CN103553994A - Method for preparing astaxanthin missible oil through phaffia rhodozyma thallus - Google Patents
Method for preparing astaxanthin missible oil through phaffia rhodozyma thallus Download PDFInfo
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- CN103553994A CN103553994A CN201310553611.4A CN201310553611A CN103553994A CN 103553994 A CN103553994 A CN 103553994A CN 201310553611 A CN201310553611 A CN 201310553611A CN 103553994 A CN103553994 A CN 103553994A
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- astaxanthin
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Abstract
The invention discloses a method for preparing astaxanthin missible oil through phaffia rhodozyma thallus. The method comprises the following steps: mixing dry powder of phaffia rhodozyma thallus with acetic acid, carrying out cell disruption under a high-temperature and high-pressure condition to obtain broken phaffia rhodozyma bacterial liquid containing astaxanthin, separating an organic phase containing the astaxanthin through a separating funnel by utilizing a mixed extraction solvent of chloroform and methanol, and then, vacuum desolventizing and drying to obtain the astaxanthin missible oil. The method disclosed by the invention has the advantages of simple and convenient operation, short time, low requirement on equipment corrosion resistance, high separation and extraction yield, and the like, the astaxanthin extraction rate is about 88.5%, which is slightly higher than that of a high-temperature and high-pressure thermal acid method, so that the method has a good industrial application prospect.
Description
Technical field
The present invention relates to a kind of preparation method of missible oil, relate in particular to a kind of method of utilizing phaffiafhodozyma thalline to prepare astaxanthin missible oil.
Background technology
Astaxanthin is a kind of of carotenoid, is extensively present in shrimp crab waste, some algae and fungi.Studies have shown that, astaxanthin has very strong resistance of oxidation; In addition, astaxanthin also has the different physiological roles such as the tumour of inhibition occurs, enhancing body immunity.Accordingly, astaxanthin is with a wide range of applications at aspects such as high-class healthy product, food, medicine and fodder additivess.
At present, astaxanthin mainly obtains by following four kinds of approach: (1) chemosynthesis; (2) from Species of Crustacea waste, extract; (3) from algae, extract; (4) utilize microorganism (as Haematocoocus Pluvialls and phaffiafhodozyma etc.) fermentative production.Because astaxanthin molecular structure is complicated, synthetic difficulty, so the astaxanthin price comparison that chemosynthesis is produced is expensive; And the method for extracting from Species of Crustacea waste is also faced with starting material source and collection difficulty, supplies unsettled problem.Therefore, current research emphasis both domestic and external has all been placed on and has utilized microorganism to produce to obtain in the approach of astaxanthin.
The ratio accounting in tens Carotenoids that research discovery astaxanthin has at wild phaffiafhodozyma is maximum, has reached 40%-95%, and therefore, phaffiafhodozyma is considered to the good potential bacterial strain that industrial fermentation is produced astaxanthin.But because astaxanthin is that the cell walls of a kind of parachrome and phaffiafhodozyma is thicker, the extraction comparison difficulty of astaxanthin.At present, the extraction process of generally taking for the astaxanthin in phaffiafhodozyma is: first after certain method broken wall, extract again.The means of existing conventional cell wall breaking mainly contain the four large classes such as mechanical process, enzyme process, Physical and chemical method, but its equal having complicated operation, expend time in and the shortcoming such as grow in various degree.
The damp and hot broken wall of acid system broken wall and high temperature is the good two kinds of wall-breaking methods of current effect, also has researchist to develop a kind of more efficient wall-breaking method---High Temperature High Pressure acid heat method.But the hydrochloric acid using in High Temperature High Pressure acid heat method has stronger corrosive nature to metal, need to use the acidproof device of a kind of high temperature, this,, having increased virtually production cost, is unfavorable for suitability for industrialized production.
Summary of the invention
For the deficiencies in the prior art, problem to be solved by this invention is to provide a kind of improved method of utilizing phaffiafhodozyma thalline to prepare astaxanthin missible oil.
