CN111139268B - Preparation method of water-soluble functional red yeast rice powder - Google Patents
Preparation method of water-soluble functional red yeast rice powder Download PDFInfo
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- 239000000843 powder Substances 0.000 title claims abstract description 94
- 238000002360 preparation method Methods 0.000 title claims abstract description 42
- 229940026314 red yeast rice Drugs 0.000 title claims description 16
- 235000007189 Oryza longistaminata Nutrition 0.000 claims abstract description 47
- 240000007594 Oryza sativa Species 0.000 claims abstract description 47
- 235000007164 Oryza sativa Nutrition 0.000 claims abstract description 47
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 46
- 238000000855 fermentation Methods 0.000 claims abstract description 42
- 230000004151 fermentation Effects 0.000 claims abstract description 42
- 238000010563 solid-state fermentation Methods 0.000 claims abstract description 36
- 229960004844 lovastatin Drugs 0.000 claims abstract description 32
- PCZOHLXUXFIOCF-UHFFFAOYSA-N Monacolin X Natural products C12C(OC(=O)C(C)CC)CC(C)C=C2C=CC(C)C1CCC1CC(O)CC(=O)O1 PCZOHLXUXFIOCF-UHFFFAOYSA-N 0.000 claims abstract description 31
- PCZOHLXUXFIOCF-BXMDZJJMSA-N lovastatin Chemical compound C([C@H]1[C@@H](C)C=CC2=C[C@H](C)C[C@@H]([C@H]12)OC(=O)[C@@H](C)CC)C[C@@H]1C[C@@H](O)CC(=O)O1 PCZOHLXUXFIOCF-BXMDZJJMSA-N 0.000 claims abstract description 31
- QLJODMDSTUBWDW-UHFFFAOYSA-N lovastatin hydroxy acid Natural products C1=CC(C)C(CCC(O)CC(O)CC(O)=O)C2C(OC(=O)C(C)CC)CC(C)C=C21 QLJODMDSTUBWDW-UHFFFAOYSA-N 0.000 claims abstract description 31
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- 239000000463 material Substances 0.000 claims description 40
- 239000000047 product Substances 0.000 claims description 40
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- 239000012528 membrane Substances 0.000 claims description 30
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- 238000001914 filtration Methods 0.000 claims description 25
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- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 claims description 10
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- 238000011081 inoculation Methods 0.000 claims description 5
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- 229910052943 magnesium sulfate Inorganic materials 0.000 claims description 5
- 235000019341 magnesium sulphate Nutrition 0.000 claims description 5
- 229940035034 maltodextrin Drugs 0.000 claims description 5
- 239000011159 matrix material Substances 0.000 claims description 5
- 229910000402 monopotassium phosphate Inorganic materials 0.000 claims description 5
- 235000019796 monopotassium phosphate Nutrition 0.000 claims description 5
- 239000002504 physiological saline solution Substances 0.000 claims description 5
- 238000005070 sampling Methods 0.000 claims description 5
- 238000002791 soaking Methods 0.000 claims description 5
- 238000003860 storage Methods 0.000 claims description 5
- 238000011068 loading method Methods 0.000 claims description 4
- GNSKLFRGEWLPPA-UHFFFAOYSA-M potassium dihydrogen phosphate Chemical compound [K+].OP(O)([O-])=O GNSKLFRGEWLPPA-UHFFFAOYSA-M 0.000 claims description 4
- LWIHDJKSTIGBAC-UHFFFAOYSA-K potassium phosphate Substances [K+].[K+].[K+].[O-]P([O-])([O-])=O LWIHDJKSTIGBAC-UHFFFAOYSA-K 0.000 claims description 4
- 235000019764 Soybean Meal Nutrition 0.000 claims description 3
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- CQIUKKVOEOPUDV-IYSWYEEDSA-N antimycin Chemical compound OC1=C(C(O)=O)C(=O)C(C)=C2[C@H](C)[C@@H](C)OC=C21 CQIUKKVOEOPUDV-IYSWYEEDSA-N 0.000 abstract description 2
- 235000013339 cereals Nutrition 0.000 abstract description 2
- CQIUKKVOEOPUDV-UHFFFAOYSA-N citrinine Natural products OC1=C(C(O)=O)C(=O)C(C)=C2C(C)C(C)OC=C21 CQIUKKVOEOPUDV-UHFFFAOYSA-N 0.000 abstract description 2
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- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 2
- 241001052560 Thallis Species 0.000 description 2
- 125000002887 hydroxy group Chemical group [H]O* 0.000 description 2
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- 208000015181 infectious disease Diseases 0.000 description 1
- 150000002632 lipids Chemical class 0.000 description 1
- 210000004185 liver Anatomy 0.000 description 1
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- 239000003960 organic solvent Substances 0.000 description 1
- PJNZPQUBCPKICU-UHFFFAOYSA-N phosphoric acid;potassium Chemical compound [K].OP(O)(O)=O PJNZPQUBCPKICU-UHFFFAOYSA-N 0.000 description 1
- 239000002244 precipitate Substances 0.000 description 1
- 231100000331 toxic Toxicity 0.000 description 1
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Abstract
The invention discloses a preparation method of water-soluble functional red rice powder, belonging to the technical field of microbial fermentation. The preparation method of the invention uses red yeast which does not produce citrinin and high-yield lovastatin as a starting strain, uses an edible fungus cultivation bag as a fermentation container, carries out solid-state fermentation on grains, and prepares the water-soluble functional red yeast powder containing lovastatin through water leaching, plate-frame filter pressing, ultrafiltration, nanofiltration and freeze-drying. The lovastatin in the water-soluble functional red rice powder is of an open-loop structure, the product has good water solubility, and the water solution prepared by the water-soluble functional red rice powder product can keep clear and transparent at room temperature, has no sediment and has good stability.
