CN113614226A - 在蛋白中高效引入赖氨酸衍生物的氨酰基-tRNA合成酶 - Google Patents

在蛋白中高效引入赖氨酸衍生物的氨酰基-tRNA合成酶 Download PDF

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Publication number
CN113614226A
CN113614226A CN202080023301.XA CN202080023301A CN113614226A CN 113614226 A CN113614226 A CN 113614226A CN 202080023301 A CN202080023301 A CN 202080023301A CN 113614226 A CN113614226 A CN 113614226A
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trna synthetase
amino acid
lysyl
acid sequence
mutant
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Pending
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CN202080023301.XA
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吴松
张振山
刘慧玲
陈卫
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Ningbo Kunpeng Biotech Co Ltd
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Ningbo Kunpeng Biotech Co Ltd
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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
    • C12N15/70—Vectors or expression systems specially adapted for E. coli
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • C07K14/575—Hormones
    • C07K14/62—Insulins
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C12N9/14—Hydrolases (3)
    • C12N9/16—Hydrolases (3) acting on ester bonds (3.1)
    • C12N9/22—Ribonucleases RNAses, DNAses
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C12N9/93—Ligases (6)
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Y—ENZYMES
    • C12Y601/00—Ligases forming carbon-oxygen bonds (6.1)
    • C12Y601/01—Ligases forming aminoacyl-tRNA and related compounds (6.1.1)
    • C12Y601/01006—Lysine-tRNA ligase (6.1.1.6)

Abstract

提供了一种突变型赖氨酰‑tRNA合成酶。与野生型赖氨酰‑tRNA合成酶相比，本发明的突变型赖氨酰‑tRNA合成酶可提高非天然氨基酸的插入量和含有非天然氨基酸的目的蛋白量，还可以提高目的蛋白的稳定性，使其不容易断裂。

Description

PCT国内申请，说明书已公开。

Claims (10)

  1. PCT国内申请，权利要求书已公开。
CN202080023301.XA 2019-03-19 2020-03-18 在蛋白中高效引入赖氨酸衍生物的氨酰基-tRNA合成酶 Pending CN113614226A (zh)

Applications Claiming Priority (3)

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CN201910209626.6A CN111718920B (zh) 2019-03-19 2019-03-19 在蛋白中高效引入赖氨酸衍生物的氨酰基-tRNA合成酶
CN2019102096266 2019-03-19
PCT/CN2020/080034 WO2020187269A1 (zh) 2019-03-19 2020-03-18 在蛋白中高效引入赖氨酸衍生物的氨酰基—tRNA合成酶

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CN202080023301.XA Pending CN113614226A (zh) 2019-03-19 2020-03-18 在蛋白中高效引入赖氨酸衍生物的氨酰基-tRNA合成酶

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US (1) US20220170030A1 (zh)
EP (1) EP3943599A4 (zh)
JP (1) JP7383306B2 (zh)
CN (2) CN111718920B (zh)
AU (1) AU2020242724B2 (zh)
BR (1) BR112021018681A2 (zh)
CA (1) CA3134163A1 (zh)
WO (1) WO2020187269A1 (zh)

Families Citing this family (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN111850020B (zh) * 2019-04-25 2021-05-07 苏州鲲鹏生物技术有限公司 利用质粒系统在蛋白中引入非天然氨基酸
CN111849929B (zh) * 2019-04-30 2021-05-11 苏州鲲鹏生物技术有限公司 高效引入赖氨酸衍生物的氨酰基—tRNA合成酶
CN113773399B (zh) * 2020-06-09 2023-05-12 宁波鲲鹏生物科技有限公司 一种甘精胰岛素衍生物及其应用
CN115701451B (zh) * 2021-08-02 2023-08-01 宁波鲲鹏生物科技有限公司 高效引入赖氨酸衍生物的氨酰基-tRNA合成酶及其应用
CN115717130A (zh) * 2022-09-02 2023-02-28 凯莱英医药集团（天津）股份有限公司 氨酰-tRNA合酶突变体及烯基酪氨酰-tRNA的制备方法

Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101910403A (zh) * 2007-10-30 2010-12-08 医药研究委员会 方法和组合物
CN104203971A (zh) * 2012-01-20 2014-12-10 医药研究委员会 多肽和方法
CN105247049A (zh) * 2013-03-15 2016-01-13 米迪缪尼有限公司 新型核酸分子
WO2018182284A1 (ko) * 2017-03-27 2018-10-04 재단법인 의약바이오컨버젼스연구단 세포 외막에 노출되는 라이실-tRNA 합성효소 N-말단 영역에 특이적으로 결합하는 항체

Family Cites Families (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
SI1666604T1 (sl) * 2003-07-07 2008-08-31 Scripps Research Inst Sestavki ortogonalnih parov lizil-tRNA in aminoacil-tRNA in aminoacil-tRNA-sintetaze in njihove uporabe
JP5590649B2 (ja) 2007-09-20 2014-09-17 独立行政法人理化学研究所 変異体ピロリジル−ｔＲＮＡ合成酵素及びこれを用いる非天然アミノ酸組み込みタンパク質の製造方法
WO2009066761A1 (ja) * 2007-11-22 2009-05-28 Riken エステル結合を含む非天然タンパク質の製造方法
CN104099360A (zh) * 2013-04-12 2014-10-15 北京大学 非天然氨基酸标记的目的蛋白或肽的制备
WO2015193897A1 (en) * 2014-06-17 2015-12-23 B G Negev Technologies And Applications Ltd At Ben Genetically expanded cell free protein synthesis systems, methods and kits
EP3309260A1 (en) * 2016-10-14 2018-04-18 European Molecular Biology Laboratory Archaeal pyrrolysyl trna synthetases for orthogonal use
WO2019010164A1 (en) * 2017-07-06 2019-01-10 President And Fellows Of Harvard College EVOLUTION OF ARNT SYNTHÉTASES

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101910403A (zh) * 2007-10-30 2010-12-08 医药研究委员会 方法和组合物
CN104203971A (zh) * 2012-01-20 2014-12-10 医药研究委员会 多肽和方法
CN105247049A (zh) * 2013-03-15 2016-01-13 米迪缪尼有限公司 新型核酸分子
WO2018182284A1 (ko) * 2017-03-27 2018-10-04 재단법인 의약바이오컨버젼스연구단 세포 외막에 노출되는 라이실-tRNA 합성효소 N-말단 영역에 특이적으로 결합하는 항체

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
NCBI GENBANK DATABASE: "poyrolysyl-tRNA ligase [Methanosarcina mazei]，NCBI Reference Sequence: WP_048047042.1， 454aa linear", NCBI GENBANK DATABASE, pages 1 *
SOON BIN KWON等: "Nucleic Acid-Dependent Structural Transition of the Intrinsically Disordered N-Terminal Appended Domain of Human tysyl-tRNA Synthetase:", INT J MOL SCI, vol. 19, no. 10, pages 1 - 16 *

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EP3943599A1 (en) 2022-01-26
AU2020242724A1 (en) 2021-11-11
CA3134163A1 (en) 2020-09-24
WO2020187269A1 (zh) 2020-09-24
CN111718920A (zh) 2020-09-29
US20220170030A1 (en) 2022-06-02
CN111718920B (zh) 2021-05-07
JP7383306B2 (ja) 2023-11-20
EP3943599A4 (en) 2023-05-10
JP2022527991A (ja) 2022-06-07
BR112021018681A2 (pt) 2021-11-23
AU2020242724B2 (en) 2023-11-30

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