The method of utilizing phaffiafhodozyma thalline to prepare astaxanthin missible oil of the present invention, step is:
(1) get phaffiafhodozyma bacterium dry powder and add in organic acid, stir;
(2) step (1) is mixed to rear feed liquid and be placed in high pressure vessel and heat, make temperature be warming up to 120 ± 2 ℃, be incubated processing;
(3) by the high pressure vessel nature decrease temperature and pressure after insulation processing, Pressure Drop in container is to normal pressure, take out feed liquid and also continue to be cooled to normal temperature, then regulate the pH of feed liquid and utilize Rotary Evaporators to remove in feed liquid 2/3 moisture, must contain the broken wall phaffiafhodozyma bacterium liquid of astaxanthin;
(4) bacterium liquid step (3) being made joins in separating funnel, then adds organic solvent in funnel, after jumping a queue and fully shaking up, and the organic phase that after standing 5～10min, sucking-off contains astaxanthin;
(5) operation of repeating step (4), until the organic phase of sucking-off becomes colorless;
(6) merge the organic phase that contains astaxanthin of collecting, centrifugal 10～15min under 3000～3500rpm condition;
(7) get the supernatant liquor after centrifugal, be placed on that in Rotary Evaporators, to carry out vacuum desolvation dry, to be driedly obtain astaxanthin missible oil after completely;
It is characterized in that:
The described organic acid of step (1) is that concentration is the acetic acid of 0.8～1.3mol/L, and phaffiafhodozyma bacterium dry powder mixes in the ratio of 1kg:18～23L with acetic acid, after mixing, in mixed liquor, adds vitamins C, and making its final concentration is 3mg/ml;
The high-temperature and high-pressure conditions of the described heating of step (2) is: 121 ℃ of temperature, saturated vapour pressure 0.1Mpa, and the time that described insulation is processed is 4～8 minutes;
PH to 5.0～7.0 of the described adjusting feed liquid of step (3);
The described organic solvent of step (4) is the mixing solutions of chloroform and methyl alcohol, wherein take volume ratio chloroform: methyl alcohol is 2～3:1; The organic phase that contains astaxanthin is chloroform layer;
The dry condition of the described vacuum desolvation of step (7) is: 45～50 ℃ of vacuum tightness 0.07～0.09MPa, temperature.
The above-mentioned phaffiafhodozyma thalline that utilizes is prepared in astaxanthin missible oil method: the described phaffiafhodozyma bacterium of step (1) dry powder is preferably purchased from Shandong extensive biotechnology Services Co., Ltd.
The above-mentioned phaffiafhodozyma thalline that utilizes is prepared in astaxanthin missible oil method: the described organic acid of step (1) acetic acid that preferably concentration is 1.0mol/L, phaffiafhodozyma bacterium dry powder preferably mixes in the ratio of 1kg:20L with acetic acid.
The above-mentioned phaffiafhodozyma thalline that utilizes is prepared in astaxanthin missible oil method: the time that the described insulation of step (2) is processed is preferably 5 minutes.
The above-mentioned phaffiafhodozyma thalline that utilizes is prepared in astaxanthin missible oil method: preferably to 6.5 of pH of the described adjusting feed liquid of step (3).
The above-mentioned phaffiafhodozyma thalline that utilizes is prepared in astaxanthin missible oil method: the mixing solutions of the described chloroform of step (4) and methyl alcohol is in volume ratio chloroform: methyl alcohol is preferably 2:1.
The method of utilizing phaffiafhodozyma thalline to prepare astaxanthin missible oil of the present invention is the organic acid heat method of a kind of High Temperature High Pressure, the method that it has improved the wall-breaking method of phaffiafhodozyma thalline and extract astaxanthin from the broken wall phaffiafhodozyma bacterium liquid that contains astaxanthin, in described method, the acetic acid that choosing is used is less to corrosion of metal, than High Temperature High Pressure acid heat method, reduced the requirement to equipment, the extraction solvent (mixing solutions of chloroform and methyl alcohol) simultaneously using has good extraction effect compared with other organic solvents (as ethanol etc.).
That the astaxanthin missible oil preparation method that the present invention proposes has is easy and simple to handle, the advantage such as short that expends time in, and its Astaxanthin extraction rate is 88.5% left and right, a little more than High Temperature High Pressure acid heat method, has good industrial applications prospect.
Embodiment
Embodiment 1
(1) get phaffiafhodozyma bacterium dry powder (purchased from Shandong extensive biotechnology Services Co., Ltd), it is in the acetic acid of 1.0mol/L that phaffiafhodozyma bacterium dry powder and acetic acid are added to concentration in the ratio of 1kg:20L, stir, after mixing, in mixed liquor, add vitamins C, making its final concentration is 3mg/ml;
(2) step (1) is mixed to rear feed liquid and be placed in high pressure vessel and heat, make temperature be warming up to 121 ℃, saturated vapour pressure is 0.1Mpa, is incubated and processes 5 minutes;
(3) by the high pressure vessel nature decrease temperature and pressure after insulation processing, Pressure Drop in container is to normal pressure, take out feed liquid and continue to be cooled to normal temperature, then regulate the pH to 6.5 of feed liquid and utilize Rotary Evaporators to remove in feed liquid 2/3 moisture, must contain the broken wall phaffiafhodozyma bacterium liquid of astaxanthin;
(4) bacterium liquid step (3) being made joins in separating funnel, again to the mixing solutions (take volume ratio chloroform: methyl alcohol is 2:1) that adds chloroform and methyl alcohol in funnel, after jumping a queue and fully shaking up, the chloroform layer (organic phase lower floor) that standing 5 rear careful sucking-offs contain astaxanthin;
(5) operation of repeating step (4), until the chloroform layer of sucking-off becomes colorless;
(6) merge the chloroform layer that contains astaxanthin of collecting, centrifugal 10min under 3500rpm condition;
(7) get the supernatant liquor after centrifugal, be placed in Rotary Evaporators, under vacuum tightness 0.08MPa, the condition of 50 ℃, carry out vacuum desolvation dry, to be driedly obtain astaxanthin missible oil after completely.
