Method for extracting and separating lutein crystals from vegetable oil resin containing lutein diester

A technology of lutein and vegetable oil, which is applied in the field of vegetable oleoresin, can solve the problems of low efficiency, poor specificity, and low product yield, and achieve the effects of high efficiency, favorable refining and purification process, and high yield

Active Publication Date: 2019-01-18
ZHEJIANG MEDICINE CO LTD XINCHANG PHAMACEUTICAL FACTORY
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  • Abstract
  • Description
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AI Technical Summary

Problems solved by technology

[0029] In order to overcome the shortcomings of producing a large amount of waste water and low product yield caused by strong alkalinity when the chemical alcoholysis method is used to produce lutein crystals, and the low efficiency and poor specificity of the product in the production of lutein crystals by using biological enzymes in the prior art Due to defects such as low yield, a suitable and easy-to-industrial production method has been invented to efficiently produce lutein crystals using a self-made esterase (lipase) derived from a metagenomic library. This lipase has been published in CN102796715B open in

Method used

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  • Method for extracting and separating lutein crystals from vegetable oil resin containing lutein diester
  • Method for extracting and separating lutein crystals from vegetable oil resin containing lutein diester
  • Method for extracting and separating lutein crystals from vegetable oil resin containing lutein diester

Examples

Experimental program
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Effect test

Embodiment 1

[0046] Take 1g of recombinant novel esterase (lipase) Est076 (C23) constructed by Professor Xie Tian of Hangzhou Normal University, and dissolve it in 95ml of water to form an enzyme solution. 1000g marigold oleoresin (total lutein content 15.8%, purchased from Qingdao Saite Perfume Co., Ltd.) mixed with 2250mL absolute ethanol, the water content in the alcohol solvent was 5.1% at the beginning of the reaction, the temperature was raised to 65 ° C, and stirred until forming A uniform flowing solution, add enzyme solution to react, monitor the progress of the reaction by thin-layer chromatography, after 18 hours, the hydrolysis of lutein fatty acid ester is complete, stop stirring and let stand for 0.5hr.

[0047] The reaction solution was cooled to 65°C, filtered while it was hot (65°C) to obtain 305.6g of a yellow filter cake, which was added to 1000ml of ethyl acetate, heated to 70°C to dissolve, washed three times with 500ml of deionized water, and separated Remove the wate...

Embodiment 2

[0051]Take 15 g of a new recombinant esterase (lipase) Est076 (C23) constructed by Professor Xie Tian of Hangzhou Normal University, and dissolve it in 988 ml of water to form an enzyme solution. 1000g of marigold oleoresin (total lutein content of 15.8%, purchased from Qingdao Saite Perfume Co., Ltd.) was mixed with 8000mL of 98.5% n-propanol, heated to 65°C, stirred until a homogeneous solution was formed, added The enzyme solution was reacted, and the progress of the reaction was monitored by thin-layer chromatography. After 6 hours, the hydrolysis of the lutein fatty acid ester was complete, and the stirring was stopped and allowed to stand for 0.5 hr. The water content in the alcohol solvent at the start of the reaction was 15.0%.

[0052] The reaction solution was heated to 30°C, filtered while hot at this temperature to obtain 321.4g of a yellow filter cake, added to 3200ml of isobutyl acetate, heated to 110°C to dissolve, and then washed three times with 800ml of deion...

Embodiment 3

[0056] Take 30 g of a new recombinant esterase (lipase) Est076 (C23) constructed by Professor Xie Tian of Hangzhou Normal University, and dissolve it in 120 ml of water to form an enzyme solution. 1000 g of marigold oleoresin (total lutein content of 15.8%, purchased from Qingdao Saite Perfume Co., Ltd.) was mixed with 3000 mL of propylene glycol containing 10.0% deionized water, heated to 30°C, and stirred until a homogeneous solution was formed, The enzyme solution was added to react, and the progress of the reaction was monitored by thin-layer chromatography. After 15 hours, the lutein fatty acid ester was hydrolyzed completely, and the stirring was stopped and allowed to stand for 0.5 hr.

[0057] The reaction solution was heated to 60°C, filtered while hot to obtain 297.6g of a yellow filter cake, which was added to 1000ml of dichloromethane, dissolved at 20°C, washed three times with 900ml of deionized water, and the water layer was separated. About 750 ml of methylene c...

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Abstract

The invention provides a method for extracting and separating lutein crystals from vegetable oil resin containing lutein diester. The method comprises the following steps: (a) dissolving lipase into deionized water, so as to form an enzyme solution; (b) dissolving lutein extract into an alcohol solvent containing deionized water, so as to form a uniform alcohol solution; (c) adding the enzyme solution into the alcohol solution for generating hydrolysis reaction, and stirring, so as to obtain a lutein solution; (d) filtering the lutein solution, and carrying out filter pressing separation, so as to obtain crystals; (e) dissolving the crystals into a non-polarity organic solvent again, and washing water-soluble impurities by virtue of the deionized water; (f) recycling the organic solvent, and cooling, so as to obtain secondary crystals; and (g) separating the secondary crystals. The method has the beneficial effects that the selectivity is relatively strong, the reaction time is relatively short, no wastewater is produced, the process is environment-friendly, the obtained lutein crystals are high in purity and yield, and the method is suitable for industrial production.

Description

technical field [0001] The present invention relates to a method for simultaneously separating and purifying lutein and zeaxanthin from plant oleoresin, especially marigold oleoresin, using esterase (lipase) catalyzed from a metagenomic library with a higher yield. technical background [0002] Carotenoids are a class of yellow to red substances widely found in nature. In different fruits and vegetables, there are different types of carotenoids, such as β-carotene in carrots, lutein in marigold flowers, zeaxanthin in strawberries, lycopene in tomatoes, capsanthin in capsicum plants and capsicum content are higher. However, there are more carotenoids in some dark fruits and vegetables, egg yolk, some fish shells, birds, seaweed and bacteria, and the content of lutein is also higher. [0003] Several animal and human trials in recent years have shown the beneficial effects of carotenoids. In general, carotenoids may be divided into two subcategories, lutein or oxygen-contai...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07C403/24
CPCC07C403/24C07C2601/16C07B2200/13C12P23/00
Inventor 许新德谢恬王升帆王秋岩郑建永曾昭武殷晓浦劳学军邵康忠
Owner ZHEJIANG MEDICINE CO LTD XINCHANG PHAMACEUTICAL FACTORY
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