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Method for efficiently and rapidly preparing free astaxanthin

A kind of astaxanthin, fast technology, applied in the biological field, can solve the problems of long reaction time, difficult to control the product, low hydrolysis efficiency of free astaxanthin, etc., and achieve the effect of improving low efficiency and increasing conversion rate

Active Publication Date: 2018-10-12
OCEAN UNIV OF CHINA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] At present, the hydrolysis of astaxanthin ester mainly adopts the saponification method. The saponification method takes a long time to react, the reaction process is too violent, and the product is difficult to control. Moreover, high-concentration alkali will damage astaxanthin and produce more similar astaxanthin and semi-astaxanthin. By-products, and the alkali and organic solvents in the reaction process will cause huge damage to the environment
Another method is enzymatic hydrolysis. At present, there are problems such as low hydrolysis efficiency, long reaction time, and complicated reaction process in the preparation of free astaxanthin by enzymatic hydrolysis. Moreover, most of the researches focus on the hydrolysis of a single esterase or lipase.

Method used

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  • Method for efficiently and rapidly preparing free astaxanthin
  • Method for efficiently and rapidly preparing free astaxanthin
  • Method for efficiently and rapidly preparing free astaxanthin

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Screening of astaxanthin ester hydrolyzed strains:

[0033] The medium formula used in this embodiment is as follows:

[0034] Screening plate solid media:

[0035] Haematococcus Pluvialls Oil, 0.05%; Triton X-100, 1.0%; KNO 3 , 0.1%; K 2 HPO 4 , 0.05%; MgSO 4 ·7H 2 O, 0.05%; NaCl, 0.05%; FeSO 4 ·7H 2 O, 0.001%; ​​peptone, 1.0%; agar powder, 2.0%; H 2 O, 100 mL; pH 7.0.

[0036] Seed medium:

[0037] Peptone, 1.0%; Yeast powder, 0.5%; NaCl, 1.0%; Water, 100mL; pH 7.0; Sterilize at 121°C for 20min.

[0038] Fermentation medium:

[0039] Peptone, 1.0%; Yeast powder, 0.5%; NaCl, 1.0%; Water, 1L; pH 7.0; Sterilize at 121°C for 20min.

[0040] Enrichment culture:

[0041] Inject the preserved glycerol tube bacteria solution into the seed medium, mix well, and culture at 180 rpm on a shaker at 37° C. for 24 hours.

[0042] Plate screening bacteria:

[0043] Take the activated bacterial solution and dilute it appropriately by 100 times, and spread it on a solid ...

Embodiment 2

[0064] Process optimization of hydrolyzed astaxanthin ester:

[0065] 1) Effect of reaction pH value on hydrolyzed astaxanthin ester

[0066] Weigh 2 mg of Haematococcus pluvialis oil, dissolve it with 0.5 mL of absolute ethanol, then weigh an appropriate amount of extracellular crude enzyme preparation, and dissolve the enzyme powder in buffer solutions with different pH values. The buffer solutions used are as follows: citric acid buffer solution (100mM, pH 5.0-6.0), sodium phosphate buffer solution (100mM, pH 6.0-8.0), Tris-HCl buffer solution (100mM, pH 7.0-9.0) and glycine-sodium hydroxide buffer solution (100mM, pH9.0- 10.0), added to the reaction bottle, filled with nitrogen, protected from light, placed in a 37°C water-bath shaker for 30 minutes, that is, absolute ethanol: buffer solution = 1:10 (v / v), a total of 5.5mL reaction system.

[0067] Such as Image 6 As shown, by adding different buffer solutions in the reaction system, the effect of different pH values ​​...

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Abstract

The invention discloses a method for efficiently and rapidly preparing free astaxanthin. The strain for producing lipase and esterase is pseudomonas aeruginosa. Conditions such as the pH, the ratio ofa reaction solution, the temperature, the enzyme addition amount and time are determined. There is no need to add any acid or alkali in a reaction process of the technology, not only is the shortcoming that byproducts including astacin and semi-astacin are easily generated in the process of preparing the astaxanthin by means of a traditional saponification method overcome, but also the problems that the efficiency of preparing the astaxanthin by means of a single enzymolysis method is low and the cost is high are solved, and a novel approach is provided for environment-friendly scale production of the natural free astaxanthin.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a method for efficiently and rapidly preparing free astaxanthin by using an extracellular crude enzyme preparation of an astaxanthin esterase production strain. Background technique [0002] Natural astaxanthin (also known as 3,3′-dihydroxy-4,4′-diketone-β,β′-carotene) is an unsaturated terpene compound and a carotenoid, the main source In crustaceans such as Haematococcus pluvialis, Phaffia rhodozyme, shrimps, and crabs, it is purple-red and easily oxidized into astaxanthin (Astacene). Astaxanthin can quench singlet oxygen and scavenge free radicals. Its antioxidant capacity is 10 times that of other carotenoids and 500 times that of vitamin E. Astaxanthin has good physiological functions, the main functions include: anti-oxidation, anti-aging, protection of optic nerve, enhancement of human immune function, reduction of cerebral infarction, anti-inflammatory effect, pr...

Claims

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Application Information

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IPC IPC(8): C12P23/00
CPCC12P23/00
Inventor 毛相朝高新炜孙建安刘振薛长湖
Owner OCEAN UNIV OF CHINA
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