Method for utilizing neutral protease to extract and purify astaxanthin in shrimp shell and crab shell

A technology of neutral protease and astaxanthin, applied in the direction of organic chemistry, can solve the problems of not obtaining high-purity astaxanthin, poor selectivity of hydrolysis, large amount of organic solvent, etc., and achieve less steps and stable astaxanthin , The effect of low equipment requirements

Active Publication Date: 2018-11-06
JINING MEDICAL UNIV
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Problems solved by technology

However, the amount of organic solvent used in the organic solvent extraction method is large, the extraction rate is low, and the production cost is high, and the astaxanthin ester needs to be hydrolyzed into monomers by strong alkali, which has great pollution to the environment
Supercritical CO 2 The extraction method is relatively safe and non-toxic, but requires high equipment requirements and high production costs, making it difficult to popularize in industrial production
[0005] CN105420326A discloses a method for extracting protein and astaxanthin by hydrolyzing shrimp shells with papain, which uses papain to hydrolyze shrimp shells to extract astaxanthin, and the final result is total carotenoids. The selectivity of its hydrolysis is poor, and no High Purity Astaxanthin

Method used

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  • Method for utilizing neutral protease to extract and purify astaxanthin in shrimp shell and crab shell
  • Method for utilizing neutral protease to extract and purify astaxanthin in shrimp shell and crab shell
  • Method for utilizing neutral protease to extract and purify astaxanthin in shrimp shell and crab shell

Examples

Experimental program
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Effect test

Embodiment 1

[0042] Astaxanthin is extracted from Argentinian red shrimp shells as follows:

[0043] Step 1: Dissolving biological enzymes

[0044] Dissolve 1g of neutral protease in 500mL of water.

[0045] Step 2: Carry out enzymatic hydrolysis reaction, inactivation and pretreatment

[0046] Crush the shells of Argentinian red shrimps, take 100 g and mix them with the above-mentioned neutral protease solution, and then enzymatically hydrolyze them for 300 min at a pH of 7 and a temperature of 35° C. After the enzymatic hydrolysis reaction, inactivate at 70°C, then centrifuge, and mix the lower layer solid with 500mL organic solvent ethanol to obtain the pretreated enzymatic hydrolysis solution.

[0047] Step 3: Extraction and Concentration

[0048] The pretreated enzymolysis solution obtained in the previous step was extracted with magnetic stirring at a constant temperature of 40°C for 120 minutes, filtered, extracted 3 times, and the extracts were combined, and then the combined ex...

Embodiment 2

[0053] Using the head of Argentinian red shrimp as raw material, astaxanthin is extracted according to the following steps:

[0054] Step 1: Dissolving biological enzymes

[0055] Get 10g neutral protease and dissolve in 2L water.

[0056]Step 2: Carry out enzymatic hydrolysis reaction, inactivation and pretreatment

[0057] Crush the head of Argentinian red shrimp, take 200g and mix it with the above-mentioned neutral protease solution, and then enzymatically hydrolyze it at pH 5 and temperature 55°C for 30min. After the enzymatic hydrolysis reaction, inactivate at 70°C, then centrifuge, and mix the solid in the lower layer with 700 mL of organic solvent ethyl acetate to obtain the pretreated enzymatic hydrolysis solution.

[0058] Step 3: Extraction and Concentration

[0059] The pretreated enzymolysis solution obtained in the previous step was mechanically stirred at a constant temperature of 50°C and 25KHz ultrasonically extracted for 20 minutes, extracted 3 times, and ...

Embodiment 3

[0064] Astaxanthin is extracted from Arctic sweet shrimp shells as follows:

[0065] Step 1: Dissolving biological enzymes

[0066] Dissolve 0.15g of neutral protease in 700mL of water.

[0067] Step 2: Carry out enzymatic hydrolysis reaction, inactivation and pretreatment

[0068] Crush the fresh shrimp shell of arctic sweet shrimp, take 100g and mix it with the above-mentioned biological enzyme solution, and then enzymolyze it for 300min under the conditions of pH 5.5 and temperature 55°C. After the enzymatic hydrolysis reaction, inactivate at 70°C, then centrifuge, and mix the lower layer solid with 900mL organic solvent acetone to obtain the pretreated enzymatic hydrolysis solution.

[0069] Step 3: Extraction and Concentration

[0070] The pretreated enzymolysis solution obtained in the previous step was subjected to ultrasonic extraction at a constant temperature of 60°C and 40KHz for 80 minutes, and extracted once, and then the extract was concentrated with a rotary ...

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Abstract

The invention discloses a method for utilizing neutral protease to extract and purify astaxanthin in shrimp shell and crab shell. Firstly fresh shrimp shell or crab shell is smashed and prepared to bean enzyme solution, the shrimp shell or crab shell is subjected to an enzymatic hydrolysis reaction with the enzyme solution, a solid is mixed with an organic solvent, the astaxanthin is extracted under the certain condition, the astaxanthin is obtained through filtration, evaporation and drying, and the astaxanthin is subjected to qualitative and quantitative analysis by using high performance liquid chromatography. The yield rate of the astaxanthin in the shrimp shell of pandalus borealis is 145 microgram / g, the purity of astaxanthin crude extract is 1.12 mg / g, and the content of the purified astaxanthin is 92%. The yield rate of the astaxanthin of shrimp shell of procambarus clarkia is 294 microgram / g, the purity of the astaxanthin crude extract is 2.06 mg / g, and the purity is as highas 95% after the crude product of the astaxanthin extract is purified. The method has the advantages that the price of the raw materials is low, the reaction condition is moderate, the method is simple to operate, easy to popularize and produce and is environmentally friendly. The method can not only rapidly extract and obtain the astaxanthin monomer, but also the relatively high extraction rate,the stability and the purity are high.

Description

technical field [0001] The invention relates to a method for extracting astaxanthin monomer, in particular to a method for extracting and purifying astaxanthin in shrimp shells and crab shells by using neutral protease. Background technique [0002] Astaxanthin (3,3′-dihydroxy-4,4′-diketone-β,β′-carotene) widely exists in the biological world, especially in shrimp, crab, fish, and algae. It is one of the main carotenoids in marine organisms. Astaxanthin has extremely strong antioxidant activity, which is more than 500 times that of vitamin E, and has multiple physiological effects such as anti-tumor, anti-oxidation and anti-aging. Not only that, astaxanthin has a unique effect on improving and protecting vision, can prevent and treat age-related macular degeneration, and improve retinal function. In addition, astaxanthin has a strong ability to scavenge free radicals, which can prevent the occurrence of cancer and cardiovascular diseases and help repair the central nervous...

Claims

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Application Information

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IPC IPC(8): C07C403/24
CPCC07C403/24C07C2601/16
Inventor 王利涛吕美胡锦霞卢尉航丁瑞芳王其宝王慧云
Owner JINING MEDICAL UNIV
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