CN103044303B - Method for using enzyme to produce astaxanthin - Google Patents
Method for using enzyme to produce astaxanthin Download PDFInfo
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- CN103044303B CN103044303B CN201210557317.6A CN201210557317A CN103044303B CN 103044303 B CN103044303 B CN 103044303B CN 201210557317 A CN201210557317 A CN 201210557317A CN 103044303 B CN103044303 B CN 103044303B
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Abstract
The invention discloses a method for using enzyme to produce astaxanthin. The method comprises the following steps: adding trichoderma koningii spore suspension into PDY fluid medium, wherein the weight of the PDY fluid medium is 9-11 times of the weight of the trichoderma koningii spore suspension, and centrifuging to obtain trichoderma koningii bacterial sludge; adding the trichoderma koningii bacterial sludge into solid fermentation medium, wherein the weight of the solid fermentation medium is 490-560 times of the weight of the bacterial sludge, first cultivating for 29-31 h at the temperature of 30-31 DEG C, then cultivating for 75-90 h at the temperature of 26-28 DEG C, and centrifuging to obtain trichoderma koningii fermentation; adding haematococcus pluvialis into BBM medium, cultivating in an enclosed photobioreactor for 2 d, and centrifuging to obtain algae sludge; adding the algae sludge into BBM medium, introducing CO2, cutivating for 2-3 d at the temperature of 32 to 35 DEG C, and centrifuging, freezing and drying to obtain algae powder; mixing the fermentation and the algae powder, centrifuging to obtain solid phase material, freezing and drying to obtain astaxanthin dry powder; and extracting the astaxanthin dry powder through corn oil to obtain astaxanthin. The method can efficiently obtain natural astaxanthin, the process is simple, the cost is low, and the environmental pollution is minimized.
Description
Technical field
The present invention relates to a kind of production method of astaxanthin, particularly relate to a kind of method of utilizing enzyme to produce astaxanthin.
Background technology
Astaxanthin is a kind of terpenes unsaturated compound, and chemical name is 3,3-dihydroxyl-4,4-diketo-β, and β-carotene, molecular formula is C
40h
52o
4, relative molecular weight is 596.86, water insoluble, tool is fat-soluble.Astaxanthin is extensively present in organism, and in the meat of Oncorhynchi, astaxanthin accounts for 70% left and right of carotenoid, have even up to 99.8%.Due to the conjugated double bond in astaxanthin molecular structure, and unsaturated ketone group and the hydroxyl of conjugated double bond end, thereby can attract free radical to there is antioxygenation, astaxanthin is in current natural antioxidants, one of compound that resistance of oxidation is the strongest, its antioxidant capacity is stronger 117 times than β-carotene, stronger 80 times than vitamin-E.
Haematocoocus Pluvialls (Haematococcus pluvialis) is a kind of fresh water unicell green alga, is subordinate to Chlorophyta volvocales haematococcus pulvialis section haematococcus.The growth of this algae is divided into obvious green swarm cell and two stages of red aplanospore b.In the abundant environment of the low light level, nitrogen phosphorus, mainly exist with green swarm cell form, in this process, haematococcus pluvialis growing is vigorous, and cell contains a small amount of astaxanthin; And under high illumination, high temperature, high salt and nutritive salt starvation conditions, exist with motionless armful of son (heavy wall is embraced son) form, now, in frustule, Chang Yin contains a large amount of astaxanthins and presents redness.
At present, mainly utilize red phaffia rhodozyma to produce astaxanthin for strain fermentation both at home and abroad.Red phaffia rhodozyma is take glucose, cellobiose as carbon source; Ammonium sulfate is nitrogenous source; Because it is facultative aerobic, be interrupted ventilation; The suitableeest culture temperature is 20～22 ℃; Optimum pH is 5.0; Oxygen supply speed will be higher than 30mmol/h; Red phaffia rhodozyma is comparatively harsh to its culture condition, and such as oxygen supply speed, during lower than 30mmol/h, the output of cell and the output of astaxanthin all obviously decline.When oxygen concentration reaches 30mmol/h, then continue to improve air flow, the output of astaxanthin improves little.
