Extraction method of astaxanthin in Pichia pastoris, astaxanthin, and cosmetics

Abstract

This application relates to the field of cosmetics. Specifically, it relates to a method for extracting astaxanthin from Pichia pastoris, astaxanthin, and cosmetics. A method for extracting astaxanthin from Pichia pastoris includes: adding an alcohol solution containing citric acid to Pichia pastoris for extraction; then taking the filtrate; wherein, in the alcohol solution containing citric acid, the concentration of citric acid is less than 0.3 mol / L. In the above technical solution, by adding an alcohol solution containing citric acid to Pichia pastoris, cell wall breaking and extraction of Pichia pastoris can be achieved, thereby effectively extracting astaxanthin from Pichia pastoris. By limiting the concentration of citric acid in the alcohol solution containing citric acid to be less than 0.3 mol / L, the extraction amount of astaxanthin in Pichia pastoris can be effectively increased. When the concentration is greater than this value, the extraction amount of astaxanthin in Pichia pastoris will decrease, which is instead not conducive to the extraction of astaxanthin from Pichia pastoris.

Description

Technical Field

[0001] This application relates to the field of cosmetics. Specifically, it relates to a method for extracting astaxanthin from Pichia pastoris, astaxanthin, and cosmetics. Background Art

[0002] The extract from Pichia pastoris has great application potential in cosmetics. However, the current development of Pichia pastoris is insufficient, which limits its application in cosmetics. Summary of the Invention

[0003] The purpose of the embodiments of this application is to provide a method for extracting astaxanthin from Pichia pastoris, astaxanthin, and cosmetics.

[0004] In a first aspect, a method for extracting astaxanthin from Pichia pastoris includes:

[0005] Adding an alcohol solution containing citric acid to Pichia pastoris for extraction; then taking the filtrate;

[0006] Wherein, in the alcohol solution containing citric acid, the concentration of citric acid is less than 0.3 mol / L.

[0007] In the above technical solution, by adding an alcohol solution containing citric acid to Pichia pastoris, cell wall breaking and extraction of Pichia pastoris can be achieved, thereby effectively extracting astaxanthin from Pichia pastoris. By limiting the concentration of citric acid in the alcohol solution containing citric acid to be less than 0.3 mol / L, the extraction amount of astaxanthin in Pichia pastoris can be effectively increased. When the concentration is greater than this value, the extraction amount of astaxanthin in Pichia pastoris will decrease, which is instead not conducive to the extraction of astaxanthin from Pichia pastoris.

[0008] Furthermore, in the above technical solution of this application, by adopting the aforementioned method for extracting astaxanthin from Pichia pastoris, the extract from Pichia pastoris in cosmetics can have great application potential.

[0009] In other embodiments of this application, in the alcohol solution containing citric acid, the concentration of citric acid is greater than or equal to 0.1 mol / L.

[0010] In other embodiments of this application, adding an alcohol solution containing citric acid to Pichia pastoris includes:

[0011] Adding according to the ratio of the weight of Pichia pastoris to the volume of alcohol being (1 g: 30 mL) to (1 g: 70 mL).

[0012] In other embodiments of this application, adding an alcohol solution containing citric acid to Pichia pastoris includes:

[0013] Adding according to the ratio of the weight of Pichia pastoris to the volume of ethanol being (1 g: 40 mL) to (1 g: 60 mL).

[0014] In other embodiments of the present application, the number of extractions includes multiple times; optionally, the number of extractions is 1 to 5 times.

[0015] In other embodiments of the present application, the number of extractions includes three times; when extracting for the first time, the concentration of citric acid in the alcohol solution containing citric acid is 0.2 mol / L; when extracting for the second time, in the alcohol solution containing citric acid, the concentration of citric acid is 0.2 mol / L; when extracting for the third time, in the alcohol solution containing citric acid, the concentration of citric acid is 0.2 mol / L.

