CN104293680A - Monascus spp., preparation method for high-activity-composition functional monascus powder, and product - Google Patents

Monascus spp., preparation method for high-activity-composition functional monascus powder, and product Download PDF

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CN104293680A
CN104293680A CN201410379473.7A CN201410379473A CN104293680A CN 104293680 A CN104293680 A CN 104293680A CN 201410379473 A CN201410379473 A CN 201410379473A CN 104293680 A CN104293680 A CN 104293680A
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sodiumnitrate
peptone
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胡文林
谢凤娇
陈影妮
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TIANYI BIOTECH Co Ltd GUANGDONG
DONGGUAN CITY TIANYI BIOLOGICAL ENGINEERING Co Ltd
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DONGGUAN CITY TIANYI BIOLOGICAL ENGINEERING Co Ltd
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Abstract

The invention discloses a monascus spp. which is named as monascus rubei TY728, is preserved in China Center for Type Culture Collection (CCTCC) since 31st, July, 2014, and the preservation number is CCTCC NO: M2014363. The invention also discloses a method of employing the monascus spp. to prepare a low-citrinin high-color-value high-activity-composition functional monascus powder and a product prepared by using the method. The method comprises: preparing a strain; performing strain culture, inoculation and fermentation; performing deep liquid-state fermentation; and processing after fermentation. The provided monascus spp. is a microbe species obtained through special culturing, is adaptive to a liquid-state fermentation technology, is strong in vital force, and generates no or low-yield citrinin during metabolism. By employing the above strain as a fermentation strain, the product, which contains no or low-yield citrinin and is high in color value and high in active functional compositions such as Monacolin K, GABA and the like, is obtained. The preparation method employs deep liquid-state fermentation, does not generate pollution, is advanced in technology and high in quality stability, is suitable for large-scale production, and reaches international sanitary standard.

Description

The method for making of red colouring agent for food, also used as a Chinese medicine bacterial classification, high active ingredient functional Monascus powder and goods
Technical field
The present invention relates to microorganism and fermentation technical field thereof, be specifically related to red colouring agent for food, also used as a Chinese medicine bacterial classification, adopt liquid state fermentation to prepare method for making and the goods of the functional Monascus powder of low citrinin High color values high active ingredient.
Background technology
China's red colouring agent for food, also used as a Chinese medicine bacterial classification aboundresources, no matter as edible or medicinal, red colouring agent for food, also used as a Chinese medicine all plays an important role.The potential value of red colouring agent for food, also used as a Chinese medicine is very large, and in order to better serve the mankind, we actively should utilize the pharmacology research and development product innovation of red colouring agent for food, also used as a Chinese medicine, and especially lovastatin class is reducing the application in cholesterol, reduces atherosclerosis and occurs, prevent and treat the cardiovascular diseasess such as coronary heart disease.Meanwhile, carry out the testing to Citrinin, the safety problem of assurance function red colouring agent for food, also used as a Chinese medicine.
After find Monacolin K isoreactivity material in red colouring agent for food, also used as a Chinese medicine, the states such as Japan utilize solid state fermentation and liquid state fermentation 2 kinds of explained hereafter, develop very abundant red rice products, particularly functional red yeast rice goods, as the functional red yeast rice (containing Monacolin K) of Red kojic rice, reducing blood-fat, hypotensive functional red yeast rice (containing GABA) and red colouring agent for food, also used as a Chinese medicine pure mellow wine, red colouring agent for food, also used as a Chinese medicine plum wine, red colouring agent for food, also used as a Chinese medicine rice wine, red colouring agent for food, also used as a Chinese medicine rice vinegar, red colouring agent for food, also used as a Chinese medicine soy sauce, red colouring agent for food, also used as a Chinese medicine bean curd, monascus tea etc., greatly widen the purposes of red colouring agent for food, also used as a Chinese medicine.After the people such as nineteen ninety-five Blanc isolate Citrinin from Monascus anka Nakazawa et sato tunning, the emphasis of applied research has also just proceeded to the control aspect of Citrinin.Current Japan maintains the leading position in this respect, and citrinin content can control at below 0.2mg/kg, and Citrinin limitation is decided to be 0.2mg/kg by current japanese product standard; And in China, though there is a small amount of correlative study to report, but the control of Citrinin is in space state substantially in actual production, existing national standards also not using Citrinin as sanitary index, Food Additives Used in China association entrusts Southern Yangtze University to generally investigate to the monascus product citrinin content index of domestic production, find that the citrinin content of most product exceeds minimum tens times of Nippon Standard, be up to more than 100 times, Citrinin has become China's red colouring agent for food, also used as a Chinese medicine and has declared the technical bottleneck that JECFA enters European & American Market.
The content of Citrinin, the functional red yeast rice product of exploitation assistant lipid-lowering step-down in current reduction monascus product, need solution three key issues: one is the security of product, two is the content improving Monacolin K, GABA isoreactivity material, and three is problems of suitability for industrialized production.Strengthen the functional red yeast rice with blood fat-reducing blood pressure-decreasing effect of exploitation no citrinin or low citrinin, extend the value of monascus product, being two main trend and the direction of current red colouring agent for food, also used as a Chinese medicine industry, is also domestic red colouring agent for food, also used as a Chinese medicine enterprise problem demanding prompt solution.
Summary of the invention
For the deficiencies in the prior art, the object of the present invention is to provide a kind of liquid state fermentation technology to obtain low citrinin, high gamma aminobutanoic acid, do not receive the method for making of functional Monascus powder of low citrinin High color values high active ingredient of Courlene K isoreactivity functional component and goods.
Above-mentioned purpose of the present invention is achieved by the following technical solution:
A kind of red colouring agent for food, also used as a Chinese medicine bacterial classification, called after Monascus anka Nakazawa et sato TY728 (Monascus rubei TY728), it is preserved in Wuhan, Hubei loujia hill belongs Wuhan University China typical culture collection center (CCTCC) on July 31st, 2014, and biological deposits is numbered: CCTCC NO:M 2014363.