Description
Technical Field
The invention belongs to the technical field of microbial fermentation, and particularly relates to a preparation method of water-soluble functional red rice powder.
Background
Red yeast, red rice and Fuqu are called as red yeast, red rice and Fuqu besides the synonym Danqu, and have extremely important significance in the development and utilization history of microbiology in China and the world. The functional red rice refers to red rice (powder) containing lovastatin as an effective component, which is produced by fermenting red rice bacteria, and has the effect of reducing blood fat; lovastatin is a hypolipidemic drug that has two chemical structures, namely an open-loop structure and a closed-loop structure. The ring-opened structure has good water solubility and is easy to be absorbed and utilized by human bodies to exert the efficacy, the ring-closed structure has poor water solubility, the ring-opened structure is only active after being taken by hydrolysis of hydroxy esterase secreted by human livers, and a small number of human endocrine hydroxy esterases have insufficient quantity or low activity, so that lovastatin with the ring-closed structure is incompletely hydrolyzed, and toxic and side effects are generated to endanger human health.
At present, the fermentation process and the preparation method of the water-soluble functional red yeast rice have been reported in patent documents, and Chinese patent publication numbers of CN100999471, CN 101773244, CN 103404785 and CN 101407825, and the documents of research on high yield lovastatin by solid state fermentation of red yeast rice bacteria, optimization of fermentation condition of the red yeast rice lovastatin and lipid lowering function, describe the fermentation process and the preparation method of the functional red yeast rice. However, the fermentation processes in the patent documents all adopt bottle type fermentation, the production workload is large, the proportion of the ring-opened lovastatin in the fermentation product is small, an organic solvent is required to be added or acid-base is adopted to adjust the pH value of the solution in the process of preparing the water-soluble functional red rice powder, the method is complicated, the extraction rate is not high, and the method is not suitable for industrial production.
Bottle type fermentation is commonly adopted in domestic functional red yeast rice production, materials are not easy to mix uniformly in the fermentation process, thalli grow slowly, production workload such as treatment of fermentation products and cleaning of bottles after fermentation is large, and lovastatin with a closed-loop structure occupies a large proportion in the fermentation products. The invention adopts the edible fungus cultivation bag as the fermentation container, solves the problems of difficult mixing of materials, cleaning of the bottle and the like in the fermentation process, ensures that the lovastatin with an open-loop structure in the fermentation product is more than 85 percent, and can directly produce the water-soluble functional red rice powder.
The extraction of the lovastatin as a functional substance in the red rice powder generally adopts an alcohol extraction or acid-base regulation process, the extraction process has high requirements on equipment and environment, is easy to cause danger, is easy to pollute the environment, has complex extraction steps and low extraction yield, causes waste of a large amount of resources, and is not suitable for being applied to scale-up production. The invention adopts edible fungi cultivation bags as the functional red rice powder produced by a fermentation container, can directly use water as solvent for extraction, and prepares the water-soluble product lovastatin after water leaching, plate and frame filter pressing, ultrafiltration, nanofiltration and freeze-drying, wherein the lovastatin in the water-soluble functional red rice powder is of an open-loop structure, the product has good water solubility, and the water solution prepared by the water-soluble functional red rice powder product can keep clear and transparent at room temperature, has no sediment and has good stability. Can be used for producing functional beverages or health-care foods, and has better economic value and application prospect.
Disclosure of Invention
The invention aims to provide a preparation method of water-soluble functional red rice powder. The method comprises the steps of taking red yeast which does not produce citrinin and high-yield lovastatin as a starting strain, taking an edible fungus cultivation bag as a fermentation container, carrying out solid-state fermentation on grains, and carrying out water leaching, plate frame filter pressing, ultrafiltration, nanofiltration and freeze-drying to obtain the water-soluble functional red yeast powder containing lovastatin. The lovastatin in the water-soluble functional red rice powder is of an open-loop structure, the product has good water solubility, and the water solution prepared by the water-soluble functional red rice powder product can keep clear and transparent at room temperature, has no sediment and has good stability.
