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Preparation method of oxidizing-type coenzyme I

An oxidized coenzyme and yeast cell technology, which is applied in chemical instruments and methods, preparation of sugar derivatives, organic chemistry, etc., can solve the problems that cannot meet the needs of industrialized large-scale production, is not suitable for industrialized large-scale production, and has unsatisfactory results. No toxic chemical residues, mild conditions, and less destructive effects

Active Publication Date: 2015-12-09
HEFEI KNATURE BIO PHARM CO LTD
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] At present, the commonly used wall-breaking methods include temperature difference method and high-pressure homogenization method. The temperature difference method (heat treatment method) mainly destroys the cell tissue structure through the force generated by water freezing and heating and melting, as well as sudden changes in temperature. The effect of this method is not ideal; The high-pressure homogenization method requires a high-pressure homogenizer and a large-capacity high-speed centrifuge. The principle of crushing is that the cells undergo shearing, collisions and changes from high pressure to normal pressure in a series of processes, resulting in cell fragmentation. The method has high production cost and low efficiency, and is not suitable for large-scale industrial production
It can be seen from the above that neither of the two methods can meet the needs of industrialized mass production.

Method used

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  • Preparation method of oxidizing-type coenzyme I

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Embodiment 1

[0030] A preparation method of oxidized coenzyme I, comprising the steps of breaking yeast cells and extracting NAD + ;

[0031] Among them, the specific steps of breaking the yeast cells are: add the yeast cells into hydrochloric acid with a concentration of 0.01mol / l and soak for 2.5 hours, heat to 85°C, keep warm for 8min, add ice cubes and cool to room temperature at a speed of 10°C / min, Centrifuge at a speed of 4000rpm for 20min to obtain supernatant A; stir for 16h, add 717 # Resin, after stirring, filter to obtain supernatant B, wherein the weight volume ratio (g / ml) of yeast cells and hydrochloric acid is 15:400, yeast cells and 717 # The weight ratio of the resin is 15:45;

[0032] Among them, the extraction of NAD + The specific steps are as follows:

[0033] S1. Acidification: adding hydrochloric acid with a concentration of 5 mol / l to the clear liquid B to adjust the pH to 2.05 to obtain a solution C;

[0034] S2. Exchange: Put the solution C obtained in S1 on...

Embodiment 2

[0041] A preparation method of oxidized coenzyme I, comprising the steps of breaking yeast cells and extracting NAD + ;

[0042] Among them, the specific steps of breaking the yeast cells are: adding the yeast cells into hydrochloric acid with a concentration of 0.12mol / l and soaking them for 0.5h, heating to 100°C, keeping the temperature for 3min, adding ice cubes and cooling to room temperature at a speed of 15°C / min, Centrifuge at a speed of 6000rpm for 10min to obtain supernatant A; stir for 12h, add 717 # Resin, filtered after stirring to obtain supernatant B, wherein the weight volume ratio (g / ml) of yeast cells and hydrochloric acid is 25:80, yeast cells and 717 # The weight ratio of the resin is 25:90;

[0043] Among them, the extraction of NAD + The specific steps are as follows:

[0044] S1. Acidification: add hydrochloric acid with a concentration of 7 mol / l to the clear liquid B to adjust the pH to 1.95 to obtain a solution C;

[0045] S2. Exchange: Put the s...

Embodiment 3

[0052] A preparation method of oxidized coenzyme I, comprising the steps of breaking yeast cells and extracting NAD + ;

[0053] Among them, the specific steps of breaking the yeast cells are: add the yeast cells into hydrochloric acid with a concentration of 0.11mol / l and soak them for 0.7h, heat them to 98°C, keep them warm for 7min, add ice cubes and cool them down to room temperature at a speed of 12°C / min, Centrifuge at a speed of 5500rpm for 18min to obtain supernatant A; stir for 13h, add 717 # Resin, after stirring, filter to obtain supernatant B, wherein the weight volume ratio (g / ml) of yeast cells and hydrochloric acid is 22:100, yeast cells and 717 # The weight ratio of the resin is 22:70;

[0054] Among them, the extraction of NAD + The specific steps are as follows:

[0055] S1. Acidification: add hydrochloric acid with a concentration of 6.5 mol / l to the clear liquid B to adjust the pH to 1.98 to obtain a solution C;

[0056] S2. Exchange: Put the solution ...

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Abstract

The invention discloses a preparation method of oxidizing-type coenzyme I, which includes the steps of disrupting yeast cells and extracting NAD+. The step of disrupting yeast cells particularly comprises: (1) soaking yeast in hydrochloric acid, heating the yeast, maintaining the temperature, adding ice cubes to cool the liquid to room temperature, and performing centrifugation to obtain a clear solution A; and (2) stirring the clear solution A for 12-16 h with addition of 717# resin, and performing filtration after stirring to obtain a clear solution B. The step of extracting NAD+ particularly comprises the following steps: acidification, exchange, elution, desalination, separation, elution and collection. The preparation method greatly increases disruption rate of yeast cells. The raw materials are low in cost and are easy to obtain. The device is simple and is convenient to use. The preparation method is suitable for industrial production.

Description

technical field [0001] The invention relates to the technical field of coenzyme I preparation, in particular to a method for preparing oxidized coenzyme I. Background technique [0002] Coenzyme I (NAD), chemically named nicotinamide adenine dinucleotide or nicotinic acid diphosphate, exists in the oxidized form in mammals (NAD + ) and reduced (NADH), which are important coenzymes in the redox reaction of the human body. Participate in the metabolism of various cell functions and play an important role in the body's immunity. In a healthy state, the concentration of coenzyme I in the human body is stable to maintain the normal functions of various cells. The concentration of coenzyme I in the body determines the process and degree of cell aging, and the decrease in concentration will accelerate the process of cell aging. [0003] Coenzyme I mainly exists in the mitochondria of yeast cells, which belongs to intracellular enzymes, and the thickness of yeast cell wall is 100...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07H19/207C07H1/08
Inventor 王康林杨杨金永红
Owner HEFEI KNATURE BIO PHARM CO LTD
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