Embodiment 2
(1) get phaffiafhodozyma bacterium dry powder (purchased from Shandong extensive biotechnology Services Co., Ltd), it is in the acetic acid of 0.8mol/L that phaffiafhodozyma bacterium dry powder and acetic acid are added to concentration in the ratio of 1kg:18L, stir, after mixing, in mixed liquor, add vitamins C, making its final concentration is 3mg/ml;
(2) step (1) is mixed to rear feed liquid and be placed in high pressure vessel and heat, make temperature be warming up to 122 ℃, saturated vapour pressure is 0.1Mpa, is incubated and processes 4 minutes;
(3) by the high pressure vessel nature decrease temperature and pressure after insulation processing, Pressure Drop in container is to normal pressure, take out feed liquid and continue to be cooled to normal temperature, then regulate the pH to 7.0 of feed liquid and utilize Rotary Evaporators to remove in feed liquid 2/3 moisture, must contain the broken wall phaffiafhodozyma bacterium liquid of astaxanthin;
(4) bacterium liquid step (3) being made joins in separating funnel, again to the mixing solutions (take volume ratio chloroform: methyl alcohol is 2:1) that adds chloroform and methyl alcohol in funnel, after jumping a queue and fully shaking up, the chloroform layer (organic phase lower floor) that standing 5 rear careful sucking-offs contain astaxanthin;
(5) operation of repeating step (4), until the chloroform layer of sucking-off becomes colorless;
(6) merge the chloroform layer that contains astaxanthin of collecting, centrifugal 15min under 3000rpm condition;
(7) get the supernatant liquor after centrifugal, be placed in Rotary Evaporators, under vacuum tightness 0.07MPa, the condition of 50 ℃, carry out vacuum desolvation dry, to be driedly obtain astaxanthin missible oil after completely.
Embodiment 3
(1) get phaffiafhodozyma bacterium dry powder (purchased from Shandong extensive biotechnology Services Co., Ltd), it is in the acetic acid of 1.3mol/L that phaffiafhodozyma bacterium dry powder and acetic acid are added to concentration in the ratio of 1kg:23L, stir, after mixing, in mixed liquor, add vitamins C, making its final concentration is 3mg/ml;
(2) step (1) is mixed to rear feed liquid and be placed in high pressure vessel and heat, make temperature be warming up to 120 ℃, saturated vapour pressure is 0.1Mpa, is incubated and processes 8 minutes;
(3) by the high pressure vessel nature decrease temperature and pressure after insulation processing, Pressure Drop in container is to normal pressure, take out feed liquid and continue to be cooled to normal temperature, then regulate the pH to 5.0 of feed liquid and utilize Rotary Evaporators to remove in feed liquid 2/3 moisture, must contain the broken wall phaffiafhodozyma bacterium liquid of astaxanthin;
(4) bacterium liquid step (3) being made joins in separating funnel, again to the mixing solutions (take volume ratio chloroform: methyl alcohol is 3:1) that adds chloroform and methyl alcohol in funnel, after jumping a queue and fully shaking up, the chloroform layer (organic phase lower floor) that standing 5 rear careful sucking-offs contain astaxanthin;
(5) operation of repeating step (4), until the chloroform layer of sucking-off becomes colorless;
(6) merge the chloroform layer that contains astaxanthin of collecting, centrifugal 10min under 30000rpm condition;
(7) get the supernatant liquor after centrifugal, be placed in Rotary Evaporators, under vacuum tightness 0.09MPa, the condition of 45 ℃, carry out vacuum desolvation dry, to be driedly obtain astaxanthin missible oil after completely.
