Proteases with modified propeptide regions

Patent metadata

Field	Value
Publication number	US-10731144-B2
Application number	US-201615575959-A
Country	US
Kind code	B2
Filing date	Jun 17, 2016
Priority date	Jun 17, 2015
Publication date	Aug 4, 2020
Grant date	Aug 4, 2020

Abstract

The present invention provides methods and compositions for the production of mature proteases in bacterial host cells. The compositions include polynucleotides encoding serine protease sequences with modified or heterologous propeptide regions; polypeptides comprising serine proteases with modified or heterologous propeptide regions; expression cassettes, DNA constructs, vectors, and chromosomes comprising such polynucleotides; and bacterial host cells comprising such polynucleotides. The methods include methods for enhancing the production of mature proteases in bacterial host cells (e.g. Bacillus sp. host cells). The produced proteases find use in the industrial production of enzymes, suitable for use in various industries, including but not limited to the cleaning, animal feed and textile processing industry.

First claim

The invention claimed is:

1. A polynucleotide encoding a modified protease, the polynucleotide comprising:
(a) optionally a first polynucleotide region encoding a signal peptide,
(b) a second polynucleotide region encoding a variant propeptide region with at least 60% identity to SEQ ID NO: 7, wherein the variant propeptide region comprises an amino acid substitution at a position corresponding to position 34 of SEQ ID NO: 7, wherein the amino acid substitution at position 34 of SEQ ID NO: 7 is selected from the group consisting of E34D, E34C, E34G, E34H, E34S, and E34V, and
(c) a third polynucleotide region encoding the mature region of a second Bacillus gibsonii -clade protease;
wherein the first polynucleotide region is operably linked to the second polynucleotide region, and the second polynucleotide region is operably linked to the third polynucleotide region.

2. The polynucleotide of claim 1 , wherein the third polynucleotide region encodes the mature region of a protease from Bacillus gibsonii.

Assignees

• Danisco Us Inc

Inventors

• Babe Lilia Maria
• Fioresi Carol Marie
• Goedegebuur Frits
• Mulder Harm

Classifications

• C07K2319/02

  containing a signal sequence · CPC title

• C12Y304/21062

  Subtilisin (3.4.21.62) · CPC title

• C12N9/54 Primary

  bacteria being Bacillus · CPC title

• C12N15/52

  Genes encoding for enzymes or proenzymes · CPC title

• C07K2319/50

  containing protease site · CPC title