In order to achieve the above purpose, the invention adopts the following technical scheme:
a preparation method of water-soluble functional red yeast rice powder specifically comprises the following steps:
the first stage solid state fermentation comprises the following specific steps:
(1) Activating strains: inoculating pure monascus strain into a flat plate culture medium, and culturing for 5-10 days at 30 ℃ to activate strains;
(2) Preparing primary seed liquid: adding 20ml of sterile physiological saline into the activated strain flat-plate culture medium, rubbing mycelia back and forth with an inoculating loop to scrub spores and mycelia to prepare flat-plate seed liquid, inoculating the flat-plate seed liquid into the seed culture medium according to an inoculum size of 2%, and culturing at 30 ℃ and 100rpm for 48 hours;
(3) Preparing a secondary seed solution: inoculating the primary seed solution into a seed culture medium (250 mL/1000 mL) according to 10% of inoculation amount under the aseptic condition, and culturing for 24 hours at 30 ℃ and 100 rpm;
(4) Solid state fermentation: inoculating the secondary seed liquid into a solid fermentation culture medium according to the weight ratio of 8-20% under the aseptic condition, uniformly stirring, and then sub-packaging into edible fungus cultivation bags according to the weight ratio of 350-380 g of each bag, wherein the bag mouth is screwed by a cover with air-permeable cotton; transferring the cultivating bag into a high-temperature room with 10 ten thousand cleanliness, culturing at 30 ℃ and 80-90% humidity, scattering every day (the edible fungus cultivating bag is not excessively severe when scattering, and the edible fungus is possibly damaged to cause bacteria contamination), transferring into a low-temperature room with 10 ten thousand cleanliness after the surface of the material starts to turn red, scattering every three days at 25 ℃ and 40-60% humidity, culturing for 30 days, discharging, transferring the fermented material into a drying room, drying at 55 ℃ until the moisture of the material is lower than 10%, crushing the material, sampling the crushed material to detect lovastatin content, and marking for warehouse storage or directly extracting;
the second stage of fermentation product separation, extraction and pulverization comprises the following specific steps:
1) Leaching with water: mixing the dried and crushed red rice powder material prepared in the first stage with purified water according to the weight ratio of 1:20-60, soaking for 1-5 hours at 55 ℃, and stirring once every 15 minutes to prepare a red rice powder aqueous solution;
2) Press filtration of the plate and the frame: filtering the aqueous solution of the red rice powder in the step 1) through high-efficiency high-density solid-liquid separation 1mm industrial filter cloth to remove insoluble substances, and collecting clear liquid;
3) Ultrafiltration: filtering the clear liquid obtained in the step 2) through a membrane, and collecting filtrate; the membrane is an organic membrane with an interception rate of 0.22 micrometers-5000 molecular mass;
4) Nanofiltration: filtering the filtrate obtained in the step 3) by a membrane, and collecting concentrated solution, wherein the membrane is an organic membrane with the interception rate of 1-2 nanometers-250 molecular mass;
5) And (3) freeze-drying: adding maltodextrin into the concentrated solution obtained in the step 4) according to the mass volume ratio of 3-7%, uniformly dissolving, and freeze-drying to obtain water-soluble functional red rice powder;
6) Pulverizing the freeze-dried powder, detecting, packaging and leaving the factory of the finished product.
In the preparation method, in the first stage solid state fermentation, the preparation of the plate culture medium comprises the following steps: 50g of potato is cut into small pieces and boiled for 30 minutes, two layers of gauze are used for filtering, 5g of glucose is added into the filtrate, 5g of agar is further boiled for 10 minutes, after cooling to room temperature, 250 milliliters of purified water is used, the temperature is 121 ℃ for 30 minutes, sterilization is carried out, and the obtained product is packaged into 8 sterile plates for standby.
In the preparation method, in the first stage solid state fermentation, the seed culture medium is prepared by the following steps: 3 to 8 percent of glucose, 3 to 5 percent of soybean powder, 0.1 to 0.3 percent of magnesium sulfate, 1 to 3 percent of peptone, 3 to 8 percent of glycerin, 0.1 to 0.5 percent of monopotassium phosphate, 0.1 to 0.3 percent of calcium sulfate, and the balance of water, wherein the pH is 3.0 to 5.0 percent of acetic acid, the mixture is packaged into 1000 milliliter triangular bottles, the liquid loading amount is 250 milliliters, and the mixture is cooled for standby after 30 minutes sterilization at 121 ℃.
In the preparation method, in the first stage of solid state fermentation, the preparation of the solid state fermentation culture medium comprises the following steps: 50-80% of indica rice flour, 10-30% of soybean powder, 5-20% of bran powder and 25-40% of nutrient solution by weight percentage of each ton of solid matrix are added and mixed uniformly; placing the mixed culture medium into a pulsating vacuum sterilizer, starting timing after the equipment temperature is 125 ℃, sterilizing for 30-60 minutes, cooling to room temperature after sterilization is finished, and crushing the culture medium again by a crusher until the water content is 35-40%, wherein the materials are loose and have no caking; crushing the indica rice powder, the soybean powder and the bran powder by a crusher respectively to 50-100 meshes for later use; the preparation of the nutrient solution comprises the following steps: 2 to 5 percent of peptone, 1 to 5 percent of glucose, 1 to 3 percent of saccharomycete extract, 0.5 to 2 percent of corn steep liquor dry powder, 0.1 percent of calcium sulfate and the balance of water, and adjusting the pH value to 5.0 by acetic acid for later use.
