Dsm IP Assets B.v.

Peter Jozef Ida Van De Vondervoort

Jan Metske Van Der Laan

Martina Beishuizen

Chantal Christis

Martine Spaans

Angela De Bruine-Paulus

Noël Nicolaas Maria Elisabeth Van Peij

Johanna Henrica Gerdina Maria Mutsaers

Noel Nicolaas Maria Elisabeth Van Peij

The present invention relates to a polypeptide having proline-specific endoprotease activity, selected from the group consisting of

Proline specific endoprotease

The present invention relates to a polypeptide having proline-specific endoprotease activity, wherein the polypeptide has less than 70% residual activity when the polypeptide has been kept at a temperature of 65&#xb0; C. for 15 min. The invention further relates to a polypeptide having proline-specific endoprotease activity comprising an amino acid sequence according to SEQ ID NO: 1, wherein SEQ ID NO: 1 comprises at least one amino acid substitution selected from the group consisting of P469A, P469C, P469D, P469E, P469F, P469G, P469H, P469I, P469K, P469L, P469M, P469N, P469Q, P469R, P469S, P469T, P469V, P469W, P469Y, a nucleic acid encoding a polypeptide having proline-specific endoprotease activity, a method of making a variant polypeptide having proline-specific endoprotease activity, a recombinant host cell and a method of producing the polypeptide and a process for the preparation of a food or feed product wherein the polypeptide is used.

Proline-specific endoprotease and use thereof

The present invention relates to a polypeptide having proline-specific endoprotease activity, wherein the polypeptide has less than 70% residual activity when the polypeptide has been kept at a temperature of 65&#xb0; C. for 15 min. The invention further relates to a polypeptide having proline-specific endoprotease activity comprising an amino acid sequence according to SEQ ID NO: 1, wherein SEQ ID NO: 1 comprises at least one combination of amino acid substitutions selected from the group consisting of a combination selected from the group consisting of (K238E, I204V, V460A), (F279S, A242V, N507I), (T145A, K424M), (T359A, F379S), (M170I, A421T), (N441S, G484S), (L470H, Q288R), (L470H, E387G), (T281S, L373I), (P304A, P469A), and (P466T, P469Q), a nucleic acid encoding a polypeptide having proline-specific endoprotease activity, a method of making a variant polypeptide having proline-specific endoprotease activity, a recombinant host cell and a method of producing the polypeptide and a process for the preparation of a food or feed product wherein the polypeptide is used.

The present invention relates to a mutant microbial host cell which is deficient in the production of the AgsE protein or in the production of an homologous thereof if compared with a parent microbial host cell which has not been modified and measured under the same conditions. It has been surprisingly found that when the mutant microbial host cell according to the invention is used in a method to produce a compound of interest, for example an enzyme, an improved yield of said compound is obtained if compared to a method in which a parent host cell which has not been modified is used when measured under the same conditions.

AgsE-deficient strain

The present invention relates to a mutant microbial host cell which has been modified, preferably in its genome, to result in a deficiency in the production of a polypeptide having &#x3b1;-amylase activity AmyC or an homologous thereof. It has been surprisingly found that when the mutant microbial host cell according to the invention is used in a method to produce a compound of interest, for example an enzyme, an improved purity of the product is obtained.

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Echt, Netherlands

Peter Jozef Ida Van De Vondervoort