阅读说明：本技术 一种叶黄素提高雨生红球藻中虾青素含量的生产工艺 (Production process for increasing astaxanthin content in haematococcus pluvialis by lutein ) 是由 赵宇翔 于 2021-09-14 设计创作，主要内容包括：本发明公开了一种叶黄素提高雨生红球藻中虾青素含量的生产工艺,该生产工艺旨在解决现有技术下采用雨生红球藻对虾青素进行提取生产时候含量不高,且在进行加工的时候,还会出现加工原料提取不完全的技术问题。该生产工艺,将雨生红球藻细胞接种进行扩增培养,对扩增培养物进行诱导培养,诱导藻细胞积累虾青素,使用雨生红球藻制备内容物料,通过增加了的叶黄素来提高雨生红球藻中虾青素的含量,同时通过加工工艺的优化,更好的进行加工生产的工作,提高工作的速度和效率,克服了皂化过程导致的虾青素损耗,使得高纯度虾青素酯产率高,工艺简单、简洁,所用分离纯化溶剂符合食品添加剂要求,且工艺操作简单,易于实现工业化。(The invention discloses a production process for increasing astaxanthin content in haematococcus pluvialis by lutein, which aims to solve the technical problems that the content is not high when astaxanthin is extracted and produced by haematococcus pluvialis in the prior art, and incomplete extraction of processing raw materials can occur during processing. The production process comprises the steps of inoculating haematococcus pluvialis cells for amplification culture, carrying out induction culture on an amplification culture, inducing the haematococcus pluvialis cells to accumulate astaxanthin, preparing a content material by using the haematococcus pluvialis, improving the content of the astaxanthin in the haematococcus pluvialis by adding lutein, optimizing a processing process, better carrying out processing production work, improving the speed and efficiency of the work, overcoming astaxanthin loss caused by a saponification process, ensuring high yield of high-purity astaxanthin ester, being simple and concise in process, enabling a used separation and purification solvent to meet the requirements of food additives, being simple in process operation and easy to realize industrialization.)

1. A production process for increasing the content of astaxanthin in haematococcus pluvialis by lutein is characterized by comprising the following steps:

the method comprises the following steps: inoculating haematococcus pluvialis cells, carrying out amplification culture to obtain an amplification culture, carrying out induction culture on the amplification culture to induce the haematococcus pluvialis cells to accumulate astaxanthin to obtain an induction culture, separating the haematococcus pluvialis cells from the induction culture to obtain the haematococcus pluvialis, and carrying out amplification culture on the haematococcus pluvialis cells in an amplification culture medium;

step two: separating algae cells from the amplified culture, transferring the algae cells to a BG11 culture medium with nitrogen deficiency for culturing for 3-5 days, and adding a biological inducer for induction culture for 24-48h to obtain an induction culture, wherein the inducer is a bacillus subtilis fermentation product of oat;

step three: preparing a content material by using haematococcus pluvialis, standing and settling collected fresh haematococcus pluvialis, removing supernatant, performing homogeneous wall breaking to obtain wall-broken algae slurry, adding lipase into the wall-broken algae slurry for enzymolysis to obtain enzymolysis wall-broken algae liquid, extracting algae oil to obtain haematococcus pluvialis algae oil and algae residues, weighing 150-600g of auxiliary materials, placing the lutein in a high-pressure reaction kettle, adding 15-25mL of emulsifier, stirring and mixing uniformly, then adding 5-40g of reaction catalyst and 20-60g of phase transfer catalyst into the reaction kettle, and reacting for 10-30h at 90-150 ℃ to obtain zeaxanthin;

step four: weighing 30-80g of giant knotweed and cassia seed respectively, grinding the giant knotweed and cassia seed into powder, adding 100mL of 40-90% ethanol solvent, refluxing, extracting and filtering the powder to obtain resveratrol, weighing 20-60g of protocatechuic acid and gallic acid respectively, mixing and stirring the mixture together, adding 10-30g of citric acid and 5-25g of alpha-glucoheptonate into the mixture, adding water, uniformly mixing, and heating in a water bath to obtain a composite antioxidant;