[0016] In other embodiments of the present application, the extraction is performed as follows:

[0017] Ultrasonic extraction is carried out under the condition that the ultrasonic power is 100 W to 300 W.

[0018] In other embodiments of the present application, for a single extraction, the extraction time is 10 min to 50 min.

[0019] In other embodiments of the present application, taking the filtrate includes: filtering the filtrate through a filter paper with a pore size of 0.2 μm to 0.5 μm and then combining them.

[0020] In a second aspect, the present application provides an astaxanthin, which is obtained by using the extraction method of astaxanthin in Pichia pastoris provided in any one of the embodiments of the first aspect.

[0021] In the above technical solution, the astaxanthin obtained by using the extraction method of astaxanthin in Pichia pastoris provided in any one of the embodiments of the first aspect

[0022] In a third aspect, the present application provides a cosmetic, which includes the astaxanthin extracted from Pichia pastoris provided in the second aspect above.

[0023] In the above technical solution, by adding the astaxanthin extracted from Pichia pastoris to the cosmetic, the antioxidant effect can be improved, the anti-aging effect can be achieved, and astaxanthin can stabilize cell membranes, improve pigmentation, and improve skin dullness. Specific Embodiments

[0024] To make the objectives, technical solutions, and advantages of the embodiments of the present application clearer, the technical solutions in the embodiments of the present application will be clearly and completely described below. Apparently, the described embodiments are some, but not all, of the embodiments of the present application.

[0025] Accordingly, the following detailed description of the embodiments of the present application is not intended to limit the scope of the claimed present application, but merely represents selected embodiments of the present application. All other embodiments obtained by those of ordinary skill in the art based on the embodiments in the present application without creative efforts fall within the scope of protection of the present application.

[0026] An extraction method of astaxanthin from Pichia pastoris provided by an embodiment of the present application includes:

[0027] Adding an alcohol solution containing citric acid to Pichia pastoris for extraction; then taking the filtrate;

[0028] Wherein, in the alcohol solution containing citric acid, the concentration of citric acid is less than 0.3 mol / L.

[0029] An extraction method of astaxanthin from Pichia pastoris includes:

[0030] Adding an alcohol solution containing citric acid to Pichia pastoris for extraction; then taking the filtrate;

[0031] Wherein, in the alcohol solution containing citric acid, the concentration of citric acid is less than 0.3 mol / L.

[0032] In the above technical solution, by adding an alcohol solution containing citric acid to Pichia pastoris, cell wall breaking and extraction of Pichia pastoris can be achieved, so as to effectively extract astaxanthin from Pichia pastoris. By limiting that in the alcohol solution containing citric acid, the concentration of citric acid is less than 0.3 mol / L, the extraction amount of astaxanthin in Pichia pastoris can be effectively increased. When the concentration is greater than this value, the extraction amount of astaxanthin in Pichia pastoris will decrease, which is instead not conducive to the extraction of astaxanthin in Pichia pastoris.

[0033] Furthermore, exemplarily, in some embodiments of the present application, in the alcohol solution containing citric acid, the concentration of citric acid is 0.29 mol / L, 0.28 mol / L, 0.26 mol / L, 0.25 mol / L, 0.23 mol / L, 0.22 mol / L, 0.20 mol / L, 0.19 mol / L, 0.18 mol / L, 0.16 mol / L, 0.15 mol / L, 0.14 mol / L, 0.13 mol / L, 0.12 mol / L, 0.11 mol / L, 0.10 mol / L, 0.09 mol / L, 0.08 mol / L, 0.07 mol / L, 0.06 mol / L, 0.05 mol / L, 0.04 mol / L, 0.03 mol / L, 0.02 mol / L, 0.01 mol / L or the range between any two of the foregoing values.

[0034] Further, in the above technical solution, the inventor creatively found that by using a compound of citric acid and alcohol, the cell wall of Pichia pastoris can be effectively broken, and astaxanthin can be extracted from Pichia pastoris by alcohol extraction, which is time-consuming and low-cost.