Utilize red phaffia rhodozyma to produce in astaxanthin process for strain fermentation, red phaffia rhodozyma is easily by extraneous mould contamination, thus during the fermentation need to be in gnotobasis, red phaffia rhodozyma is aerobic bacteria simultaneously, fermentation need to pass into aseptic oxygen, greatly raises the cost; In fermenting process, to strictly control temperature, pH value and oxygen supply speed, any one fermentation condition all directly affects the production efficiency of astaxanthin; Can produce toxic byproduct to fermentation later stage thalline, the follow-up purification of astaxanthin has been brought to difficulty, also contaminate environment of while.
Summary of the invention
Based on this, the invention provides a kind of method of utilizing enzyme to produce astaxanthin.
The concrete technical scheme solving the problems of the technologies described above is as follows:
Utilize enzyme to produce a method for astaxanthin, comprise the following steps:
(1) cultivation of koning trichoderma: by koning trichoderma spore suspension access PDY liquid nutrient medium, cultivate 1.5-2.5d, the centrifugal supernatant liquor that goes, obtains koning trichoderma bacterium bacterium mud;
(2) cultivation of koning trichoderma fermented product: get in the koning trichoderma bacterium bacterium mud access solid fermentation substratum of step (1) gained, at 30-32 ℃, cultivate 29-31h, after at 26-28 ℃, cultivate 75-90h, the centrifugal supernatant liquor that goes, obtains koning trichoderma fermented product; By regulation and control fermentation condition, make rich cellulose enzyme in the koning trichoderma fermented product of gained;
(3) spreading cultivation of Haematocoocus Pluvialls: Haematocoocus Pluvialls is accessed to BBM substratum and is placed in closed photo bioreactor, and blowing air is cultivated 1.5-2.5d, centrifuging and taking Haematocoocus Pluvialls algae mud;
(4) preparation of the Haematocoocus Pluvialls algae powder of enrichment astaxanthin: by the Haematocoocus Pluvialls algae mud access BBM substratum of step (3) gained, pass into CO
2, air flow be 0.5～1.5L/ (Lmin) and in 32-35 ℃ cultivate 2-3d, the centrifugal supernatant liquor that goes, lyophilize obtains the Haematocoocus Pluvialls algae powder of enrichment astaxanthin; Cultivate enrichment astaxanthin Haematocoocus Pluvialls by regulation and control illumination and temperature;
(5) preparation of astaxanthin dry powder: the Haematocoocus Pluvialls algae powder of the enrichment astaxanthin of the koning trichoderma fermented product of step (2) gained and step (4) gained is mixed, homogeneous, centrifugal, cross leaching solid-phase, lyophilize, obtains astaxanthin dry powder; Due to the cell walls of rich cellulose enzyme degradable Haematocoocus Pluvialls in koning trichoderma fermented product, and homogeneous can further make frustule fragmentation, discharge astaxanthin, because astaxanthin is water insoluble, so centrifugal rear astaxanthin is present in non-dissolving phase, be solid-phase, this solid-phase after drying, obtains astaxanthin dry powder.
(6) extraction of astaxanthin: with the dry powder that is rich in astaxanthin of Semen Maydis oil extraction step (5) gained, get final product to obtain astaxanthin; Because astaxanthin is fat-soluble, can effectively extract astaxanthin with Semen Maydis oil.
Therein in some embodiment, composition and the weight part proportioning of the described PDY liquid nutrient medium of step (1) are as follows: potato 18-22wt%, and glucose 1.5-2.5wt%, yeast extract paste 0.8-1.2wt%, surplus is water.
Therein in some embodiment, the described koning trichoderma spore suspension access amount of step (1) is: the weight part proportioning of koning trichoderma spore suspension and PDY liquid nutrient medium is 1:9-11, and in described koning trichoderma spore suspension, the concentration of koning trichoderma spore is 0.04-0.06mg/L.
Therein in some embodiment, the condition of the described cultivation of step (1) is temperature 23-27 ℃, rotating speed 120-150r/min.
Therein in some embodiment, the access amount of the described koning trichoderma bacterium mud of step (2) is: the weight part proportioning of koning trichoderma bacterium mud and solid fermentation substratum is 1:490-560.
Therein in some embodiment, composition and the weight part of the described solid fermentation substratum of step (2) are joined as follows: dregs of beans 0.9-1.1 part, straw 11-13 part, wheat bran 9-11 part, Xylo-Mucine 0.75-0.85 part, tween-80 0.009-0.011 part, (NH
4)
2sO
40.07-0.09 part, water 28-30 part.
Therein in some embodiment, the parameter of the described closed photo bioreactor of step (3) is: temperature 25-28 ℃, illumination 55-65 μ molm
-2s
-1.
