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1. (WO2018221892) TECHNIQUE FOR MASS PRODUCTION OF RECOMBINANT BOTULINUM TOXIN PROTEIN THROUGH FUSION PROTEIN CONFIGURATION OF HIGHLY SOLUBLE GREEN FLUORESCENT PROTEIN AND BOTULINUM TOXIN PROTEIN
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Pub. No.: WO/2018/221892 International Application No.: PCT/KR2018/005947
Publication Date: 06.12.2018 International Filing Date: 25.05.2018
IPC:
C07K 14/33 (2006.01) ,C12N 9/52 (2006.01)
C CHEMISTRY; METALLURGY
07
ORGANIC CHEMISTRY
K
PEPTIDES
14
Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
195
from bacteria
33
from Clostridium (G)
C CHEMISTRY; METALLURGY
12
BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
N
MICRO-ORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICRO-ORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
9
Enzymes, e.g. ligases (6.); Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating, or purifying enzymes
14
Hydrolases (3.)
48
acting on peptide bonds, e.g. thromboplastin, leucine aminopeptidase (3.4)
50
Proteinases
52
derived from bacteria
Applicants:
바이오위더스 주식회사 BIOWITHUS, INC. [KR/KR]; 서울시 성동구 광나루로6길 35, 508호 #508, 35, Gwangnaru-ro 6-gil, Seongdong-gu, Seoul 04799, KR
Inventors:
권오중 KWON, Oh Joong; KR
Agent:
윤대웅 YOON, Dae Woong; KR
Priority Data:
10-2017-006632429.05.2017KR
Title (EN) TECHNIQUE FOR MASS PRODUCTION OF RECOMBINANT BOTULINUM TOXIN PROTEIN THROUGH FUSION PROTEIN CONFIGURATION OF HIGHLY SOLUBLE GREEN FLUORESCENT PROTEIN AND BOTULINUM TOXIN PROTEIN
(FR) TECHNIQUE DE PRODUCTION EN MASSE DE PROTÉINE RECOMBINANTE DE TOXINE BOTULIQUE PAR CONFIGURATION DE PROTÉINE DE FUSION À PARTIR DE PROTÉINE FLUORESCENTE VERTE HAUTEMENT SOLUBLE ET DE PROTÉINE DE TOXINE BOTULIQUE
(KO) 고용해성 녹색형광 단백질과 보툴리늄 독소 단백질의 융합단백질 구성을 통한 재조합 보툴리늄 독소 단백질의 대량생산 기술
Abstract:
(EN) The present invention relates to a technique for mass production of a recombinant botulinum toxin protein through the fusion protein configuration of a highly soluble green fluorescent protein and a botulinum toxin protein. A recombinant botulinum toxin protein according to the present invention in which a highly soluble GFP-HS1 tag and a botulinum toxin protein are fused to each other exhibits excellent production yield thanks to the enhanced solubility in the cytoplasm upon the expression of the recombinant protein and is expressed as a holotoxin that is non-toxic. After protein expression, a protease region included within the fusion protein makes it easy not only to separate and purify the fusion protein, but also to obtain an active form of botulinum toxin from the fusion protein.
(FR) La présente invention concerne une technique de production en masse d'une protéine recombinante de toxine botulique par la configuration de protéine de fusion à partir d'une protéine fluorescente verte hautement soluble et d'une protéine de toxine botulique. Une protéine recombinante de toxine botulique selon la présente invention dans laquelle une étiquette GFP-HS1 hautement soluble et une protéine de toxine botulique sont fusionnées l'une à l'autre présente un excellent rendement de production grâce à la solubilité améliorée dans le cytoplasme lors de l'expression de la protéine recombinante et est exprimée sous la forme d'une holotoxine non toxique. Après l'expression de la protéine, une région de protéase incluse dans la protéine de fusion permet non seulement de séparer et de purifier la protéine de fusion, mais permet également d'obtenir une forme active de toxine botulique à partir de la protéine de fusion.
(KO) 본 발명은 고용해성 녹색형광 단백질과 보툴리늄 독소 단백질의 융합단백질 구성을 통한 재조합 보툴리늄 독소 단백질의 대량생산 기술에 관한 것이다. 본 발명의 재조합 보툴리늄 독소 단백질은 고용해성의 GFP-HS1 태그와 보툴리늄 독소 단백질을 융합한 것으로, 재조합 단백질의 발현 시 세포질 내 용해도가 향상되어 생산수율이 우수하며, 비-독성인 완전독소(holotoxin)로서 발현되고, 프로테아제 영역을 포함하여 단백질 발현 후 분리 정제뿐만 아니라 활성화 형태의 보툴리늄 독소를 수득하는 데에도 용이하다.
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Designated States: AE, AG, AL, AM, AO, AT, AU, AZ, BA, BB, BG, BH, BN, BR, BW, BY, BZ, CA, CH, CL, CN, CO, CR, CU, CZ, DE, DJ, DK, DM, DO, DZ, EC, EE, EG, ES, FI, GB, GD, GE, GH, GM, GT, HN, HR, HU, ID, IL, IN, IR, IS, JO, JP, KE, KG, KH, KN, KP, KW, KZ, LA, LC, LK, LR, LS, LU, LY, MA, MD, ME, MG, MK, MN, MW, MX, MY, MZ, NA, NG, NI, NO, NZ, OM, PA, PE, PG, PH, PL, PT, QA, RO, RS, RU, RW, SA, SC, SD, SE, SG, SK, SL, SM, ST, SV, SY, TH, TJ, TM, TN, TR, TT, TZ, UA, UG, US, UZ, VC, VN, ZA, ZM, ZW
African Regional Intellectual Property Organization (ARIPO) (BW, GH, GM, KE, LR, LS, MW, MZ, NA, RW, SD, SL, ST, SZ, TZ, UG, ZM, ZW)
Eurasian Patent Office (AM, AZ, BY, KG, KZ, RU, TJ, TM)
European Patent Office (EPO) (AL, AT, BE, BG, CH, CY, CZ, DE, DK, EE, ES, FI, FR, GB, GR, HR, HU, IE, IS, IT, LT, LU, LV, MC, MK, MT, NL, NO, PL, PT, RO, RS, SE, SI, SK, SM, TR)
African Intellectual Property Organization (BF, BJ, CF, CG, CI, CM, GA, GN, GQ, GW, KM, ML, MR, NE, SN, TD, TG)
Publication Language: Korean (KO)
Filing Language: Korean (KO)