The present invention discloses modified Staphylococcus aureus HIa proteins which show reduced tendency to aggregate, improving protein stability and yield. Said modified HIa proteins optionally also contain glycosylation site consensus sequences. The invention also discloses a conjugate comprising a modified HIa protein and an antigen (for example a Staphylococcus aureus saccharide antigen), wherein the antigen is linked to an amino acid residue of the modified HIa protein.

Provided herein, in some aspects, are methods and compositions for cell-free production of ribonucleic acid.

The present invention discloses modified Staphylococcus aureus HIa proteins which show reduced tendency to aggregate, improving protein stability and yield. Said modified HIa proteins optionally also contain glycosylation site consensus sequences. The invention also discloses a conjugate comprising a modified HIa protein and an antigen (for example a Staphylococcus aureus saccharide antigen), wherein the antigen is linked to an amino acid residue of the modified HIa protein.

A method for producing a fibroin-like protein is provided. A fibroin-like protein is produced by a method of culturing Escherichia coli having a gene encoding the fibroin-like protein in a medium, inducing expression of the gene encoding the fibroin-like protein, and collecting the fibroin-like protein, wherein accumulation of an organic acid at the time of inducing the expression is reduced, and wherein the gene is expressed under control of a tryptophan promoter.

Provided herein are specific CD137 receptor agonist proteins, nucleic acids encoding the same, and methods of treating a subject having a CD137L-associated disease or disorder. The CD137 receptor agonist proteins provided herein comprise three soluble CD137L domains and an Fe fragment. The CD137 receptor agonist proteins are substantially non-aggregating and suitable for therapeutic, diagnostic and/or research applications.

The present invention is directed to a bioconjugate vaccine, such as an O 1-bioconjugate vaccine, comprising: a protein carrier comprising a protein carrier containing at least one consensus sequence, DIE-X-N-Z-S/T, wherein X and Z may be any natural amino acid except proline; at least one antigenic polysaccharide from at least one pathogenic bacterium, linked to the protein carrier; and, optionally, an adjuvant. In another aspect, the present invention is directed to a method of producing an O 1-bioconjugate in a bioreactor comprising a number steps.

In alternative embodiments, provided are non-natural or genetically engineered vinylisomerase-dehydratase enzymes, including alkenol dehydratases, linalool dehydratases and crotyl alcohol dehydratases. In alternative embodiments, provided are non-natural or genetically engineered polypeptides having an activity comprising, for example, a vinylisomerase-dehydratase, an alkenol dehydratase, a linalool dehydratase and/or a crotyl alcohol dehydratase activity, or a combination thereof. In alternative embodiments, also provided are non-natural or genetically engineered nucleic acids (polynucleotides) encoding polypeptides described herein, expression or cloning vehicles comprising or having contained therein nucleic acids as described herein, and non-natural or genetically engineered cells comprising or having contained therein nucleic acids as described herein. In alternative embodiments, also provided are methods for making various organic compounds, including methyl vinyl carbinol and butadiene.

The present invention discloses modified Staphylococcus aureus HIa proteins which show reduced tendency to aggregate, improving protein stability and yield. Said modified HIa proteins optionally also contain glycosylation site consensus sequences. The invention also discloses a conjugate comprising a modified HIa protein and an antigen (for example a Staphylococcus aureus saccharide antigen), wherein the antigen is linked to an amino acid residue of the modified HIa protein.

Provided herein are specific CD137 receptor agonist proteins, nucleic acids encoding the same, and methods of treating a subject having a CD137L-associated disease or disorder. The CD137 receptor agonist proteins provided herein comprise three soluble CD137L domains and an Fe fragment. The CD137 receptor agonist proteins are substantially non-aggregating and suitable for therapeuctic, diagnostic and/or research applications.

The present invention is directed to a bioconjugate vaccine, such as an O1-bioconjugate vaccine, comprising: a protein carrier comprising a protein carrier containing at least one consensus sequence, D/E-X-N-Z-S/T, wherein X and Z may be any natural amino acid except proline; at least one antigenic polysaccharide from at least one pathogenic bacterium, linked to the protein carrier; and, optionally, an adjuvant. In another aspect, the present invention is directed to a method of producing an O1-bioconjugate in a bioreactor comprising a number steps.