Therein in some embodiment, composition and the weight part proportioning of the described BBM substratum of step (3) are as follows: SODIUMNITRATE 245-255 part; Potassium primary phosphate 170-180 part; Dipotassium hydrogen phosphate 70-80 part; Magnesium sulfate heptahydrate 70-80 part; Calcium dichloride dihydrate 23-38 part; Sodium-chlor 23-38 part; EDTA48-53 part; Potassium hydroxide 29-33 part; Iron vitriol 4.8-5 part; Boric acid 11-12 part; Seven water cure zinc 8.5-9 parts; Manganous chloride tetrahydrate 1.4-1.5 part; Molybdenum oxide 0.7-0.8 part; Cupric sulfate pentahydrate 1.5-1.6 part; Cobalt nitrate hexahydrate 0.7-0.8 part, distilled water 1000ml.
Therein in some embodiment, the access amount of the described Haematocoocus Pluvialls of step (3) is: in every 100mLBBM substratum, access Haematocoocus Pluvialls 3-5g.
Therein in some embodiment, composition and the weight part proportioning of the described BBM substratum of step (4) are as follows: magnesium sulfate heptahydrate 70-80 part, Calcium dichloride dihydrate 23-38 part, sodium-chlor 23-38 part, EDTA48-53 part, potassium hydroxide 29-33 part, iron vitriol 4.8-5 part, boric acid 11-12 part, seven water cure zinc 8.5-9 parts, Manganous chloride tetrahydrate 1.4-1.5 part, molybdenum oxide 0.7-0.8 part, cupric sulfate pentahydrate 1.5-1.6 part, distilled water 1000ml.
Therein in some embodiment, the access amount of the described Haematocoocus Pluvialls algae mud of step (4) is: in every 100mL BBM substratum, access Haematocoocus Pluvialls algae mud 20-25g.
A kind of method of utilizing enzyme to produce astaxanthin of the present invention has the following advantages and beneficial effect:
The present invention can obtain natural astaxanthin efficiently, and technique is simple, and cost is low, low in the pollution of the environment, and does not produce poisonous by product during the fermentation, and later purification method of purification is simple and easy, is applicable to industrialization promotion.
Embodiment
The present invention adopts koning trichoderma solid fermentation to prepare the fermented product of rich cellulose enzyme, cultivate enrichment astaxanthin Haematocoocus Pluvialls by regulation and control illumination and temperature, fermented product can efficiently decompose haematococcus pluvialis cell wall, further destroy cytolemma through homogeneous, the astaxanthin of enrichment in Haematocoocus Pluvialls body is discharged, and centrifuging and taking is not dissolved phase, lyophilize, finally utilize Semen Maydis oil extraction, obtain astaxanthin.
Below with reference to specific embodiment, the present invention is further elaborated.
Embodiment 1
Utilize enzyme to produce a method for astaxanthin, comprise the following steps:
(1) cultivation of koning trichoderma: with under aseptic washing, the concentration of making koning trichoderma spore is 0.05mg/L spore suspension by the koning trichoderma spore on slant medium; At the PDY of 1000ml substratum, (this substratum is containing potato 20wt%, glucose 2wt%, yeast extract paste 1wt%, surplus is water, above-mentioned substance amounts to 100wt%) middle access 100ml koning trichoderma spore suspension, under 25 ℃, 120～150r/min condition, cultivate 2d, the centrifugal supernatant liquor that goes, collects bacterium mud;
(2) cultivation of koning trichoderma fermented product: the cultured koning trichoderma bacterium of step (1) bacterium mud is got to 100g and be inoculated into solid fermentation substratum (this substratum is containing dregs of beans 1kg, straw 12kg, wheat bran 10kg, CMC-Na0.8kg, tween-80 0.01kg, (NH
4)
2sO
40.08kg, water 29kg), get in the solid fermentation substratum of koning trichoderma bacterium mud access 52.89kg of 100g step (1) gained, at 30-32 ℃, cultivate 30h, after at 26-28 ℃, cultivate 88h, the centrifugal supernatant liquor that goes, and with the centrifugal gained solid-phase of sterile water wash, obtain koning trichoderma fermented product;
