The disclosure discloses recombinant Bacillus subtilis for synthesizing guanosine diphosphate fucose and a construction method and application thereof. The recombinant Bacillus subtilis is obtained by intensively expressing guanylate kinase and nucleotide diphosphokinase genes and expressing exogenous fucokinase and phosphate guanylyltransferase genes in a genome of Bacillus subtilis 168. According to the disclosure, a bacterial strain for synthesizing the guanosine diphosphate fucose is obtained by reconstructing the Bacillus subtilis 168, with a volume of intracellular accumulation up to 196.15 g/L. According to the disclosure, by intensively expressing the guanylate kinase and nucleotide diphosphokinase genes, and enhancing the supply of intracellular GDP-L-fucose composition cofactors, the synthesis of the guanosine diphosphate fucose is promoted. The construction method for the recombinant Bacillus subtilis of the disclosure is simple and convenient to use, thus having good application prospects.

The invention relates a method for producing a stable recombinant protein, comprising growing a non-naturally occurring host cell in a culture medium to produce a recombinant protein, and making a composition comprising the recombinant protein and a polysorbate. The production of endogenous lipoprotein lipase by the host cell is reduced. The endogenous lipoprotein lipase is present in the composition in a small amount, and is capable of degrading the polysorbate. The invention also relates to the relevant host cells and compositions, and preparation thereof.

The present invention relates to novel C-glycosyltransferase variants and a use thereof. The C-glycosyltransferase variants according to the present invention have improved glycosidic bond-forming ability as compared with wild-type C-glycosyltransferase, and thus can increase the glycoside production effects of polyketide groups and pseudo-natural products, particularly type I, II, III polyketide, nonribosomal peptides, phenylpropanoids, and other aromatic natural products, and thus can be useful for the preparation of a drug, a food additive, a nutritional supplement, and the like containing a C-glycoside compound as a constituent ingredient.

The invention provides modular cell-free de-novo synthesis of glycans with immobilized bionanocatalysts. The invention provides materials, and in particular, magnetic materials, for producing glycans of defined length and sequences using one or more enzymes that are immobilized within bionanocatalysts (BNCs) which in turn are embedded within scaffolds to control the synthesis in batch or continuous processes manufacturing. In some embodiments, the scaffolds are high magnetism and high porosity composite blends of thermoplastics or thermosets comprising magnetic particles that form powders. In some embodiments, Selective Laser Sintering (SLS) is used to design and produce objects via 3D printing by sintering composite magnetic powders. The modular flow cells may be mixed and matched for a highly customizable and highly efficient cell-free manufacturing process. In some embodiments the elementary and system modules provided by the invention are employed. In preferred embodiments, human milk oligosaccharides (HMOs) are produced.

The present disclosure provides enzymatic methods for the production of 3-phosphate-nucleoside-5-triphosphate (NQP). The present disclosure further provides enzymatic methods for the production of nucleotide-5-triphosphates.

Provided herein, in some embodiments, are methods and composition for the production of nucleoside triphosphates and ribonucleic acids.

The present application discloses inhibitors of NME family of proteins.

The present application discloses inhibitors of NME family of proteins.

Provided is a method for producing circular DNA in which a region that is sandwiched by a region Ha and a region Hb in circular double-stranded DNA is substituted with the entirety or a portion of a linear-DNA fragment, wherein the region Hb is located downstream of the region Ha in the circular double-stranded DNA; the linear-DNA fragment is single-stranded or double-stranded linear DNA that has a homologous region that corresponds to the region Ha and a homologous region that corresponds to the region Hb, the latter homologous region being positioned downstream of the former homologous region; and the method comprising: preparing a reaction solution that contains the circular double-stranded DNA, the linear-DNA fragment, and a protein that has RecA-family-recombinase activity, and performing homologous recombination reaction by incubating the reaction solution for a predetermined period of time, thereby producing circular DNA in which the region that is from the region Ha to the region Hb in the circular double-stranded DNA is substituted with the region that is from the homologous region that corresponds to the region Ha to the homologous region that corresponds to the region Hb in the linear-DNA fragment.

The invention relates a method for producing a stable recombinant protein, comprising growing a non-naturally occurring host cell in a culture medium to produce a recombinant protein, and making a composition comprising the recombinant protein and a polysorbate. The production of endogenous lipoprotein lipase by the host cell is reduced. The endogenous lipoprotein lipase is present in the composition in a small amount, and is capable of degrading the polysorbate. The invention also relates to the relevant host cells and compositions, and preparation thereof.