Preparation method of reduced nicotinamide adenine dinucleotide

A technology of nicotinamide adenine and dinucleotides, applied in biochemical equipment and methods, oxidoreductases, enzymes, etc., can solve problems such as unfavorable purification, achieve reduced NADH loss, high product yield, and simplified purification process Effect

Pending Publication Date: 2022-03-01
NANJING NUOYUN BIOLOGICAL TECH CO LTD
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Problems solved by technology

However, the reduction of NAD by formate dehydrogenase will generate carbon dioxide, which escapes from the reaction mixture only

Method used

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  • Preparation method of reduced nicotinamide adenine dinucleotide
  • Preparation method of reduced nicotinamide adenine dinucleotide
  • Preparation method of reduced nicotinamide adenine dinucleotide

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Embodiment Construction

[0032] The technical solutions in the embodiments of the present invention will be clearly and completely described below in conjunction with the accompanying drawings in the embodiments of the present invention. Obviously, the described embodiments are only some of the embodiments of the present invention, not all of them. Based on the embodiments of the present invention, all other embodiments obtained by persons of ordinary skill in the art without making creative efforts belong to the protection scope of the present invention.

[0033] (1) Acquisition of formate dehydrogenase

[0034] Wild-type formate dehydrogenase derived from Candida boidinii (Candida boidinii) was purchased from Nanjing Langen Biotechnology Co., Ltd., product number FDH010. Three formate dehydrogenases were purchased from Nanjing Langen Biotechnology Co., Ltd., product numbers FDH015, FDH018, and FDH019.

[0035] (2) Screening of formate dehydrogenase activity under high temperature conditions

[003...

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Abstract

The invention relates to the technical field of industrial catalysis, in particular to a preparation method of reduced nicotinamide adenine dinucleotide, which comprises the following steps: 1) screening the activity of formate dehydrogenase at high temperature; 2) screening the activity of formate dehydrogenase under an acidic condition; (3) according to the results of the step (1) and the step (2), formate dehydrogenase with relatively high activity under a high-temperature condition and an acidic condition is selected for later use; respectively weighing ammonium formate and NAD (Nicotinamide Adenine Dinucleotide) into a flask, adding pure water, stirring and dissolving in a water bath, and regulating the pH to be acidic by using sodium hydroxide; finally, formate dehydrogenase obtained through screening is added for reaction, sodium hydroxide is used for maintaining the pH value to be acidic, and the reduced nicotinamide adenine dinucleotide is prepared. According to the method, the reaction can be completed in a short time, so that hydrolysis of a product under an acidic condition is avoided, and carbonate generated under an alkaline condition is also avoided; after the reaction, no large amount of carbonate exists, and the purification cost is low.

Description

technical field [0001] The invention relates to the technical field of industrial catalysis, specifically a method for preparing reduced nicotinamide adenine dinucleotide. Background technique [0002] Reduced nicotinamide adenine dinucleotide (NADH) is a coenzyme of various redox enzymes, which plays an important role in industrial catalysis. In recent years, researchers have found that NADH also has a variety of human health functions, which may be a potential protective Health products / drugs for liver protection and hangover. [0003] The mainstream production method of NADH is prepared by extracting NAD from yeast and reducing it to NADH. At present, there are many enzymatic methods for reducing NAD to NADH, such as alcohol dehydrogenase, formate dehydrogenase, glucose dehydrogenase and 6-phosphate glucose dehydrogenase, etc., but different preparation processes affect the purification of the product larger. [0004] Currently, the production of NADH mainly uses alcoh...

Claims

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Application Information

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IPC IPC(8): C12P19/36C12N9/96C12N9/02
CPCC12P19/36C12N9/96C12N9/0008C12Y102/01002
Inventor 丁雪峰钱明李佳松李帅杰刘珊珊
Owner NANJING NUOYUN BIOLOGICAL TECH CO LTD
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