(3) spreading cultivation of Haematocoocus Pluvialls: get 40g Haematocoocus Pluvialls access BBM substratum (SODIUMNITRATE 250mg, potassium primary phosphate 175mg, dipotassium hydrogen phosphate 75mg, magnesium sulfate heptahydrate 75mg, Calcium dichloride dihydrate 25mg, sodium-chlor 25mg, EDTA50mg, potassium hydroxide 31mg, iron vitriol 4.98mg, boric acid 11.42mg, seven water cure zinc 8.82mg, Manganous chloride tetrahydrate 1.44mg, molybdenum oxide 0.71mg, cupric sulfate pentahydrate 1.57mg, cobalt nitrate hexahydrate 0.75mg, distilled water 1000ml) and be placed in closed photo bioreactor, the parameter of closed photo bioreactor is: 27 ℃ of temperature, continuous illumination under 60 μ mol.m-2.s-1, blowing air is cultivated 2d simultaneously, centrifuging and taking Haematocoocus Pluvialls algae mud,
(4) preparation of the Haematocoocus Pluvialls algae powder of enrichment astaxanthin: by Haematocoocus Pluvialls algae mud 230g access BBM substratum (magnesium sulfate heptahydrate 75mg, Calcium dichloride dihydrate 25mg, the sodium-chlor 25mg of step (3) gained, EDTA50mg, potassium hydroxide 31mg, iron vitriol 4.98mg, boric acid 11.42mg, seven water cure zinc 8.82mg, Manganous chloride tetrahydrate 1.44mg, molybdenum oxide 0.71mg, cupric sulfate pentahydrate 1.57mg, distilled water 1000ml) in, pass into CO
2air flow be 0.5～1.5L/ (Lmin) and in 32-35 ℃ cultivate 2.5d, obtain algae liquid, algae liquid removes supernatant liquor through sedimentation, centrifugal concentrating, lyophilize obtains the Haematocoocus Pluvialls algae powder of enrichment astaxanthin;
(5) preparation of the dry powder of astaxanthin: the Haematocoocus Pluvialls algae powder of the enrichment astaxanthin of the koning trichoderma fermented product of step (2) gained and step (4) gained is mixed, homogeneous, centrifugal, cross leaching solid-phase, lyophilize, obtains the dry powder of astaxanthin;
(6) extraction of astaxanthin: with the dry powder that is rich in astaxanthin of Semen Maydis oil extraction step (5) gained, get final product to such an extent that Astaxanthin extraction rate is 0.97% ~ 0.98%.
Embodiment 2
Utilize enzyme to produce a method for astaxanthin, comprise the following steps:
(1) cultivation of koning trichoderma: with under aseptic washing, the concentration of making koning trichoderma spore is 0.04mg/L spore suspension by the koning trichoderma spore on slant medium; At the PDY of 1000ml substratum, (this substratum is containing potato 18wt%, glucose 1.5wt%, yeast extract paste 0.8wt%, surplus is water, above-mentioned substance amounts to 100wt%) middle access 100ml koning trichoderma spore suspension, under 23 ℃, 120～150r/min condition, cultivate 1.5d, the centrifugal supernatant liquor that goes, collects bacterium mud;
(2) cultivation of koning trichoderma fermented product: the cultured koning trichoderma of step (1) is got to 100g and be inoculated into solid fermentation substratum (this substratum is containing dregs of beans 0.9kg, straw 11kg, wheat bran 9kg, CMC-Na0.75kg, tween-80 0.009kg, (NH
4)
2sO
40.07kg, water 28kg), get in the solid fermentation substratum of koning trichoderma bacterium mud access 49.73kg of 100g step (1) gained, at 30-32 ℃, cultivate 29h, after at 26-28 ℃, cultivate 76h, the centrifugal supernatant liquor that goes, and with the centrifugal gained solid-phase of sterile water wash, obtain koning trichoderma fermented product;
(3) spreading cultivation of Haematocoocus Pluvialls: by 31g Haematocoocus Pluvialls access BBM substratum (SODIUMNITRATE 245mg, potassium primary phosphate 170mg, dipotassium hydrogen phosphate 70mg, magnesium sulfate heptahydrate 70mg, Calcium dichloride dihydrate 23mg, sodium-chlor 23mg part, EDTA48mg, potassium hydroxide 29mg, iron vitriol 4.8mg, boric acid 11mg, seven water cure zinc 8.5mg, Manganous chloride tetrahydrate 1.4mg, molybdenum oxide 0.7mg, cupric sulfate pentahydrate 1.5mg, cobalt nitrate hexahydrate 0.7mg, , distilled water 1000ml) and be placed in closed photo bioreactor, the parameter of closed photo bioreactor is: 25 ℃ of temperature, continuous illumination under 55 μ mol.m-2.s-1, blowing air is cultivated 1.5d simultaneously, centrifuging and taking Haematocoocus Pluvialls algae mud,
(4) preparation of the Haematocoocus Pluvialls algae powder of enrichment astaxanthin: by Haematocoocus Pluvialls algae mud 210g access BBM substratum (Calcium dichloride dihydrate 23mg, sodium-chlor 23mg part of step (3) gained; EDTA48mg; Potassium hydroxide 29mg; Iron vitriol 4.8mg; Boric acid 11mg; Seven water cure zinc 8.5mg; Manganous chloride tetrahydrate 1.4mg; Molybdenum oxide 0.7mg; Cupric sulfate pentahydrate 1.5mg; Distilled water 1000ml) in, pass into CO
