The present invention relates to an enzyme-catalyzed process for producing UDP-galactose from low-cost substrates uridine monophosphate and D-galactose in a single reaction mixture. Said process can be operated (semi)continuously or in batch mode. Said process can be extended to uridine as starting material instead of uridine monophosphate. Further, said process can be adapted to produce galactosylated molecules and biomolecules including saccharides, proteins, peptides, glycoproteins or glycopeptides, particularly human milk oligosaccharides (HMO) and (monoclonal) antibodies.

The invention pertains to a PPA from a microorganism belonging to the family Halobacteriaceae (HPPA), for example, a PPA from Haloferax volcanii. The HPPA provided by the invention is soluble, thermostable and active at high concentrations of salt and/or organic solvent. An embodiment of the invention provides a method of increasing the rate of a reaction by adding an HPPA to the reaction mixture, wherein the reaction produces PPi, for example, an enzymatic reaction, and wherein the reaction is carried out at moderately high temperature and/or low water activity. Further embodiments of the invention provide an assay to detect the PPi released during a reaction which produces PPi by adding an HPPA to convert the PPi in to Pi and measuring the resultant Pi. The invention further pertains to an assay to monitor a reaction which produces PPi in the presence or the absence of an HPPA.

The present invention relates to an enzyme-catalyzed process for producing UDP-galactose from low-cost substrates uridine monophosphate and D-galactose in a single reaction mixture. The process can be operated (semi)continuously or in batch mode. The process can be extended to uridine as starting material instead of uridine monophosphate. Further, the process can be adapted to produce galactosylated molecules and biomolecules including saccharides, proteins, peptides, glycoproteins or glycopeptides, particularly human milk oligosaccharides (HMO) and (monoclonal) antibodies.

The present invention relates to oligomeric nucleic acids and uses thereof for the treatment of tumors. The oligomeric nucleic acid for tumor treatment provided by the present application can be small activating nucleic acid molecules. A small activating nucleic acid molecule of the present invention can be a double-stranded or single-stranded RNA molecule targeting the promoter region of an LHPP gene comprising a first nucleic acid strand and a second nucleic acid strand. The double-stranded RNA molecule targeting the promoter region of the LHPP gene comprises two nucleic acid strands of 16 to 35 nucleotides in length, wherein one of the nucleic acid strands has at least 75% homology or complementarity to a target selected from the promoter region of the LHPP gene. The present invention also relates to pharmaceutical compositions comprising the small activating nucleic acids and optional pharmaceutically acceptable carriers, and methods for upregulating the expression of the LHPP gene in a cell and methods for treating diseases or conditions related to insufficient or decreased expression of LHPP gene by using the small activating nucleic acid molecules or the pharmaceutical compositions.

The present invention provides a method for producing a target substance, the biosynthetic pathway of which is ATP-dependent, such as, for example, amino acids, nucleosides, nucleotides, isoprenoids, and peptides, by fermentation of a bacterium which has been modified to overexpress a gene encoding a protein having H.sup.+-translocating membrane-bound pyrophosphatase activity, such as, for example, the hppA gene native to R. rubrum or a variant thereof.

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 invention provides a method for producing a target substance, the biosynthetic pathway of which is ATP-dependent, for example, amino acids, nucleosides, nucleotides, isoprenoids, and peptides, by fermentation of a bacterium which has been modified to overexpress a gene encoding a protein having H.sup.+-translocating membrane-bound pyrophosphatase activity, for example, the hppA gene native to R. rubrum or a variant thereof.

The present invention relates to an inorganic pyrophosphatase (PPase), methods of producing the same and uses thereof. Further disclosed are an enzyme reactor and a kit comprising the PPase.

The invention pertains to a PPA from a microorganism belonging to the family Halobacteriaceae (HPPA), for example, a PPA from Haloferax volcanii. The HPPA provided by the invention is soluble, thermostable and active at high concentrations of salt and/or organic solvent. An embodiment of the invention provides a method of increasing the rate of a reaction by adding an HPPA to the reaction mixture, wherein the reaction produces PPi, for example, an enzymatic reaction, and wherein the reaction is carried out at moderately high temperature and/or low water activity. Further embodiments of the invention provide an assay to detect the PPi released during a reaction which produces PPi by adding an HPPA to convert the PPi in to Pi and measuring the resultant Pi. The invention further pertains to an assay to monitor a reaction which produces PPi in the presence or the absence of an HPPA.

The present invention relates to a method for producing cytidine 5-monophospho-N-acetyl-neuraminic acid (CMP-Neu5Ac, 1) from low-cost substrates N-acetyl-D-glucosamine (GlcNAc), pyruvate, cytidine and polyphosphate in a single reaction mixture with a set of optionally immobilized or optionally co-immobilized enzymes comprising N-acylglucoamine 2-epimerase (AGE), an N-acetylneuraminate lyase (NAL), an N-acylneuraminate cytidylyltransferase (CSS), a uridine kinase (UDK), a uridine monophosphate kinase and a polyphosphate kinase 3 (PPK3). Further, said process may be adapted to produce Neu5Acylated i.e. sialylated biomolecules and biomolecules including a saccharide, a peptide, a protein, a glycopeptide, a glycoprotein, a glycolipid, a glycan, an antibody, and a glycoconjugate, in particular, an antibody drug conjugate, and a carbohydrate conjugate vaccine, or a flavonoid.