Abstract: The present invention relates to a method for producing at least one polypeptide of interest in a Bacillus host cell, said method comprising the steps of a) providing a Bacillus host cell comprising a polynucleotide encoding the at least one polypeptide of interest, b) cultivating the Bacillus host cell under conditions which allow for intracellular accumulation of said at least one polypeptide of interest, thereby producing said at least one polypeptide of interest, c) stop of aeration and addition of feed, and d) incubating the cells for at least hours. Moreover, the present invention relates to a Bacillus host cell comprising a polynucleotide encoding at least one polypeptide of interest lacking a functional secretion sequence, wherein the ispA gene has been inactivated in the Bacillus host cell.

Abstract: The present invention relates to a Bacillus host cell for increased production of biological compounds. Specifically, the invention relates to a Bacillus host with genetic modifications that lead to decreased production of components of biofilm and increased levels of phosphorylated De-gll. Said Bacillus host comprises at least one gene expression cassette with a polynucleotide encoding at least one polypeptide of interest, operably linked to a promoter for the production of compounds. The present invention further relates to a method for increased production of at least one polypeptide of interest based on cultivating the bacterial host cell of the present invention.

Abstract: The present invention relates to a Bacillus licheniformis host cell for production of biological compounds with increased purity. Specifically, the invention relates to a Bacillus licheniformis host with one or more genetic modifications selected from a) a genetic modification causing compared to an unmodified control cell an increased expression of at least one of the genes selected from the group consisting of degQ, degll, degS, degR, and phrG, b) a genetic modification causing compared to an unmodified control cell an increased autophosphorylation activity of the DegS protein, and c) a genetic modification causing compared to an unmodified control cell a reduced phosphatase activity of the DegS protein. The present invention further relates to a method for production of at least one compound of interest with increased purity based on cultivating the bacterial host cell of the present invention.

Abstract: The present invention relates to a Bacillus host cell for increased production of biological compounds. Specifically, the invention relates to a Bacillus host with genetic modifications in the remA and/or remB gene. The present invention further relates to a method for increased production of at least one polypeptide of interest based on cultivating the bacterial host cell of the present invention.

Abstract: The present invention is directed to a nucleic acid construct comprising a polynucleotide operably linked to one or more control sequence that directs the expression of the polynucleotide in a host cell, wherein at least one control sequence comprises a promoter sequence of an operon comprising a secA gene or a functional fragment or functional variant thereof and wherein said promoter sequence is heterologous to the polynucleotide. In a further embodiment, the invention is directed to an expression vector and a host cell comprising the nucleic acid construct comprising the promoter sequence described herein and a method of expressing a polynucleotide in a host cell.

Abstract: The present invention generally relates to the field of fermentation technology and microorganisms useful for such fermentations. The invention also relates to materials including nucleic acids and proteins useful for altering fermentation characteristics of microorganisms, and to microorganisms comprising such nucleic acids and/or proteins.

Abstract: The present invention provides a modified Bacillus cell for the production of a compound of interest, wherein the Bacillus cell comprises a genetic modification that leads to decreased production of enzymes with lipase and/or esterase activity. The use of compounds of interest produced with said improved Bacillus host cells reduces the formation of malodour and/or off-flavour when used for applications such as washing or cleaning or as feed or food additives.

Abstract: The present invention generally relates to the field of fermentation technology and microorganisms useful for such fermentations. The invention also relates to materials including nucleic acids and proteins useful for altering fermentation characteristics of microorganisms, and to microorganisms comprising such nucleic acids and/or proteins.

Abstract: The present invention relates to a bacterial host cell in which a first chromosomal gene encoding a first alanine racemase and a second chromosomal gene encoding a second alanine racemase have been inactivated. Said bacterial host cell comprises – either on a plasmid comprising at least one autonomous replication sequence or present as multiple copies in the chromosome – a gene expression cassette comprising a polynucleotide encoding at least one polypeptide of interest, operably linked to a promoter, and a polynucleotide encoding a third alanine racemase, operably linked to a promoter. The present invention further relates to a method for producing at least one polypeptide of interest based on cultivating the bacterial host cell of the present invention.

