CN105503683A - Method for extracting astaxanthin by hydrolyzing shrimp shell through papain - Google Patents
Method for extracting astaxanthin by hydrolyzing shrimp shell through papain Download PDFInfo
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- CN105503683A CN105503683A CN201511017390.4A CN201511017390A CN105503683A CN 105503683 A CN105503683 A CN 105503683A CN 201511017390 A CN201511017390 A CN 201511017390A CN 105503683 A CN105503683 A CN 105503683A
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- C07C403/24—Derivatives of cyclohexane or of a cyclohexene or of cyclohexadiene, having a side-chain containing an acyclic unsaturated part of at least four carbon atoms, this part being directly attached to the cyclohexane or cyclohexene or cyclohexadiene rings, e.g. vitamin A, beta-carotene, beta-ionone having side-chains substituted by six-membered non-aromatic rings, e.g. beta-carotene
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- C12P23/00—Preparation of compounds containing a cyclohexene ring having an unsaturated side chain containing at least ten carbon atoms bound by conjugated double bonds, e.g. carotenes
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Abstract
The invention provides a method for extracting astaxanthin by hydrolyzing shrimp a shell through papain, which comprises the steps of cleaning and smashing the shrimp shell, adding distilled water, performing enzymolysis of papain, performing ice-water bath for cooling and enzyme deactivation, filtering, adding acetone into a sediment for extracting, filtering, and spinning-evaporating filtrate to obtain the astaxanthin. The selected papain is protein enzyme capable of keeping higher activity in acid, neutral and alkali environments, and has characteristics of wide application range, strong operability and optimized, simple and convenient process, and the like; the papain can selectively degrade the GIcNAc-GleN glucosidic bond of chitin, which is favorable for extracting the astaxanthin, and the method for recycling the astaxanthin is economical, feasible, small in pollution, and can realize industrialized production.
Description
Technical field
The present invention relates to a kind of method extracting astaxanthin, be specifically related to a kind of method utilizing papain hydrolysis shrimp shell to extract astaxanthin.
Background technology
Astaxanthin (astaxanthin), formal name used at school 3,3 ＇-dihydroxyl-4,4 '-diketo-β, β ' carotene, belongs to Ye Huang race (xanthohylls) keto-acid carotenoid, is soluble in organic solvent, be insoluble in water, have that unique painted, anti-oxidant, anti-cancer, enhancing antibody produce, resist ultraviolet, are with vitamin A-active, improve eyesight, immunizing power, chromogenesis are communicated with nerve and improve the functions such as fertility.Astaxanthin is extensively present in occurring in nature, as in: most of Crustacean and salmon fishes body, in the spending of plant and the feather of flamingo medium, play coloration.
Procambius clarkii is commonly called as small lobsters, and in China, distribution is wide, and amount of fishing is large, and the waste after processing accounts for 70% ~ 85% of raw material, is that the one extracting astaxanthin is well originated.But in small lobsters processing at home, shrimp shell is except extraction chitosan, and the overwhelming majority goes out of use, and causes the wasting of resources and environmental pollution, is unfavorable for that it fully utilizes.
Summary of the invention
The object of this invention is to provide a kind of method utilizing papain hydrolysis shrimp shell to extract astaxanthin, the GIcNAc-GleN glycosidic link of papoid alternative degraded chitin, is conducive to the extraction of astaxanthin.
In order to achieve the above object, the technical scheme that the present invention takes is:
Utilize papain hydrolysis shrimp shell to extract a method for astaxanthin, comprise the steps:
(1) take the shrimp shell of certain part by weight, clean up with distilled water, drain excessive moisture and smash to pieces to 5 ~ 10mm for subsequent use with tissue mashing machine;
(2) in the shrimp shell smashed to pieces, add distilled water to mix, regulate pH=5.5 ~ 6.5, heating, then adds papoid, water bath with thermostatic control vibration papain enzymolysis;
(3) after enzymolysis terminates, ice-water bath cools the enzyme that goes out, and filters, repeatedly uses distilled water flushing throw out, be washed till neutrality;
(4) in throw out, add acetone, ultrasonic 10 ~ 30min, shaking table normal temperature vibrates, at the centrifugal 5 ~ 10min of refrigerated centrifuge, filtering separation liquid and solid, namely filtrate obtain astaxanthin through rotary evaporation, and revolving the acetone after steaming can again for extracting astaxanthin.