The invention has the advantages that:
(1) Bottle type fermentation is commonly adopted in domestic functional red yeast rice production, materials are not easy to mix uniformly in the fermentation process, thalli grow slowly, production workload such as treatment of fermentation products and cleaning of bottles after fermentation is large, and lovastatin with a closed-loop structure occupies a large proportion in the fermentation products. The invention adopts the edible fungus cultivation bag as the fermentation container, solves the problems of difficult mixing of materials, cleaning of the bottle and the like in the fermentation process, ensures that the lovastatin with an open-loop structure in the fermentation product is more than 85 percent, and the produced red rice powder is suitable for directly producing water-soluble functional red rice powder.
The extraction of the lovastatin as a functional substance in the red rice powder generally adopts an alcohol extraction or acid-base regulation process, the extraction process has high requirements on equipment and environment, is easy to cause danger, is easy to pollute the environment, has complicated extraction steps and low extraction yield, can cause waste of a large amount of resources, and is not suitable for being applied to scale-up production. The invention adopts edible fungi cultivation bags as the functional red rice powder produced by a fermentation container, can directly use water as solvent for extraction, and prepares the water-soluble product lovastatin after water leaching, plate and frame filter pressing, ultrafiltration, nanofiltration and freeze-drying, wherein the lovastatin in the water-soluble functional red rice powder is of an open-loop structure, the product has good water solubility, and the water solution prepared by the water-soluble functional red rice powder product can keep clear and transparent at room temperature, has no sediment and has good stability.
Drawings
FIG. 1 is a schematic flow chart of a preparation method of water-soluble functional red yeast rice powder.
FIG. 2 shows an HPLC detection spectrum of the solid state fermentation product red rice powder.
FIG. 3 shows the HPLC detection spectrum of the finished product of the water-soluble functional red yeast rice powder in the second stage.
The specific embodiment is as follows:
in order to make the contents of the present invention easier to understand, the technical solutions of the present invention will be further described with reference to the specific embodiments, but the following examples are only examples of the present invention and do not represent the scope of the present invention defined by the claims.
And (3) strain: monascus is a strain purchased from China industry microbiological culture collection center (CICC) with a strain collection number of 41649.
Example 1
A preparation method of water-soluble functional red yeast rice powder specifically comprises the following steps:
the first stage solid state fermentation comprises the following specific steps:
(1) Activating strains: inoculating pure Monascus to plate culture medium, and culturing at 30deg.C for 5 days to activate strain;
(2) Preparing primary seed liquid: adding 20mL of sterile physiological saline into the activated strain flat-plate culture medium, rubbing mycelia back and forth with an inoculating loop to scrub spores and mycelia to prepare flat-plate seed liquid, inoculating the flat-plate seed liquid into the seed culture medium according to an inoculum size of 2%, and culturing at 30 ℃ and 100rpm for 48 hours;
(3) Preparing a secondary seed solution: inoculating the primary seed solution into a seed culture medium (250 mL/1000 mL) according to 10% of inoculation amount under the aseptic condition, and culturing for 24 hours at 30 ℃ and 100 rpm;
(4) Solid state fermentation: inoculating the secondary seed liquid into a solid fermentation culture medium according to the weight ratio of 8% under the aseptic condition, uniformly stirring, and then sub-packaging into edible fungus cultivation bags according to the weight ratio of 350g per bag, wherein the bag mouth is tightly screwed by a cover filled with breathable cotton; transferring the cultivating bag into a high-temperature room with 10 ten thousand cleanliness, culturing at 30 ℃ and 80% humidity, scattering every day (taking care not to be too severe when scattering, and the edible fungus cultivating bag is possibly damaged to cause bacteria contamination), transferring into a low-temperature room with 10 ten thousand cleanliness after the surface of the material starts to turn red, culturing at 25 ℃ and 40% humidity, scattering every three days, culturing for 30 days, discharging, transferring the fermented material into a drying room, drying at 55 ℃, waiting until the water content of the material is lower than 10%, crushing the material, sampling the crushed material to detect lovastatin content, and marking, warehousing for storage or directly extracting;
the second stage of fermentation product separation, extraction and pulverization comprises the following specific steps:
1) Leaching with water: weighing 1kg of the dried and crushed red rice powder material prepared in the first stage, mixing with purified water according to the weight ratio of 1:20, soaking for 5 hours at 55 ℃, and stirring once every 15 minutes to prepare a red rice powder aqueous solution;
2) Press filtration of the plate and the frame: filtering the aqueous solution of the red rice powder in the step 1) through high-efficiency high-density solid-liquid separation 1mm industrial filter cloth to remove insoluble substances, and collecting clear liquid;
3) Ultrafiltration: filtering the clear liquid obtained in the step 2) through a membrane, and collecting filtrate; the membrane is an organic membrane with an interception rate of 0.22 micrometers-5000 molecular mass;
4) Nanofiltration: filtering the filtrate obtained in the step 3) by a membrane, and collecting concentrated solution, wherein the membrane is an organic membrane with the interception rate of 1-2 nanometers-250 molecular mass;
5) And (3) freeze-drying: adding maltodextrin into the concentrated solution obtained in the step 4) according to the mass volume ratio of 3%, uniformly dissolving, and freeze-drying to obtain water-soluble functional red rice powder;
6) Pulverizing the freeze-dried powder, detecting, packaging and leaving the factory of the finished product.