Adopt above-mentioned bacterial classification to produce a method for the functional Monascus powder of low citrinin High color values high active ingredient through liquid state fermentation, it comprises the steps:
(1) bacterial classification is produced:
Produce red colouring agent for food, also used as a Chinese medicine bacterial classification as fermented bacterium;
(2) bacterial classification is through culture medium culturing:
The bacterial classification that step (1) is produced is cultivated with following media respectively: slant medium, seed liquor substratum, liquid Shake flask medium;
(21) slant medium described in is: Zulkovsky starch 3-5%, maltose 3-5%, peptone 2-4%, and the water of agar 2-4%, SODIUMNITRATE 0.5-0.8% and residual content mixes; Loaded in triangular flask by this slant medium, through sterilizing, cooling, inoculation step (1) bacterial classification also carries out the cultivation of slant medium, is cultured to mycelia and covers with inclined-plane in 30 DEG C of incubators;
(22) the seed liquor substratum described in is: glucose 20-24g/L, peptone 30-34g/L, glycerine 20-22g/L, SODIUMNITRATE 2-4g/L, zinc sulfate 2-4g/L, potassium primary phosphate 1-2g/L, through sterilizing, cooling; With the slant culture liquid of gained after 40-60mL sterilized water washing step (21) triangular flask slant pore, the 50-60% being this seed liquor substratum according to volume ratio is inoculated in the triangular flask that this seed liquor substratum of 100ml is housed, 30 DEG C, liquidly on the rotary shaker of 180r/min to cultivate 1 day;
(23) the liquid Shake flask medium described in is: starch 16-18%, SODIUMNITRATE 0.4-0.6%, yeast extract paste 0.6-0.8%, corn steep liquor 0.8-1% mix with the water of residual content, regulate pH value to be 4.0 to make, through sterilizing, cooling with lactic acid; The seed liquor of step (22) gained is cultivated and plants, be that the amount of the 50-60% of this liquid Shake flask medium is inoculated in the triangular flask that this liquid Shake flask medium of 100mL is housed according to volume ratio, at 28 DEG C, liquid shake-flask culture 2 days under the condition of 180r/min;
(3) fermentative processing:
Using rice meal-glycerine as carbon source, soya bean is as major nitrogen source, peptone, corn steep liquor are auxiliary material as nitrogenous source, regulate pH value to make fermention medium with lactic acid, this fermention medium is loaded in triangular flask, normal temperature is cooled to after high-temperature sterilization, the liquid shaking flask kind that step (23) is produced, by volume per-cent was that the inoculum size of 6-8% is inoculated in this fermention medium of 50ml, in 26 DEG C of fermentations 12 days;
(4) deep layer liquid state fermentation:
Based on the liquid shaking flask kind fermented liquid of step (3) gained, the optimization adopt fermentor tank, regulating through batch fermentation in batches, submerged fermentation, setting fermentating culturing process, carries out deep layer liquid state fermentation, obtains functional red yeast rice fermented liquid;
(5) fermentation aftertreatment:
By the functional red yeast rice fermented liquid of step (4) gained, pot group type continuous countercurrent extraction process is adopted to extract, adopt Chamber Type Diaphragm Filter Press to carry out purifying, concentrate, refine afterwards, and carry out spraying dry, obtain the functional Monascus powder of low citrinin High color values high active ingredient.
In described step (3), fermention medium is rice meal 10-12g/L, glycerine 8-10g/L, soya bean 6-8g/L, peptone 1-3g/L, corn steep liquor 2-4g/L, magnesium sulfate 4.5-6.0g/L, potassium primary phosphate 1-2g/L, SODIUMNITRATE 0.4-0.6g/L, yeast extract paste 0.6-0.8g/L, and lactic acid regulates pH value to be 3.5.
Fermentating culturing process in described step (4) is that fermentor tank adopts 30M 3fermentor tank, effective volume of just surely fermenting is 22.0 cubic metres, growth temperature 32-35 DEG C during fermentation, and ventilating ratio is 1:0.3-0.6, tank pressure 0.4-0.7MPa, time 65-85h;
Pot group type continuous countercurrent extraction process in described step (5) is that three stage countercurrents extract, and wherein Extracting temperature is 25-55 DEG C, Extraction solvent ethanol consumption is 5-15L/kg, extraction time is 40-60min.
Ultrafiltration MWCO10000, nanofiltration MWCO200 in Chamber Type Diaphragm Filter Press in described step (5).
A kind of functional Monascus powder of the low citrinin High color values high active ingredient adopting above-mentioned method to prepare, it is by described red colouring agent for food, also used as a Chinese medicine bacterial classification respectively through slant medium, seed liquor substratum, liquid Shake flask medium, the inoculation culture of fermention medium, fermentation, after extract through deep layer liquid state fermentation, three stage countercurrents, ultrafiltration-nanofiltration concentrates refining, spraying dry and obtains.
Described slant medium is Zulkovsky starch 3-5%, maltose 3-5%, peptone 2-4%, and the water of agar 2-4%, SODIUMNITRATE 0.5-0.8% and residual content mixes; Described seed liquor substratum is glucose 20-24g/L, peptone 30-34g/L, glycerine 20-22g/L, SODIUMNITRATE 2-4g/L, zinc sulfate 2-4g/L, potassium primary phosphate 1-2g/L; Described liquid Shake flask medium is: starch 16-18%, SODIUMNITRATE 0.4-0.6%, yeast extract paste 0.6-0.8%, corn steep liquor 0.8-1% mix with the water of residual content, regulate pH value to be 4.0 with lactic acid; Described fermention medium is rice meal 10-12g/L, glycerine 8-10g/L, soya bean 6-8g/L, peptone 1-3g/L, corn steep liquor 2-4g/L, magnesium sulfate 4.5-6.0g/L, potassium primary phosphate 1-2g/L, SODIUMNITRATE 0.4-0.6g/L, yeast extract paste 0.6-0.8g/L, and lactic acid regulates pH value to be 3.5.
Described deep layer liquid state fermentation adopts 30M 3fermentor tank, effective volume of just surely fermenting is 22.0 cubic metres, and growth temperature 32-35 DEG C during fermentation, ventilating ratio is 1:0.3-0.6, tank pressure 0.4-0.7MPa, time 65-85h.
Extracting temperature during described three stage countercurrents extract is 25-55 DEG C, Extraction solvent ethanol consumption is 5-15L/Kg, extraction time is 40-60min; Described ultrafiltration-nanofiltration adopts Chamber Type Diaphragm Filter Press, wherein ultrafiltration MWCO10000, nanofiltration MWCO200.
