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1. (WO2017131279) INSOLUBLE FUSION PROTEIN COMPRISING ANTIMICROBIAL PEPTIDE AND METHOD FOR PRODUCING ANTIMICROBIAL PEPTIDE USING SAME
Latest bibliographic data on file with the International Bureau

Pub. No.: WO/2017/131279 International Application No.: PCT/KR2016/001940
Publication Date: 03.08.2017 International Filing Date: 26.02.2016
IPC:
C07K 14/195 (2006.01) ,C07K 14/37 (2006.01) ,C12N 15/62 (2006.01) ,C12N 15/70 (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
C CHEMISTRY; METALLURGY
07
ORGANIC CHEMISTRY
K
PEPTIDES
14
Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
37
from fungi
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
15
Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
09
Recombinant DNA-technology
11
DNA or RNA fragments; Modified forms thereof
62
DNA sequences coding for fusion proteins
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
15
Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
09
Recombinant DNA-technology
63
Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
70
Vectors or expression systems specially adapted for E. coli
Applicants:
건국대학교 산학협력단 KONKUK UNIVERSITY INDUSTRIAL COOPERATION CORP [KR/KR]; 서울시 광진구 능동로 120 120 Neungdong-ro Gwangjin-gu Seoul 05029, KR
Inventors:
박찬규 PARK, Chan Kyu; KR
Agent:
김정수 KIM, Jung Su; KR
Priority Data:
10-2016-000997027.01.2016KR
Title (EN) INSOLUBLE FUSION PROTEIN COMPRISING ANTIMICROBIAL PEPTIDE AND METHOD FOR PRODUCING ANTIMICROBIAL PEPTIDE USING SAME
(FR) PROTÉINE DE FUSION INSOLUBLE COMPRENANT UN PEPTIDE ANTIMICROBIEN ET PROCÉDÉ DE PRODUCTION DE PEPTIDE ANTIMICROBIEN UTILISANT CETTE DERNIÈRE
(KO) 항균 펩타이드를 포함하는 불용성 융합단백질 및 이를 이용한 항균 펩타이드의 제조 방법
Abstract:
(EN) The present invention relates to a method for producing an antimicrobial peptide, and more specifically, includes a method for mass-producing an antimicrobial peptide through a process in which an antimicrobial peptide gene is fused with a green fluorescent protein gene which is expressed as insoluble in Escherichia coli, and same is inserted and expressed in E. coli, then the green fluorescent protein is removed, and the antimicrobial peptide is obtained. According to the present invention, a method for producing an antimicrobial peptide is provided, the method having a simple production process, being economical, and having a high production yield, and thus the present invention has the advantage of providing a natural antibiotic that may replace conventional antibiotics to the pharmaceutical and feed industries which have an urgent need to develop an antibiotic having a new mechanism of action that may eradicate resistant strains since such multidrug-resistant microorganisms are rapidly increasing. Or, according to the present invention, a method for cleaving amino acids by means of acid treatment, instead of using the conventional cyanogen bromide, is used, and thus, in purifying a target protein from an insoluble protein, costs may be cut, danger in the process may be reduced, and the process may be more quickly carried out.
(FR) La présente invention concerne un procédé de production d'un peptide antimicrobien, et plus particulièrement, comprend un procédé de production en masse d'un peptide antimicrobien par l'intermédiaire d'un processus dans lequel un gène de peptide antimicrobien est fusionné avec un gène de protéine fluorescente verte qui est exprimé sous forme insoluble dans Escherichia coli, et ce dernier est inséré et exprimé dans E. coli, la protéine fluorescente verte est éliminée, et le peptide antimicrobien est obtenu. La présente invention procure un procédé de production d'un peptide antimicrobien, le procédé présentant un processus de production simple, étant économique et présentant un haut rendement de production, et ainsi, la présente invention présente l'avantage de fournir un antibiotique naturel qui peut remplacer des antibiotiques classiques dans les industries pharmaceutiques et alimentaires qui connaissent un besoin urgent de mettre au point un antibiotique présentant un nouveau mécanisme d'action qui peut éradiquer des souches résistantes puisque ces micro-organismes résistants à de multiples médicaments augmentent rapidement. Ou, selon la présente invention, un procédé permettant de cliver des acides aminés au moyen d'un traitement acide, au lieu d'utiliser le bromure de cyanogène classique, est utilisé, et ainsi, lors de la purification d'une protéine cible à partir d'une protéine insoluble, les coûts peuvent être réduits, le danger du procédé peut être réduit, et le procédé peut être effectué plus rapidement.
(KO) 본 발명은 항균 펩타이드의 제조방법에 관한 것으로서, 더욱 상세하게는 항균 펩타이드 유전자를 대장균 내에서 불용성으로 발현되는 녹색형광단백질 유전자와 융합시켜 대장균에 도입 및 발현시킨 후 녹색형광단백질을 제거하고 항균 펩타이드를 수득하는 과정을 통하여 항균 펩타이드를 대량으로 제조하는 방법을 포함한다. 상기와 같은 본 발명에 따르면, 제조공정이 간단하고 경제적이며 생산 수율이 높은 항균 펩타이드의 제조방법을 제공함으로써, 다중약물내성 미생물이 급증하고 있어, 이들 내성 균주를 퇴치할 수 있는 새로운 작용 메커니즘을 가지는 항생제의 개발이 시급한 제약 및 사료 산업에 기존의 항생제를 대체할 천연 항생제를 제공하는 효과가 있다. 또한 본 발명에 따르면, 기존의 시안화브롬을 이용하는 대신 산처리를 통해 아미노산을 절단하는 방법을 이용함으로써 불용성 단백질로부터 목적 단백질을 정제하는 데 있어 비용을 절감하고 공정의 위험성을 낮추며 더욱 신속하게 진행할 수 있는 효과가 있다.
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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, DK, DM, DO, DZ, EC, EE, EG, ES, FI, GB, GD, GE, GH, GM, GT, HN, HR, HU, ID, IL, IN, IR, IS, JP, KE, KG, KN, KP, 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 Organization (AM, AZ, BY, KG, KZ, RU, TJ, TM)
European Patent Office (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)
Also published as:
CN108779152EP3409685