In the preparation method, in the first stage solid state fermentation, the preparation of the plate culture medium comprises the following steps: 50g of potato is cut into small pieces and boiled for 30 minutes, two layers of gauze are used for filtering, 5g of glucose is added into the filtrate, 5g of agar is further boiled for 10 minutes, after cooling to room temperature, 250 milliliters of purified water is used, the temperature is 121 ℃ for 30 minutes, sterilization is carried out, and the obtained product is packaged into 8 sterile plates for standby.
In the preparation method, in the first stage solid state fermentation, the seed culture medium is prepared by the following steps: glucose 8%, soybean meal 3%, magnesium sulfate 0.3%, peptone 3%, glycerol 8%, monopotassium phosphate 0.5%, calcium sulfate 0.3%, and water in balance, wherein pH is 3.0, the mixture is prepared by acetic acid, the mixture is packaged into 1000ml triangular flasks, the liquid loading amount is 250ml, and the mixture is cooled for standby after sterilization at 121 ℃ for 30 minutes.
In the preparation method, in the first stage of solid state fermentation, the preparation of the solid state fermentation culture medium comprises the following steps: 50% of indica rice flour, 30% of soybean powder and 20% of bran powder, and adding 25% of nutrient solution by weight per ton of solid matrix, and uniformly mixing; placing the mixed culture medium into a pulse vacuum sterilizer, starting timing after the equipment temperature is 125 ℃, sterilizing for 30 minutes, cooling to room temperature after sterilization is finished, and crushing the culture medium again by using a crusher, wherein the materials are loose and have no caking; pulverizing the indica rice powder, the soybean powder and the bran powder to 50 meshes respectively by a pulverizer for later use; the preparation of the nutrient solution comprises the following steps: 2% of peptone, 5% of glucose, 3% of saccharomycete extract, 2% of corn steep liquor dry powder, 0.1% of calcium sulfate and the balance of water, and adjusting pH to 5.0 with acetic acid for later use.
The flow chart of the preparation method of the water-soluble functional red yeast rice powder is shown in fig. 1. First stage of this embodiment
The HPLC detection spectrum of the product red rice powder obtained by solid state fermentation is shown in figure 2, and from figure 2, the red rice powder obtained by solid state fermentation in the first stage contains lovastatin in two forms of ring opening and ring closing, and the content of the ring opening lovastatin is more than 85%. In the second stage of this embodiment, the HPLC detection diagram of the finished product of the water-soluble functional red yeast powder obtained by separating and extracting the red yeast powder of the fermentation product obtained in the first stage is shown in fig. 3, and as shown in fig. 3, the finished product obtained by separating and purifying the fermentation product of the red Qu Jungu field of the first stage only contains ring-opened lovastatin and does not contain ring-closed lovastatin.
