Method for preparing astaxanthin monomer

A technology of astaxanthin and astaxanthin ester, applied in the biological field, can solve the problems of low yield, difficult conditions, etc., and achieve the effects of simple operation, energy saving, and avoiding the loss of astaxanthin

Active Publication Date: 2010-11-24
秦皇岛惠恩生物技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the hydrolysis method of astaxanthin ester is mainly alkali saponification, which requires low temperatur

Method used

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  • Method for preparing astaxanthin monomer
  • Method for preparing astaxanthin monomer
  • Method for preparing astaxanthin monomer

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] The preparation of embodiment 1Penicillium cyclopium var.albus alkaline lipase enzyme liquid

[0021] With the cDNA of Penicillium cyclopium var.albus (CGMCC 3.4041) alkaline lipase as a template, primers (P1: gaattc gcaactgctgacgccgct, P2: gcggccg tcagctcagatagccaca) for PCR amplification. The target band was recovered from the PCR product, and connected to the vector pMD18-T-Simple (purchased from TaKaRa Company); then double-digested pMD18-T-alip and pKIC9K with restriction endonuclease SnaBI and Not 1 respectively , and then the target fragment was recovered and ligated with T4 ligase to obtain the expression plasmid pPIC9K-alip. Digest the carrier PIC9K-alip with restriction endonuclease Sal I, then concentrate the linearized fragment by ethanol precipitation, transform Pichia pastoris GS115 competent cells by electroshock method, and spread the histidine-free cells after transformation Minimal Medium MD Plates. After the plates were cultured at 30°C for 2-3 ...

Embodiment 2

[0023] Example 2 Emulsion Preparation and Enzymolysis Reaction Condition Optimization

[0024] Take 0.1g Haematococcus pluvialis astaxanthin crude extract oil (natural L-astaxanthin 5% oil, purchased from Jingzhou Natural Astaxanthin Co., Ltd.) Warm at 80°C, grind and emulsify, and add 0.1M sodium phosphate buffer (pH5.0-8.0) to make up to 10ml; add 2-12U lipase per microgram of total carotenoids and add emulsified product (1ml) and enzyme, Then add 0.1M sodium phosphate buffer solution with pH 5.0-8.0 to a total volume of 10ml; shake the reaction at 30°C at 180rpm, take samples every 1-2h, detect with TLC and HPLC, and stop the reaction after 12h.

[0025] The results show that Tween 80 can be effectively emulsified by adding 1 to 3 times the weight of the oil agent of the crude extract of Haematococcus pluvialis, but because the amount of Tween 80 will increase the amount of acetone in the extract, considering economic factors, the amount of Tween 80 is preferred It is 1 ti...

Embodiment 3

[0026] The hydrolysis of embodiment 3 astaxanthin esters

[0027] Take 0.1g of Haematococcus pluvialis astaxanthin crude extract oil (purchased from Jingzhou Natural Astaxanthin Co., Ltd.) in a mortar, add 1 times the mass of Tween 80, grind and emulsify, and add 0.1M sodium phosphate Buffer (pH7.0) to 10ml; add emulsification (1ml) and enzyme at the amount of 4U lipase per microgram of total carotenoids, then add 0.1M sodium phosphate buffer at pH7.0 to the total volume 10ml; 28°C, 150rpm shaking reaction for 7 hours, the yield of astaxanthin was 63.2%. After the reaction, add the same volume of acetone as the sample, shake with a vortex shaker for 30s at room temperature, then add an equal volume of n-hexane, shake for 30s in the same way as above, 12000rpm, centrifuge for 1min, and take the supernatant for TLC and HPLC detection .

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Abstract

The invention provides a new method for preparing an astaxanthin monomer. In the method, a free astaxanthin monomer is obtained by hydrolyzing astaxanthin ester with lipase. The recovery rate of the astaxanthin monomer reaches 63.2 percent by using the method. The method is simple to operate, namely a desired product can be obtained by one reaction and extraction; and the method saves the energy consumption and can avoid the loss of the astaxanthin during long-term reaction to a certain extent. The method lays the foundation of the industrial application for extracting the astaxanthin monomer from crude extract of haematococcus pluvialis by an enzyme method.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a method for hydrolyzing astaxanthin esters to obtain astaxanthin monomers. Background technique [0002] Astaxanthin (3,3'-dihydroxy-β,β'-carotene-4,4'-diketone carotenoid) is an oxygenated derivative of carotenoids. It has a strong antioxidant capacity, and its free radical quenching activity is 10 times that of β-carotene and 100-500 times that of vitamin E. It can be used as a natural coloring agent in aquaculture; it not only has the activity of enhancing immune response and anticancer, but also can be used in food, medicine, cosmetics and other industries. [0003] Astaxanthin can be chemically synthesized or extracted from natural samples. When astaxanthin is synthesized by chemical means, impurity chemical substances, such as unnatural by-products produced during the synthesis process, will inevitably be introduced, which will reduce its bioavailability safety. ...

Claims

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Application Information

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IPC IPC(8): C12P23/00
Inventor 李颖赵莹莹关菲菲王贵利关国华王珍芳
Owner 秦皇岛惠恩生物技术有限公司
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