[0035] Further, the inventor found that when the concentration of citric acid is less than 0.3 mol / L, the extraction amount of astaxanthin in Pichia pastoris can be effectively increased. When the concentration is greater than this value, it will instead consume the astaxanthin in Pichia pastoris, resulting in a decrease in the extraction amount of astaxanthin in Pichia pastoris, which is instead not conducive to the extraction of astaxanthin in Pichia pastoris.

[0036] Further optionally, in some embodiments of the present application, the above-mentioned alcohol is ethanol.

[0037] Further, in some embodiments of the present application, in the alcohol solution containing citric acid, the concentration of citric acid is greater than or equal to 0.1 mol / L.

[0038] In the above technical solution, in the alcohol solution containing citric acid, when the concentration of citric acid is greater than or equal to 0.1 mol / L, the cell wall breaking treatment of Pichia pastoris can be further improved, and the astaxanthin in Pichia pastoris will not be excessively consumed, thereby achieving a balance point between cell wall breaking and consumption, and achieving the effect of increasing the extraction amount of astaxanthin in Pichia pastoris.

[0039] Further optionally, in some embodiments of the present application, in the alcohol solution containing citric acid, the concentration of citric acid is 0.1 mol / L to 0.2 mol / L.

[0040] Further, in some embodiments of the present application, the above-mentioned Pichia pastoris powder is the dry powder from Best Pharmaceutical (Guangzhou) Co., Ltd., batch number: XQS20240528.

[0041] Further, in some embodiments of the present application, adding an alcohol solution containing citric acid to Pichia pastoris includes:

[0042] Adding according to the ratio of the weight of Pichia pastoris to the volume of alcohol being (1 g: 30 mL) to (1 g: 70 mL).

[0043] In the above technical solution, by setting the addition according to the ratio of the weight of Pichia pastoris to the volume of alcohol being (1 g: 30 mL) to (1 g: 70 mL), it is beneficial to increase the content of astaxanthin obtained by extraction from Pichia pastoris.

[0044] Exemplarily, in some embodiments of the present application, adding an alcohol solution containing citric acid to Pichia pastoris includes:

[0045] Add according to the ratio of the weight of Pichia pastoris to the volume of alcohol being 1 g:30 mL, 1 g:32 mL, 1 g:35 mL, 1 g:38 mL, 1 g:40 mL, 1 g:42 mL, 1 g:45 mL, 1 g:48 mL, 1 g:50 mL, 1 g:55 mL, 1 g:58 mL, 1 g:60 mL, 1 g:62 mL, 1 g:65 mL, 1 g:68 mL, 1 g:70 mL or within the ratio range of any two of the foregoing values.

[0046] Further, in some embodiments of the present application, adding an alcohol solution containing citric acid to Pichia pastoris includes:

[0047] Add according to the ratio of the weight of Pichia pastoris to the volume of ethanol being (1 g:40 mL) to (1 g:60 mL).

[0048] In the above technical solution, through further limitation, adding according to the ratio of the weight of Pichia pastoris to the volume of ethanol being (1 g:40 mL) to (1 g:60 mL) is further beneficial to increasing the content of astaxanthin obtained by extraction from Pichia pastoris.

[0049] Exemplarily, in some embodiments of the present application, adding an alcohol solution containing citric acid to Pichia pastoris includes:

[0050] Add according to the ratio of the weight of Pichia pastoris to the volume of ethanol being 1 g:40 mL, 1 g:42 mL, 1 g:45 mL, 1 g:48 mL, 1 g:49 mL, 1 g:50 mL, 1 g:52 mL, 1 g:55 mL, 1 g:58 mL, 1 g:60 mL or within the ratio range of any two of the foregoing values.

[0051] Further, in some embodiments of the present application, the number of extractions includes multiple times; optionally, the number of extractions is 1 to 5 times.

