Bacillus host
AssigneeDsm IP Assets B.V.
InventorsEvert Tjeerd Van Rij, Martine Spaans, Hendrikus Bernardus Carolus Hillebrand
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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.
Claims20
A recombinant Bacillus host cell modified in its genome to be deficient in the production of a neutral protease and an 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 Spounalikevirus Bacillus phage SPO1 (SPO1) promoter sequence, and wherein the bacteriophage Spounalikevirus Bacillus phage SPO1 (SPO1) promoter sequence is part of an expression module comprising the bacteriophage SPO1 promoter, a modified riboflavin biosynthetic genes (rib) leader and a sequence coding for a ribosome binding site (RBS), wherein the expression module further comprises a mRNA stabilizing element, and wherein the modified rib leader is located downstream of the promoter and upstream of the sequence coding for the RBS.
A recombinant Bacillus host cell modified in its genome to be deficient in the production of a neutral protease and an 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 Spounalikevirus Bacillus phage SPO1 (SPO1) promoter sequence wherein the host cell is a Bacillus subtilis and wherein the neutral protease in which the host cell is deficient has an amino acid sequence according to SEQ ID NO:
- 8 or an amino acid sequence at least 90% identical to SEQ ID NO:
- 8 and wherein the alkaline protease has an amino acid sequence according to SEQ ID NO:
- 6 or an amino acid sequence at least 90% identical to SEQ ID NO:
- 6, wherein the Bacillus subtilis host cell is further deficient in the production of neutral amylase, wherein the neutral amylase has an amino acid sequence according to SEQ ID NO:
- 20 or an amino acid sequence at least 90% identical to SEQ ID NO:
- 20.
Dependencies are inferred from the claim text.