step five: preparing a BBM culture medium with sufficient nitrogen, adding sodium acetate or magnesium acetate into the BBM culture medium, performing high-pressure moist heat sterilization on the culture medium, wherein the adding amount of the sodium acetate is 1-5g/L, the adding amount of the magnesium acetate is 1-2.5 g/L, inoculating haematococcus pluvialis seed liquid cultured to the later stage of logarithmic phase growth into the culture medium for culture, ensuring the concentration of the haematococcus pluvialis to be 2.0 x 105-2.5 x 105cells/mL, sampling every other day to measure the biomass of the haematococcus pluvialis, making a growth curve of the haematococcus pluvialis in the culture medium, and collecting the haematococcus pluvialis with the maximum biomass;

step six: weighing 0.5-2.4g of halogenating agent, dissolving with 1-15g of chloroform, slowly adding into a reactor, reacting for 1-15h, and washing with water for three times to remove alkali and salt to obtain a primary solution;

step seven: and adding the composite antioxidant obtained in the fourth step into the primary liquid, adding a silane coupling agent into the primary liquid, uniformly mixing, heating in a water bath, and filtering to obtain the astaxanthin.

2. The production process of lutein for increasing astaxanthin content in haematococcus pluvialis according to claim 1, wherein the pressure for homogeneous wall breaking in the first step is 0MPa-100MPa, and the wall breaking temperature is 40 ℃ to 60 ℃.

3. The production process of lutein for increasing the astaxanthin content in haematococcus pluvialis according to claim 1, wherein the lipase dosage in the first step accounts for 5% -10% of the mass of the wall-broken algae pulp, the enzymolysis temperature is 35-45 ℃, the enzymolysis time is 18-24h, and the enzymolysis pH value is 6.0-8.0.

4. The production process of lutein for increasing astaxanthin content in haematococcus pluvialis according to claim 1, wherein in the first step, 1-15 parts by weight of haematococcus pluvialis oil, 1-5 parts by weight of algae residues, 1-10 parts by weight of lutein ester, 5-15 parts by weight of vitamin C and 20-40 parts by weight of vegetable oil are mixed, crushed, mixed uniformly and sieved by a 180-mesh sieve to obtain the content material.

5. The production process of lutein for increasing astaxanthin content in Haematococcus pluvialis according to claim 1, wherein the conditions of high pressure wet heat sterilization in the third step are 90-150 ℃, and the treatment time is 10-30 min.

6. The production process of lutein for increasing astaxanthin content in Haematococcus pluvialis according to claim 1, wherein the emulsifier is sodium dodecyl sulfate.

7. The production process of lutein for increasing astaxanthin content in Haematococcus pluvialis according to claim 1, wherein the catalyst is phosphorous acid.

8. The production process of lutein for increasing astaxanthin content in haematococcus pluvialis according to claim 1, wherein the water bath heating temperature in the fourth step is 50-90 ℃, and the water bath time is 10-40 min.

9. The production process of lutein for increasing astaxanthin content in Haematococcus pluvialis according to claim 1, wherein the stirring speed in the fourth step is 150-600r/min, and the stirring time is 10-50 min.

Technical Field

The invention belongs to the field of astaxanthin production, and particularly relates to a production process for increasing the content of astaxanthin in haematococcus pluvialis by using lutein.

Background

Astaxanthin is a ketone carotenoid, red solid powder, is fat-soluble, insoluble in water, soluble in organic solvents, widely exists in the biological world, particularly in feathers of aquatic animals such as shrimps, crabs, fish and birds, has the function of developing color, has extremely strong oxidation resistance, can remove nitrogen dioxide, sulfide, disulfide and the like, can also reduce lipid peroxidation, and effectively inhibits lipid peroxidation caused by free radicals, and the main production method of the astaxanthin comprises two modes of artificial synthesis and biological acquisition. The artificially synthesized astaxanthin is only expensive, while the biological source of natural astaxanthin is most suitable for large-scale production of astaxanthin accumulated by haematococcus pluvialis, but the astaxanthin is slowly grown and easily polluted by other organisms in the culture process, and the astaxanthin in the haematococcus pluvialis cannot be completely extracted.