2air flow be 0.8～1.2L/ (Lmin) and in 32-35 ℃ cultivate 2d, obtain algae liquid, algae liquid removes supernatant liquor through sedimentation, centrifugal concentrating, lyophilize obtains the Haematocoocus Pluvialls algae powder of enrichment astaxanthin; In described BBM substratum, do not contain NaNO
3;
(5) preparation of the dry powder of astaxanthin: the Haematocoocus Pluvialls algae powder of the enrichment astaxanthin of the koning trichoderma fermented product of step (2) gained and step (4) gained is mixed, homogeneous, centrifugal, cross leaching solid-phase, lyophilize, obtains the dry powder of astaxanthin;
(6) extraction of astaxanthin: with the dry powder that is rich in astaxanthin of Semen Maydis oil extraction step (5) gained, get final product to such an extent that Astaxanthin extraction rate is 0.95% ~ 0.96%.
Embodiment 3
Utilize enzyme to produce a method for astaxanthin, comprise the following steps:
(1) cultivation of koning trichoderma: with under aseptic washing, the concentration of making koning trichoderma spore is 0.06mg/L spore suspension by the koning trichoderma spore on slant medium; At the PDY of 1000ml substratum, (this substratum is containing potato 22wt%, glucose 2.5wt%, yeast extract paste 1.2wt%, surplus is water, above-mentioned substance amounts to 100wt%) middle access 100ml koning trichoderma spore suspension, under 27 ℃, 120～150r/min condition, cultivate 1.5d, the centrifugal supernatant liquor that goes, collects bacterium mud;
(2) cultivation of koning trichoderma fermented product: the cultured koning trichoderma of step (1) is got to 100g and be inoculated into solid fermentation substratum (this substratum is containing dregs of beans 1.1kg, straw 13kg, wheat bran 11kg, CMC-Na0.85kg, tween-80 0.011kg, (NH
4)
2sO
40.09kg, water 30kg), get in the solid fermentation substratum of koning trichoderma bacterium mud access 56.05kg of 100g step (1) gained, at 30-32 ℃, cultivate 31h, after at 26-28 ℃, cultivate 81h, the centrifugal supernatant liquor that goes, and with the centrifugal gained solid-phase of sterile water wash, obtain koning trichoderma fermented product;
(3) spreading cultivation of Haematocoocus Pluvialls: by 49g Haematocoocus Pluvialls access BBM substratum (SODIUMNITRATE 255mg, potassium primary phosphate 180mg, dipotassium hydrogen phosphate 80mg, magnesium sulfate heptahydrate 80mg, Calcium dichloride dihydrate 38mg, sodium-chlor 38mg, EDTA53mg, potassium hydroxide 33mg, iron vitriol 5mg, boric acid 12mg, seven water cure zinc 9mg, Manganous chloride tetrahydrate 1.5mg, molybdenum oxide 0.8mg, cupric sulfate pentahydrate 1.6mg, cobalt nitrate hexahydrate 0.8mg, distilled water 1000ml) and be placed in closed photo bioreactor, the parameter of closed photo bioreactor is: 28 ℃ of temperature, continuous illumination under 65 μ mol.m-2.s-1, blowing air is cultivated 2.5d simultaneously, centrifuging and taking Haematocoocus Pluvialls algae mud,
(4) preparation of the Haematocoocus Pluvialls algae powder of enrichment astaxanthin: the Haematocoocus Pluvialls algae mud 248g of step (3) gained is met to BBM substratum (magnesium sulfate heptahydrate 80mg, Calcium dichloride dihydrate 38mg, sodium-chlor 38mg, EDTA53mg, potassium hydroxide 33mg, iron vitriol 5mg, boric acid 12mg, seven water cure zinc 9mg, Manganous chloride tetrahydrate 1.5mg, molybdenum oxide 0.8mg, cupric sulfate pentahydrate 1.6mg, distilled water 1000ml) in, pass into CO
2air flow be 1～1.4L/ (Lmin) and in 32-35 ℃ cultivate 3d, obtain algae liquid, algae liquid removes supernatant liquor through sedimentation, centrifugal concentrating, lyophilize obtains the Haematocoocus Pluvialls algae powder of enrichment astaxanthin; In described BBM substratum, do not contain NaNO
3;
(5) preparation of the dry powder of astaxanthin: the Haematocoocus Pluvialls algae powder of the enrichment astaxanthin of the koning trichoderma fermented product of step (2) gained and step (4) gained is mixed, homogeneous, centrifugal, cross leaching solid-phase, lyophilize, obtains the dry powder of astaxanthin;
(6) extraction of astaxanthin: with the dry powder that is rich in astaxanthin of Semen Maydis oil extraction step (5) gained, get final product to such an extent that Astaxanthin extraction rate is 0.95% ~ 0.96%.
