Extraction, separation and purification method of astaxanthin in rhodotorula mucilaginosa
A technology of Rhodotorula colloides and astaxanthin, which is applied in the direction of organic chemistry, can solve the problems of increasing the cost of separation and purification, reducing the activity of astaxanthin, and low yield of astaxanthin, and achieves good saponification effect, low cost, and easy extraction high efficiency effect
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Embodiment 1
[0061] Embodiment 1 A kind of preparation method of Rhodotorula gum dry bacterium somatic cell
[0062] A preparation method of rhodotorula gelatin dried bacterium somatic cells, comprising the following steps:
[0063] (1) Inoculate the preserved strain of Rhodotorula japonicus ZTHY2 on the slant surface of Rhodotorula japonicus seed culture medium, shake culture at 30°C and 140r / min for 48h, and obtain the rhodotorula japonicus seed liquid;
[0064] (2) Inoculate the Rhodotorula japonicus seed solution at 5% in the Rhodotorula japonicus fermentation medium, and vibrate at 30° C. and 140 r / min for 72 hours to obtain the Rhodotorula japonicus fermentation broth;
[0065] (3) Centrifuge the fermented liquid of Rhodotorula japonicus at 4000r / min for 20min, discard the supernatant, wash twice with sterilized distilled water, dry the precipitated cells in an incubator at 50°C to constant weight, and obtain dry bacteria of Rhodotorula japonicus somatic cells.
Embodiment 2
[0066] Embodiment 2 A method for extracting and separating and purifying astaxanthin in Rhodotorula gelatinosa
[0067] 1. A method for extracting and separating and purifying astaxanthin in Rhodotorula gelatinus, comprising the following steps:
[0068] (1) Weigh 0.1g of the dried Rhodotorula gum cells of Example 1, add 5mL of 3mol / L hydrochloric acid, mix well, let stand at room temperature for 40min, place in a boiling water bath for 3min, cool rapidly, centrifuge at 4000r / min for 10min, discard The supernatant was washed twice with distilled water; 7 mL of a mixed solution of ethyl acetate and ethanol (the volume ratio of ethyl acetate and ethanol was 2:1), extracted in a water bath at 50°C for 30 min, centrifuged at 4000 r / min for 15 min, and repeated for 2 times, to obtain astaxanthin crude extract;
[0069] (2) Take 5mL of astaxanthin concentrate, put it in a separatory funnel, add 5mL of ethyl acetate, then add 10mL of 20g / L potassium hydroxide ethanol solution, let i...
Embodiment 3
[0076] Embodiment 3 The selection and optimization of wall-breaking method and extracting party
[0077] 1. Wall breaking method and extraction method
[0078] The acid-heat method, ultrasonic cracking method, and dimethyl sulfoxide (DMSO)+absolute ethanol extraction method were used to break the wall of Rhodotorula gluei respectively. There are three kinds of extracts in the acid-heat method, namely: acetone solution , Ethyl acetate + ethanol (2:1) mixed solution, acetonitrile + methanol (7.5:2.5) mixed solution.
[0079] The specific steps of the three wall-breaking extraction methods are as follows:
[0080] (1) Acid-heat method: Weigh 0.1 g of dried bacterial cells into a centrifuge tube, add 5 mL of 3 mol / L hydrochloric acid, mix well, let stand at room temperature for 40 min, place in a boiling water bath for 3 min, cool rapidly, centrifuge at 4000 r / min for 10 min, discard Wash the supernatant with distilled water twice, add 7mL extract, extract in a water bath at 50°...
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