The invention discloses a method of using an in-vitro enzyme method to catalyze a heparin precursor to prepare heparin, and belongs to the field of biomedicine. A method of using a pure biological enzyme method to synthesize heparin pentasaccharide with biological functions is provided. By fusing maltose fusion protein or amphiphilic short peptide sequences to 5' terminal of a gene, the expression amount and enzymatic activity of corresponding sulfate transferase or allosteric enzymes are increased, and single blood clotting drugs with a stable structure is obtained.

Method of using in-vitro enzyme method to catalyze heparin precursor to prepare heparin

The invention discloses a recombinant bacterium for expressing a chondroitin 6-sulfate transferase gene and application of the recombinant bacterium, and belongs to the technical field of bioengineering. According to the invention, escherichia coli or bacillus subtilis is used as a host to express the chondroitin 6-sulfate transferase gene with the amino acid sequence as shown in SEQ ID NO. 1, four recombinant strains are constructed, high-efficiency expression of the chondroitin 6-sulfate transferase is realized, and the final enzyme activities respectively reach 36.3 U/mL, 40.2 U/mL, 38.4 U/mL and 51.6 U/mL. Chondroitin sulfate C is synthesized through one-step catalysis by taking enzyme as a catalyst and chondroitin as a substrate, and the final conversion efficiency reaches 80%. The method of the invention has potential and quite wide value industrially for the synthesis of chondroitin sulfate C.

Recombinant bacterium for expressing chondroitin 6-sulfate transferase gene and application of recombinant bacterium

The invention belongs to the technical field of pharmaceutical and chemical industry, discloses zymochemistry synthesis and applications of a heparan sulfate and FK506 conjugate, and provides a smallmolecular tool or probe for studying the embryo development regulating action mechanism of heparan sulfate and understanding the structure-activity relationship of heparan sulfate in cell nucleus, wherein the heparan sulfate refers to a hexuronic acid and glucosamine repeat disaccharide unit with characteristics of defined structure and diversification, and the isotope C labeling radioactivityanalysis results show that the conjugate has the greatly-improved transmembrane and nuclear import ability.

Zymochemistry synthesis and applications of heparan sulfate and FK506 conjugate

The invention discloses a chondroitinase ABC producing recombinant yeast strain and application thereof and belongs to the technical field of bioengineering. According to the chondroitinase ABC producing recombinant yeast strain, chondroitinase ABC from Proteus vulgaris ATCC33420 is subjected to heterogeneous expression, and with pPIC9K carriers and through induction of methanol, secretory expression of the chondroitinase ABC in pichia pastoris GS115 can be achieve; by means of a designed chondroitinase ABC-enzymatic membrane reactor, efficient and continuous production of small-molecular chondroitin sulfate can be achieved, the chondroitinase ABC can be recycled and reutilized, and the production cost can be greatly reduced. By taking food-grade yeast as host strains, the chondroitinase ABC producing recombinant yeast strain can be safe and reliable, provide effective references for industrialized and green production of small-molecular chondroitin sulfate A, B and C, and meanwhile, save energy, reduce emission and achieve significant economic and social benefits.

Chondroitinase ABC producing recombinant yeast strain and structuring and applying methods thereof

The invention discloses recombinant strains expressing a chondroitin 4-sulfotransferase gene and an applicationof the recombinant strain, and relates to the technical field of bioengineering. Chondroitin 4-sulfotransferase gene with the amino acid sequence shown in SEQ ID NO.1 is expressed with Escherichia coli or bacillus subtilis as a host, four recombinant strains are constituted, the high-efficiency expression of chondroitin 4-sulforansferase gene is realized, and final enzyme activity respectively reaches 41.3 U/mL, 32.8 U/mL, 36.5 U/mL and 50.2 U/mL. By using the enzyme as a catalyst, chondroitin sulfate A is catalyzed and synthesis in one step with chondroitin as a substrate, the final conversion efficiency reaches 80%. The method has potential and quite wide values in industrial synthesis of chondroitin sulfate A.

Recombinant strains expressing chondroitin 4-sulfotransferase gene and application of recombinant strain

The invention discloses an enzymatic method for preparing chondroitin sulfate and belongs to the medical field. Chondroitin is used as a substrate, chondroitin sulfate A with bioactivity is formed through catalysis of chondroitin 4-sulfate transferase for microbial cell heterologous expression or chondroitin sulfate C with bioactivity is formed through catalysis of chondroitin 6-sulfate transferase, and a PAPS sulfuric donor and a regeneration system are provided by acyl sulfate transferase. A new way is provided for synthesis and industrial production of biological active substances, namely, chondroitin sulfate A and chondroitin sulfate C.

Du Guocheng

Papers

Enzymatic method for preparing chondroitin sulfate

Method of using in-vitro enzyme method to catalyze heparin precursor to prepare heparin