The above embodiment has only expressed several embodiment of the present invention, and it describes comparatively concrete and detailed, but can not therefore be interpreted as the restriction to the scope of the claims of the present invention.It should be pointed out that for the person of ordinary skill of the art, without departing from the inventive concept of the premise, can also make some distortion and improvement, these all belong to protection scope of the present invention.Therefore, the protection domain of patent of the present invention should be as the criterion with claims.
Claims (7)
1. utilize enzyme to produce a method for astaxanthin, it is characterized in that, comprise the following steps:
(1) cultivation of koning trichoderma: by koning trichoderma spore suspension access PDY liquid nutrient medium, cultivate 1.5-2.5d, the centrifugal supernatant liquor that goes, obtains koning trichoderma bacterium bacterium mud;
(2) cultivation of koning trichoderma fermented product: get in the koning trichoderma bacterium bacterium mud access solid fermentation substratum of step (1) gained, at 30-32 ℃, cultivate 29-31h, after at 26-28 ℃, cultivate 75-90h, the centrifugal supernatant liquor that goes, obtains koning trichoderma fermented product; The access amount of described koning trichoderma bacterium bacterium mud is: the weight part proportioning of koning trichoderma bacterium bacterium mud and solid fermentation substratum is 1:490-560; Composition and the weight part of described solid fermentation substratum are joined as follows: dregs of beans 0.9-1.1 part, straw 11-13 part, wheat bran 9-11 part, Xylo-Mucine 0.75-0.85 part, tween-80 0.009-0.011 part, (NH
4)
2sO
40.07-0.09 part, water 28-30 part;
(3) spreading cultivation of Haematocoocus Pluvialls: Haematocoocus Pluvialls is accessed to BBM substratum and is placed in closed photo bioreactor, and blowing air is cultivated 1.5-2.5d, and the centrifugal supernatant liquor that goes, obtains Haematocoocus Pluvialls algae mud; The parameter of described closed photo bioreactor is: temperature 25-28 ℃, illumination 55-65 μ molm
-2s
-1;
(4) preparation of the Haematocoocus Pluvialls algae powder of enrichment astaxanthin: by the Haematocoocus Pluvialls algae mud access BBM substratum of step (3) gained, pass into CO
2, air flow be 0.5～1.5L/ (Lmin) and in 32-35 ℃ cultivate 2-3d, the centrifugal supernatant liquor that goes, lyophilize obtains the Haematocoocus Pluvialls algae powder of enrichment astaxanthin;
(5) preparation of astaxanthin dry powder: the Haematocoocus Pluvialls algae powder of the enrichment astaxanthin of the koning trichoderma fermented product of step (2) gained and step (4) gained is mixed, homogeneous, centrifugal, cross leaching solid-phase, lyophilize, obtains astaxanthin dry powder;
(6) extraction of astaxanthin: with the dry powder that is rich in astaxanthin of Semen Maydis oil extraction step (5) gained, get final product to obtain astaxanthin.
