Patent ID: 11939615
Assignee: ANHUI GSH BIO-TECH CO., LTD.
Field: Chemical engineering (Chemistry)
Classification: CPC C  B  Y | IPC C

Claim 1:
2. A method of producing a product from an enzymatic reaction requiring adenosine triphosphate (ATP), the method comprising the following steps:
(1) adding ATP regeneration enzymes, AK (adenosine kinase) enzyme, enzyme that produces the product, and adenosine to an enzymatic reaction system to carry out the enzymatic reaction, wherein:
the ATP generation enzymes are a combination of two or three enzymes selected from the group consisting of: polyphosphate kinase (PPK), adenylate kinase (ADK), and polyphosphate: AMP phosphotransferase (PAP);
adenosine is used instead of ATP and adenosine monophosphate (AMP);
the product is fructose 1,6-diphosphate;
the enzyme that produces the product is a transferase for transferring a phosphate group selected from fructokinase (FK) and phosphofructokinase (PFK);
the enzymatic reaction requiring ATP is a synthesis reaction of fructose 1,6-diphosphate; and
the enzymatic reaction system is a 100 L enzymatic reaction system that comprises a reaction solution comprising:
1.8 kg of fructose;
0.3 kg of adenosine;
2.0 kg of sodium hexametaphosphate;
0.5 kg of ammonium chloride;
0.5 kg of sodium chloride;
0.5 kg of magnesium sulfate heptahydrate;
0.2 kg of manganese sulfate; and
0.85 kg of disodium hydrogen phosphate,
wherein the pH of the reaction solution is adjusted to 6.8 and brought to a temperature of 35-40° C.;
the ATP regeneration enzymes, the AK enzyme, and catalytic enzymes are immobilized in the enzymatic reaction system;
the ATP generation enzymes are a combination of the ADK enzyme and the PAP enzyme;
the catalytic enzymes are fructokinase (FK) and phosphofructokinase (PFK);
the FK enzyme, the PFK enzyme, the ADK enzyme, the PAP enzyme, and the AK enzyme in the enzymatic reaction system are in an activity ratio of 2:2:1:1:1;
the AK enzyme activity in the enzyme solution is 2000 U/L;
the reaction solution is passed through the enzymatic reaction system at a constant flow rate of 30 L/h; and
the enzymatic reaction system is maintained at 37° C. for the synthesis of fructose 1,6-diphosphate;

(2) directly separating the ATP regeneration enzymes, the AK enzyme, the FK enzyme, and the PFK enzyme from the enzymatic reaction system to produce a filtrate comprising fructose 1,6-diphosphate, ATP, ADP, and AMP; and
(3) separating and purifying the filtrate of step (2) to obtain the fructose 1,6-diphosphate.