[0052] Exemplarily, in some embodiments of the present application, the number of extractions includes multiple times; optionally, the number of extractions is 1 time, 2 times, 3 times, 4 times or 5 times.

[0053] In the above technical solution, by setting the number of extractions to include multiple times, it is beneficial to extract the content of astaxanthin from Pichia pastoris.

[0054] Further, in some embodiments of the present application, the number of extractions is three times; when performing the first extraction, the concentration of citric acid in the alcohol solution containing citric acid is 0.1 mol / L to 0.2 mol / L; when performing the second extraction, the concentration of citric acid in the alcohol solution containing citric acid is 0.1 mol / L to 0.2 mol / L; when performing the third extraction, the concentration of citric acid in the alcohol solution containing citric acid is 0.1 mol / L to 0.2 mol / L.

[0055] In the above technical solution, by setting the number of extractions to three times and limiting that when performing the first extraction, the second extraction, and the third extraction, the concentration of citric acid in the alcohol solution containing citric acid is within the above range, it is beneficial to increase the content of astaxanthin obtained by extraction from Pichia pastoris.

[0056] Exemplarily, in some embodiments of the present application, the number of extractions is three times; when performing the first extraction, the concentration of citric acid in the alcohol solution containing citric acid is 0.1 mol / L, 0.11 mol / L, 0.12 mol / L, 0.13 mol / L, 0.14 mol / L, 0.15 mol / L, 0.16 mol / L, 0.17 mol / L, 0.18 mol / L, 0.19 mol / L or 0.2 mol / L; when performing the second extraction, the concentration of citric acid in the alcohol solution containing citric acid is 0.1 mol / L, 0.11 mol / L, 0.12 mol / L, 0.13 mol / L, 0.14 mol / L, 0.15 mol / L, 0.16 mol / L, 0.17 mol / L, 0.18 mol / L, 0.19 mol / L or 0.2 mol / L; when performing the third extraction, the concentration of citric acid in the alcohol solution containing citric acid is 0.1 mol / L, 0.11 mol / L, 0.12 mol / L, 0.13 mol / L, 0.14 mol / L, 0.15 mol / L, 0.16 mol / L, 0.17 mol / L, 0.18 mol / L, 0.19 mol / L or 0.2 mol / L.

[0057] Further, in some embodiments of the present application, performing the extraction includes:

[0058] Performing ultrasonic extraction under the condition that the ultrasonic power is 100 W to 300 W.

[0059] In the above technical solution, by setting that performing the extraction includes performing ultrasonic extraction under the condition that the ultrasonic power is 100 W to 300 W; it is beneficial to increase the content of astaxanthin obtained by extraction from Pichia pastoris.

[0060] Exemplarily, in some embodiments of the present application, performing the extraction includes:

[0061] Under the condition of ultrasonic power of 100W, 105W, 110W, 115W, 120W, 125W, 130W, 135W, 140W, 145W, 150W, 155W, 160W, 165W, 170W, 175W, 180W, 185W, 190W, 195W, 200W, 210W, 220W, 230W, 240W, 250W, 280W, 300W or the power within the range between any two of the foregoing values, ultrasonic extraction is carried out.

[0062] Further, in some embodiments of the present application, the single extraction time is 10 min to 50 min.

[0063] In the above technical solution, by setting the single extraction time to 10 min to 50 min, it is beneficial to increase the content of astaxanthin obtained by extraction from Pichia pastoris.

[0064] Exemplarily, in some embodiments of the present application, the single extraction time is 10 min, 15 min, 20 min, 25 min, 30 min, 35 min, 40 min, 45 min, 50 min, 40 min or the extraction time within the range between any two of the foregoing values for extraction.

[0065] Further, in some embodiments of the present application, taking the filtrate includes: filtering the filtrate through a filter paper with a pore size of 0.2 μm to 0.5 μm and then combining them.

[0066] In the above technical solution, by setting taking the filtrate, including: filtering the filtrate through a filter paper with a pore size of 0.2 μm to 0.5 μm and then combining them, it is beneficial to obtain astaxanthin with better comprehensive performance.