Abstract: The present invention relates to a Bacillus host cell belonging to the species Bacillus lichemformis or Bacillus pumilus in which the chromosomal alr gene has been inactivated. Said bacterial host cell comprises a plasmid comprising at least one autonomous replication sequence, a first polynucleotide encoding at least one polypeptide of interest, wherein said first polynucleotide is operably linked to a promoter, and a second polynucleotide encoding an alanine racemase which is not native to the host cell, wherein said second polynucleotide is operably linked to a promoter. The present invention further relates to a method for producing at least one polypeptide of interest based on cultivating the bacterial host cell of the present invention.

Abstract: Described herein are materials and methods for user-controlled adjustment of plasmid copy numbers.

Abstract: Disclosed herein are methods for the production of low to medium expressing constitutive promoters in bacteria and promoters produced therewith.

Abstract: Methods for modification of target nucleic acids. The method involves a construct in which guide RNA is covalently linked to donor RNA (fusion NA) to be introduced into the target nucleic acid by homologous recombination and is based on the introduction of a nuclease, e.g. CRISPR or TALEN, into the cell containing the target nucleic acid. The fusion NA may be introduced as a DNA vector.

Abstract: Disclosed herein is a shuttle vector for use in E. coli and Bacilli including a high copy replication origin functional in E. coli, a low to medium copy ORI functional in Bacilli, and a synthetic constitutive regulatory nucleic acid conferring reduced constitutive expression compared to a respective starting regulatory nucleic acid molecule in a bacterial cell.

Abstract: Methods for modification of target nucleic acids. The method involves a construct in which guide RNA is covalently linked to donor RNA (fusion NA) to be introduced into the target nucleic acid by homologous recombination and is based on the introduction of a nuclease, e.g. CRISPR or TALEN, into the cell containing the target nucleic acid. The fusion NA may be introduced as a DNA vector.

Abstract: Methods for modification of target nucleic acids. The method involves a construct in which guide RNA is covalently linked to donor RNA (fusion NA) to be introduced into the target nucleic acid by homologous recombination and is based on the introduction of a nuclease, e.g. CRISPR or TALEN, into the cell containing the target nucleic acid. The fusion NA may be introduced as a DNA vector.

Abstract: Methods for modification of target nucleic acids. The method involves a construct in which guide RNA is covalently linked to donor RNA (fusion NA) to be introduced into the target nucleic acid by homologous recombination and is based on the introduction of a nuclease, e.g. CRISPR or TALEN, into the cell containing the target nucleic acid. The fusion NA may be introduced as a DNA vector.

Abstract: Methods for modification of target nucleic acids. The method involves a construct in which guide RNA is covalently linked to donor RNA (fusion NA) to be introduced into the target nucleic acid by homologous recombination and is based on the introduction of a nuclease, e.g. CRISPR or TALEN, into the cell containing the target nucleic acid. The fusion NA may be introduced as a DNA vector.

Abstract: The present invention is directed to an industrial fermentation process for cultivating a Bacillus cell in a chemically defined fermentation medium and a method for producing a protein of inter-est comprising the steps of providing a chemically defined fermentation medium, inoculating the fermentation medium with a Bacillus cell comprising a gene encoding a protein of interest, cultivating the Bacillus cell in the fermentation medium under conditions conductive for the growth of the Bacillus cell and the expression of the protein of interest, wherein the cultivation of the Bacillus cell comprises the addition of one or more feed solutions comprising one or more chemically defined carbon sources and magnesium ions to the fermentation medium.

Abstract: The present invention is directed to an industrial fermentation process for cultivating a Bacillus cell in a chemically defined fermentation medium and a method for producing a protein of interest comprising the steps of providing a chemically defined fermentation medium, inoculating the fermentation medium with a Bacillus cell comprising a gene encoding a protein of interest, cultivating the Bacillus cell in the fermentation medium under conditions conductive for the growth of the Bacillus cell and the expression of the protein of interest, wherein the cultivation of the Bacillus cell comprises the addition of one or more feed solutions comprising one or more chemically defined carbon sources and one or more trace element ions to the fermentation medium.