At present, the invention patent with the patent number of CN202110275002.1 discloses a method for extracting astaxanthin from haematococcus pluvialis, firstly, an ultrasonic cell disruptor can be used for effectively disrupting cell walls of the haematococcus pluvialis, then an organic solvent is used for extracting astaxanthin, and the extraction efficiency of the astaxanthin is obviously improved by adjusting and improving the extraction solvent, the extraction time, the extraction temperature, the extraction times, the feed-liquid ratio and the like in a targeted manner, so that the comprehensive benefit ratio is strong. The astaxanthin content produced by the extraction method is not high, only the processing speed and efficiency are improved, the working yield is not high, and different processing and production requirements cannot be met.

Therefore, in order to solve the problem of low extraction content of astaxanthin, it is necessary to improve the use scenario of astaxanthin.

Disclosure of Invention

(1) Technical problem to be solved

Aiming at the defects of the prior art, the invention aims to provide a production process for increasing the content of astaxanthin in haematococcus pluvialis by lutein, which aims to solve the technical problems that the content is not high when the haematococcus pluvialis is used for extracting astaxanthin in the prior art, and the extraction of processing raw materials is incomplete when the haematococcus pluvialis is processed.

(2) Technical scheme

In order to solve the technical problems, the invention provides a production process for increasing the content of astaxanthin in haematococcus pluvialis by lutein, which comprises the following steps:

the method comprises the following steps: inoculating haematococcus pluvialis cells, carrying out amplification culture to obtain an amplification culture, carrying out induction culture on the amplification culture to induce the haematococcus pluvialis cells to accumulate astaxanthin to obtain an induction culture, separating the haematococcus pluvialis cells from the induction culture to obtain the haematococcus pluvialis, and carrying out amplification culture on the haematococcus pluvialis cells in an amplification culture medium;

step two: separating algae cells from the amplified culture, transferring the algae cells to a BG11 culture medium with nitrogen deficiency for culturing for 3-5 days, and adding a biological inducer for induction culture for 24-48h to obtain an induction culture, wherein the inducer is a bacillus subtilis fermentation product of oat;

step three: preparing a content material by using haematococcus pluvialis, standing and settling collected fresh haematococcus pluvialis, removing supernatant, performing homogeneous wall breaking to obtain wall-broken algae slurry, adding lipase into the wall-broken algae slurry for enzymolysis to obtain enzymolysis wall-broken algae liquid, extracting algae oil to obtain haematococcus pluvialis algae oil and algae residues, weighing 150-600g of auxiliary materials, placing the lutein in a high-pressure reaction kettle, adding 15-25mL of emulsifier, stirring and mixing uniformly, then adding 5-40g of reaction catalyst and 20-60g of phase transfer catalyst into the reaction kettle, and reacting for 10-30h at 90-150 ℃ to obtain zeaxanthin;

step four: weighing 30-80g of giant knotweed and cassia seed respectively, grinding the giant knotweed and cassia seed into powder, adding 100mL of 40-90% ethanol solvent, refluxing, extracting and filtering the powder to obtain resveratrol, weighing 20-60g of protocatechuic acid and gallic acid respectively, mixing and stirring the mixture together, adding 10-30g of citric acid and 5-25g of alpha-glucoheptonate into the mixture, adding water, uniformly mixing, and heating in a water bath to obtain a composite antioxidant;

step five: preparing a BBM culture medium with sufficient nitrogen, adding sodium acetate or magnesium acetate into the BBM culture medium, performing high-pressure moist heat sterilization on the culture medium, wherein the adding amount of the sodium acetate is 1-5g/L, the adding amount of the magnesium acetate is 1-2.5 g/L, inoculating haematococcus pluvialis seed liquid cultured to the later stage of logarithmic phase growth into the culture medium for culture, ensuring the concentration of the haematococcus pluvialis to be 2.0 x 105-2.5 x 105cells/mL, sampling every other day to measure the biomass of the haematococcus pluvialis, making a growth curve of the haematococcus pluvialis in the culture medium, and collecting the haematococcus pluvialis with the maximum biomass;

step six: weighing 0.5-2.4g of halogenating agent, dissolving with 1-15g of chloroform, slowly adding into a reactor, reacting for 1-15h, and washing with water for three times to remove alkali and salt to obtain a primary solution;

step seven: and adding the composite antioxidant obtained in the fourth step into the primary liquid, adding a silane coupling agent into the primary liquid, uniformly mixing, heating in a water bath, and filtering to obtain the astaxanthin.