Preferably, the consumption of described shrimp shell and the amount ratio of distilled water are 0.5 ~ 2.5:1.
Preferably, the papoid enzyme-to-substrate in described step (2) is than being 2000U/g ~ 3000U/g.
Preferably, in described step (4), the consumption of acetone is 2 ~ 6 times of shrimp shell consumption.
Preferably, in described step (2), the time of enzymolysis is 45 ~ 65min.
Preferably, in described step (3), the time of ice-water bath cooling is 10 ~ 30min.
Preferably, in described step (4), the time of shaking table normal temperature vibration is 2 ~ 5h.
Compared with prior art, beneficial effect of the present invention:
The present invention utilizes papain hydrolysis shrimp shell to extract astaxanthin, and papoid is a kind of proteolytic enzyme that all can keep greater activity under acidity, neutrality, alkaline environment, has the features such as wide accommodation, workable, process optimization is easy; And the GIcNAc-GleN glycosidic link of papoid alternative degraded chitin, be conducive to the extraction of astaxanthin, utilize this method to reclaim astaxanthin, economically feasible, pollutes little, can industrialization produce.
Embodiment
Below in conjunction with specific embodiment, the present invention is further described.
Embodiment 1
(1) take 5g shrimp shell, clean up with distilled water, drain excessive moisture and smash to pieces to 5 ~ 10mm for subsequent use with tissue mashing machine;
(2) in the shrimp shell smashed to pieces, add 10mL distilled water to mix, regulate pH=5.5 with pH damping fluid, be heated to 35 DEG C, then add enzyme-to-substrate than the papoid for 2000U/g, water bath with thermostatic control vibration papain enzymolysis 45min;
(3), after enzymolysis terminates, ice-water bath cooling 10min goes out enzyme, filters, repeatedly uses distilled water flushing throw out, be washed till neutrality;
(4) in throw out, 10mL acetone is added, ultrasonic 10min, shaking table normal temperature vibration 2h, at refrigerated centrifuge 10000r/min, centrifugal 10min at 4 DEG C, filtering separation liquid and solid, namely filtrate obtain astaxanthin through rotary evaporation, and revolving the acetone after steaming can again for extracting astaxanthin.
Embodiment 2
(1) take 8g shrimp shell, clean up with distilled water, drain excessive moisture and smash to pieces to 5 ~ 10mm for subsequent use with tissue mashing machine;
(2) in the shrimp shell smashed to pieces, add 8mL distilled water to mix, regulate pH=5.8 with pH damping fluid, be heated to 40 DEG C, then add enzyme-to-substrate than the papoid for 2250U/g, water bath with thermostatic control vibration papain enzymolysis 50min;
(3), after enzymolysis terminates, ice-water bath cooling 15min goes out enzyme, filters, repeatedly uses distilled water flushing throw out, be washed till neutrality;
(4) in throw out, 24mL acetone is added, ultrasonic 15min, shaking table normal temperature vibration 3h, at refrigerated centrifuge 10000r/min, centrifugal 8min at 4 DEG C, filtering separation liquid and solid, namely filtrate obtain astaxanthin through rotary evaporation, and revolving the acetone after steaming can again for extracting astaxanthin.