[0067] Exemplarily, in some embodiments of the present application, taking the filtrate includes: filtering the filtrate through a filter paper with a pore size of 0.2 μm, 0.21 μm, 0.22 μm, 0.23 μm, 0.25 μm, 0.26 μm, 0.28 μm, 0.3 μm, 0.32 μm, 0.4 μm, 0.42 μm, 0.45 μm, 0.48 μm, 0.5 μm or the pore size within the range between any two of the foregoing values and then combining them.

[0068] The extraction method of the present application uses citric acid to break the cell wall of Pichia pastoris and extracts astaxanthin from Pichia pastoris with ethanol, which is time-consuming and low-cost.

[0069] The extraction method of the present application extracts astaxanthin from Pichia pastoris for the first time and optimizes its process. The reagents used, citric acid and ethanol, are included in the cosmetics use catalog, and subsequent application in cosmetics is convenient, safe and green.

[0070] Some embodiments of the present application provide an astaxanthin, which is obtained by extracting the astaxanthin in Pichia pastoris provided by any of the foregoing embodiments.

[0071] Some embodiments of the present application provide a cosmetic, which includes the astaxanthin provided by any of the foregoing embodiments.

[0072] In the above technical solution, by adding the astaxanthin extracted from Pichia pastoris to the cosmetic, the antioxidant effect can be improved, playing an anti-aging role. Moreover, astaxanthin can stabilize cell membranes, improve pigmentation, and improve skin dullness.

[0073] The features and properties of the present application will be further described in detail with reference to the following examples:

[0074] Example 1

[0075] A method for extracting astaxanthin from Pichia pastoris is provided, which is carried out according to the following steps:

[0076] (1) Weigh the Pichia pastoris powder, add an ethanol solution containing citric acid for ultrasonic extraction, centrifuge, filter, and separate the filtrate and the filter residue. The pore size of the filter paper used for filtration is 0.2 μm.

[0077] Among them, for the concentration of citric acid in the ethanol solution containing citric acid, the material-liquid ratio of the Pichia pastoris powder to the ethanol solution containing citric acid, the ultrasonic power, the ultrasonic time, and the number of extraction times, see Table 1 for details.

[0078] Examples 2 - 15

[0079] A method for extracting astaxanthin from Pichia pastoris is provided. The difference from Example 1 is that: some of the parameter values of the concentration of citric acid in the ethanol solution containing citric acid, the material-liquid ratio of the Pichia pastoris powder to the ethanol solution containing citric acid, the ultrasonic power, the ultrasonic time, or the number of extraction times are different. See Table 1 for details.

[0080] Comparative Example 1

[0081] A method for extracting astaxanthin from Pichia pastoris is provided. The difference from Example 1 is that the extraction agent used is an ethanol solution, that is, it does not contain citric acid. See Table 1 for details.

[0082] Comparative Examples 2 - 3

[0083] A method for extracting astaxanthin from Pichia pastoris is provided. The difference from Example 1 is that the concentration of the ethanol solution containing citric acid added is different. See Table 1 for details.

[0084] Comparative Examples 4 - 6

[0085] A method for extracting astaxanthin from Pichia pastoris is provided. The difference from Example 5 is that different extraction agents are used. See Table 1 for details.

[0086] Examples 16 - 19

[0087] A method for extracting astaxanthin from Pichia pastoris is provided and is carried out according to the following steps:

[0088] (1) Weigh the Pichia pastoris powder, add an ethanol solution containing citric acid, and perform the first extraction by ultrasonic treatment. Then centrifuge and filter to separate the filtrate and the filter residue. The pore size of the filter paper used for filtration is 0.2 μm.

[0089] (2) Take the Pichia pastoris filter residue obtained in step (1), add the ethanol solution containing citric acid again, and perform the second extraction by ultrasonic treatment. Then centrifuge and filter to collect the filtrate. The pore size of the filter paper used for filtration is 0.2 μm.

