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Method for improving extraction rate of marigold lutein by combined use of multiple enzymes

A marigold and lutein technology, applied in the direction of microorganism-based methods, biochemical equipment and methods, microorganisms, etc., can solve the problems of hindering the contact between extraction solvent and lutein and low extraction efficiency, so as to increase the content and reduce the Loss, the effect of improving extraction efficiency

Pending Publication Date: 2021-01-15
JIANGXI BONTAC GREEN BIOCATALYSIS ECOIND PARK CO LTD
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AI Technical Summary

Problems solved by technology

The membrane of cell wall and organelle hinders the contact between extraction solvent and lutein, resulting in low extraction efficiency

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] (1) Spray the multi-enzyme combination solution evenly on the surface of 1 kg of fresh marigold according to the dosage of 10 mL / kg, compact it, and put it in a closed environment at 25 degrees Celsius for 8 days;

[0031] (2) The ratio of the multi-enzyme combination solution is: 20 parts of β-1,4-endo-xylanase, 20 parts of endo-β-glucanase, 45 parts of pectinase, 8 parts of cutinase, protease 5 parts, 2 parts antioxidant;

[0032] (3) Dry the fresh marigolds after 8 days.

[0033] (4) Add 3-5L of n-hexane to 1 kg of dried marigold flowers, add n-hexane, extract statically at room temperature for 30 minutes, and collect the extract;

[0034] (5) Use a vacuum rotary dryer to concentrate the extract under the conditions of 45°C, -0.06MPa, and 30rpm until there is no distillate. The extract contains lutein and its precursor substance α-carotene;

[0035] (6) Pick a single colony BL21-pETDuet-CitHYb-CitCYP97C in 5ml LB liquid medium containing 50ug / ml ampicillin, and act...

Embodiment 2

[0038] Conventional extraction process: use n-hexane to extract, filter to obtain the marigold lutein extract, use 20% n-hexane by weight, and saline with a mass percentage concentration of 1.0-1.2% to extract the marigold lutein Separating and removing impurities to obtain lutein, the content of lutein was determined to be 0.36‰ (W / DW).

Embodiment 3

[0040] The difference between this example and Example 1 is that the BL21-pETDuet-CitHYb-CitCYP97C engineering bacteria is not used to continue the catalysis, and the rest is the same as Example 1. The content of lutein was determined to be 0.46‰ (W / DW).

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Abstract

The present invention belongs to the field of extraction of traditional Chinese medicinal materials, and particularly relates to a multi-enzyme combination capable of improving an extraction rate of lutein in traditional Chinese medicine marigold and a method for extracting the lutein in the marigold by using the multi-enzyme combination. 20 parts of beta-1,4-endoxylanase, 20 parts of endo-beta-glucanase, 45 parts of pectinase, 8 parts of cutinase, 5 parts of protease and 2 parts of an antioxidant are combined to form combined enzymes which can effectively degrade cell walls and cell membranesof petal tissues and lutein-protein complexes in cells, so that lutein molecules are in contact with extraction solvent molecules to the maximum extent, extraction efficiency is improved, then BL21-pETDuet-CitHYb-CitCYP97C engineering bacteria are used for continuous catalysis, alpha-carotene can be effectively converted into lutein through beta-carotene hydroxylase and carotenoid epsilon hydroxylase, and the lutein content is increased.

Description

technical field [0001] The invention belongs to the field of extracting Chinese medicinal materials, and in particular relates to a multi-enzyme combination capable of improving the extraction efficiency of lutein from marigolds, a traditional Chinese medicinal material, and a method for extracting lutein from marigolds by using the multi-enzyme group. Background technique [0002] Marigold, the name of Chinese medicine. It is the flower or root of Tagetes erecta L. in the family Asteraceae. The plant marigold is cultivated all over my country. Marigold has the effects of clearing heat and detoxifying, reducing phlegm and relieving cough, and the roots are detoxifying and detumescence. Flowers are used to treat upper respiratory tract infection, pertussis, tracheitis, keratitis, pharyngitis, stomatitis, toothache; external use is used to treat mumps, mastitis, carbuncle and sore. The external application of fresh herbs is mainly used to treat mastitis, unknown swelling, a...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P23/00C12R1/19
CPCC12P23/00
Inventor 傅荣昭宋鹏
Owner JIANGXI BONTAC GREEN BIOCATALYSIS ECOIND PARK CO LTD
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