Compared with prior art, the invention has the beneficial effects as follows:
A kind of red colouring agent for food, also used as a Chinese medicine bacterial classification provided by the invention, called after Monascus anka Nakazawa et sato TY728 (Monascus rubei TY728), biological deposits is numbered: CCTCC NO:M 2014363, be through the microorganism kind of special cultivation and acquisition, adapt to adopt liquid-state fermentation technology, vitality is strong, efficiency is high, do not produce or low-yield citrinin in metabolic process, through cultivation, the zymotechnique of preferred substratum, the functional red yeast rice powder product of acquisition possess not containing or low-yield citrinin, High color values, high gamma aminobutanoic acid, Gao Mona Courlene K isoreactivity functional component.
The method for making of the functional Monascus powder of low citrinin High color values high active ingredient provided by the invention adopts fermentor tank to carry out deep layer liquid state fermentation, and do not produce pollution, its production technique is advanced, quality stability is high, suitability for scale production, reaches international hygiene standards.
Describe the present invention in detail below in conjunction with specific embodiment, but be not to further restriction of the present invention.
Embodiment
Embodiment 1:
A kind of red colouring agent for food, also used as a Chinese medicine bacterial classification that the present embodiment provides, called after Monascus anka Nakazawa et sato TY728 (Monascus rubei TY728), it is preserved in Wuhan, Hubei loujia hill belongs Wuhan University China typical culture collection center (CCTCC) on July 31st, 2014, and biological deposits is numbered: CCTCC NO:M 2014363.
Adopt above-mentioned bacterial classification to prepare a method for the functional Monascus powder of low citrinin High color values high active ingredient, it comprises the following steps:
(1) bacterial classification is produced:
Produce red colouring agent for food, also used as a Chinese medicine bacterial classification as fermented bacterium;
(2) bacterial classification is through culture medium culturing:
The bacterial classification that step (1) is produced is cultivated with following media respectively: slant medium, seed liquor substratum, liquid Shake flask medium;
(21) slant medium described in is: Zulkovsky starch 3-5%, maltose 3-5%, peptone 2-4%, and the water of agar 2-4%, SODIUMNITRATE 0.5-0.8% and residual content mixes; Loaded in triangular flask by this slant medium, through sterilizing, cooling, inoculation step (1) bacterial classification also carries out the cultivation of slant medium, is cultured to mycelia and covers with inclined-plane in 30 DEG C of incubators;
(22) the seed liquor substratum described in is: glucose 20-24g/L, peptone 30-34g/L, glycerine 20-22g/L, SODIUMNITRATE 2-4g/L, zinc sulfate 2-4g/L, potassium primary phosphate 1-2g/L, through sterilizing, cooling; With the slant culture liquid of gained after 40-60mL sterilized water washing step (21) triangular flask slant pore, the 50-60% being this seed liquor substratum according to volume ratio is inoculated in the triangular flask that this seed liquor substratum of 100ml is housed, 30 DEG C, liquidly on the rotary shaker of 180r/min to cultivate 1 day;
(23) the liquid Shake flask medium described in is: starch 16-18%, SODIUMNITRATE 0.4-0.6%, yeast extract paste 0.6-0.8%, corn steep liquor 0.8-1% mix with the water of residual content, regulate pH value to be 4.0 to make, through sterilizing, cooling with lactic acid; The seed liquor of step (22) gained is cultivated and plants, be that the amount of the 50-60% of this liquid Shake flask medium is inoculated in the triangular flask that this liquid Shake flask medium of 100mL is housed according to volume ratio, at 28 DEG C, liquid shake-flask culture 2 days under the condition of 180r/min;
(3) fermentative processing:
Using rice meal-glycerine as carbon source, soya bean is as major nitrogen source, peptone, corn steep liquor are auxiliary material as nitrogenous source, regulate pH value to make fermention medium with lactic acid, this fermention medium is loaded in triangular flask, normal temperature is cooled to after high-temperature sterilization, the liquid shaking flask kind that step (23) is produced, by volume per-cent was that the inoculum size of 6-8% is inoculated in this fermention medium of 50ml, in 26 DEG C of fermentations 12 days;
(4) deep layer liquid state fermentation:
Based on the liquid shaking flask kind fermented liquid of step (3) gained, the optimization adopt fermentor tank, regulating through batch fermentation in batches, submerged fermentation, setting fermentating culturing process, carries out deep layer liquid state fermentation, obtains functional red yeast rice fermented liquid;
(5) fermentation aftertreatment:
By the functional red yeast rice fermented liquid of step (4) gained, pot group type continuous countercurrent extraction process is adopted to extract, adopt Chamber Type Diaphragm Filter Press to carry out purifying, concentrate, refine afterwards, and carry out spraying dry, obtain the functional Monascus powder of low citrinin High color values high active ingredient.
In described step (3), fermention medium is rice meal 10-12g/L, glycerine 8-10g/L, soya bean 6-8g/L, peptone 1-3g/L, corn steep liquor 2-4g/L, magnesium sulfate 4.5-6.0g/L, potassium primary phosphate 1-2g/L, SODIUMNITRATE 0.4-0.6g/L, yeast extract paste 0.6-0.8g/L, and lactic acid regulates pH value to be 3.5.
Fermentating culturing process in described step (4) is that fermentor tank adopts 30M 3fermentor tank, effective volume of just surely fermenting is 22.0 cubic metres, growth temperature 32-35 DEG C during fermentation, and ventilating ratio is 1:0.3-0.6, tank pressure 0.4-0.7MPa, time 65-85h;
Pot group type continuous countercurrent extraction process in described step (5) is that three stage countercurrents extract, and wherein Extracting temperature is 25-55 DEG C, Extraction solvent ethanol consumption is 5-15L/kg, extraction time is 40-60min.
Ultrafiltration MWCO10000, nanofiltration MWCO200 in Chamber Type Diaphragm Filter Press in described step (5).
A kind of functional Monascus powder of the low citrinin High color values high active ingredient adopting above-mentioned bacterial classification and method to prepare, it is by described red colouring agent for food, also used as a Chinese medicine bacterial classification respectively through slant medium, seed liquor substratum, liquid Shake flask medium, the inoculation culture of fermention medium, fermentation, after through deep layer liquid state fermentation, three stage countercurrents extract, ultrafiltration-nanofiltration concentrates refining, spraying dry and the powder that obtains.