Example 2
A preparation method of water-soluble functional red yeast rice powder specifically comprises the following steps:
the first stage solid state fermentation comprises the following specific steps:
(1) Activating strains: inoculating pure Monascus to plate culture medium, and culturing at 30deg.C for 10 days to activate strain;
(2) Preparing primary seed liquid: adding 20mL of sterile physiological saline into the activated strain flat-plate culture medium, rubbing mycelia back and forth with an inoculating loop to scrub spores and mycelia to prepare flat-plate seed liquid, inoculating the flat-plate seed liquid into the seed culture medium according to an inoculum size of 2%, and culturing at 30 ℃ and 100rpm for 48 hours;
(3) Preparing a secondary seed solution: inoculating the primary seed solution into a seed culture medium (250 mL/1000 mL) according to 10% of inoculation amount under the aseptic condition, and culturing for 24 hours at 30 ℃ and 100 rpm;
(4) Solid state fermentation: inoculating the secondary seed liquid into a solid fermentation culture medium according to the weight ratio of 20% under the aseptic condition, uniformly stirring, and then sub-packaging 380g of the secondary seed liquid into edible fungus cultivation bags according to each bag of material, wherein the bag mouth is tightly screwed by a cover filled with breathable cotton; transferring the cultivating bag into a high-temperature room with 10 ten thousand cleanliness, culturing at 30 ℃ and 90% humidity, scattering every day (taking care not to be too severe when scattering, and the edible fungus cultivating bag is possibly damaged to cause bacteria contamination), transferring into a low-temperature room with 10 ten thousand cleanliness after the surface of the material starts to turn red, culturing at 25 ℃ and 60% humidity, scattering every three days, culturing for 30 days, discharging, transferring the fermented material into a drying room, drying at 55 ℃, waiting until the water content of the material is lower than 10%, crushing the material, sampling the crushed material to detect lovastatin content, and marking, warehousing for storage or directly extracting;
the second stage of fermentation product separation, extraction and pulverization comprises the following specific steps:
1) Leaching with water: weighing 1kg of the dried and crushed red rice powder material prepared in the first stage, mixing with purified water according to the weight ratio of 1:40, soaking for 1 hour at 55 ℃, and stirring once every 15 minutes to prepare a red rice powder aqueous solution;
2) Press filtration of the plate and the frame: filtering the aqueous solution of the red rice powder in the step 1) through high-efficiency high-density solid-liquid separation 1mm industrial filter cloth to remove insoluble substances, and collecting clear liquid;
3) Ultrafiltration: filtering the clear liquid obtained in the step 2) through a membrane, and collecting filtrate; the membrane is an organic membrane with an interception rate of 0.22 micrometers-5000 molecular mass;
4) Nanofiltration: filtering the filtrate obtained in the step 3) by a membrane, and collecting concentrated solution, wherein the membrane is an organic membrane with the interception rate of 1-2 nanometers-250 molecular mass;
5) And (3) freeze-drying: adding maltodextrin into the concentrated solution obtained in the step 4) according to the mass volume ratio of 5%, uniformly dissolving, and freeze-drying to obtain water-soluble functional red rice powder;
6) Pulverizing the freeze-dried powder, detecting, packaging and leaving the factory of the finished product.
In the preparation method, in the first stage solid state fermentation, the preparation of the plate culture medium comprises the following steps: 50g of potato is cut into small pieces and boiled for 30 minutes, two layers of gauze are used for filtering, 5g of glucose is added into the filtrate, 5g of agar is further boiled for 10 minutes, after cooling to room temperature, 250 milliliters of purified water is used, the temperature is 121 ℃ for 30 minutes, sterilization is carried out, and the obtained product is packaged into 8 sterile plates for standby.
In the preparation method, in the first stage solid state fermentation, the seed culture medium is prepared by the following steps: glucose 3%, soybean powder 5%, magnesium sulfate 0.1%, peptone 1%, glycerol 8%, potassium dihydrogen phosphate 0.5%, calcium sulfate 0.3%, and water in balance, wherein pH is 5.0, and acetic acid is used for preparing pH, packaging into 1000ml triangular flask, filling 250ml, sterilizing at 121deg.C for 30 min, and cooling for use.
In the preparation method, in the first stage of solid state fermentation, the preparation of the solid state fermentation culture medium comprises the following steps: 80% of indica rice flour, 10% of soybean powder and 10% of bran powder, and adding 40% of nutrient solution by weight per ton of solid matrix, and uniformly mixing; the mixed culture medium is placed into a pulsation vacuum sterilizer, timing is started after the temperature of the equipment is displayed at 125 ℃, and sterilization is carried out for 60 minutes. Cooling to room temperature after sterilization, wherein the moisture content is 40%, and crushing the culture medium again by a crusher, wherein the materials are loose and have no caking; pulverizing the indica rice powder, the soybean powder and the bran powder to 100 meshes respectively by a pulverizer for later use; the preparation of the nutrient solution comprises the following steps: 5% of peptone, 1% of glucose, 3% of saccharomycete extract, 0.5% of corn steep liquor dry powder, 0.1% of calcium sulfate and the balance of water, and adjusting pH to 5.0 with acetic acid for later use.