When the production process of the technical scheme is used, haematococcus pluvialis cells are inoculated for amplification culture to obtain an amplification culture, the amplification culture is subjected to induction culture, astaxanthin is accumulated in the induced haematococcus pluvialis cells to obtain an induced culture, haematococcus pluvialis is separated from the induced culture and is subjected to amplification culture in an amplification culture medium, haematococcus pluvialis cells are separated from the amplification culture and are transferred to a nitrogen-deficient BG11 culture medium for culture for 3-5 days, a biological inducer is added for induction culture for 24-48 hours to obtain the induced culture, the inducer is a bacillus subtilis fermentation product of oat, the haematococcus pluvialis is used for preparing a content material, the collected fresh haematococcus pluvialis is subjected to standing and sedimentation, the supernatant is removed, the wall-broken algae slurry is subjected to homogenization and wall breaking, lipase is added into the algae slurry for enzymolysis to obtain an enzymolysis algae solution, used for extracting algae oil to obtain haematococcus pluvialis algae oil and algae residues, weighing 600g of lutein, placing the lutein into a high-pressure reaction kettle, adding 15-25mL of emulsifier, stirring and mixing uniformly, then adding 5-40g of reaction catalyst and 20-60g of phase transfer catalyst into the reaction kettle, reacting for 10-30h at 90-150 ℃ to obtain zeaxanthin, weighing 30-80g of polygonum cuspidatum and cassia seed respectively, grinding the polygonum cuspidatum and cassia seed into powder, adding 100mL of ethanol solvent with the concentration of 40-90%, refluxing, extracting and filtering the mixture to obtain resveratrol, weighing 20-60g of protocatechuic acid and gallic acid respectively, mixing and stirring the protocatechuic acid and the gallic acid together, adding 10-30g of citric acid and 5-25g of alpha-glucoheptonate into the mixture, mixing uniformly, adding water, heating in a water bath to obtain a composite antioxidant, preparing BBM culture medium with sufficient nitrogen, adding sodium acetate or magnesium acetate, performing high-pressure moist heat sterilization on the culture medium, wherein the adding amount of sodium acetate is 1-5g/L, the adding amount of magnesium acetate is 1-2.5 g/L, inoculating Haematococcus pluvialis seed liquid cultured to the later logarithmic phase of growth into the culture medium for culturing, ensuring the algae concentration to be 2.0 x 105-2.5 x 105cells/mL, sampling every other day to measure the biomass, making the growth curve of the Haematococcus pluvialis in the culture medium, collecting the Haematococcus pluvialis with the maximum biomass, weighing 0.5-2.4g of halogenating agent, dissolving with 1-15g of chloroform, slowly adding into a reactor for reaction for 1-15h, removing alkali and salt through three times of water washing to obtain primary liquid, adding the composite antioxidant in the fourth step into the primary liquid, adding silane coupling agent into the primary liquid, after being mixed evenly, the astaxanthin is obtained after being heated in water bath and filtered.

Preferably, the pressure used for homogenizing wall breaking in the first step is 0MPa-100MPa, the wall breaking temperature is 40-60 ℃, and the wall breaking rate is more than or equal to 95%.

Preferably, the lipase in the first step accounts for 5-10% of the mass of the wall-broken algae slurry, the enzymolysis temperature is 35-45 ℃, the enzymolysis time is 18-24h, and the enzymolysis pH value is 6.0-8.0.

Preferably, in the first step, 1-15 parts by weight of haematococcus pluvialis oil, 1-5 parts by weight of algae residue, 1-10 parts by weight of lutein ester, 5-15 parts by weight of vitamin C and 20-40 parts by weight of vegetable oil are mixed, crushed, mixed uniformly and sieved by a 180-mesh sieve to obtain the content material.