Described slant medium is Zulkovsky starch 3-5%, maltose 3-5%, peptone 2-4%, and the water of agar 2-4%, SODIUMNITRATE 0.5-0.8% and residual content mixes; Described seed liquor substratum is glucose 20-24g/L, peptone 30-34g/L, glycerine 20-22g/L, SODIUMNITRATE 2-4g/L, zinc sulfate 2-4g/L, potassium primary phosphate 1-2g/L; Described liquid Shake flask medium is: starch 16-18%, SODIUMNITRATE 0.4-0.6%, yeast extract paste 0.6-0.8%, corn steep liquor 0.8-1% mix with the water of residual content, regulate pH value to be 4.0 with lactic acid; Described fermention medium is rice meal 10-12g/L, glycerine 8-10g/L, soya bean 6-8g/L, peptone 1-3g/L, corn steep liquor 2-4g/L, magnesium sulfate 4.5-6.0g/L, potassium primary phosphate 1-2g/L, SODIUMNITRATE 0.4-0.6g/L, yeast extract paste 0.6-0.8g/L, and lactic acid regulates pH value to be 3.5.
Described deep layer liquid state fermentation adopts 30M 3fermentor tank, effective volume of just surely fermenting is 22.0 cubic metres, and growth temperature 32-35 DEG C during fermentation, ventilating ratio is 1:0.3-0.6, tank pressure 0.4-0.7MPa, time 65-85h.
Extracting temperature during described three stage countercurrents extract is 25-55 DEG C, Extraction solvent ethanol consumption is 5-15L/Kg, extraction time is 40-60min; Described ultrafiltration-nanofiltration adopts Chamber Type Diaphragm Filter Press, wherein ultrafiltration MWCO10000, nanofiltration MWCO200.
Embodiment 2:
The method for making of the functional Monascus powder of the bacterial classification that the present embodiment provides, low citrinin High color values high active ingredient and goods, all substantially the same manner as Example 1, its difference is:
Adopt bacterial classification of the present invention to produce a method for the functional Monascus powder of low citrinin High color values high active ingredient through liquid state fermentation, comprise the steps, its difference is:
(2) bacterial classification is through culture medium culturing:
The bacterial classification that step (1) is produced is cultivated with following media respectively: slant medium, seed liquor substratum, liquid Shake flask medium;
(21) slant medium described in is: Zulkovsky starch 4%, maltose 3%, peptone 2%, agar 2%, and SODIUMNITRATE 0.5% mixes with the water of residual content; Loaded in triangular flask by this slant medium, through sterilizing, cooling, inoculation step (1) bacterial classification also carries out the cultivation of slant medium, is cultured to mycelia and covers with inclined-plane in 30 DEG C of incubators;
(22) the seed liquor substratum described in is: glucose 20g/L, peptone 34g/L, glycerine 20g/L, SODIUMNITRATE 2g/L, zinc sulfate 4g/L, potassium primary phosphate 1g/L, through sterilizing, cooling; With the slant culture liquid of gained after 60mL sterilized water washing step (21) triangular flask slant pore, be that 60% of this seed liquor substratum is inoculated in the triangular flask that this seed liquor substratum of 100ml is housed according to volume ratio, 30 DEG C, liquidly on the rotary shaker of 180r/min to cultivate 1 day;
(23) the liquid Shake flask medium described in is: starch 16%, SODIUMNITRATE 0.6%, yeast extract paste 0.6%, corn steep liquor 0.8% mix with the water of residual content, regulate pH value to be 4.0 to make, through sterilizing, cooling with lactic acid; The seed liquor of step (22) gained cultivated kind, the amount being 50% of this liquid Shake flask medium according to volume ratio is inoculated in the triangular flask that this liquid Shake flask medium of 100mL is housed, at 28 DEG C, liquid shake-flask culture 2 days under the condition of 180r/min;
(3) fermentative processing:
Using rice meal-glycerine as carbon source, soya bean is as major nitrogen source, peptone, corn steep liquor are auxiliary material as nitrogenous source, regulate pH value to make fermention medium with lactic acid, this fermention medium is loaded in triangular flask, normal temperature is cooled to after high-temperature sterilization, the liquid shaking flask kind that step (23) is produced, by volume per-cent be 6% inoculum size be inoculated in this fermention medium of 50ml, in 26 DEG C fermentation 12 days;
In described step (3), fermention medium is rice meal 10g/L, glycerine 10g/L, soya bean 6g/L, peptone 3g/L, corn steep liquor 4g/L, magnesium sulfate 4.5g/L, potassium primary phosphate 1g/L, SODIUMNITRATE 0.4g/L, yeast extract paste 0.6g/L, and lactic acid regulates pH value to be 3.5.
Fermentating culturing process in described step (4) is that fermentor tank adopts 30M 3fermentor tank, effective volume of just surely fermenting is 22.0 cubic metres, growth temperature 32 DEG C during fermentation, and ventilating ratio is 1:0.3, tank pressure 0.4MPa, time 85h;
Pot group type continuous countercurrent extraction process in described step (5) is that three stage countercurrents extract, and wherein Extracting temperature is 25 DEG C, Extraction solvent ethanol consumption is 5L/kg, extraction time is 60min.
A functional Monascus powder for the low citrinin High color values high active ingredient adopting above-mentioned method to prepare, its difference is:
Described slant medium is Zulkovsky starch 4%, maltose 3%, peptone 2%, agar 2%, and SODIUMNITRATE 0.5% mixes with the water of residual content; Described seed liquor substratum is glucose 20g/L, peptone 34g/L, glycerine 20g/L, SODIUMNITRATE 2g/L, zinc sulfate 4g/L, potassium primary phosphate 1g/L; Described liquid Shake flask medium is: starch 16%, SODIUMNITRATE 0.6%, yeast extract paste 0.6%, corn steep liquor 0.8% mix with the water of residual content, regulate pH value to be 4.0 with lactic acid; Described fermention medium is rice meal 10g/L, glycerine 10g/L, soya bean 6g/L, peptone 3g/L, corn steep liquor 4g/L, magnesium sulfate 4.5g/L, potassium primary phosphate 1g/L, SODIUMNITRATE 0.4g/L, yeast extract paste 0.6g/L, and lactic acid regulates pH value to be 3.5;
Described deep layer liquid state fermentation adopts 30M 3fermentor tank, effective volume of just surely fermenting is 22.0 cubic metres, growth temperature 32 DEG C during fermentation, and ventilating ratio is 1:0.3, tank pressure 0.4MPa, time 85h;
Extracting temperature during described three stage countercurrents extract is 25 DEG C, Extraction solvent ethanol consumption is 5L/kg, extraction time is 60min.