Claims (6)
1. utilize phaffiafhodozyma thalline to prepare a method for astaxanthin missible oil, step is:
(1) get phaffiafhodozyma bacterium dry powder and add in organic acid, stir;
(2) step (1) is mixed to rear feed liquid and be placed in high pressure vessel and heat, make temperature be warming up to 120 ± 2 ℃, be incubated processing;
(3) by the high pressure vessel nature decrease temperature and pressure after insulation processing, Pressure Drop in container is to normal pressure, take out feed liquid and also continue to be cooled to normal temperature, then regulate the pH of feed liquid and utilize Rotary Evaporators to remove in feed liquid 2/3 moisture, must contain the broken wall phaffiafhodozyma bacterium liquid of astaxanthin;
(4) bacterium liquid step (3) being made joins in separating funnel, then adds organic solvent in funnel, after jumping a queue and fully shaking up, and the organic phase that after standing 5～10min, sucking-off contains astaxanthin;
(5) operation of repeating step (4), until the organic phase of sucking-off becomes colorless;
(6) merge the organic phase that contains astaxanthin of collecting, centrifugal 10～15min under 3000～3500rpm condition;
(7) get the supernatant liquor after centrifugal, be placed on that in Rotary Evaporators, to carry out vacuum desolvation dry, to be driedly obtain astaxanthin missible oil after completely;
It is characterized in that:
The described organic acid of step (1) is that concentration is the acetic acid of 0.8～1.3mol/L, and phaffiafhodozyma bacterium dry powder mixes in the ratio of 1kg:18～23L with acetic acid, after mixing, in mixed liquor, adds vitamins C, and making its final concentration is 3mg/ml;
The high-temperature and high-pressure conditions of the described heating of step (2) is: 121 ℃ of temperature, saturated vapour pressure 0.1Mpa, and the time that described insulation is processed is 4～8 minutes;
PH to 5.0～7.0 of the described adjusting feed liquid of step (3);
The described organic solvent of step (4) is the mixing solutions of chloroform and methyl alcohol, wherein take volume ratio chloroform: methyl alcohol is 2～3:1; The organic phase that contains astaxanthin is chloroform layer;
The dry condition of the described vacuum desolvation of step (7) is: 45～50 ℃ of vacuum tightness 0.07～0.09MPa, temperature.
2. utilize according to claim 1 phaffiafhodozyma thalline to prepare the method for astaxanthin missible oil, it is characterized in that: the described phaffiafhodozyma bacterium of step (1) dry powder is purchased from Shandong extensive biotechnology Services Co., Ltd.
3. utilize according to claim 1 phaffiafhodozyma thalline to prepare the method for astaxanthin missible oil, it is characterized in that: the described organic acid of step (1) is that concentration is the acetic acid of 1.0mol/L, and phaffiafhodozyma bacterium dry powder mixes in the ratio of 1kg:20L with acetic acid.
4. utilize according to claim 1 phaffiafhodozyma thalline to prepare the method for astaxanthin missible oil, it is characterized in that: the time that the described insulation of step (2) is processed is 5 minutes.
5. utilize according to claim 1 phaffiafhodozyma thalline to prepare the method for astaxanthin missible oil, it is characterized in that: the pH to 6.5 of the described adjusting feed liquid of step (3).
6. utilize according to claim 1 phaffiafhodozyma thalline to prepare the method for astaxanthin missible oil, it is characterized in that: the mixing solutions of the described chloroform of step (4) and methyl alcohol be take volume ratio chloroform: methyl alcohol is 2:1.
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Cited By (5)
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| CN103848769A (en) * | 2014-02-21 | 2014-06-11 | 集美大学 | Method of separating and purifying astaxanthin from Phaffia rhodozyma |
| CN105624256A (en) * | 2016-01-29 | 2016-06-01 | 山东百龙创园生物科技有限公司 | Method for producing astaxanthin missible oil by aid of yeast |
| CN105732452A (en) * | 2016-02-05 | 2016-07-06 | 厦门汇盛生物有限公司 | Method for extracting phaffia rhodozyma intracellular astaxanthin |
| CN115417749A (en) * | 2022-09-19 | 2022-12-02 | 杭州时光肌生物科技有限公司 | Extraction method of bakuchiol |
| CN116508996A (en) * | 2023-05-23 | 2023-08-01 | 华南理工大学 | Carotenoid emulsion and preparation method and application thereof |
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Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103848769A (en) * | 2014-02-21 | 2014-06-11 | 集美大学 | Method of separating and purifying astaxanthin from Phaffia rhodozyma |
| CN103848769B (en) * | 2014-02-21 | 2016-06-01 | 集美大学 | A kind of method of separation and purification astaxanthin from Fife's yeast |
| CN105624256A (en) * | 2016-01-29 | 2016-06-01 | 山东百龙创园生物科技有限公司 | Method for producing astaxanthin missible oil by aid of yeast |
| CN105732452A (en) * | 2016-02-05 | 2016-07-06 | 厦门汇盛生物有限公司 | Method for extracting phaffia rhodozyma intracellular astaxanthin |
| CN115417749A (en) * | 2022-09-19 | 2022-12-02 | 杭州时光肌生物科技有限公司 | Extraction method of bakuchiol |
| CN115417749B (en) * | 2022-09-19 | 2024-04-16 | 杭州时光肌生物科技有限公司 | Method for extracting bakuchiol |
| CN116508996A (en) * | 2023-05-23 | 2023-08-01 | 华南理工大学 | Carotenoid emulsion and preparation method and application thereof |
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Application publication date: 20140205 |