公开(公告)号：US10202594B2

公开(公告)日：2019-02-12

申请号：US15314892

申请日：2015-06-03

Applicant: DSM IP ASSETS B.V.

Inventor： Jan Metske Van Der Laan ,  Peter Jozef Ida Van De Vondervoort ,  Chantal Christis ,  Martine Spaans ,  Angela De Bruine-Paulus

IPC: C12N9/50 ,  C12N9/62

Abstract: 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° 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.

公开(公告)号：US10450554B2

公开(公告)日：2019-10-22

申请号：US15312171

申请日：2015-05-19

Applicant: DSM IP ASSETS B.V.

Inventor： Jan Metske Van Der Laan ,  Peter Jozef Ida Van De Vondervoort ,  Chantal Christis ,  Martine Spaans ,  Angela De Bruine-Paulus ,  Johanna Henrica Gerdina Maria Mutsaers

IPC: A61K38/48 ,  C12N1/20 ,  C12N9/62 ,  A23L2/84 ,  A23J3/34 ,  C12N9/50 ,  A23L2/66 ,  A23L7/10 ,  A23L33/18 ,  A23L33/195

Abstract: The present invention relates to a polypeptide having proline-specific endoprotease activity, selected from the group consisting of i. a polypeptide comprising a mature polypeptide sequence of SEQ ID NO: 2; ii. a polypeptide that has least 70%, 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to the mature polypeptide sequence of SEQ ID NO: 2; iii. a polypeptide encoded by a nucleic acid that hybridizes under medium stringency, preferably under high stringency conditions to the complementary strand of the mature polypeptide coding sequence of SEQ ID NO:1; iv. a polypeptide encoded by a nucleic acid that has at least 70%, 75% 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identity to the mature polypeptide coding sequence of SEQ ID NO: 1. The invention further relates to a method of producing the polypeptide and a process for the preparation of a food or feed product wherein the polypeptide is used.

公开(公告)号：US10294467B2

公开(公告)日：2019-05-21

申请号：US15314825

申请日：2015-06-03

Applicant: DSM IP ASSETS B.V.

Inventor： Jan Metske Van Der Laan ,  Peter Jozef Ida Van De Vondervoort ,  Chantal Christis ,  Martine Spaans ,  Angela De Bruine-Paulus

IPC: C12N9/62 ,  C12C11/00

Abstract: 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° 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.

公开(公告)号：US10287592B2

公开(公告)日：2019-05-14

申请号：US15116933

申请日：2015-02-06

Applicant: DSM IP Assets B.V.

Inventor： Evert Tjeerd Van Rij ,  Martine Spaans ,  Hendrikus Bernardus Carolus Hillebrand

IPC: C12N15/63 ,  C12N15/75 ,  C12N9/54 ,  C12P21/02 ,  C12N9/26 ,  C12N9/24 ,  C12P21/00

Abstract: The invention provides Bacillus host cell deficient in the production of a neutral protease and of a alkaline protease which host cell comprises a nucleic acid construct, wherein the nucleic acid construct comprises a polynucleotide encoding a compound of interest or encoding a compound involved in the synthesis of a compound of interest operably linked to a promoter sequence allowing for expression of the polynucleotide in the host cell, wherein the promoter comprises a bacteriophage promoter sequence, more preferably a bacteriophage SPO1 promoter sequence. Said host cell can advantageously be used in a method of production of a compound of interest.