Embodiment 3:
The method for making of the functional Monascus powder of the bacterial classification that the present embodiment provides, low citrinin High color values high active ingredient and goods, all substantially identical with embodiment 1,2, its difference is:
Adopt bacterial classification of the present invention to produce a method for the functional Monascus powder of low citrinin High color values high active ingredient through liquid state fermentation, comprise the steps: that its difference is:
(2) bacterial classification is through culture medium culturing:
The bacterial classification that step (1) is produced is cultivated with following media respectively: slant medium, seed liquor substratum, liquid Shake flask medium;
(21) slant medium described in is: Zulkovsky starch 3%, maltose 5%, peptone 3%, agar 4%, and SODIUMNITRATE 0.8% mixes with the water of residual content; Loaded in triangular flask by this slant medium, through sterilizing, cooling, inoculation step (1) bacterial classification also carries out the cultivation of slant medium, is cultured to mycelia and covers with inclined-plane in 30 DEG C of incubators;
(22) the seed liquor substratum described in is: glucose 22g/L, peptone 30g/L, glycerine 22g/L, SODIUMNITRATE 4g/L, zinc sulfate 3g/L, potassium primary phosphate 1.5g/L, through sterilizing, cooling; With the slant culture liquid of gained after 40mL sterilized water washing step (21) triangular flask slant pore, be that 55% of this seed liquor substratum is inoculated in the triangular flask that this seed liquor substratum of 100ml is housed according to volume ratio, 30 DEG C, liquidly on the rotary shaker of 180r/min to cultivate 1 day;
(23) the liquid Shake flask medium described in is: starch 17%, SODIUMNITRATE 0.4%, yeast extract paste 0.8%, corn steep liquor 0.9% mix with the water of residual content, regulate pH value to be 4.0 to make, through sterilizing, cooling with lactic acid; The seed liquor of step (22) gained cultivated kind, the amount being 55% of this liquid Shake flask medium according to volume ratio is inoculated in the triangular flask that this liquid Shake flask medium of 100mL is housed, at 28 DEG C, liquid shake-flask culture 2 days under the condition of 180r/min;
(3) fermentative processing:
Using rice meal-glycerine as carbon source, soya bean is as major nitrogen source, peptone, corn steep liquor are auxiliary material as nitrogenous source, regulate pH value to make fermention medium with lactic acid, this fermention medium is loaded in triangular flask, normal temperature is cooled to after high-temperature sterilization, the liquid shaking flask kind that step (23) is produced, by volume per-cent be 7% inoculum size be inoculated in this fermention medium of 50ml, in 26 DEG C fermentation 12 days;
In described step (3), fermention medium is rice meal 11g/L, glycerine 9g/L, soya bean 7g/L, peptone 2g/L, corn steep liquor 3g/L, magnesium sulfate 5.5/L, potassium primary phosphate 1.5g/L, SODIUMNITRATE 0.6g/L, yeast extract paste 0.8g/L, and lactic acid regulates pH value to be 3.5.
Fermentating culturing process in described step (4) is that fermentor tank adopts 30M 3fermentor tank, effective volume of just surely fermenting is 22.0 cubic metres, growth temperature 33.5 DEG C during fermentation, and ventilating ratio is 1:0.45, tank pressure 0.55MPa, time 75h;
Pot group type continuous countercurrent extraction process in described step (5) is that three stage countercurrents extract, and wherein Extracting temperature is 40 DEG C, Extraction solvent ethanol consumption is 10L/kg, extraction time is 50min.
A functional Monascus powder for the low citrinin High color values high active ingredient adopting above-mentioned method to prepare, its difference is:
Described slant medium is Zulkovsky starch 3%, maltose 5%, peptone 3%, agar 4%, and SODIUMNITRATE 0.8% mixes with the water of residual content; Described seed liquor substratum is glucose 22g/L, peptone 30g/L, glycerine 22g/L, SODIUMNITRATE 4g/L, zinc sulfate 3g/L, potassium primary phosphate 1.5g/L; Described liquid Shake flask medium is: starch 17%, SODIUMNITRATE 0.4%, yeast extract paste 0.8%, corn steep liquor 0.9% mix with the water of residual content, regulate pH value to be 4.0 with lactic acid; Described fermention medium is rice meal 11g/L, glycerine 9g/L, soya bean 7g/L, peptone 2g/L, corn steep liquor 3g/L, magnesium sulfate 5.5/L, potassium primary phosphate 1.5g/L, SODIUMNITRATE 0.6g/L, yeast extract paste 0.8g/L, and lactic acid regulates pH value to be 3.5.
Described deep layer liquid state fermentation adopts 30M 3fermentor tank, effective volume of just surely fermenting is 22.0 cubic metres, growth temperature 33.5 DEG C during fermentation, and ventilating ratio is 1:0.45, tank pressure 0.55MPa, time 75h;
Extracting temperature during described three stage countercurrents extract is 40 DEG C, Extraction solvent ethanol consumption is 10L/kg, extraction time is 50min.