Example 3
A preparation method of water-soluble functional red yeast rice powder specifically comprises the following steps:
the first stage solid state fermentation comprises the following specific steps:
(1) Activating strains: inoculating pure Monascus to plate culture medium, and culturing at 30deg.C for 8 days to activate strain;
(2) Preparing primary seed liquid: adding 20mL of sterile physiological saline into the activated strain flat-plate culture medium, rubbing mycelia back and forth with an inoculating loop to scrub spores and mycelia to prepare flat-plate seed liquid, inoculating the flat-plate seed liquid into the seed culture medium according to an inoculum size of 2%, and culturing at 30 ℃ and 100rpm for 48 hours;
(3) Preparing a secondary seed solution: inoculating the primary seed solution into a seed culture medium (250 mL/1000 mL) according to 10% of inoculation amount under the aseptic condition, and culturing for 24 hours at 30 ℃ and 100 rpm;
(4) Solid state fermentation: inoculating the secondary seed liquid into a solid fermentation culture medium according to the weight ratio of 15% under the aseptic condition, uniformly stirring, and then sub-packaging into edible fungus cultivation bags according to 370g of each bag, wherein the bag mouth is tightly screwed by a cover with air-permeable cotton; transferring the cultivating bag into a high-temperature room with 10 ten thousand cleanliness, culturing at 30 ℃ and with 85% humidity, scattering every day (taking care of not being too severe when scattering, and the edible fungi cultivating bag may be damaged to cause bacteria infection), transferring into a low-temperature room with 10 ten thousand cleanliness after the surface of the material starts to turn red, culturing at 25 ℃ and with 50% humidity, scattering every three days, culturing for 30 days, discharging, transferring the fermented material into a drying room, drying at 55 ℃, until the water content of the material is lower than 10%, crushing the material, sampling the crushed material, detecting lovastatin content, and marking, warehousing for storage or directly extracting;
the second stage of fermentation product separation, extraction and pulverization comprises the following specific steps:
1) Leaching with water: weighing 1kg of the dried and crushed red rice powder material prepared in the first stage, mixing with purified water according to the weight ratio of 1:60, soaking for 3 hours at 55 ℃, and stirring once every 15 minutes to prepare a red rice powder aqueous solution;
2) Press filtration of the plate and the frame: filtering the aqueous solution of the red rice powder in the step 1) through high-efficiency high-density solid-liquid separation 1mm industrial filter cloth to remove insoluble substances, and collecting clear liquid;
3) Ultrafiltration: filtering the clear liquid obtained in the step 2) through a membrane, and collecting filtrate; the membrane is an organic membrane with an interception rate of 0.22 micrometers-5000 molecular mass;
4) Nanofiltration: filtering the filtrate obtained in the step 3) by a membrane, and collecting concentrated solution, wherein the membrane is an organic membrane with the interception rate of 1-2 nanometers-250 molecular mass;
5) And (3) freeze-drying: adding maltodextrin into the concentrated solution obtained in the step 4) according to the mass volume ratio of 7%, uniformly dissolving, and freeze-drying to obtain water-soluble functional red rice powder;
6) Pulverizing the freeze-dried powder, detecting, packaging and leaving the factory of the finished product.
In the preparation method, in the first stage solid state fermentation, the preparation of the plate culture medium comprises the following steps: 50g of potato is cut into small pieces and boiled for 30 minutes, two layers of gauze are used for filtering, 5g of glucose is added into the filtrate, 5g of agar is further boiled for 10 minutes, the mixture is cooled to room temperature, then 250mL of purified water is used for setting the temperature to 121 ℃, and after 30 minutes of sterilization, the mixture is packaged into 8 sterile plates for standby.
In the preparation method, in the first stage solid state fermentation, the seed culture medium is prepared by the following steps: glucose 6%, soybean meal 4%, magnesium sulfate 0.2%, peptone 2%, glycerol 5%, monopotassium phosphate 0.3%, calcium sulfate 0.2%, and water in balance, wherein pH is 4.0, the mixture is packaged into 1000mL triangular flasks with acetic acid, 250mL of liquid loading volume, 121 ℃ and cooled after 30 minutes sterilization.
In the preparation method, in the first stage of solid state fermentation, the preparation of the solid state fermentation culture medium comprises the following steps: 65% of indica rice flour, 30% of soybean powder and 5% of bran powder, and adding 35% of nutrient solution by weight per ton of solid matrix, and uniformly mixing; the mixed culture medium is placed into a pulsation vacuum sterilizer, timing is started after the temperature of the equipment is displayed at 125 ℃, and sterilization is carried out for 45 minutes. Cooling to room temperature after sterilization, wherein the moisture content is 38%, and crushing the culture medium again by a crusher, wherein the materials are loose and have no caking; pulverizing the indica rice powder, the soybean powder and the bran powder to 80 meshes respectively by a pulverizer for later use; the preparation of the nutrient solution comprises the following steps: 3% of peptone, 3% of glucose, 2% of saccharomycete extract, 1.2% of corn steep liquor dry powder, 0.1% of calcium sulfate and the balance of water, and adjusting pH to 5.0 with acetic acid for later use.
In the above examples, 2g of the finished product obtained by separating, extracting and pulverizing the fermentation product of the second stage is dissolved in 100ml of purified water to prepare an aqueous solution, and the aqueous solution can be kept clear and transparent at room temperature, has no precipitate and has better stability.
The detection is carried out according to the method of the light industry standard QB/T2847-2007 of the people's republic of China, and the related indexes of the products obtained by the embodiment are as follows:
TABLE-lovastatin content and ratio in fermentation products of examples
Table II results of product yields in examples
The foregoing description is only of the preferred embodiments of the invention, and all changes and modifications that come within the meaning and range of equivalency of the claims are therefore intended to be embraced therein.