[0090] And so on, extract multiple times. See Table 2 for the specific number of extraction times in each example; see Table 2 for the citric acid concentration in each extraction in each example; the ultrasonic power, ultrasonic time, and number of extraction times in each example are the same. See Table 2 for details.

[0091] Experimental Example 1

[0092] Test the astaxanthin extraction amount of the extraction methods of each example and comparative example.

[0093] Conduct tests using the Pichia pastoris extracts prepared in each example and comparative example respectively.

[0094] 1. Method principle

[0095] After Pichia pastoris is acid-hydrolyzed by citric acid, an ethanol solution is used as a solvent for ultrasonic extraction. After centrifugation and filtration, take the filtrate.

[0096] 2. Reagents and equipment

[0097] 2.1 Reagents

[0098] Ethanol (analytical pure), astaxanthin standard, citric acid (analytical pure)

[0099] 2.2 Instruments and equipment

[0100] Microplate reader, 96-well microplate, ultrasonic cleaner.

[0101] 3. Analysis steps

[0102] 3.1 Preparation of standard series solutions

[0103] Take a certain amount of astaxanthin standard, dissolve and dilute it with an ethanol solution to the corresponding concentration to obtain standard series solutions with concentrations of 5 μg / mL, 10 μg / mL, 15 μg / mL, 20 μg / mL, and 25 μg / mL. Using the ethanol solution as the blank, measure the absorbance values of the astaxanthin standard solutions at 474 nm, and repeat the experiment three times. Plot the astaxanthin standard curve with the concentration of astaxanthin (μg / mL) as the abscissa and the absorbance (OD474) as the ordinate.

[0104] 3.2 Sample treatment

[0105] Weigh an appropriate amount of Pichia pastoris powder, acid-hydrolyze it with citric acid, extract it twice by ultrasonic treatment with an ethanol solution and then centrifuge it, and take the supernatant. Combine the two filtrates, filter them through a 0.45 μm filter membrane, and use the filtrate as the test solution.

[0106] 4. Determination

[0107] 4.1 Standard curve determination

[0108] Take the standard series solutions under "3.1" and measure their absorbance values at 474 nm respectively. Plot the standard curve with the concentration of the standard series solutions as the abscissa and the absorbance values as the ordinate.

[0109] 4.2 Sample determination

[0110] Take the test solution of the sample under "3.2" and measure its absorbance value at 474 nm. Obtain the concentration of astaxanthin in the test solution of the sample according to the standard curve. Calculate the content of astaxanthin in the sample according to "5.1".

[0111] 5. Expression of analysis results

[0112] 5.1 Data calculation

[0113]

[0114] Where:

[0115] OD 474 —— Absorbance value at 474 nm;

[0116] V —— Volume of solvent used for the sample, mL;

[0117] D —— Dilution factor;

[0118] a —— Conversion coefficient between astaxanthin concentration and OD 474 value;

[0119] W —— Mass of Pichia pastoris, g.

[0120] 5.2 Precision

[0121] The linear range of this method is 5 μg / mL - 25 μg / mL. Under repeatability conditions, the absolute value of the two independent test results obtained shall not exceed 10% of the arithmetic mean.

[0122] The test results are shown in Table 1 and Table 2.

[0123] Table 1

[0124]

[0125]

[0126]

[0127] It can be seen from the test results in Table 1 above:

[0128] In Comparative Example 1, a single ethanol extraction was used, and the astaxanthin extraction amount in Pichia pastoris was relatively low, significantly lower than that in each example. When a too high citric acid concentration was used in Comparative Examples 2 - 3, the astaxanthin extraction amount in Pichia pastoris showed a downward trend, significantly lower than that in each example.

[0129] In Comparative Example 4, DMSO (dimethyl sulfoxide) was used. Although the astaxanthin extraction amount in Pichia pastoris was relatively high, however, DMSO (dimethyl sulfoxide) is highly toxic and cannot be used in cosmetics.

