Disclosed herein are recombinant nucleic acids encoding tumor antigens fused to immunogenic polypeptides and recombinant Listeria strains comprising the same, methods of preparing same, and methods of inducing an immune response, and treating, inhibiting, or suppressing cancer or tumors comprising administering same.

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.

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.

The present disclosure provides microbial organisms having decreased production of unwanted by-products (e.g, pyruvate-, CO.sub.2—, TCA-derived by-products; acetate; ethanol; and/or, alanine) to enhance carbon flux through acetyl-CoA, which can increase production of acetyl-CoA derived compounds (e.g, 1,3-BDO, MMA, and (3R)-hydroxybutyl (3R)-hydroxybutyrate, or any other acetyl-CoA derived compounds), and products made from any of these compounds. Also provided are one or more exogenous nucleic acids encoding enzymes that can decrease production of unwanted by-products (e.g, aldehyde dehydrogenase, acetyl-CoA synthase, amino acid dehydrogenase, alanine racemase, and/or citrate synthase), and/or one or more gene attenuations occurring in genes (e.g., acetolactate synthase) that result in decreased production of unwanted by-products. Various combinations of the exogenous nucleic acids and gene deletions are also provided in the present disclosure. Methods of making and using the same, including methods for culturing cells, and for the production of the various products are also provided.

The present disclosure provides recombinant Staphylococcus bacterium (e.g. S. epidermidis) that are dependent on D-alanine for growth. In one aspect, the disclosure features a recombinant Staphylococcus bacterium comprising two inactivated alanine racemase genes (Δalr1Δalr2); and an inactivated D-alanine aminotransferase (dat) gene. In another aspect, the disclosure features a method of making the recombinant Staphylococcus bacterium.

The present invention pertains to a probiotic comprising a generally recognized as safe (GRAS) microbiological organism, which GRAS microbiological organism comprises a food-grade expression vector, which vector comprises in functional linkage a nucleic acid sequence encoding for a soluble form of Amuc_1100 or a functionally equivalent fragment of said soluble form of Amuc_1100, wherein said GRAS microbiological organism is capable of expressing and secreting said soluble form of Amuc_1100 or said fragment thereof, as further defined in the claims. Methods for treating a disease in a patient, comprising oral administration of the probiotic as defined herein are also described, as well as methods of preparing the probiotic disclosed herein.

The present disclosure provides recombinant Staphylococcus bacterium (e.g. S. epidermidis) that are dependent on D-alanine for growth. In one aspect, the disclosure features a recombinant Staphylococcus bacterium comprising two inactivated alanine racemase genes (Δalr1Δalr2); and an inactivated D-alanine aminotransferase (dat) gene. In another aspect, the disclosure features a method of making the recombinant Staphylococcus bacterium.

The present disclosure provides recombinant Staphylococcus bacterium (e.g. S. epidermidis) that are dependent on D-alanine for growth. In one aspect, the disclosure features a recombinant Staphylococcus bacterium comprising two inactivated alanine racemase genes (Δalr1Δalr2); and an inactivated D-alanine aminotransferase (dat) gene. In another aspect, the disclosure features a method of making the recombinant Staphylococcus bacterium. In another aspect, the disclosure features a method of treating or preventing a rash in a subject, comprising administering to the subject a population of the recombinant Staphylococcus bacterium of any one of the aspects or embodiments described herein, in an effective amount to treat or prevent the rash in the subject.

Disclosed herein are bacteria engineered to treat diseases associated with hyperammonemia and methods of use thereof. Specifically, the bacteria are engineered to comprise a racemase to enable the detection of non-naturally occurring metabolites as an indication that the engineered bacteria are effectively converting ammonia.

The present invention provides Listeria vaccine strains that express a heterologous antigen and a metabolic enzyme, and methods of generating same.