Embodiment 4:
The method for making of the functional Monascus powder of the bacterial classification that the present embodiment provides, low citrinin High color values high active ingredient and goods, substantially identical with embodiment 1,2,3, its difference is:
Adopt bacterial classification of the present invention to produce a method for the functional Monascus powder of low citrinin High color values high active ingredient through liquid state fermentation, comprise the steps: that its difference is:
(2) bacterial classification is through culture medium culturing:
The bacterial classification that step (1) is produced is cultivated with following media respectively: slant medium, seed liquor substratum, liquid Shake flask medium;
(21) slant medium described in is: Zulkovsky starch 5%, maltose 4%, peptone 4%, agar 3%, and SODIUMNITRATE 0.6% mixes with the water of residual content; Loaded in triangular flask by this slant medium, through sterilizing, cooling, inoculation step (1) bacterial classification also carries out the cultivation of slant medium, is cultured to mycelia and covers with inclined-plane in 30 DEG C of incubators;
(22) the seed liquor substratum described in is: glucose 24g/L, peptone 32g/L, glycerine 21g/L, SODIUMNITRATE 3g/L, zinc sulfate 2g/L, potassium primary phosphate 2g/L, through sterilizing, cooling; With the slant culture liquid of gained after 50mL sterilized water washing step (21) triangular flask slant pore, be that 50% of this seed liquor substratum is inoculated in the triangular flask that this seed liquor substratum of 100ml is housed according to volume ratio, 30 DEG C, liquidly on the rotary shaker of 180r/min to cultivate 1 day;
(23) the liquid Shake flask medium described in is: starch 18%, SODIUMNITRATE 0.5%, yeast extract paste 0.7%, corn steep liquor 1% mix with the water of residual content, regulate pH value to be 4.0 to make, through sterilizing, cooling with lactic acid; The seed liquor of step (22) gained cultivated kind, the amount being 60% of this liquid Shake flask medium according to volume ratio is inoculated in the triangular flask that this liquid Shake flask medium of 100mL is housed, at 28 DEG C, liquid shake-flask culture 2 days under the condition of 180r/min;
(3) fermentative processing:
Using rice meal-glycerine as carbon source, soya bean is as major nitrogen source, peptone, corn steep liquor are auxiliary material as nitrogenous source, regulate pH value to make fermention medium with lactic acid, this fermention medium is loaded in triangular flask, normal temperature is cooled to after high-temperature sterilization, the liquid shaking flask kind that step (23) is produced, by volume per-cent be 8% inoculum size be inoculated in this fermention medium of 50ml, in 26 DEG C fermentation 12 days;
In described step (3), fermention medium is rice meal 12g/L, glycerine 8g/L, soya bean 8g/L, peptone 1g/L, corn steep liquor 4g/L, magnesium sulfate 6.0g/L, potassium primary phosphate 2g/L, SODIUMNITRATE 0.5g/L, yeast extract paste 0.7g/L, and lactic acid regulates pH value to be 3.5.
Fermentating culturing process in described step (4) is that fermentor tank adopts 30M 3fermentor tank, effective volume of just surely fermenting is 22.0 cubic metres, growth temperature 35 DEG C during fermentation, and ventilating ratio is 1:0.6, tank pressure 0.7MPa, time 65h;
Pot group type continuous countercurrent extraction process in described step (5) is that three stage countercurrents extract, and wherein Extracting temperature is 55 DEG C, Extraction solvent ethanol consumption is 15L/kg, extraction time is 40min.
A functional Monascus powder for the low citrinin High color values high active ingredient adopting above-mentioned method to prepare, its difference is:
Described slant medium is Zulkovsky starch 5%, maltose 4%, peptone 4%, agar 3%, and SODIUMNITRATE 0.6% mixes with the water of residual content; Described seed liquor substratum is glucose 24g/L, peptone 32g/L, glycerine 21g/L, SODIUMNITRATE 3g/L, zinc sulfate 2g/L, potassium primary phosphate 2g/L; Described liquid Shake flask medium is: starch 18%, SODIUMNITRATE 0.5%, yeast extract paste 0.7%, corn steep liquor 1% mix with the water of residual content, regulate pH value to be 4.0 with lactic acid; Described fermention medium is rice meal 12g/L, glycerine 8g/L, soya bean 8g/L, peptone 1g/L, corn steep liquor 4g/L, magnesium sulfate 6.0g/L, potassium primary phosphate 2g/L, SODIUMNITRATE 0.5g/L, yeast extract paste 0.7g/L, and lactic acid regulates pH value to be 3.5.
Described deep layer liquid state fermentation adopts 30M 3fermentor tank, effective volume of just surely fermenting is 22.0 cubic metres, growth temperature 35 DEG C during fermentation, and ventilating ratio is 1:0.6, tank pressure 0.7MPa, time 65h;
Extracting temperature during described three stage countercurrents extract is 55 DEG C, Extraction solvent ethanol consumption is 15L/kg, extraction time is 40min.
Embodiment 5:
The method for making of the functional Monascus powder of the bacterial classification that the present embodiment provides, low citrinin High color values high active ingredient and goods, all substantially identical with embodiment 1,2,3,4, its difference is:
Adopt bacterial classification of the present invention to produce a method for the functional Monascus powder of low citrinin High color values high active ingredient through liquid state fermentation, comprise the steps: that its difference is:
(2) bacterial classification is through culture medium culturing:
The bacterial classification that step (1) is produced is cultivated with following media respectively: slant medium, seed liquor substratum, liquid Shake flask medium;
(21) slant medium described in is: Zulkovsky starch 4%, maltose 3%, peptone 4%, agar 3%, and SODIUMNITRATE 0.5% mixes with the water of residual content; Loaded in triangular flask by this slant medium, through sterilizing, cooling, inoculation step (1) bacterial classification also carries out the cultivation of slant medium, is cultured to mycelia and covers with inclined-plane in 30 DEG C of incubators;
(22) the seed liquor substratum described in is: glucose 24g/L, peptone 30-34g/L, glycerine 20g/L, SODIUMNITRATE 3g/L, zinc sulfate 3g/L, potassium primary phosphate 1g/L, through sterilizing, cooling; With the slant culture liquid of gained after 55mL sterilized water washing step (21) triangular flask slant pore, be that 60% of this seed liquor substratum is inoculated in the triangular flask that this seed liquor substratum of 100ml is housed according to volume ratio, 30 DEG C, liquidly on the rotary shaker of 180r/min to cultivate 1 day;
(23) the liquid Shake flask medium described in is: starch 17%, SODIUMNITRATE 0.6%, yeast extract paste 0.6%, corn steep liquor 1% mix with the water of residual content, regulate pH value to be 4.0 to make, through sterilizing, cooling with lactic acid; The seed liquor of step (22) gained cultivated kind, the amount being 55% of this liquid Shake flask medium according to volume ratio is inoculated in the triangular flask that this liquid Shake flask medium of 100mL is housed, at 28 DEG C, liquid shake-flask culture 2 days under the condition of 180r/min;
(3) fermentative processing:
Using rice meal-glycerine as carbon source, soya bean is as major nitrogen source, peptone, corn steep liquor are auxiliary material as nitrogenous source, regulate pH value to make fermention medium with lactic acid, this fermention medium is loaded in triangular flask, normal temperature is cooled to after high-temperature sterilization, the liquid shaking flask kind that step (23) is produced, by volume per-cent be 6% inoculum size be inoculated in this fermention medium of 50ml, in 26 DEG C fermentation 12 days;
In described step (3), fermention medium is rice meal 10g/L, glycerine 8g/L, soya bean 8g/L, peptone 3g/L, corn steep liquor 4g/L, magnesium sulfate 5.5g/L, potassium primary phosphate 1g/L, SODIUMNITRATE 0.4g/L, yeast extract paste 0.6g/L, and lactic acid regulates pH value to be 3.5.