Claims (1)
1. The preparation method of the water-soluble functional red yeast rice powder is characterized by comprising the following steps of:
the first stage solid state fermentation comprises the following specific steps:
(1) Activating strains: inoculating pure monascus strain into a flat plate culture medium, and culturing for 5-10 days at 30 ℃ to activate strains;
(2) Preparing primary seed liquid: adding 20ml of sterile physiological saline into the activated strain flat-plate culture medium, rubbing mycelia back and forth with an inoculating loop to scrub spores and mycelia to prepare flat-plate seed liquid, inoculating the flat-plate seed liquid into the seed culture medium according to an inoculum size of 2%, and culturing at 30 ℃ and 100rpm for 48 hours;
(3) Preparing a secondary seed solution: inoculating the first-stage seed solution into a seed culture medium according to 10% of inoculation amount under the aseptic condition, and culturing for 24 hours at 30 ℃ and 100 rpm;
(4) Solid state fermentation: inoculating the secondary seed liquid into a solid fermentation culture medium according to the weight ratio of 8-20% under the aseptic condition, uniformly stirring, and then sub-packaging into edible fungus cultivation bags according to the weight ratio of 350-380 g of each bag, wherein the bag mouth is screwed by a cover with air-permeable cotton; transferring the cultivation bag into a high-temperature room with 10 ten thousand cleanliness, culturing at 30 ℃ and 80-90% humidity, scattering every day, transferring into a low-temperature room with 10 ten thousand cleanliness after the surface of the material starts to turn red, culturing at 25 ℃ and 40-60% humidity, scattering every three days, culturing for 30 days, discharging, transferring the fermented material into a drying room, drying at 55 ℃ until the moisture of the material is lower than 10%, crushing the material, sampling the crushed material, detecting the lovastatin content, and marking, warehousing for storage or directly extracting;
the second stage of fermentation product separation, extraction and pulverization comprises the following specific steps:
1) Leaching with water: mixing the dried and crushed red rice powder material prepared in the first stage with purified water according to the weight ratio of 1:20-60, soaking for 1-5 hours at 55 ℃, and stirring once every 15 minutes to prepare a red rice powder aqueous solution;
2) Press filtration of the plate and the frame: filtering the aqueous solution of the red rice powder in the step 1) through high-efficiency high-density solid-liquid separation 1mm industrial filter cloth to remove insoluble substances, and collecting clear liquid;
3) Ultrafiltration: filtering the clear liquid obtained in the step 2) through a membrane, and collecting filtrate; the membrane is an organic membrane with an interception rate of 0.22 micrometers-5000 molecular mass;
4) Nanofiltration: filtering the filtrate obtained in the step 3) by a membrane, and collecting concentrated solution, wherein the membrane is an organic membrane with the interception rate of 1-2 nanometers-250 molecular mass;
5) And (3) freeze-drying: adding maltodextrin into the concentrated solution obtained in the step 4) according to the mass volume ratio of 3-7%, uniformly dissolving, and freeze-drying to obtain water-soluble functional red rice powder;
6) Crushing, detecting, packaging and leaving a factory of finished products of the freeze-dried powder;
the monascus is deposited with the China center for type culture Collection of microorganisms with the accession number 41649;
in the first stage solid state fermentation, the preparation of the plate culture medium comprises the following steps: cutting 50g of potato into small pieces, boiling for 30 minutes, filtering with two layers of gauze, adding 5g of glucose into the filtrate, continuously boiling 5g of agar for 10 minutes, cooling to room temperature, fixing to 250 milliliters with purified water, sterilizing at 121 ℃ for 30 minutes, and packaging into 8 sterile plates for later use;
in the first stage solid state fermentation, the seed culture medium is prepared by: 3 to 8 percent of glucose, 3 to 5 percent of soybean meal, 0.1 to 0.3 percent of magnesium sulfate, 1 to 3 percent of peptone, 3 to 8 percent of glycerol, 0.1 to 0.5 percent of monopotassium phosphate, 0.1 to 0.3 percent of calcium sulfate, and the balance of water, wherein the pH is 3.0 to 5.0 percent of acetic acid, the mixture is packaged into 1000 milliliter triangular bottles, the liquid loading amount is 250 milliliters, and the mixture is cooled for standby after sterilization for 30 minutes at 121 ℃;
in the first stage solid state fermentation, the preparation of the solid state fermentation culture medium: 50-80% of indica rice flour, 10-30% of soybean powder, 5-20% of bran powder and 25-40% of nutrient solution by weight percentage of each ton of solid matrix are added and mixed uniformly; placing the mixed culture medium into a pulsating vacuum sterilizer, starting timing after the equipment temperature is 125 ℃, sterilizing for 30-60 minutes, cooling to room temperature after the sterilization is finished, and crushing the culture medium again by a crusher until the moisture content is 35-40%, wherein the materials are loose and have no caking; crushing the indica rice powder, the soybean powder and the bran powder by a crusher respectively to 50-100 meshes for later use; the preparation of the nutrient solution comprises the following steps: 2 to 5 percent of peptone, 1 to 5 percent of glucose, 1 to 3 percent of saccharomycete extract, 0.5 to 2 percent of corn steep liquor dry powder, 0.1 percent of calcium sulfate and the balance of water, and adjusting the pH value to 5.0 by acetic acid for later use.
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