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1. WO2020221365 - AMINOACYL-TRNA SYNTHETASE EFFICIENTLY INTRODUCING LYSINE DERIVATIVES

Publication Number WO/2020/221365
Publication Date 05.11.2020
International Application No. PCT/CN2020/088506
International Filing Date 30.04.2020
IPC
C12N 9/00 2006.1
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9Enzymes, e.g. ligases (6.); Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating, or purifying enzymes
C12N 15/63 2006.1
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15Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
09Recombinant DNA-technology
63Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
CPC
C07K 14/62
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07ORGANIC CHEMISTRY
KPEPTIDES
14Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
435from animals; from humans
575Hormones
62Insulins
C07K 2319/00
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2319Fusion polypeptide
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15Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
09Recombinant DNA-technology
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62DNA sequences coding for fusion proteins
C12N 15/70
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15Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
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70Vectors or expression systems specially adapted for E. coli
C12N 9/93
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21Preparation of peptides or proteins
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Applicants
  • 苏州鲲鹏生物技术有限公司 SUZHOU KUNPENG BIOTECH CO., LTD. [CN]/[CN]
Inventors
  • 刘慧玲 LIU, Huiling
  • 吴松 WU, Song
  • 张振山 ZHANG, Zhenshan
  • 陈卫 CHEN, Wei
Agents
  • 上海一平知识产权代理有限公司 XU & PARTNERS, LLC.
Priority Data
201910364788.730.04.2019CN
Publication Language Chinese (zh)
Filing Language Chinese (ZH)
Designated States
Title
(EN) AMINOACYL-TRNA SYNTHETASE EFFICIENTLY INTRODUCING LYSINE DERIVATIVES
(FR) AMINOACYL-ARNT SYNTHÉTASE INTRODUISANT EFFICACEMENT DES DÉRIVÉS DE LYSINE
(ZH) 高效引入赖氨酸衍生物的氨酰基—tRNA合成酶
Abstract
(EN) Provided by the present invention is a mutant lysyl-tRNA synthetase, which is deleted at one or more sites selected from the following group corresponding to the amino acid sequence of wild-type aminoacyl-tRNA synthetase: 102nd, 128th-140th and 159th-179th amino acid residues. Compared with wild-type lysyl-tRNA synthetase, the mutant lysyl-tRNA synthetase according to the present invention has high activity, a high level of expression and good solubility, and may significantly increase the amount of an inserted unnatural amino acid and the level of expression of a target protein containing an unnatural amino acid.
(FR) La présente invention concerne une lysyl-ARNt synthétase mutante, étant supprimée en un ou plusieurs sites choisis dans le groupe suivant correspondant à la séquence d'acides aminés de l'aminoacyl-ARNt synthétase de type sauvage : 102 ème, 128 ème à 140 ème et 159 ème à 179 ème résidus d'acides aminés. Par comparaison avec la lysyl-ARNt synthétase de type sauvage, la lysyl-ARNt synthétase mutante selon la présente invention présente une activité élevée, un niveau élevé d'expression et une bonne solubilité, et peut augmenter significativement la quantité d'un acide aminé non naturel inséré et le niveau d'expression d'une protéine cible contenant un acide aminé non naturel.
(ZH) 本发明提供了一种突变型赖氨酰-tRNA合成酶,其在对应于野生型氨酰基-tRNA合成酶的氨基酸序列的选自下组的一个或多个位点发生缺失:第102位、第128-140位和第159-179位氨基酸残基。相比野生型赖氨酰-tRNA合成酶,本发明突变型赖氨酰-tRNA合成酶活性高,表达量高,可溶性好,可显著提高非天然氨基酸插入量和含有非天然氨基酸的目的蛋白的表达量。
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