CN103865798A - Method for wall breaking of phaffia rhodozyma through enzymolysis and application thereof - Google Patents
Method for wall breaking of phaffia rhodozyma through enzymolysis and application thereof Download PDFInfo
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- CN103865798A CN103865798A CN201410099783.3A CN201410099783A CN103865798A CN 103865798 A CN103865798 A CN 103865798A CN 201410099783 A CN201410099783 A CN 201410099783A CN 103865798 A CN103865798 A CN 103865798A
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Abstract
The invention discloses a method for wall breaking of phaffia rhodozyma through enzymolysis. The method comprises the following steps: firstly, carrying out solid-liquid separation on a phaffia rhodozyma yeast fermentation liquor containing astaxanthin, and concentrating into bacteria slurry; carrying out efficient wall breaking in a composite enzyme enzymolysis mode, wherein the composite enzyme is compounded by cellulase and alkaline protease according to the ratio of 3:7; neutralizing the bacteria slurry subjected to wall breaking through enzymolysis to pH of 7.0+/-0.5 and evenly mixing with a complex antioxidant and an antitackiness agent; drying by adopting a spray drying process, so as to prepare phaffia rhodozyma powder; directly extracting and leaching the bacteria slurry subjected to wall breaking through enzymolysis by using an organic solvent, and carrying out low-temperature vacuum concentration to obtain an astaxanthin concentrated liquor; spray-drying, so as to prepare astaxanthin powder. By adopting the method, the wall breaking rate can be effectively maintained over 80%; after the wall-broken phaffia rhodozyma powder or astaxanthin powder is added to aquaculture and poultry feeds, individual growth can be effectively facilitated, the seedling survival is improved by 10-20%, and significant economic benefits are achieved.
Description
Technical field
The present invention relates to a kind of red phaffia rhodozyma enzymolysis wall-breaking method and application thereof.
Background technology
Red phaffia rhodozyma is (Phaff etc., 1972) that first 20 century 70s are separated in the exudate of Japan and mountain area, U.S. Alaska deciduous trees by people such as Phaff.It is unique one of mycota, Mycophyta, Acarasiales subphylum, Cryptococcaceae, phaffia rhodozyma genus.Its modes of reproduction is asexual budding.This bacterium fermentation energy produces 10 multiple types carotene, mainly contains astaxanthin, β-carotene, gamma carotene etc., and wherein astaxanthin accounts for 60%～85%.It can utilize several kinds of carbon source to ferment, and is suitable for 20～28 ℃ of growth temperatures most, optimal pH 5.0～6.9, and red phaffia rhodozyma produces acid in aerobic fermentation process, release of carbon dioxide simultaneously synthesizing astaxanthin.
Astaxanthin (astaxathin) is 3,3 '-dihydroxyl-4,4 '-diketo-β, and β '-carotene, is a kind of keto-acid carotenoid.The main source of astaxanthin comprises two kinds, synthetic and organism source.Synthetic astaxanthin due to purity reason and side effect many, use range narrower (only for the dyeing of some fish), comes from wide material sources (red phaffia rhodozyma, patter of rain haematococcus pulvialis, ocean rhodotorula etc.), the natural health of astaxanthin of organism.Therefore be subject to the mankind's attention and widespread use.The domestic research to biotechnology production astaxanthin starts from the beginning of this century, and starts to carry out large quantity research from the beginning of the eighties in last century abroad.Because astaxanthin is natural red-purple, be used as the additive of bait (in August, 2000, FDA ratified the fodder additives of red phaffia rhodozyma as Oncorhynchi) of water industry at first as pigment in American-European countries.Popularized and add in more aquaculture bait successively afterwards, within 1998, U.S. FDA official approval patter of rain haematococcus pulvialis astaxanthin is for sector applications such as feed, food, medicine, makeup.
Current research showed that astaxanthin had extremely strong antioxidation, also has effects such as the generation of the body antibody of promotion, strengthening immunity and ultra-violet radiation resisting in recent years.Thereby aspect foodstuff additive, aquaculture, makeup, healthcare products and medicine industry, there iing more wide application prospect.
