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Stable composition containing NAD+ or NADH

A composition and stable technology, applied in sugar derivatives, sugar derivatives, organic chemistry, etc., can solve problems such as inaccurate measurement results, and achieve the effect of wide application value

Inactive Publication Date: 2013-01-09
上海执诚生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] The inventors noticed that the mere hydrolysis of the coenzymes NAD and NADH by ambient humidity would lead to inaccurate measurements even under less severe conditions such as in aqueous solutions

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Application of the present invention in lactate dehydrogenase test kit:

[0024] First prepare kit reagent 1: 40 mmol / l lactic acid, 100 mmol / l Tris solution, adjust pH to 9.2 with 1 mol / l hydrochloric acid or sodium hydroxide.

[0025] Re-preparation kit reagent 2: containing trehalose 10g / l, glycerol 1g / l, Tris 0.1mol / l, Tween 20 0.1g / l solution, then use 1mol / l hydrochloric acid or sodium hydroxide to adjust its pH to 7.0, then weigh some NAD + Dissolve in 10ml of the above protective agent to make the concentration reach 20g / l, and stir evenly.

[0026] The above-mentioned reagent 1 and reagent 2 constitute the lactate dehydrogenase kit, and the storage period of this kit can reach 18 months at 2-8°C. Through comparative experiments, the lactate dehydrogenase kit without stabilizers can only be kept for twelve months under the same conditions.

Embodiment 2

[0028] Application of the present invention in α-hydroxybutyrate dehydrogenase kit:

[0029] First prepare the kit reagent 1: a solution containing trehalose 5g / l, glycerol 10g / l, Tris 0.1mol / l, Tween 20 0.5g / l, then adjust its pH to 7.0, then weigh 10mg NAD and dissolve it in 10ml of the above protective agent, stir evenly.

[0030] Reconstitute kit reagent 2: nitric acid 50mmol / l solution, α-oxobutyric acid 3mmol / l. Adjust the pH to 7.5.

[0031] The above-mentioned reagent 1 and reagent 2 constitute the α-hydroxybutyrate dehydrogenase kit, and the storage period of this kit can reach 18 months at 2-8°C. Through comparative experiments, the lactate dehydrogenase kit without stabilizers can only be kept for twelve months under the same conditions.

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Abstract

The invention provides a stable composition containing NAD+ or NADH. Bioactivity of NAD+ or NADH can be stably maintained for a long time. By the use of a stabilizer which contains trehalose, glycerol, a buffer solution and a surfactant, stability of NAD+ and NADH dissolved state is raised. The invention also provides a method for stabilizing NAD+ or NADH, and also provides an application of a composition composed of trehalose, glycerol, the buffer solution, the surfactant and a pH regulator in increasing stability of NAD+ or NADH. The composition can be used for a lactate dehydrogenase reagent, an alanine aminotransferase reagent, an aspartate aminotransferase reagent, a urea kid and the like, and is widely applied.

Description

technical field [0001] The invention belongs to the field of biological technology, and relates to a + or NADH stable compositions, and methods of increasing the stability of NAD+ or NADH. Background technique [0002] Nicotinamide adenine dinucleotide (NAD for short) is a coenzyme that transfers electrons, and it appears in many metabolic reactions of cells. NADH is its reduced form. NAD is the coenzyme of dehydrogenases, such as alcohol dehydrogenase (ADH), used to oxidize ethanol. It plays an irreplaceable role in glycolysis, gluconeogenesis, tricarboxylic acid cycle and respiratory chain. The intermediate product will pass the hydrogen off to NAD, making it NADH. NADH will act as a carrier of hydrogen to synthesize ATP through chemiosmotic coupling in the respiratory chain. [0003] In terms of light absorption, NADH has an absorption peak at 260nm and 340nm, while NAD has only one absorption peak at 260nm, which is an important attribute to distinguish the two. Th...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07H21/02C07H1/00
Inventor 王辉王军峰吴静孟菲王宇飞
Owner 上海执诚生物科技有限公司
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