Preferably, the conditions of the high-pressure damp-heat sterilization in the third step are 90-150 ℃, and the treatment time is 10-30 min.

Preferably, the emulsifier is sodium lauryl sulfate.

Preferably, the catalyst is phosphorous acid.

Preferably, the water bath heating temperature in the fourth step is 50-90 ℃, and the water bath time is 10-40 min.

Preferably, the stirring speed in the fourth step is 150-600r/min, and the stirring time is 10-50 min.

(3) Advantageous effects

Compared with the prior art, the invention has the beneficial effects that: the astaxanthin content produced by the production process is obviously higher than that of the original production process, the astaxanthin content in haematococcus pluvialis is improved by adding lutein, the processing and production work is better performed by optimizing the processing process, the working speed and efficiency are improved, the astaxanthin loss caused by the saponification process is overcome, the high-purity astaxanthin ester yield is high, the process is simple and concise, the used separation and purification solvent meets the requirements of food additives, the process operation is simple, and the industrialization is easy to realize.

Detailed Description

In order to make the technical means, the original characteristics, the achieved purposes and the effects of the invention easily understood and obvious, the technical solutions in the embodiments of the present invention are clearly and completely described below to further illustrate the invention, and obviously, the described embodiments are only a part of the embodiments of the present invention, but not all the embodiments.

Example 1

The specific embodiment is a production process for improving the astaxanthin content in haematococcus pluvialis, comprising the following steps:

the method comprises the following steps: inoculating haematococcus pluvialis cells, carrying out amplification culture to obtain an amplification culture, carrying out induction culture on the amplification culture to induce the haematococcus pluvialis cells to accumulate astaxanthin to obtain an induction culture, separating the haematococcus pluvialis cells from the induction culture to obtain the haematococcus pluvialis, and carrying out amplification culture on the haematococcus pluvialis cells in an amplification culture medium;

step two: separating algae cells from the amplified culture, transferring the algae cells to a BG11 culture medium with nitrogen deficiency for culturing for 5 days, and adding a biological inducer for induction culture for 48 hours to obtain an induced culture, wherein the inducer is a bacillus subtilis fermentation product of oat;

step three: preparing a content material by using haematococcus pluvialis, standing and settling collected fresh haematococcus pluvialis, removing supernatant, homogenizing and breaking walls to obtain wall-broken algae slurry, adding lipase into the wall-broken algae slurry for enzymolysis to obtain enzymolysis wall-broken algae liquid, extracting algae oil to obtain haematococcus pluvialis algae oil and algae residues, weighing 600g of lutein, placing the lutein in a high-pressure reaction kettle, adding 25mL of emulsifier, stirring and mixing uniformly, then adding 40g of a reaction catalyst and 60g of a phase transfer catalyst into the reaction kettle, and reacting for 30 hours at 150 ℃ to obtain zeaxanthin;

step four: weighing 80g of giant knotweed and cassia seed respectively, grinding the giant knotweed and cassia seed into powder, adding 100mL of 90% ethanol solvent, performing reflux extraction and filtration on the powder to obtain resveratrol, weighing 60g of protocatechuic acid and gallic acid respectively, mixing and stirring the protocatechuic acid and the gallic acid together, adding 30g of citric acid and 25g of alpha-glucoheptonate into the mixture, adding water, uniformly mixing, and heating in a water bath to obtain a composite antioxidant;

step five: preparing a BBM culture medium with sufficient nitrogen, adding sodium acetate or magnesium acetate, performing high-pressure moist heat sterilization on the culture medium, wherein the adding amount of the sodium acetate is 5g/L, the adding amount of the magnesium acetate is 2.5g/L, inoculating haematococcus pluvialis seed liquid cultured to the later logarithmic phase of growth into the culture medium for culturing, so that the concentration of the haematococcus pluvialis is 2.5 multiplied by 105cells/mL, sampling every other day to measure the biomass of the haematococcus pluvialis, making a growth curve of the haematococcus pluvialis in the culture medium, and collecting the haematococcus pluvialis with the maximum biomass;

step six: weighing 2.4g of halogenating agent, dissolving with 15g of chloroform, slowly adding into a reactor, reacting for 15h, and washing for three times to remove alkali and salt to obtain a primary solution;

step seven: and adding the composite antioxidant obtained in the fourth step into the primary liquid, adding a silane coupling agent into the primary liquid, uniformly mixing, heating in a water bath, and filtering to obtain the astaxanthin.