Preparation production of astaxanthin method mainly contains 3 kinds at present: chemosynthesis, shrimp shell class are extracted, microorganism fermentation.Chemical synthesis process complexity, and in shrimp shell class waste, content astaxanthin is low, cost is higher.Therefore first-elected microorganism fermentation, research is red Phaffia Rhodozyma and Haematocoocus Pluvialls the most widely at present.Determination of Astaxanthin in Haematococcus Pluvialis content is up to 4% of its dry cell weight, but its culture condition harshness.Along with the development of biotechnology, people can obtain astaxanthin by corresponding biotechnology.The patent of extracting at present astaxanthin and related application both at home and abroad from microorganism has many pieces of writing (as application number CN03105314.9, CN03117031.5, CN200410013634.7, CN200710149906.X, CN200910238464.5, CN201010268910.X, CN201010570151, CN201110282683.0, US5744502, US5679567, US5972642, PCT/CN2011/00168 etc.), be all to adopt the modes such as chemistry, physics, biotechnology class to carry out broken wall, extraction or lixiviate mode to prepare astaxanthin substantially.
Phaffia Rhodozyma cell walls is comparatively tough and tensile, in order to promote the emission and absorption of parachrome, and must smudge cells before feeding animals.The present invention is intended to research and develop specially a kind of red phaffia rhodozyma enzymolysis wall-breaking method.
Summary of the invention
The object of the invention is to provide a kind of red phaffia rhodozyma enzymolysis wall-breaking method and application thereof, and it can be simplified and compress rear road and extract the astaxanthin process time, and cost is low, and suitability for industrialized is produced.
In order to reach above-mentioned purpose, solution of the present invention is:
A kind of red phaffia rhodozyma enzymolysis wall-breaking method, its step is as follows:
The first step, first by be rich in astaxanthin red phaffia rhodozyma fermented liquid solid-liquid separation, be condensed into bacterium slurry body;
Second step, above-mentioned bacterium slurry body is carried out to high-efficiency wall breaking by complex enzyme zymohydrolysis mode, and prozyme adopts cellulase and Sumizyme MP to form in 3:7 ratio is composite, can obtain more than 80% astaxanthin, the highest even can reaching more than 90%, wherein total pigment, astaxanthin loss are less than 5%.
Described second step, by bacterium for slurry body alkaline solution (sodium hydroxide or potassium hydroxide) adjust pH control to 7.0～10.0, prozyme dosage accounts for bacterium slurry body solution gross weight 0.1～5.0%, and carries out enzymolysis broken wall in 35～60 ℃ of intervals, broken time 5.0～13 hours.
Described second step, adds compound antioxidant in broken wall process, and compound antioxidant is Ascorbyl Palmitate or sodium ascorbate etc., and the bacterium slurry body after broken wall is carried out to corresponding effective protection.
A kind of preparation method of red phaffia rhodozyma dry powder, adopt the pH of bacterium slurry body after above-mentioned enzymolysis broken wall to be neutralized to 7.0 ± 0.5, and after mixing with compound antioxidant, anticaking agent (as silicon-dioxide, monocalcium phosphate), employing drying process with atomizing is dry, make red phaffia rhodozyma dry powder, wherein pigment, astaxanthin loss amount are less than 5%.
One after described broken wall in the pH of bacterium slurry body and in employing citric acid and lactic acid.
The described dry red phaffia rhodozyma dry powder of making is directly used aluminium foil bag vacuum packaging, and lucifuge preservation under 2～8 ℃ of environment.
A kind of preparation method of astaxanthin dry powder, after adopting above-mentioned enzymolysis broken wall, bacterium slurry body directly carries out organic solvent (ethanol, acetone, ethyl acetate etc.) extraction, extraction, cryogenic vacuum is concentrated can obtain astaxanthin concentrated solution, the dry astaxanthin dry powder of making of spraying, powder is amaranth or garnet, and its content astaxanthin is at 5000～12000mg/kg dry powder, and total pigment is at 8000～20000mg/kg dry powder, dry powder moisture≤6%, crude protein >=10～35%.
Adopt after such scheme; the red phaffia rhodozyma enzymolysis of the present invention wall-breaking method can effectively maintain 80% above sporoderm-broken rate; through spraying dry after, dry powder pigment, astaxanthin loss amount be lower than in 5%, can effectively the accomplish scale production red phaffia rhodozyma dry powder of broken wall and astaxanthin dry powder.The red phaffia rhodozyma dry powder that broken wall of the present invention makes is added in the aquatic products such as rainbow trout, cabrilla, prawn and poultry farming basal feed, or add the astaxanthin dry powder that makes of 0.8～2.0% broken wall of the present invention, can effectively promote individual growth, the survival of growing seedlings improves 10～20%, and economic benefit is obvious.