Fermentating culturing process in described step (4) is that fermentor tank adopts 30M 3fermentor tank, effective volume of just surely fermenting is 22.0 cubic metres, growth temperature 33 DEG C during fermentation, and ventilating ratio is 1:0.5, tank pressure 0.6MPa, time 70h;
Pot group type continuous countercurrent extraction process in described step (5) is that three stage countercurrents extract, and wherein Extracting temperature is 35 DEG C, Extraction solvent ethanol consumption is 12L/kg, extraction time is 55min.
A functional Monascus powder for the low citrinin High color values high active ingredient adopting above-mentioned method to prepare, its difference is:
Described slant medium is Zulkovsky starch 4%, maltose 3%, peptone 4%, agar 3%, and SODIUMNITRATE 0.5% mixes with the water of residual content; Described seed liquor substratum is glucose 24g/L, peptone 30-34g/L, glycerine 20g/L, SODIUMNITRATE 3g/L, zinc sulfate 3g/L, potassium primary phosphate 1g/L; Described liquid Shake flask medium is: starch 17%, SODIUMNITRATE 0.6%, yeast extract paste 0.6%, corn steep liquor 1% mix with the water of residual content, regulate pH value to be 4.0 with lactic acid; Described fermention medium is rice meal 10g/L, glycerine 8g/L, soya bean 8g/L, peptone 3g/L, corn steep liquor 4g/L, magnesium sulfate 5.5g/L, potassium primary phosphate 1g/L, SODIUMNITRATE 0.4g/L, yeast extract paste 0.6g/L, and lactic acid regulates pH value to be 3.5.
Described deep layer liquid state fermentation adopts 30M 3fermentor tank, effective volume of just surely fermenting is 22.0 cubic metres, growth temperature 33 DEG C during fermentation, and ventilating ratio is 1:0.5, tank pressure 0.6MPa, time 70h;
Extracting temperature during described three stage countercurrents extract is 35 DEG C, Extraction solvent ethanol consumption is 12L/kg, extraction time is 55min.
Detect according to international standard, the index of correlation of above-described embodiment products obtained therefrom is as following table:
Red colouring agent for food, also used as a Chinese medicine bacterial classification provided by the invention, called after Monascus anka Nakazawa et sato TY728 (Monascus rubei TY728), biological deposits is numbered: CCTCC NO:M 2014363, be through the microorganism kind of special cultivation and acquisition, be suitable for adopting liquid-state fermentation technology, vitality is strong, efficiency is high, do not produce or low-yield citrinin in metabolic process, through cultivation, the zymotechnique of preferred substratum, the functional red yeast rice powder product of acquisition possess not containing or low-yield citrinin, High color values, high gamma aminobutanoic acid, Gao Mona Courlene K isoreactivity functional component.
The method for making of the functional Monascus powder of low citrinin High color values high active ingredient provided by the invention adopts fermentor tank to carry out deep layer liquid state fermentation, and do not produce pollution, its production technique is advanced, quality stability is high, suitability for scale production, reaches international hygiene standards.
The present invention is not limited to above-mentioned embodiment; adopt the raw material identical or approximate with the above embodiment of the present invention and preparation method; and the method for making of the functional Monascus powder of other the similar low citrinin High color values high active ingredient obtained and goods, all within protection scope of the present invention.

Claims (10)

1. a red colouring agent for food, also used as a Chinese medicine bacterial classification, called after Monascus anka Nakazawa et sato TY728 (Monascus rubei TY728), it is preserved in China typical culture collection center (CCTCC) on July 31st, 2014, and biological deposits is numbered: CCTCC NO:M 2014363.
2. adopt bacterial classification according to claim 1 to produce a method for high active ingredient functional Monascus powder, it is characterized in that, comprise the steps:
(1) bacterial classification is produced:
Produce red colouring agent for food, also used as a Chinese medicine bacterial classification as fermented bacterium;
(2) bacterial classification is through culture medium culturing:
The bacterial classification that step (1) is produced is cultivated with following media respectively: slant medium, seed liquor substratum, liquid Shake flask medium;
(21) slant medium described in is: Zulkovsky starch 3-5%, maltose 3-5%, peptone 2-4%, and the water of agar 2-4%, SODIUMNITRATE 0.5-0.8% and residual content mixes; Loaded in triangular flask by this slant medium, through sterilizing, cooling, inoculation step (1) bacterial classification also carries out the cultivation of slant medium, is cultured to mycelia and covers with inclined-plane in 30 DEG C of incubators;
(22) the seed liquor substratum described in is: glucose 20-24g/L, peptone 30-34g/L, glycerine 20-22g/L, SODIUMNITRATE 2-4g/L, zinc sulfate 2-4g/L, potassium primary phosphate 1-2g/L, through sterilizing, cooling; With the slant culture liquid of gained after 40-60mL sterilized water washing step (21) triangular flask slant pore, the 50-60% being this seed liquor substratum according to volume ratio is inoculated in the triangular flask that this seed liquor substratum of 100ml is housed, 30 DEG C, liquidly on the rotary shaker of 180r/min to cultivate 1 day;
(23) the liquid Shake flask medium described in is: starch 16-18%, SODIUMNITRATE 0.4-0.6%, yeast extract paste 0.6-0.8%, corn steep liquor 0.8-1% mix with the water of residual content, regulate pH value to be 4.0 to make, through sterilizing, cooling with lactic acid; The seed liquor of step (22) gained is cultivated and plants, be that the amount of the 50-60% of this liquid Shake flask medium is inoculated in the triangular flask that this liquid Shake flask medium of 100mL is housed according to volume ratio, at 28 DEG C, liquid shake-flask culture 2 days under the condition of 180r/min;
(3) fermentative processing:
Using rice meal-glycerine as carbon source, soya bean is as major nitrogen source, peptone, corn steep liquor are auxiliary material as nitrogenous source, regulate pH value to make fermention medium with lactic acid, this fermention medium is loaded in triangular flask, normal temperature is cooled to after high-temperature sterilization, the liquid shaking flask kind that step (23) is produced, by volume per-cent was that the inoculum size of 6-8% is inoculated in this fermention medium of 50ml, in 26 DEG C of fermentations 12 days;
(4) deep layer liquid state fermentation:
Based on the liquid shaking flask kind fermented liquid of step (3) gained, the optimization adopt fermentor tank, regulating through batch fermentation in batches, submerged fermentation, setting fermentating culturing process, carries out deep layer liquid state fermentation, obtains functional red yeast rice fermented liquid;
(5) fermentation aftertreatment:
By the functional red yeast rice fermented liquid of step (4) gained, pot group type continuous countercurrent extraction process is adopted to extract, adopt Chamber Type Diaphragm Filter Press to carry out purifying, concentrate, refine afterwards, and carry out spraying dry, obtain the functional Monascus powder of low citrinin High color values high active ingredient.