Example 2

The specific embodiment is a production process for improving the astaxanthin content in haematococcus pluvialis, comprising the following steps:

the method comprises the following steps: inoculating haematococcus pluvialis cells, carrying out amplification culture to obtain an amplification culture, carrying out induction culture on the amplification culture to induce the haematococcus pluvialis cells to accumulate astaxanthin to obtain an induction culture, separating the haematococcus pluvialis cells from the induction culture to obtain the haematococcus pluvialis, and carrying out amplification culture on the haematococcus pluvialis cells in an amplification culture medium;

step two: separating algae cells from the amplified culture, transferring the algae cells to a BG11 culture medium with nitrogen deficiency for culturing for 3 days, and adding a biological inducer for induction culture for 24 hours to obtain an induced culture, wherein the inducer is a bacillus subtilis fermentation product of oat;

step three: preparing a content material by using haematococcus pluvialis, standing and settling collected fresh haematococcus pluvialis, removing supernatant, performing homogeneous wall breaking to obtain wall-broken algae slurry, adding lipase into the wall-broken algae slurry for enzymolysis to obtain enzymolysis wall-broken algae liquid, extracting algae oil to obtain haematococcus pluvialis algae oil and algae residues, weighing 150g of lutein, placing the lutein in a high-pressure reaction kettle, adding 15mL of emulsifier, stirring and mixing uniformly, then adding 5g of a reaction catalyst and 20g of a phase transfer catalyst into the reaction kettle, and reacting for 30 hours at 150 ℃ to obtain zeaxanthin;

step four: weighing 80g of giant knotweed and cassia seed respectively, grinding the giant knotweed and cassia seed into powder, adding 100mL of 90% ethanol solvent, carrying out reflux extraction and filtration on the powder to obtain resveratrol, weighing 20g of protocatechuic acid and gallic acid respectively, mixing and stirring the protocatechuic acid and the gallic acid together, adding 10g of citric acid and 25g of alpha-glucoheptonate into the mixture, adding water, uniformly mixing, and heating in a water bath to obtain a composite antioxidant;

step five: preparing a BBM culture medium with sufficient nitrogen, adding sodium acetate or magnesium acetate, performing high-pressure moist heat sterilization on the culture medium, wherein the adding amount of the sodium acetate is 5g/L, the adding amount of the magnesium acetate is 1g/L, inoculating haematococcus pluvialis seed liquid cultured to the later logarithmic phase of growth into the culture medium for culturing, enabling the concentration of the haematococcus pluvialis to be 2.0 x 105cells/mL, sampling every other day to measure the biomass of the haematococcus pluvialis, taking a growth curve of the haematococcus pluvialis in the culture medium, and collecting the haematococcus pluvialis with the maximum biomass;

step six: weighing 2.4g of halogenating agent, dissolving with 15g of chloroform, slowly adding into a reactor, reacting for 15h, and washing for three times to remove alkali and salt to obtain a primary solution;

step seven: and adding the composite antioxidant obtained in the fourth step into the primary liquid, adding a silane coupling agent into the primary liquid, uniformly mixing, heating in a water bath, and filtering to obtain the astaxanthin.

Example 3

The specific embodiment is a production process for improving the astaxanthin content in haematococcus pluvialis, comprising the following steps:

the method comprises the following steps: inoculating haematococcus pluvialis cells, carrying out amplification culture to obtain an amplification culture, carrying out induction culture on the amplification culture to induce the haematococcus pluvialis cells to accumulate astaxanthin to obtain an induction culture, separating the haematococcus pluvialis cells from the induction culture to obtain the haematococcus pluvialis, and carrying out amplification culture on the haematococcus pluvialis cells in an amplification culture medium;

step two: separating algae cells from the amplified culture, transferring the algae cells to a BG11 culture medium with nitrogen deficiency for culturing for 3 days, and adding a biological inducer for induction culture for 24 hours to obtain an induced culture, wherein the inducer is a bacillus subtilis fermentation product of oat;