Accompanying drawing explanation
Fig. 1 is process flow diagram of the present invention.
Embodiment
As shown in Figure 1, technical process of the present invention is:
First by be rich in astaxanthin red phaffia rhodozyma fermented liquid solid-liquid separation, be condensed into bacterium slurry body;
Again by bacterium for slurry body alkaline solution (sodium hydroxide or potassium hydroxide) adjust pH control to 7.0～10.0, prozyme dosage accounts for bacterium slurry body solution gross weight 0.1～5.0%, and carries out enzymolysis broken wall in 35～60 ℃ of intervals, broken time 5.0～13 hours; Prozyme adopts cellulase and Sumizyme MP to form in 3:7 ratio is composite, in broken wall process, adds compound antioxidant, and compound antioxidant is Ascorbyl Palmitate or sodium ascorbate etc., and the bacterium slurry body after broken wall is carried out to corresponding effective protection;
Then the pH of bacterium slurry body after above-mentioned enzymolysis broken wall is neutralized to 7.0 ± 0.5, and after mixing with compound antioxidant, anticaking agent, adopts drying process with atomizing dry, make red phaffia rhodozyma dry powder; Or bacterium slurry body after above-mentioned enzymolysis broken wall is directly carried out to organic solvent extraction, extraction, and cryogenic vacuum is concentrated can obtain astaxanthin concentrated solution, the dry astaxanthin dry powder of making of spraying.
Embodiment 1 adds 1.5% broken wall astaxanthin dry powder (amounting to astaxanthin 70～90mg/kg) in rainbow trout bait, feeds 6 weeks, and individuality has increased more than 20%.The rainbow trout nursing of growing seedlings, surviving rate and control group contrast improve respectively 15～20%.
Embodiment 2 adds and contains 2.0% broken wall astaxanthin dry powder (amounting to astaxanthin 60～80 mg/kg) in cabrilla (porgy section class) bait, feeds 4 weeks, individual growth and control group contrast 18～22%.In cabrilla is grown seedlings bait, feed, surviving rate and control group contrast improve respectively 15～18%.
Embodiment 3 adds 1.5% broken wall astaxanthin dry powder (amounting to astaxanthin 60～80 mg/kg) in shrimp seedling bait, feeds 13 days, and the surviving rate of shrimp seedling and control group contrast are increased to 20～23%; Individual growth more than 20.0%.
Embodiment 4. adds 2% broken wall astaxanthin dry powder in poultry basal feed, feeds after 4 weeks, and skin, pin, beak present golden yellow, wherein astaxanthin can also be enriched among birds, beasts and eggs, and these can both improve the commodity value of birds, beasts and eggs, meat greatly.In addition, astaxanthin can also promote the growth of poultry and improve laying rate as nutritive substance.
Embodiment 5, in aquarium fish (pigmentation 5.5～6.0 grammes per square metres) bait, add 1.5～2.0% broken wall astaxanthin dry powder, feed continuously 4 weeks, body colour coloring effect and control group contrast improve more than 25%.
The present invention is economical and practical, can effectively maintain 80% above sporoderm-broken rate, and astaxanthin loss is less than 5%; Not only can produce in batches, and can effectively compress the subsequent extracted astaxanthin process time, be applicable to industrial scaleization and produce.With existing other tradition
red Phaffia Rhodozyma broken wallmode contrasts, and can effectively avoid the astaxanthin loss causing in broken wall and powder drying process.The dry mash of producing directly adopts the preservation of aluminium foil bag lucifuge, and the storage rack time is longer, and therefore power-product related application can be more extensive.
Claims (8)
1. a red phaffia rhodozyma enzymolysis wall-breaking method, is characterized in that step is as follows:
The first step, first by be rich in astaxanthin red phaffia rhodozyma fermented liquid high speed centrifugation solid-liquid separation, be condensed into bacterium slurry body;
Second step, carries out high-efficiency wall breaking by above-mentioned bacterium slurry body by complex enzyme zymohydrolysis mode, and prozyme adopts cellulase and Sumizyme MP to form in 3:7 ratio is composite.
