ALANINE RACEMASE SINGLE DELETION AND TRANSCOMPLEMENTATION

    公开(公告)号:US20230272358A1

    公开(公告)日:2023-08-31

    申请号:US18017426

    申请日:2021-07-23

    Applicant: BASF SE

    CPC classification number: C12N9/48 C12N15/75 C12N9/90 C12N1/20 C12R2001/10

    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.

    METHOD FOR IMPROVED PRODUCTION OF INTRACELLULAR PROTEINS IN BACILLUS

    公开(公告)号:US20250051822A1

    公开(公告)日:2025-02-13

    申请号:US18720123

    申请日:2022-12-19

    Applicant: BASF SE

    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.

    PROMOTER FOR HETEROLOGOUS EXPRESSION
    6.
    发明申请

    公开(公告)号:US20200181627A1

    公开(公告)日:2020-06-11

    申请号:US16631999

    申请日:2018-07-11

    Applicant: BASF SE

    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.

    ALANINE RACEMASE DOUBLE DELETION AND TRANSCOMPLEMENTATION

    公开(公告)号:US20230295603A1

    公开(公告)日:2023-09-21

    申请号:US18017430

    申请日:2021-07-23

    Applicant: BASF SE

    CPC classification number: C12N9/90 C12N15/75 C12Y501/01001 C12N2800/101

    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.

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