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1. (WO2018220032) OPTIMIZATION OF THE EXPRESSION OF SERINE PROTEASES IN HOST CELLS
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Pub. No.: WO/2018/220032 International Application No.: PCT/EP2018/064216
Publication Date: 06.12.2018 International Filing Date: 30.05.2018
IPC:
C12N 9/64 (2006.01) ,C07K 14/81 (2006.01) ,C12N 15/81 (2006.01) ,C12N 15/67 (2006.01) ,C12P 21/02 (2006.01) ,C12N 1/19 (2006.01)
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
64
derived from animal tissue, e.g. rennin
C CHEMISTRY; METALLURGY
07
ORGANIC CHEMISTRY
K
PEPTIDES
14
Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
81
Protease inhibitors
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
79
Vectors or expression systems specially adapted for eukaryotic hosts
80
for fungi
81
for yeasts
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
67
General methods for enhancing the expression
C CHEMISTRY; METALLURGY
12
BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
P
FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
21
Preparation of peptides or proteins
02
having a known sequence of two or more amino acids, e.g. glutathione
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
1
Micro-organisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving micro-organisms or compositions thereof; Processes of preparing or isolating a composition containing a micro-organism; Culture media therefor
14
Fungi; Culture media therefor
16
Yeasts; Culture media therefor
19
modified by introduction of foreign genetic material
Applicants:
B.R.A.I.N. BIOTECHNOLOGY RESEARCH AND INFORMATION NETWORK AG [DE/DE]; Darmstädter Str. 34-36 64673 Zwingenberg, DE
Inventors:
PELZER, Alexander; DE
KELETY, Bela; DE
Agent:
WACHENFELD, Joachim; VOSSIUS & PARTNER Siebertstraße 3 81675 München, DE
Priority Data:
17173813.131.05.2017EP
Title (EN) OPTIMIZATION OF THE EXPRESSION OF SERINE PROTEASES IN HOST CELLS
(FR) OPTIMISATION DE L'EXPRESSION DE SÉRINE PROTÉASES DANS DES CELLULES HÔTES
Abstract:
(EN) The present invention relates to a method for the recombinant production of a serine protease comprising (a) culturing a host cell comprising one or more vectors, wherein the one or more vectors encode in expressible form the serine protease and a proteinaceous inhibitor of the serine protease, wherein the proteinaceous inhibitor of the serine protease is heterologous with respect to the serine protease, under conditions wherein the serine protease and the proteinaceous inhibitor of the serine protease are expressed; or (a') culturing a host cell the genome of which encodes in expressible form the serine protease and a proteinaceous inhibitor of the serine protease, wherein the proteinaceous inhibitor of the serine protease is heterologous with respect to the serine protease, and wherein the coding sequences of the serine protease and/or the proteinaceous inhibitor have been introduced into the host cell genome by applying a CRISPR technology, under conditions wherein the serine protease and the proteinaceous inhibitor of the serine protease are expressed; and (b) isolating the serine protease expressed in step (a) or (a') from the host cell. The present invention also relates to a host cell comprising one or more vectors, wherein the one or more vectors encode in expressible form a serine protease and a proteinaceous inhibitor of the serine protease.
(FR) La présente invention concerne une méthode de production recombinante d'une sérine protéase comprenant (a) la culture d'une cellule hôte comprenant un ou plusieurs vecteurs, le ou les vecteurs codant sous forme exprimable la sérine protéase et un inhibiteur protéique de la sérine protéase, l'inhibiteur protéique de la sérine protéase étant hétérologue par rapport à la sérine protéase, dans des conditions dans lesquelles la sérine protéase et l'inhibiteur protéique de la sérine protéase sont exprimés; ou (a') cultiver une cellule hôte dont le génome code sous forme exprimable la sérine protéase et un inhibiteur protéique de la sérine protéase, l'inhibiteur protéique de la sérine protéase étant hétérologue par rapport à la sérine protéase, et les séquences codantes de la sérine protéase et/ou de l'inhibiteur protéique ayant été introduites dans le génome de la cellule hôte par application d'une technologie CRISPR, dans des conditions dans lesquelles la sérine protéase et l'inhibiteur protéique de la sérine protéase sont exprimés; et (b) isoler la sérine protéase exprimée à l'étape (a) ou (a') de la cellule hôte. La présente invention concerne également une cellule hôte comprenant un ou plusieurs vecteurs, le ou les vecteurs codant sous forme exprimable la sérine protéase et un inhibiteur protéique de la sérine protéase.
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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, KR, 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: English (EN)
Filing Language: English (EN)