2. the red phaffia rhodozyma enzymolysis of one as claimed in claim 1 wall-breaking method, it is characterized in that: second step, adjust pH to control to 7.0～10.0 with alkaline solution bacterium slurry body, prozyme dosage accounts for bacterium slurry body solution gross weight 0.1～5.0%, and carry out enzymolysis broken wall in 35～60 ℃ of intervals, broken time 5.0～13 hours.
3. the red phaffia rhodozyma enzymolysis of one as claimed in claim 1 wall-breaking method, is characterized in that: second step, in broken wall process, add compound antioxidant, and compound antioxidant is Ascorbyl Palmitate or sodium ascorbate.
4. the preparation method of a red phaffia rhodozyma dry powder, it is characterized in that: after employing claim 1 second step enzymolysis broken wall, the pH of bacterium slurry body is neutralized to 7.0 ± 0.5, and after mixing with compound antioxidant, anticaking agent, spray-dried technique is dry, makes red phaffia rhodozyma dry powder.
5. the preparation method of a kind of red phaffia rhodozyma dry powder as claimed in claim 4, is characterized in that: the one after described broken wall in the pH of bacterium slurry body and in employing citric acid and lactic acid.
6. the preparation method of a kind of red phaffia rhodozyma dry powder as claimed in claim 4, is characterized in that: the described dry red phaffia rhodozyma dry powder of making is directly used aluminium foil bag vacuum packaging, and lucifuge preservation under 2～8 ℃ of environment.
7. a preparation method for astaxanthin dry powder, is characterized in that: after adopting claim 1 second step enzymolysis broken wall, bacterium slurry body directly carries out organic solvent extraction, extraction, and cryogenic vacuum is concentrated, obtains astaxanthin concentrated solution.
8. the preparation method of a kind of astaxanthin dry powder as claimed in claim 7, it is characterized in that: the dry astaxanthin dry powder of making of astaxanthin concentrated solution spraying, powder is amaranth or garnet, its content astaxanthin is at 5000～12000mg/kg dry powder, total pigment is at 8000～20000mg/kg dry powder, dry powder moisture≤6%, crude protein >=10～35%.
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Cited By (15)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105028996A (en) * | 2015-06-18 | 2015-11-11 | 宁德市南海水产科技有限公司 | Bait additive capable of increasing raised fry survival rate of epinephelus and use method thereof |
| CN105211673A (en) * | 2015-09-18 | 2016-01-06 | 南京理工大学 | For the compound anti-caking agent in food-grade potassium chloride |
| CN105348165A (en) * | 2015-10-08 | 2016-02-24 | 华南理工大学 | Method for extracting astaxanthin yeast, yeast extract and glucan from yeast |
| CN105434319A (en) * | 2015-12-07 | 2016-03-30 | 华南理工大学 | Cosmetics based on phaffia rhodozyma raw materials and preparation method thereof |
| CN105639204A (en) * | 2016-01-18 | 2016-06-08 | 山东宝来利来生物工程股份有限公司 | Biological color enhancing agent and application thereof to aquatic animal breeding |
| 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 |
| CN107821813A (en) * | 2017-11-13 | 2018-03-23 | 广州傲农生物科技有限公司 | A kind of feed addictive for improving the pig colour of skin and hair and preparation method and application |
| CN110804638A (en) * | 2019-10-31 | 2020-02-18 | 厦门昶科生物工程有限公司 | Production method for improving astaxanthin yield |
| CN111450969A (en) * | 2020-04-10 | 2020-07-28 | 山东畜牧兽医职业学院 | Bacterial sludge enzymolysis wall breaking equipment for industrial production of high-yield astaxanthin |
| CN112423770A (en) * | 2018-06-25 | 2021-02-26 | (株)库恩生物 | Composition for improving diabetes or enhancing antioxidant activity comprising yeast extract and method for preparing yeast extract |
| CN112515053A (en) * | 2020-11-02 | 2021-03-19 | 无锡泓宝微生物科技有限公司 | A QINGSU bacterium for enhancing pigment and immunity of aquatic animal |