Embodiment 3
(1) take 10g shrimp shell, clean up with distilled water, drain excessive moisture and smash to pieces to 5 ~ 10mm for subsequent use with tissue mashing machine;
(2) in the shrimp shell smashed to pieces, add 5mL distilled water to mix, regulate pH=6.0 with pH damping fluid, be heated to 45 DEG C, then add enzyme-to-substrate than the papoid for 2500U/g, water bath with thermostatic control vibration papain enzymolysis 60min;
(3), after enzymolysis terminates, ice-water bath cooling 20min goes out enzyme, filters, repeatedly uses distilled water flushing throw out, be washed till neutrality;
(4) in throw out, 40mL acetone is added, ultrasonic 20min, shaking table normal temperature vibration 3h, at refrigerated centrifuge 10000r/min, centrifugal 5min at 4 DEG C, filtering separation liquid and solid, namely filtrate obtain astaxanthin through rotary evaporation, and revolving the acetone after steaming can again for extracting astaxanthin.
Embodiment 4
(1) take 15g shrimp shell, clean up with distilled water, drain excessive moisture and smash to pieces to 5 ~ 10mm for subsequent use with tissue mashing machine;
(2) in the shrimp shell smashed to pieces, add 10mL distilled water to mix, regulate pH=6.3 with pH damping fluid, be heated to 50 DEG C, then add enzyme-to-substrate than the papoid for 2750U/g, water bath with thermostatic control vibration papain enzymolysis 55min;
(3), after enzymolysis terminates, ice-water bath cooling 25min goes out enzyme, filters, repeatedly uses distilled water flushing throw out, be washed till neutrality;
(4) in throw out, 75mL acetone is added, ultrasonic 25min, shaking table normal temperature vibration 4h, at refrigerated centrifuge 10000r/min, centrifugal 7min at 4 DEG C, filtering separation liquid and solid, namely filtrate obtain astaxanthin through rotary evaporation, and revolving the acetone after steaming can again for extracting astaxanthin.
Embodiment 5
(1) take 20g shrimp shell, clean up with distilled water, drain excessive moisture and smash to pieces to 5 ~ 10mm for subsequent use with tissue mashing machine;
(2) in the shrimp shell smashed to pieces, add 8mL distilled water to mix, regulate pH=6.5 with pH damping fluid, be heated to 55 DEG C, then add enzyme-to-substrate than the papoid for 3000U/g, water bath with thermostatic control vibration papain enzymolysis 65min;
(3), after enzymolysis terminates, ice-water bath cooling 30min goes out enzyme, filters, repeatedly uses distilled water flushing throw out, be washed till neutrality;
(4) in throw out, 120mL acetone is added, ultrasonic 30min, shaking table normal temperature vibration 5h, at refrigerated centrifuge 10000r/min, centrifugal 10min at 4 DEG C, filtering separation liquid and solid, namely filtrate obtain astaxanthin through rotary evaporation, and revolving the acetone after steaming can again for extracting astaxanthin.
The performance that embodiment 1,2,3,4,5 extracts the astaxanthin obtained tested, result is as shown in table 1 below.
Table 1
The above, it is only present pre-ferred embodiments, not technical scope of the present invention is imposed any restrictions, thus every above embodiment is done according to technical spirit of the present invention any trickle amendment, equivalent variations and modification, all still belong in the scope of technical solution of the present invention.
Claims (7)
1. utilize papain hydrolysis shrimp shell to extract a method for astaxanthin, it is characterized in that, comprise the steps:
(1) take the shrimp shell of certain part by weight, clean up with distilled water, drain excessive moisture and smash to pieces to 5 ~ 10mm for subsequent use with tissue mashing machine;
(2) in the shrimp shell smashed to pieces, add distilled water to mix, regulate pH=5.5 ~ 6.5, heating, then adds papoid, water bath with thermostatic control vibration papain enzymolysis;
(3) after enzymolysis terminates, ice-water bath cools the enzyme that goes out, and filters, repeatedly uses distilled water flushing throw out, be washed till neutrality;
(4) in throw out, add acetone, ultrasonic 10 ~ 30min, shaking table normal temperature vibrates, at the centrifugal 5 ~ 10min of refrigerated centrifuge, filtering separation liquid and solid, namely filtrate obtain astaxanthin through rotary evaporation, and revolving the acetone after steaming can again for extracting astaxanthin.
2. the method utilizing papain hydrolysis shrimp shell to extract astaxanthin according to claim 1, it is characterized in that, the consumption of described shrimp shell and the amount ratio of distilled water are 0.5 ~ 2.5:1.
