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
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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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