The present disclosure relates to nucleobase editors and methods of use thereof. Disclosed herein are fusion proteins, systems, and compositions for editing disease-associated mutations and methods of use thereof. In some aspects, disclosed herein is a fusion protein comprising a Cas9 nickase and a nucleotide deaminase, wherein the Cas9 nickase comprises a first amino acid substitution at a position selected from the group consisting of 262, 324, 409, 480, 543, 694, and 1219 when compared to SEQ ID NO: 11, and wherein the Cas9 nickase comprises a second amino acid substitution at a position selected from the group consisting of 1111, 1135, 1218, 1219, 1322, 1335, and 1337 when compared to SEQ ID NO: 11.

The present invention provides an enzymatic means for the biosynthetic production of caffeine. The present invention provides biosynthetic methods for production of caffeine comprising: providing guanine, a guanine deaminase, at least one methyl transferase, and a methyl donor; contacting the guanine with the gtheuanine deaminase to produce xanthine; contacting the xanthine with the methyl transferase and a methyl donor, under conditions wherein the xanthine is methylated, to produce a monomethylxanthine; contacting the monomethylxanthine with the methyl transferase and a methyl donor, under conditions wherein the monomethylxanthine is methylated, to produce a dimethylxanthine; and contacting the dimethylxanthine with the methyl transferase and a methyl donor, under conditions wherein the dimethylxanthine is methylated, to produce caffeine (i.e., 1,3,7-trimethylxanthine).

Disclosed are transformed cells and related nucleotide and protein sequences, and fermentation compositions and methods, all of which are related to providing selective advantage in fermentation. For example, a selective advantage results from transformation of a cell with a nucleic acid that allows a transformed cell to metabolize one or more nitrogen-, phosphorous-, and/or sulfur-containing compounds that a native cell of the same species as the transformed cell cannot metabolize, and from fermentation of the transformed cell using one or more feedstocks, such as fractioned grain, which are depleted in or free of conventional nitrogen-, phosphorous-, and/or sulfur-containing compounds that a native cell of the same species as the transformed cell can metabolize. Also disclosed are methods for improved oxygen transfer in an aerobic or microaerobic fermentation.

The present invention relates to methods of metabolic engineering cells to increase their ability to compete with contaminating microorganisms without the need for antibiotics. More particularly, the invention provides methods to engineer cyanobacteria to utilize melamine as nitrogen source, phosphite as phosphorous source, optionally also utilizing NADP+ over NAD+, and also provides genetically engineered cells made using such methods.

An object of the present invention is to provide an enzyme preparation useful for reducing purine bodies, especially, in beer or beer-based beverages, and use thereof. Provided is a nucleosidase preparation having an activity ratio (U/U), which is guanine deaminase activity per nucleosidase activity, of 0.4 or less.

An object of the present invention is to provide an enzyme preparation useful for reducing purine bodies, especially, in beer or beer-based beverages, and use thereof. Provided is a nucleosidase preparation having an activity ratio (U/U), which is guanine deaminase activity per nucleosidase activity, of 0.4 or less.

The present invention relates to a new process for the preparation of ammeline and/or ammelide from melamine by a solid-to-solid reaction in an aqueous reaction mixture comprising a biocatalyst, wherein the biocatalyst comprises at least one enzyme belonging to the amidohydrolase superfamily and having aminohydrolase activity towards 1,3,5-triazine compounds. The invention further relates a product obtainable by the process according to the invention, wherein the product comprises ammeline and/or ammelide.

Disclosed herein are methodologies and kits for dynamic targeted hypermutation that harness the enzymatic activity of a polynucleic acid-binding protein fused to a nucleobase-editing enzyme to specifically target mutations across a region of interest. These methodologies and kits facilitate the rapid creation of diverse DNA libraries in vivo or in vitro.

The present invention relates to a new process for the preparation of ammeline and/or ammelide from melamine by a solid-to-solid reaction in an aqueous reaction mixture comprising a biocatalyst, wherein the biocatalyst comprises at least one enzyme belonging to the amidohydrolase superfamily and having aminohydrolase activity towards 1,3,5-triazine compounds. The invention further relates a product obtainable by the process according to the invention, wherein the product comprises ammeline and/or ammelide.

Disclosed are transformed cells and related nucleotide and protein sequences, and fermentation compositions and methods, all of which are related to providing selective advantage in fermentation. For example, a selective advantage results from transformation of a cell with a nucleic acid that allows a transformed cell to metabolize one or more nitrogen-, phosphorous-, and/or sulfur-containing compounds that a native cell of the same species as the transformed cell cannot metabolize, and from fermentation of the transformed cell using one or more feedstocks, such as fractioned grain, which are depleted in or free of conventional nitrogen-, phosphorous-, and/or sulfur-containing compounds that a native cell of the same species as the transformed cell can metabolize. Also disclosed are methods for improved oxygen transfer in an aerobic or microaerobic fermentation.