step three: preparing a content material by using haematococcus pluvialis, standing and settling collected fresh haematococcus pluvialis, removing supernatant, homogenizing and breaking walls to obtain wall-broken algae slurry, adding lipase into the wall-broken algae slurry for enzymolysis to obtain enzymolysis wall-broken algae liquid, extracting algae oil to obtain haematococcus pluvialis algae oil and algae residues, weighing 150g of lutein, placing the lutein in a high-pressure reaction kettle, adding 25mL of emulsifier, stirring and mixing uniformly, then adding 5g of a reaction catalyst and 60g of a phase transfer catalyst into the reaction kettle, and reacting for 10 hours at 90 ℃ to obtain zeaxanthin;

step four: weighing 30g of giant knotweed and cassia seed respectively, grinding the giant knotweed and cassia seed into powder, adding 100mL of 90% ethanol solvent, carrying out reflux extraction and filtration on the powder to obtain resveratrol, weighing 20g of protocatechuic acid and gallic acid respectively, mixing and stirring the protocatechuic acid and the gallic acid together, adding 30g of citric acid and 25g of alpha-glucoheptonate into the mixture, adding water, uniformly mixing, and heating in a water bath to obtain a composite antioxidant;

step five: preparing a BBM culture medium with sufficient nitrogen, adding sodium acetate or magnesium acetate, performing high-pressure moist heat sterilization on the culture medium, wherein the adding amount of the sodium acetate is 1g/L, the adding amount of the magnesium acetate is 1g/L, inoculating haematococcus pluvialis seed liquid cultured to the later logarithmic phase of growth into the culture medium for culturing, enabling the concentration of the haematococcus pluvialis to be 2.5 multiplied by 105cells/mL, sampling every other day to measure the biomass of the haematococcus pluvialis, taking a growth curve of the haematococcus pluvialis in the culture medium, and collecting the haematococcus pluvialis with the maximum biomass;

step six: weighing 0.5g of halogenating agent, dissolving the halogenating agent by using 15g of chloroform, slowly adding the solution into a reactor, reacting for 1h, and washing for three times to remove alkali and salt to obtain a primary solution;

step seven: and adding the composite antioxidant obtained in the fourth step into the primary liquid, adding a silane coupling agent into the primary liquid, uniformly mixing, heating in a water bath, and filtering to obtain the astaxanthin.

Wherein the pressure adopted for homogenizing wall breaking is 100MPa, the wall breaking temperature is 60 ℃, the wall breaking rate is more than or equal to 95%, the lipase amount accounts for 10% of the mass of the wall-broken algae slurry, the enzymolysis temperature is 45 ℃, the enzymolysis time is 24h, the enzymolysis pH value is 8.0, 15 parts by weight of haematococcus pluvialis algae oil, 5 parts by weight of algae residue, 10 parts by weight of lutein ester, 15 parts by weight of vitamin C and 40 parts by weight of vegetable oil are mixed, crushed, uniformly mixed and then sieved by a 180-mesh sieve to obtain a content material, the conditions of high-pressure wet heat sterilization are 150 ℃, the treatment time is 30min, the emulsifier is sodium dodecyl sulfate, the catalyst is phosphorous acid, the water bath heating temperature is 90 ℃, the water bath time is 40min, the stirring speed is 600r/min, and the stirring time is 50 min.

