Dsm IP Assets B.v.

Dsm N.v.

Danisco US Inc.

Dyadic Nederland B.v.

Jan Wery

Jan Cornelis Verdoes

Jacob B Visser

Peter Jan Punt

Mark Aaron Emalfarb

Richard Paul Burlingame

Christine Marie Pynnonen

Albert Johannes Joseph Van Ooijen

Johannes Heinrich Visser

Phillip T Olson

Albert Johannes Van Ooijen

The present invention provides a new fungal production system comprising a fungal host strain ofwherein the endogenous cellulase secretion is less than 20% of the endogenous cellulase secretion ofstrain UV 18-25. Preferably, also the secretion of endogenous protease, endogenous &#x3b2;-glucanase and endogenous cellobiohydrolase is less than 20% of the secretion ofstrain 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.

Fungal production system

The present invention provides recombinant DNA comprising a transcription promoter and a downstream sequence to be expressed, in operable linkage therewith, wherein the transcription promoter comprises a region found upstream of the open reading frame of a highly expressedgene, preferably a glycolytic pathway gene, more preferably the gene coding for Glyceraldehyde-3-Phosphate Dehydrogenase. Further preferred recombinant DNAs according to the invention contain promoters of ribosomal protein encoding genes, more preferably wherein the transcription promoter comprises a region found upstream of the open reading frame encoding a protein as represented by one of the amino acid sequences depicted in any one of SEQIDNOs: 24 to 50. According to a further aspect of the invention an isolated DNA sequence coding for an enzyme involved in the carotenoid biosynthetic pathway ofis provided, preferably wherein said enzyme has an activity selected from isopentenyl pyrophosphate isomerase activity, geranylgeranyl pyrophosphate synthase activity, phytoene synthase activity, phytoene desaturase activity and lycopene cyclase activity, still more preferably those coding for an enzyme having an amino acid sequence selected from the one represented by SEQIDNO: 13, SEQIDNO: 15, SEQIDNO: 17, SEQIDNO: 19, SEQIDNO: 21 or SEQIDNO: 23. Further embodiments concern vectors, transformed host organisms, methods for making proteins and/or carotenoids, such as astaxanthin, and methods for isolating highly expressed promoters from

Recombinant materials for carotenoid production

The present invention provides recombinant DNA comprising a transcription promoter and a downstream sequence to be expressed, in operable linkage therewith, wherein the transcription promoter comprises a region found upstream of the open reading frame of a highly expressed Phaffia gene, preferably a glycolytic pathway gene, more preferably the gene coding for Glyceraldehyde-3-Phosphate Dehydrogenase. Further preferred recombinant DNAs according to the invention contain promoters of ribosomal protein encoding genes, more preferably wherein the transcription promoter comprises a region found upstream of the open reading frame encoding a protein as represented by one of the disclosed amino acid sequences. According to a further aspect of the invention an isolated DNA sequence coding for an enzyme involved in the carotenoid biosynthetic pathway of,is provided.

Methods for transforming Phaffia strains, transformed Phaffia strains so obtained and recombinant DNA in said methods

Growing community of inventors

Gorssel, Netherlands

Jan Wery