3. the method utilizing papain hydrolysis shrimp shell to extract astaxanthin according to claim 1, is characterized in that, the papoid enzyme-to-substrate in described step (2) is than being 2000U/g ~ 3000U/g.
4. the method utilizing papain hydrolysis shrimp shell to extract astaxanthin according to claim 1, is characterized in that, in described step (4), the consumption of acetone is 2 ~ 6 times of shrimp shell consumption.
5. the method utilizing papain hydrolysis shrimp shell to extract astaxanthin according to claim 1, it is characterized in that, in described step (2), the time of enzymolysis is 45 ~ 65min.
6. the method utilizing papain hydrolysis shrimp shell to extract astaxanthin according to claim 1, is characterized in that, in described step (3), the time of ice-water bath cooling is 10 ~ 30min.
7. the method utilizing papain hydrolysis shrimp shell to extract astaxanthin according to claim 1, is characterized in that, in described step (4), the time of shaking table normal temperature vibration is 2 ~ 5h.
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107033049A (en) * | 2017-06-09 | 2017-08-11 | 中国水产科学研究院南海水产研究所 | A kind of method that combined-enzyme method extracts krill astaxanthin |
| CN108850778A (en) * | 2018-06-19 | 2018-11-23 | 佛山皖阳生物科技有限公司 | A kind of preparation method of long-acting stable type biomass antioxidant |
| CN115108865A (en) * | 2022-06-21 | 2022-09-27 | 苏州益植生物科技有限公司 | Green composite biological stimulator and preparation method thereof |
| CN115466145A (en) * | 2022-08-12 | 2022-12-13 | 苏州益植生物科技有限公司 | Coated biostimulant and preparation method and application thereof |
| CN115477602A (en) * | 2022-09-28 | 2022-12-16 | 湖北兴祥生物科技有限公司 | Extraction process of astaxanthin ester in crayfish waste |
Citations (4)
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| CN1644073A (en) * | 2005-01-25 | 2005-07-27 | 江南大学 | Method for processing fent by comprehensive utilizing shrimp by enzyme method |
| CN101176549A (en) * | 2007-11-29 | 2008-05-14 | 湖北东方天琪生物工程股份有限公司 | Low energy- wasting and environment protection method for extracting chitin as well as biologically active substance thereof from shrimp shell |
| EP2085381A1 (en) * | 2008-01-30 | 2009-08-05 | Bioptik Technology, Inc. | Method for obtaining natural astaxanthin from eggs and gonads of snails |
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2015
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Patent Citations (4)
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| CN1644073A (en) * | 2005-01-25 | 2005-07-27 | 江南大学 | Method for processing fent by comprehensive utilizing shrimp by enzyme method |
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| EP2085381A1 (en) * | 2008-01-30 | 2009-08-05 | Bioptik Technology, Inc. | Method for obtaining natural astaxanthin from eggs and gonads of snails |
| KR20100001773A (en) * | 2008-06-27 | 2010-01-06 | 인만진 | Method of extraction for astaxanthin from haematococcus pluvialis using second enzyme reaction |
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107033049A (en) * | 2017-06-09 | 2017-08-11 | 中国水产科学研究院南海水产研究所 | A kind of method that combined-enzyme method extracts krill astaxanthin |
| CN108850778A (en) * | 2018-06-19 | 2018-11-23 | 佛山皖阳生物科技有限公司 | A kind of preparation method of long-acting stable type biomass antioxidant |
| CN115108865A (en) * | 2022-06-21 | 2022-09-27 | 苏州益植生物科技有限公司 | Green composite biological stimulator and preparation method thereof |
| CN115466145A (en) * | 2022-08-12 | 2022-12-13 | 苏州益植生物科技有限公司 | Coated biostimulant and preparation method and application thereof |
| CN115477602A (en) * | 2022-09-28 | 2022-12-16 | 湖北兴祥生物科技有限公司 | Extraction process of astaxanthin ester in crayfish waste |
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