Provided are a method for preparing an immobilized multi-enzyme system, and a method for producing tagatose by the immobilized multi-enzyme system. The immobilized multi-enzyme system is formed by uniformly mixing a porous dopamine microsphere with a multi-enzyme mixture which is used for producing tagatose. Five enzymes in an enzymatic catalysis path for converting starch to tagatose are co-immobilized by means of a porous microsphere to obtain an immobilized multi-enzyme system, the immobilized multi-enzyme system is used to catalyze conversion of starch into tagatose, and thus, enzymes can be recycled, thereby greatly reducing the amount of enzymes required for preparation of tagatose, and reducing the production cost.

In various aspects and embodiments, the present disclosure provides methods for making glycosylated products, as well as bacterial cells and uridine diphosphate (UDP)-dependent glycosyltransferase (UGT) enzymes useful for the same. In other aspects and embodiments, the disclosure provides methods for high yield and/or high purity recovery of such glycoside products from microbial cultures or cell free reactions. In various aspects and embodiments, the disclosure provides for whole cell bioconversion processes involving the glycosylation of a desired substrate, and/or the recovery of the glycosylated product at high yield and/or high purity.

The current disclosure provides a process for enzymatically converting a saccharide into allulose. The invention also relates to a process for preparing allulose where the process involves converting fructose 6-phosphate (F6P) to allulose 6-phosphate (A6P), catalyzed by allulose 6-phosphate 3-epimerase (A6PE), and converting the A6P to allulose, catalyzed by allulose 6-phosphate phosphatase (A6PP).

The present disclosure relates to a non-naturally occurring microorganism that includes an endogenous genetic deletion that eliminates the expression of at least a pyruvate kinase, where the genetically modified prokaryotic microorganism is capable of producing 3-deoxy-D-arabino-heptulosonate-7-phosphate.

The invention discloses a method for improving the yield of Bacillus subtilis acetylglucosamine, which belongs to the technical field of genetic engineering. In the invention, the recombinant Bacillus subtilis S5 (S5-PxylA-glmS-P43-GNA1) is taken as a starting strain, and the glmS ribozyme is integrated into the mid of rbs and the promoter sequence of the glmM and pfkA gene, respectively. The ribozyme mutant has the advantage of prolonging the stability of the mRNA and integrated into the mid of rbs and the promoter sequence of the pgi gene. The yield of GlcNAc of the recombinant strain reaches 11.79-20.05 g/L. This laid the foundation for the further metabolic engineering of Bacillus subtilis to produce GlcNAc.

A nanoplatelet serves as a substrate for immobilizing enzymes involved in consecutive reactions as a cascade. This results in a significant increase in the rate of catalysis as well as final product yield compared to non-immobilized enzymes or enzymes immobilized to quantum dots.

The invention relates to artificial forisome bodies including a fusion protein of at least one SEO-F protein or an at least 50-amino acid portion thereof, and at least one additional protein or peptide, with the exception of GFP and the Venus protein, wherein in an embodiment the forisome body further includes an unfused SEO-F protein or a form of the protein having C-terminal deletions of up to 45 amino acids and/or N-terminal deletions of up to 13 amino acids, wherein the unfused SEO-F protein is selected from proteins having the property of being capable of forming homomeric forisome bodies in the absence of additional SEO-F proteins.

Artificial forisome bodies include a fusion protein of at least one SEO-F protein or an at least 50-amino acid portion of an SEO-F protein, and at least one additional protein or peptide, with the exception of GFP and the Venus protein. The additional protein or peptide has a mass of at most 30 kDa, or the forisome body further includes an unfused SEO-F protein or a form of the protein having C-terminal deletions of up to 45 amino acids and/or N-terminal deletions of up to 13 amino acids, in which the unfused SEO-F protein is a protein capable of forming homomeric forisome bodies in the absence of additional SEO-F proteins.

Various aspects and embodiments herein relate to recombinant proteins with at least one protease recognition sequence, wherein the recombinant proteins can be inactivated by a cognate protease and methods of preparing such proteins. In some embodiments, recombinant phosphoglucose isomerase (Pgi) proteins are provided. In other embodiments, recombinant phosphotransacetylase (Pta) proteins are provided. In yet other embodiments, recombinant transketolase A (TktA) proteins are provided.

Disclosed herein are methods of producing hexoses from saccharides by enzymatic processes. The methods utilize fructose 6-phosphate and at least one enzymatic step to convert it to a hexose.