2. enzyme according to claim 1 is produced the method for astaxanthin, it is characterized in that, composition and the weight part proportioning of the described PDY liquid nutrient medium of step (1) are as follows: potato 18-22wt%, glucose 1.5-2.5wt%, yeast extract paste 0.8-1.2wt%, surplus is water.
3. enzyme according to claim 1 is produced the method for astaxanthin, it is characterized in that, the described koning trichoderma spore suspension access amount of step (1) is: the weight part proportioning of koning trichoderma spore suspension and PDY liquid nutrient medium is 1:9-11, in described koning trichoderma spore suspension, the concentration of koning trichoderma spore is 0.04-0.06mg/L, and described culture condition is temperature 23-27 ℃, rotating speed 120-150r/min.
4. enzyme according to claim 1 is produced the method for astaxanthin, it is characterized in that, composition and the weight part proportioning of the BBM substratum described in step (3) are as follows: SODIUMNITRATE 245-255 part, potassium primary phosphate 170-180 part, dipotassium hydrogen phosphate 70-80 part, magnesium sulfate heptahydrate 70-80 part, Calcium dichloride dihydrate 23-38 part, sodium-chlor 23-38 part, EDTA48-53 part, potassium hydroxide 29-33 part, iron vitriol 4.8-5 part, boric acid 11-12 part, seven water cure zinc 8.5-9 parts, Manganous chloride tetrahydrate 1.4-1.5 part, molybdenum oxide 0.7-0.8 part, cupric sulfate pentahydrate 1.5-1.6 part, cobalt nitrate hexahydrate 0.7-0.8 part, distilled water 1000ml.
5. enzyme according to claim 1 is produced the method for astaxanthin, it is characterized in that, the access amount of the described Haematocoocus Pluvialls of step (3) is: in every 100mL BBM substratum, access Haematocoocus Pluvialls 3-5g.
6. enzyme according to claim 1 is produced the method for astaxanthin, it is characterized in that, composition and the weight part proportioning of the described BBM substratum of step (4) are as follows: magnesium sulfate heptahydrate 70-80 part, Calcium dichloride dihydrate 23-38 part, sodium-chlor 23-38 part, EDTA48-53 part, potassium hydroxide 29-33 part, iron vitriol 4.8-5 part, boric acid 11-12 part, seven water cure zinc 8.5-9 parts, Manganous chloride tetrahydrate 1.4-1.5 part, molybdenum oxide 0.7-0.8 part, cupric sulfate pentahydrate 1.5-1.6 part, distilled water 1000ml.
7. enzyme according to claim 1 is produced the method for astaxanthin, it is characterized in that, the access amount of the described Haematocoocus Pluvialls algae mud of step (4) is: in every 100mL BBM substratum, access Haematocoocus Pluvialls algae mud 20-25g.
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| CN104232721A (en) * | 2014-08-26 | 2014-12-24 | 中国科学院天津工业生物技术研究所 | Method for inducing haematococcus pluvialis to quickly accumulate astaxanthin by using tungstate |
| CN104529851A (en) * | 2014-12-06 | 2015-04-22 | 黑龙江众生生物工程有限公司 | New technology for extracting astaxanthin from Haematococcus pluvialis through enzyme method |
| CN104856051B (en) * | 2015-04-10 | 2017-07-14 | 杭州鑫伟低碳技术研发有限公司 | A kind of method of utilization Haematococcus pluvialis production micro-capsule astaxanthin powder |
| CN105028636A (en) * | 2015-07-07 | 2015-11-11 | 南京师范大学 | Compound pearl barley-millet flavor functional yoghurt and preparation method thereof |
| AU2016381993B2 (en) | 2015-12-28 | 2019-04-04 | Eneos Corporation | Method for producing fermented carotenoid using carotenoid-producing bacteria obtained by using cobalt-containing culturing medium |
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| CN1181184C (en) * | 2002-07-26 | 2004-12-22 | 中国科学院武汉植物研究所 | Method for producing astaxanthin by cultivating haematococcus pulvialis |
| CN1181209C (en) * | 2003-02-24 | 2004-12-22 | 天津大学 | Algae with astaxanthin production and astaxanthin production by yeast mixed culture fermentation |
| EP1749890A1 (en) * | 2004-05-26 | 2007-02-07 | Yamaha Motor Co., Ltd. | Method of producing xanthophyll |
| DK1681060T3 (en) * | 2005-01-15 | 2010-09-06 | Cognis Ip Man Gmbh | Improved method for obtaining carotene and carotenoids from algae or cyanobacteria |
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