| CN113265439A (en) * | 2021-06-08 | 2021-08-17 | 江苏海飞生物科技有限公司 | Method for preparing trans-astaxanthin |
| CN113292469A (en) * | 2021-06-28 | 2021-08-24 | 浙江皇冠科技有限公司 | Method for extracting phaffia rhodozyma intracellular astaxanthin by using beta-glucanase and chitinase |
| CN114369049A (en) * | 2022-01-12 | 2022-04-19 | 威海东巽生物科技有限公司 | Preparation method of quaternized astaxanthin and antioxidant biological agent |
| CN114437950A (en) * | 2021-12-24 | 2022-05-06 | 青岛尚德生物技术有限公司 | Phaffia rhodozyma GOY1 for improving aquatic animal body color and preparation method and application of culture of phaffia rhodozyma GOY1 |
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Cited By (20)
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| CN105028996A (en) * | 2015-06-18 | 2015-11-11 | 宁德市南海水产科技有限公司 | Bait additive capable of increasing raised fry survival rate of epinephelus and use method thereof |
| CN105211673A (en) * | 2015-09-18 | 2016-01-06 | 南京理工大学 | For the compound anti-caking agent in food-grade potassium chloride |
| CN105348165A (en) * | 2015-10-08 | 2016-02-24 | 华南理工大学 | Method for extracting astaxanthin yeast, yeast extract and glucan from yeast |
| CN105348165B (en) * | 2015-10-08 | 2017-04-19 | 华南理工大学 | Method for extracting astaxanthin yeast, yeast extract and glucan from yeast |
| CN105434319A (en) * | 2015-12-07 | 2016-03-30 | 华南理工大学 | Cosmetics based on phaffia rhodozyma raw materials and preparation method thereof |
| CN105434319B (en) * | 2015-12-07 | 2018-09-14 | 华南理工大学 | A kind of cosmetics and preparation method thereof based on phaffiafhodozyma raw material |
| CN105639204A (en) * | 2016-01-18 | 2016-06-08 | 山东宝来利来生物工程股份有限公司 | Biological color enhancing agent and application thereof to aquatic animal breeding |
| CN105639204B (en) * | 2016-01-18 | 2019-11-22 | 山东宝来利来生物工程股份有限公司 | A kind of biology toner and its application in aquaculture of aquatic animal |
| 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 |
| CN107821813A (en) * | 2017-11-13 | 2018-03-23 | 广州傲农生物科技有限公司 | A kind of feed addictive for improving the pig colour of skin and hair and preparation method and application |
| CN112423770A (en) * | 2018-06-25 | 2021-02-26 | (株)库恩生物 | Composition for improving diabetes or enhancing antioxidant activity comprising yeast extract and method for preparing yeast extract |
| CN110804638A (en) * | 2019-10-31 | 2020-02-18 | 厦门昶科生物工程有限公司 | Production method for improving astaxanthin yield |
| CN111450969A (en) * | 2020-04-10 | 2020-07-28 | 山东畜牧兽医职业学院 | Bacterial sludge enzymolysis wall breaking equipment for industrial production of high-yield astaxanthin |
| CN112515053A (en) * | 2020-11-02 | 2021-03-19 | 无锡泓宝微生物科技有限公司 | A QINGSU bacterium for enhancing pigment and immunity of aquatic animal |
| CN113265439A (en) * | 2021-06-08 | 2021-08-17 | 江苏海飞生物科技有限公司 | Method for preparing trans-astaxanthin |
| CN113292469A (en) * | 2021-06-28 | 2021-08-24 | 浙江皇冠科技有限公司 | Method for extracting phaffia rhodozyma intracellular astaxanthin by using beta-glucanase and chitinase |
| CN114437950A (en) * | 2021-12-24 | 2022-05-06 | 青岛尚德生物技术有限公司 | Phaffia rhodozyma GOY1 for improving aquatic animal body color and preparation method and application of culture of phaffia rhodozyma GOY1 |
| CN114437950B (en) * | 2021-12-24 | 2023-11-24 | 青岛尚德生物技术有限公司 | Phaffia rhodozyma GOY1 for improving body color of aquatic animals and preparation method and application of culture of Phaffia rhodozyma GOY |
| CN114369049A (en) * | 2022-01-12 | 2022-04-19 | 威海东巽生物科技有限公司 | Preparation method of quaternized astaxanthin and antioxidant biological agent |
| CN114369049B (en) * | 2022-01-12 | 2023-10-03 | 威海东巽生物科技有限公司 | Preparation method of quaternized astaxanthin and antioxidation biological agent |
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Application publication date: 20140618 |