New fungal production system

C - Chemistry – Metallurgy – 12 – N

Patent

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Details

C12N 15/113 (2010.01) C12N 1/14 (2006.01) C12N 9/24 (2006.01) C12N 9/58 (2006.01) C12N 15/00 (2006.01) C12N 15/80 (2006.01) C12P 21/02 (2006.01) C12Q 1/04 (2006.01) C12Q 1/37 (2006.01)

Patent

CA 2755417

The present invention provides a new fungal production system comprising a fungal host strain of Chrysosporium lucknowense wherein the endogenous cellulase secretion is less than 20 % of the endogenous cellulase secretion of Chrysosporium lucknowense strain UV 18-25. Preferably, also the secretion of endogenous protease, endogenous .beta.- glucanase and endogenous cellobiohydrolase is less than 20 % of the secretion of Chrysosporium lucknowense strain UV 18-25. Furthermore, fungal host strains are provided wherein several genes have been disrupted. According to another aspect of the invention a method for homologous and/or heterologous production of a pure protein with a purity of higher than 75 %, comprising expressing a gene encoding said protein in the strains according to the invention have been described. Furthermore, a method for production of artificial protein mixes comprising expressing a gene encoding each of said proteins in a strain according to the invention have been disclosed. Finally a method for simplified screening of strains functionally expressing a desired enzyme by application of said strains have been provided.

La présente invention concerne un nouveau système de production fongique qui comprend une souche hôte fongique de Chrysosporium lucknowense, la sécrétion de cellulase endogène étant inférieure à 20 % de la sécrétion de cellulase endogène de la souche UV 18-25 de Chrysosporium lucknowense. De préférence, la sécrétion de protéase endogène, de ß-glucanase endogène et de cellobiohydrolase endogène est également inférieure à 20 % de la sécrétion de la souche UV 18-25 de Chrysosporium lucknowense. L'invention concerne également des souches hôtes fongiques dont plusieurs gènes ont été perturbés. Selon un autre aspect, l'invention concerne un procédé de production homologue et/ou hétérologue d'une protéine pure ayant une pureté supérieure à 75 %, qui comprend l'expression d'un gène codant pour ladite protéine dans les souches selon l'invention. L'invention concerne également un procédé de production de mélanges artificiels de protéines qui comprend l'expression d'un gène codant pour chacune desdites protéines dans une souche selon l'invention. L'invention concerne enfin un procédé de détection simplifiée de souches exprimant fonctionnellement une enzyme souhaitée par application desdites souches.

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