When the cleaning device of the technical scheme is used, haematococcus pluvialis cells are inoculated for amplification culture to obtain an amplification culture, the amplification culture is subjected to induction culture, astaxanthin is accumulated in the induced haematococcus pluvialis cells to obtain an induction culture, haematococcus pluvialis is separated from the induction culture to obtain the haematococcus pluvialis, the haematococcus pluvialis is subjected to amplification culture in an amplification culture medium, the homogeneous wall breaking in the first step is performed at the pressure of 100MPa and the wall breaking temperature of 60 ℃ and the wall breaking rate of more than or equal to 95%, the lipase dosage in the first step accounts for 5% -10% of the mass of the algal wall breaking pulp, the enzymolysis temperature is 35 ℃ -45 ℃, the enzymolysis time is 18-24h, and the enzymolysis pH value is 6.0-8.0, in the first step, 1-15 parts by weight of haematococcus pluvialis oil, 1-5 parts by weight of algal residues, 1-10 parts by weight of lutein esters, 5-15 parts by weight of vitamin C and 20-40 parts by weight of vegetable oil are mixed, crushing, uniformly mixing, sieving by a 180-mesh sieve to obtain a content material, separating algae cells from an amplification culture, transferring to a BG11 culture medium with nitrogen deficiency for culturing for 3-5 days, adding a biological inducer for induction culture for 24-48 hours to obtain an induction culture, wherein the inducer is a bacillus subtilis fermentation product of oat, preparing the content material by using haematococcus pluvialis, standing and settling collected fresh haematococcus pluvialis, removing supernatant, homogenizing and breaking walls to obtain wall-broken algae slurry, adding lipase into the wall-broken algae slurry for enzymolysis to obtain enzymolysis wall-broken algae liquid, extracting algae oil to obtain haematococcus pluvialis oil and algae residues, weighing 150g of lutein, placing the lutein in a high-pressure reaction kettle, adding 15-25mL of emulsifier, stirring and uniformly mixing, then adding 5-40g of a reaction catalyst and 20-60g of a phase transfer catalyst into the reaction kettle, reacting at 90-150 ℃ for 10-30h to obtain zeaxanthin, wherein the conditions of high-pressure damp-heat sterilization in the third step are 90-150 ℃, the treatment time is 10-30min, 30-80g of giant knotweed and cassia seed are respectively weighed, the giant knotweed and cassia seed are ground into powder, 100mL of ethanol solvent with the concentration of 40-90% is added, the resveratrol is obtained by refluxing, extracting and filtering, 20-60g of protocatechuic acid and gallic acid are respectively weighed, the protocatechuic acid and the gallic acid are mixed and stirred together, 10-30g of citric acid and 5-25g of alpha-glucoheptonate are added, the water is added and mixed uniformly, the mixture is heated in a water bath to obtain the composite antioxidant, the heating temperature in the water bath in the fourth step is 50-90 ℃, the stirring speed in the water bath time is 10-40min is 150-phase 600r/min, and the stirring time is 10-50min, preparing BBM culture medium with sufficient nitrogen, adding sodium acetate or magnesium acetate, performing high-pressure moist heat sterilization on the culture medium, wherein the adding amount of sodium acetate is 1-5g/L, the adding amount of magnesium acetate is 1-2.5 g/L, inoculating Haematococcus pluvialis seed liquid cultured to the later logarithmic phase of growth into the culture medium for culturing, ensuring the algae concentration to be 2.0 x 105-2.5 x 105cells/mL, sampling every other day to measure the biomass, making the growth curve of the Haematococcus pluvialis in the culture medium, collecting the Haematococcus pluvialis with the maximum biomass, weighing 0.5-2.4g of halogenating agent, dissolving with 1-15g of chloroform, slowly adding into a reactor for reaction for 1-15h, removing alkali and salt through three times of water washing to obtain primary liquid, adding the composite antioxidant in the fourth step into the primary liquid, adding silane coupling agent into the primary liquid, after being mixed evenly, the astaxanthin is obtained after being heated in water bath and filtered.

Having thus described the principal technical features and basic principles of the invention, and the advantages associated therewith, it will be apparent to those skilled in the art that the invention is not limited to the details of the foregoing illustrative embodiments, but is capable of other specific forms without departing from the spirit or essential characteristics thereof. The present embodiments are therefore to be considered in all respects as illustrative and not restrictive, the scope of the invention being indicated by the appended claims rather than by the foregoing description, and all changes which come within the meaning and range of equivalency of the claims are therefore intended to be embraced therein.

Furthermore, it should be understood that although the present description is described in terms of various embodiments, not every embodiment includes only a single embodiment, and such descriptions are provided for clarity only, and those skilled in the art will recognize that the embodiments described herein can be combined as a whole to form other embodiments as would be understood by those skilled in the art.