[0130] In Comparative Example 5, hydrochloric acid was used for extraction. However, the astaxanthin extraction amount in Pichia pastoris was relatively low because the hydrochloric acid was too acidic, which was instead unfavorable to astaxanthin and would decompose astaxanthin, greatly reducing the astaxanthin extraction amount in Pichia pastoris.

[0131] In Comparative Example 6, lactic acid was used for extraction. However, lactic acid is an organic solvent that cannot be removed in cosmetics and has an irritating effect on the skin, so it cannot be applied to cosmetics.

[0132] Comparing Example 15 with Example 14 and Example 13, it can be seen that with long-term air exposure and light exposure, the astaxanthin extraction amount in Pichia pastoris decreased. This may be because light will decompose part of the astaxanthin, and oxygen in the air will also oxidize part of the astaxanthin. It can be seen that an extraction time of greater than or equal to 30 min and less than 60 min is more conducive to extraction.

[0133] Comparing Examples 5, 7, 8, 9, and 10, it can be seen that both too high and too low ultrasonic power will cause a decrease in the astaxanthin extraction amount in Pichia pastoris; this may be because when the power is too high, the temperature is too high and affects its stability; while when the power is low, the extraction is insufficient. It can be seen that when the ultrasonic power is 200 - 250 W, the astaxanthin extraction amount in Pichia pastoris is higher.

[0134] Table 2

[0135]

[0136]

[0137]

[0138] It can be seen from the test results in Table 1 and Table 2 that:

[0139] Compared with Example 5, when the extraction times are increased, the astaxanthin extraction amount shows an increasing trend.

[0140] The above are only the preferred embodiments of the present application and are not used to limit the present application. For those skilled in the art, various changes and modifications can be made to the present application. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present application shall be included in the protection scope of the present application.

Claims

1. A method for extracting astaxanthin from Pichia pastoris, characterized in that: include: adding an alcohol solution containing citric acid to Pichia pastoris for extraction; and then taking a filtrate; Wherein, in the alcohol solution containing citric acid, the concentration of citric acid is 0.1 mol / L to 0.2 mol / L; The extracting comprises: Ultrasonic extraction is carried out under the condition of ultrasonic power of 200W~250W; the extraction time is 30min~50min.

2. The method for extracting astaxanthin in Pichia pastoris according to claim 1, characterized in that: The step of adding an alcohol solution containing citric acid to Pichia pastoris comprises: The ratio of the weight of Pichia pastoris to the volume of alcohol was (1 g: 30 mL) to (1 g: 70 mL).

3. The method for extracting astaxanthin in Pichia pastoris according to claim 1, characterized in that: The step of adding an alcohol solution containing citric acid to Pichia pastoris comprises: The Pichia pastoris was added at a weight to volume ratio of ethanol of (1 g: 40 mL) to (1 g: 60 mL).

4. The method for extracting astaxanthin in Pichia pastoris according to claim 1, characterized in that: The number of extractions includes multiple times.

5. The method for extracting astaxanthin in Pichia pastoris according to claim 1, characterized in that: The extraction is performed 1 to 5 times.

6. The method for extracting astaxanthin from Pichia pastoris according to claim 1, characterized in that: The number of extractions includes three times; during the first extraction, the concentration of citric acid in the alcohol solution containing citric acid is 0.1mol / L~0.2mol / L; during the second extraction, the concentration of citric acid in the alcohol solution containing citric acid is 0.1mol / L~0.2mol / L; during the third extraction, the concentration of citric acid in the alcohol solution containing citric acid is 0.1mol / L~0.2mol / L.

7. The method for extracting astaxanthin from Pichia pastoris according to any one of claims 1 to 6, characterized in that: The method of taking the filtrate comprises: passing the filtrate through filter paper with a diameter of 0.2 μm to 0.5 μm and then combining the filtrate.

Images