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Low-temperature superoxide dismutase protecting agent and preparation method thereof

A superoxide and dismutase technology, applied in the field of bioengineering, can solve the problems of loss of SOD enzyme activity, high production cost, restricting the industrial application of superoxide dismutase, etc., and achieves the effect of maintaining enzyme activity and low production cost.

Inactive Publication Date: 2017-08-22
DALIAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Because superoxide dismutase is sensitive to heat and easily oxidized, most factories choose freeze-drying as the drying process, but the freeze-drying process has high energy consumption, high investment, low efficiency, and high production costs; and the freeze-drying process will The loss of about 10% or more of the SOD enzyme activity, and the additives added during freeze-drying severely restrict the industrial application of the superoxide dismutase industry

Method used

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  • Low-temperature superoxide dismutase protecting agent and preparation method thereof
  • Low-temperature superoxide dismutase protecting agent and preparation method thereof
  • Low-temperature superoxide dismutase protecting agent and preparation method thereof

Examples

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Effect test

Embodiment 1

[0033] A low-temperature superoxide dismutase protective agent, comprising the following raw materials in parts by weight, low-temperature superoxide dismutase 300mL, polyethylene glycol 100g, sodium hyaluronate 11g, DTT 2g, imidazolidinyl urea 1g, trehalose 5g, 122g of deionized water.

[0034] The preparation method of described low-temperature superoxide dismutase is:

[0035] (1) Preparation of crude enzyme solution: Aseptic operation, inoculate the activated Pseudomonas marinosa into the liquid fermentation medium, ferment at 20°C, 160r / min for 48h, and ferment the bacteria liquid at 4°C, 6 Centrifuge at 000r / min for 30min to obtain wet cells, centrifuge at 6000r / min for 30min at 4°C, dissolve the wet cells in phosphate buffer, and crush them in a 480W ultrasonic ice bath for 30min (crushing for 5s with an interval of 5s). After crushing, 4 Centrifuge at 12 000r / min for 15min, and the supernatant is the crude enzyme solution. The formula percentage of the above liquid fe...

Embodiment 2

[0045] A low-temperature superoxide dismutase protective agent, comprising the following raw materials in parts by weight, low-temperature superoxide dismutase 400mL, polyethylene glycol 200g, sodium hyaluronate 30g, DTT 4g, imidazolidinyl urea 2g, trehalose 10g, deionized water 312g.

[0046] The preparation method of the low-temperature superoxide dismutase is the same as in Example 1.

[0047] The preparation method of described low-temperature superoxide dismutase protective agent, comprises the following steps:

[0048] (1) Trehalose and polyethylene glycol were added to deionized water, stirred evenly, and subjected to high-temperature moist heat sterilization. During sterilization, the temperature was controlled at 120°C, and sterilized at this temperature for 25 minutes to obtain the base liquid;

[0049] (2) Add sodium hyaluronate and DTT to low-temperature superoxide dismutase, and stir evenly, then use a filter to filter and sterilize the mixed solution of low-temp...

Embodiment 3

[0052] A low-temperature superoxide dismutase protective agent, comprising the following raw materials in parts by weight, low-temperature superoxide dismutase 500mL, polyethylene glycol 300g, sodium hyaluronate 54g, DTT 6g, imidazolidinyl urea 3g, trehalose 15g, 582g of deionized water.

[0053] The preparation method of the low-temperature superoxide dismutase is the same as in Example 1.

[0054] The preparation method of described low-temperature superoxide dismutase protective agent, comprises the following steps:

[0055] (1) Trehalose and polyethylene glycol were added to deionized water, stirred evenly, and subjected to high-temperature moist heat sterilization. During sterilization, the temperature was controlled at 125°C, and sterilized at this temperature for 30 minutes to obtain the base liquid;

[0056] (2) Add sodium hyaluronate and DTT to low-temperature superoxide dismutase, and stir evenly, then use a filter to filter and sterilize the mixed solution of low-t...

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Abstract

The invention relates to a low-temperature superoxide dismutase protecting agent and a preparation method thereof. The protecting agent is composed of the following components: low-temperature superoxide dismutase, polyethylene glycol, dithiothreitol (DTT), sodium hyaluronate, imidazolidinyl urea, trehalose and deionized water. The preparation method comprises the following steps of: adding trehalose and polyethylene glycol into the deionized water to sterilize to obtain a base liquid; adding sodium hyaluronate and DTT into the low-temperature superoxide dismutase, uniformly stirring and filtering the mixture to degerm to obtain a low-temperature superoxide dismutase mixed solution; adding the low-temperature superoxide dismutase mixed solution into the base liquid in a sterile condition and then adding imidazolidinyl urea, uniformly mixing and stirring the mixture, and adding the deionized water to adjust to obtain the low-temperature superoxide dismutase protecting agent. According to the low-temperature superoxide dismutase protecting agent, the enzyme activity of the low-temperature superoxide dismutase is kept stable, and the preparation cost is low, the loss of the enzyme activity can be greatly reduced, and the enzyme service life is prolonged and the application effect of the enzyme is enhanced.

Description

technical field [0001] The invention relates to the field of bioengineering, in particular to a low-temperature superoxide dismutase protective agent and a preparation method thereof. Background technique [0002] Superoxide dismutase (superoxide dismutase, EC 1.15.1.1, abbreviated as SOD) is a kind of metalloenzyme widely present in organisms, which catalyzes superoxide anion free radical (O 2 - .) and hydrogen ions to generate hydrogen peroxide and molecular oxygen, thereby removing oxygen free radicals in the organism and protecting the organism from damage. According to the types of metal ions contained, SOD is divided into four categories: one is blue-green copper-zinc superoxide dismutase (Cu / Zn-SOD), which mainly exists in the cytosol and chloroplast of eukaryotic cells, and is the most studied, The most in-depth category; the second category is purple-red nickel superoxide dismutase (Ni-SOD) and manganese superoxide dismutase (Mn-SOD), which mainly exist in the mat...

Claims

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Application Information

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IPC IPC(8): C12N9/96C12N9/02
CPCC12N9/0089C12N9/96C12Y115/01001
Inventor 迟乃玉张庆芳王晓辉
Owner DALIAN UNIV
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