C - Chemistry – Metallurgy – 12 – N
Patent
C - Chemistry, Metallurgy
12
N
C12N 15/56 (2006.01) A01H 5/00 (2006.01) A23K 1/165 (2006.01) C12C 7/04 (2006.01) C12C 11/00 (2006.01) C12N 9/24 (2006.01) C12N 9/42 (2006.01) C12N 15/82 (2006.01)
Patent
CA 2166777
This invention relates to a (1 ? 3, 1 ? 4)-.beta.-glucanase (glucanase EII endohyrolase) enzyme, whose amino acid sequence has been modified in order to provide an enzyme whose three-dimensional structure confers improved improved thermostability and/or pH stability. Specific modifications are based upon a comparison between the three-dimensional structure of (1 ? 3, 1 ? 4)-.beta.-glucanase and that of (1 ? 3)-.beta.-glucanase. The (1 ? 3, 1 ? 4)-.beta.-glucanase gene has been modified by site-directed mutagenesis, and modified enzymes have been expressed in Escherichia coli. Modified sequences, DNA molecules encoding them, plasmids, expression vectors and transgenic plants are disclosed and claimed.
L'invention concerne un enzyme (1 } 3, 1 } 4)- beta -glucanase (glucanase EII endohydrolase), dont la séquence d'acides aminés a été modifiée, afin de produire un enzyme dont la structure tridimensionnelle confère une thermostabilité et/ou une stabilité de pH améliorées. Des modifications spécifiques se basent sur une comparaison entre la structure tridimensionnelle de (1 } 3, 1 } 4)- beta -glucanase et celle de (1 } 3)- beta -glucanase. Le gène de (1 } 3, 1 } 4)- beta -glucanase a été modifié par une mutagène ciblée et des enzymes modifiés ont été exprimés dans Escherichia coli. L'invention concerne également des séquences modifiées, des molécules d'ADN les codant, des plasmides, des vecteurs d'expression et des plantes transgéniques.
Chen Lin
Fincher Geoffrey Bruce
Garrett Thomas Peter John
Hõj Peter Bordier
Varghese Joseph Noozhumurry
Biomolecular Research Institute Ltd.
Borden Ladner Gervais Llp
Latrobe University
Luminis Pty. Ltd.
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