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1. (WO1987004461) THERMALLY STABLE AND pH STABLE SUBTILISIN ANALOGS AND METHOD FOR PRODUCTION THEREOF
Latest bibliographic data on file with the International Bureau

Pub. No.: WO/1987/004461 International Application No.: PCT/US1987/000027
Publication Date: 30.07.1987 International Filing Date: 07.01.1987
IPC:
C12N 15/90 (2006.01) ,C12N 9/56 (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
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
87
Introduction of foreign genetic material using processes not otherwise provided for, e.g. co-transformation
90
Stable introduction of foreign DNA into chromosome
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
54
bacteria being Bacillus
56
Bacillus subtilis or Bacillus licheniformis
Applicants:
AMGEN [US/US]; 1900 Oak Terrace Lane Thousand Oaks, CA 91320, US
Inventors:
STABINSKY, Yitzhak; US
ZUKOWSKI, Mark, M.; US
Agent:
BORUN, Michael, F.; Marshall, O'Toole, Gerstein, Murray & Bicknell Two First National Plaza, Suite 2100 Chicago, IL 60603, US
Priority Data:
819,24115.01.1986US
Title (EN) THERMALLY STABLE AND pH STABLE SUBTILISIN ANALOGS AND METHOD FOR PRODUCTION THEREOF
(FR) ANALOGUES DE SUBTILISINE THERMIQUEMENT STABLE ET STABLE AU pH, ET LEUR PROCEDE DE PRODUCTION
Abstract:
(EN) A mutated subtilisin suitable for admixture to washing compositions and exhibiting substantially improved stability over naturally occuring $i(Bacillus) serine proteases is prepared by expressing a modified gene encoding subtilisin in $i(Bacillus subtilis). A preferred subtilisin analog product differs from wild-type $i(Bacillus) alkaline proteases by having any amino acid, and preferably serine, at position (218) in place of asparagine. The product is preferably produced in a strain of $i(B. subtilis) which is mutated to block synthesis of endogenous proteases. The method of replacing an Asn or Gly in an Asn-Gly sequence in order to improve pH and thermal stability may be applied to other sites in substilisin and to other proteins as well.
(FR) De la subtilisine mutée appropriée à être mélangée à des compositions de lavage et présentant une stabilité bien plus grande par rapport à des sérine protéases du $i(Bacillus) d'origine naturelle est préparée en exprimant un gène modifié codant la subtilisine dans le $i(Bacillus subtilis). Un produit analogue de subtilisine préféré diffère des protéases alcalines de $i(Bacillus) de type sauvage par le fait d'avoir un acide aminé quelconque, et de préférence la sérine, en position 218 au lieu de l'asparagine. Le produit est obtenu de préférence dans une souche de $i(B. subtilis) qui est muté pour bloquer la synthèse de protéases endogènes. Le procédé de remplacement d'un Asn ou un Gly dans une séquence Asn-Gly de manière à améliorer la stabilité thermique au pH peut être appliqué à d'autres sites dans la subtilisine et à d'autres protéines également.
Designated States: AU, DK, FI, JP, NO
European Patent Office (AT, BE, CH, DE, FR, GB, IT, LU, NL, SE)
Publication Language: English (EN)
Filing Language: English (EN)
Also published as:
FI873980NO19873839EP0254735JP2599946AU1987069398DKPR 175409
DK175409