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

Claim 2:
3. 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 carnosine;
the enzyme that the produces the product is a transferase for transferring a phosphate group and is carnosine synthetase;
the enzymatic reaction requiring ATP is a synthesis reaction of carnosine; and
the enzymatic reaction system is a 100 L enzymatic reaction system that comprises:
a solution comprising:
1.8 kg of L-histidine;
1.0 kg of β-alanine;
0.25 kg of adenosine;
1.0 kg of sodium polyphosphate;
0.3 kg of sodium chloride;
0.38 kg of potassium chloride;
1.0 kg of magnesium chloride hexahydrate; and
0.6 kg of disodium hydrogen phosphate, wherein the pH of the solution is adjusted to 8.0;

ATP regeneration enzymes comprising:
ADK enzyme 600 U/L; and
PAP enzyme 600 U/L;

AK enzyme 800 U/L; and
carnosine synthetase 1000 U/L;
wherein the enzymatic reaction system is maintained at a temperature of 37° C. and a pH of 8.0;

(2) separating the ATP regeneration enzymes, the AK enzyme, and the carnosine synthetase from the enzymatic reaction system to produce a filtrate comprising carnosine, ATP, ADP, and AMP, wherein:
the ATP regeneration enzymes, the AK enzyme, and the carnosine synthase are separated from the enzymatic reaction system with an ultrafiltration membrane in a filter;
the activity of the separated ATP regeneration enzymes, the separated AK enzyme, and the separated carnosine synthase are reduced by 10%-20% compared with that before the reaction; and
the separated ATP regeneration enzyme, the separated AK enzyme, and the separated carnosine synthase are recycled for use in step (1) for the enzymatic reaction; and

(3) separating and purifying the filtrate of step (2) to obtain the carnosine.