3. the method producing the functional Monascus powder of low citrinin High color values high active ingredient according to claim 2, be characterised in that, in described step (3), fermention medium is rice meal 10-12g/L, glycerine 8-10g/L, soya bean 6-8g/L, peptone 1-3g/L, corn steep liquor 2-4g/L, magnesium sulfate 4.5-6.0g/L, potassium primary phosphate 1-2g/L, SODIUMNITRATE 0.4-0.6g/L, yeast extract paste 0.6-0.8g/L, and lactic acid regulates pH value to be 3.5.
4. the method producing the functional Monascus powder of low citrinin High color values high active ingredient according to claim 2, is characterised in that, the fermentating culturing process in described step (4) is that fermentor tank adopts 30M 3fermentor tank, effective volume of just surely fermenting is 22.0 cubic metres, and growth temperature 32-35 DEG C during fermentation, ventilating ratio is 1:0.3-0.6, tank pressure 0.4-0.7MPa, time 65-85h.
5. the method producing the functional Monascus powder of low citrinin High color values high active ingredient according to claim 2, be characterised in that, pot group type continuous countercurrent extraction process in described step (5) is that three stage countercurrents extract, and wherein Extracting temperature is 25-55 DEG C, Extraction solvent ethanol consumption is 5-15L/kg, extraction time is 40-60min.
6. the method producing the functional Monascus powder of low citrinin High color values high active ingredient according to claim 2, is characterised in that, ultrafiltration MWCO10000, nanofiltration MWCO200 in the Chamber Type Diaphragm Filter Press in described step (5).
7. the high active ingredient functional Monascus powder adopting the described method of one of claim 1-6 to prepare, it is characterized in that, it is by described red colouring agent for food, also used as a Chinese medicine bacterial classification respectively through slant medium, seed liquor substratum, liquid Shake flask medium, the inoculation culture of fermention medium, fermentation, after through deep layer liquid state fermentation, three stage countercurrents extract, ultrafiltration-nanofiltration concentrates refining, spraying dry and the powder that obtains.
8. high active ingredient functional Monascus powder according to claim 7, is characterized in that, described slant medium is Zulkovsky starch 3-5%, maltose 3-5%, peptone 2-4%, the water of agar 2-4%, SODIUMNITRATE 0.5-0.8% and residual content mixes; Described seed liquor substratum is glucose 20-24g/L, peptone 30-34g/L, glycerine 20-22g/L, SODIUMNITRATE 2-4g/L, zinc sulfate 2-4g/L, potassium primary phosphate 1-2g/L; Described liquid Shake flask medium is: starch 16-18%, SODIUMNITRATE 0.4-0.6%, yeast extract paste 0.6-0.8%, corn steep liquor 0.8-1% mix with the water of residual content, regulate pH value to be 4.0 with lactic acid; Described fermention medium is rice meal 10-12g/L, glycerine 8-10g/L, soya bean 6-8g/L, peptone 1-3g/L, corn steep liquor 2-4g/L, magnesium sulfate 4.5-6.0g/L, potassium primary phosphate 1-2g/L, SODIUMNITRATE 0.4-0.6g/L, yeast extract paste 0.6-0.8g/L, and lactic acid regulates pH value to be 3.5.
9. high active ingredient functional Monascus powder according to claim 7, is characterized in that, described deep layer liquid state fermentation adopts 30M 3fermentor tank, effective volume of just surely fermenting is 22.0 cubic metres, and growth temperature 32-35 DEG C during fermentation, ventilating ratio is 1:0.3-0.6, tank pressure 0.4-0.7MPa, time 65-85h.
10. high active ingredient functional Monascus powder according to claim 7, is characterized in that, the Extracting temperature during described three stage countercurrents extract is 25-55 DEG C, Extraction solvent ethanol consumption is 5-15L/Kg, extraction time is 40-60min; Described ultrafiltration-nanofiltration adopts Chamber Type Diaphragm Filter Press, wherein ultrafiltration MWCO10000, nanofiltration MWCO200.
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CN111019843B (en) * 2020-01-06 2024-02-13 广东天益生物科技有限公司 Method for preparing functional red rice powder by spray drying method and method for effectively maintaining MK total content and acid MK stability
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CN115197854A (en) * 2022-07-08 2022-10-18 贵州省生物技术研究所(贵州省生物技术重点实验室、贵州省马铃薯研究所、贵州省食品加工研究所) Method for screening red yeast rice fermented summer and autumn tea strains and adaptive substrate
CN115197854B (en) * 2022-07-08 2024-01-30 贵州省生物技术研究所(贵州省生物技术重点实验室、贵州省马铃薯研究所、贵州省食品加工研究所) Screening method of strain of red yeast fermented summer and autumn tea and adaptive matrix
CN115386440A (en) * 2022-08-31 2022-11-25 广东天益生物发展有限公司 Health-care red yeast rice sweet wine and preparation method thereof
CN115386440B (en) * 2022-08-31 2024-03-26 广东天益生物发展有限公司 Health care red Qu Tianjiu and preparation method thereof

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