Unilever Patent Holdings B.v.

Other

Johannes M Verbakel

Wouter Musters

Hein Stam

Jacob B Visser

Maria E Suykerbuyk

Cornelis T Verrips

Roelf J Planta

Maria T Lopes

Marco F Giuseppin

Jan Maat

Robertus Johannes Gouka

Peter J Van De Vonderwoort

Peter J Schaap

Cornelis Antonius Van Den Hondel

A process is disclosed for preparing a protein by a eukaryote transformed by multicopy integration of an expression vector into the genome of a yeast, such as Saccharomyces, Hansenula, and Kluyveromyces, or of a mould such as Aspergillus, Rbizopus and Trichoderma, the expression vector containing both an 'expressible gene' encoding the protein and a so-called 'deficient selection marker meeded for the growth of the yeast or mould in a specific medium', such as the LEU2d, TRP1d or URA3d gene, in combination witha ribosomal DNA sequence, resulting in stable high copy integration of 100-300 copies per cell. This multicopy integration results in an increased production of the desired protein, which can be guar .alpha.-galactosidase, an oxidase or a hydrolytic enzyme such as a lipase.

Process for preparing a protein by a fungus transformed by multicopy

The invention relates to isolation of an Aspergillus gene encoding rhamnogalacturonase (RG-ase) and the construction of recombinant Aspergillus strains with overexpression of RG-ase. These strains can be used for the commercial production of RG-ase. RG-ase is an important enzyme in processes requiring the degradation and/or modification of pectin or modification of pectin-containing vegetable or plant cell wall material. RG-ase may be used in various applications, including the processing of fruits and vegetables, in the extraction of components from vegetable material or for improving the functionality of pectin or pectin-containing vegetable material, food material or plant cell wall material.

Cloning and expression of DNA encoding a ripening form of a polypeptide

A process is disclosed for preparing a protein by a eukaryote transformed by multicopy integration of an expression vector into the genome of a yeast, such as Saccharomyces, Hansenula, and Kluyveromyces, or of a mould such as Aspergillus, Rhizopus and Trichoderma, the expression vector containing both an 'expressible gene' encoding the protein and a so-called 'deficient selection marker needed for the growth of the yeast or mould in a specific medium', such as the LEU2d, TRP1d or URA3d gene, in combination with a ribosomal DNA sequence, resulting in stable high copy integration of 100-300 copies per cell. This multicopy integration results in an increased production of the desired protein, which can be guar .alpha.-galactosidase, an oxidase or a hydrolytic enzyme such as a lipase.

Process for producing/secreting a protein by a transformed mould using

Methods of detecting and isolating a ripening form of a polypeptide

The invention relates to recombinant DNA technology for the production of an enzyme having sulfhydryl oxidase ('SOX') activity. This SOX-enzyme can be used where the oxidation of free sulfhydryl groups (thio compounds) to the corresponding disulfides is desirable. SOX enzyme may be used for treatment of bakery products or for removal of off-flavour from milk or beer.

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Maasland, Netherlands

Johannes M Verbakel