Recombinant Proteins

Номер: US20240150416A1
Автор: Carlos Gustavo Pesce, Daniel M. WIDMAIER, David N. Breslauer, Joshua KITTLESON, Rena Hill, Thomas Stevens
Принадлежит: Bolt Threads Inc

The present disclosure relates to methods for producing recombinant proteins, as well as compositions used in and produced by such methods. Specifically, the present disclosure relates to methods for producing high secreted yields of recombinant proteins, and the compositions provided herein include expression constructs, recombinant vectors, and recombinant host cells that comprise polynucleotide sequences encoding proteins operably linked to recombinant secretion signals that comprise the leader peptide of the α-mating factor (αMF) of Saccharomyces cerevisiae and a non-αMF signal peptide.

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Номер записи: 15
08-05-2018 дата публикации

Methods and compositions for synthesizing improved silk fibers

Номер: US09963554B2
Автор: Brendan TURNER, Daniel M. WIDMAIER, David N. Breslauer, Joshua KITTLESON, Lindsay WRAY
Принадлежит: Bolt Threads Inc

The present disclosure provides methods and compositions for directed to synthetic block copolymer proteins, expression constructs for their secretion, recombinant microorganisms for their production, and synthetic fibers (including advantageously, microfibers) comprising these proteins that recapitulate many properties of natural silk. The recombinant microorganisms can be used for the commercial production of silk-like fibers.

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Номер записи: 16