CN1242068C - Method for producing somatic powder containing lovastatin by liquid submerged fermentation - Google Patents
Method for producing somatic powder containing lovastatin by liquid submerged fermentation Download PDFInfo
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- CN1242068C CN1242068C CN 200410038293 CN200410038293A CN1242068C CN 1242068 C CN1242068 C CN 1242068C CN 200410038293 CN200410038293 CN 200410038293 CN 200410038293 A CN200410038293 A CN 200410038293A CN 1242068 C CN1242068 C CN 1242068C
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Abstract
The present invention relates to a method for obtaining monascus thallus powder containing 1.2% of lovastatin from monascus mutation thalli T-18 with high content of lovastatin by submerged liquid fermentation. The monascus thallus powder can be used to prepare various foods and medicines for lowering blood fat. The production of the monascus thallus powder comprises the following five steps: (1) culturing pure seeds on a slant; (2) culturing the seeds in a two-level shaking bottle; (3) culturing the seeds in a seeding tank; (4) fermenting and culturing the seeds in a fermentation tank; (5) pulping, homogenizing and spray-drying monascus thalli to obtain monascus thallus powder containing 1.2% of lovastatin. The method has the advantages of simple technology, easy control of reaction parameters, sealed production, no pollution, high thallus purity, low labor intensity, good product quality and high content of lovastatin.
Description
Technical field:
The present invention relates to a kind of preparation method of bacterial classification powder, relate in particular to a kind of liquid submerged fermentation method and produce the method that contains 1.2% lovastatin thalline powder.
Background technology:
Monascus is to be used for producing the main bacteria seed of wine of rice fermented with red yeast and monascorubin at the food color of works such as the historical Compendium of Material Medica of the existing more than one thousand years of China, " animal doctor this chapter ", the Exploitation of the Works of Nature to its fermented product red colouring agent for food, also used as a Chinese medicine, and the application of food antiseptic and drug medical aspect is existing to load.After Japan rattan chapter professor far away isolated cholesterol synthesis inhibitor from red monascus in 1979, the function of monascus and application are subjected to domestic and international extensive concern, obtained comprehensive deep research, the existing how tame food company of Japan drop into product that a large amount of manpower and materials carry out the listing of health red rice development of food can reducing blood-fat be popular and for inventor China of red colouring agent for food, also used as a Chinese medicine not show song protective foods take advantage.The functional Monascus product has only several places such as Fujian, Zhejiang, Hebei that production is arranged in China, the red colouring agent for food, also used as a Chinese medicine made by the monascus fermentation such as the zhibituo in Pekinese's Xuezhikang, Chengdu also occupies certain market as blood lipid-lowering medicine in addition.But generally speaking, the shared market share of functional Monascus product is also little, the lovastatin that monascus ruber produces is very big and be free from side effects to reducing blood lipid, and the functional Monascus that China produces now is still with the ancient method of koji tray or aerated koji making, labour intensity is big, easy pollution of products, it is low that product contains lovastatin.
Summary of the invention:
Effects such as technical problem to be solved by this invention provides a kind of liquid submerged fermentation method for preparing and produces the method that contains 1.2% lovastatin thalline powder, and it is simple that this method has production technique, and the health-care effect of product purity and product is obvious.
The present invention selects tens strain monascus rubers through separating, screening, and obtains the higher red monascus of strain product lovastatin and passes through ultraviolet ray and cobalt 60 (Co again
60) shine repeatedly and carry out mutagenesis.Obtain the mutagenic strain T-180 that a plant height produces the red monascus of lovastatin.
The applicant is deposited in China Committee for Culture Collection of Microorganisms common micro-organisms center on May 19th, 2004, and it abbreviates CGMCC as, and deposit number is 1148.The address of China Committee for Culture Collection of Microorganisms's common micro-organisms center (it abbreviates CGMCC as) is: No. 13, North No.1 Row, Zhongguancun, Haidian District, Beijing City, Institute of Microorganism, Academia Sinica (100080).
The mutagenic strain T-180 classification called after of the red monascus of this high yield lovastatin: belong to the red monascus bacterial classification.
The content that the monascus thalline powder that the present invention utilizes this this bacterium to make contains lovastatin is 1.2%, the content that is higher than the functional red yeast rice lovastatin of present China far away, and I am the production of adopting the automatic controlled liq submerged fermentation of a complete set of sealing, the purity height, produce and stablize, be difficult for the modern production technology of polluting.
The present invention adopts a plant height of my mutagenesis to produce the red monascus mutagenic strain T-180 of lovastatin through slant culture one-level shake-flask culture, the secondary shake-flask culture, again through seed tank culture, arrive fermentor cultivation again through defibrination, the homogeneous spraying drying powder-forming, promptly contain 1.2% lovastatin monascus thalline powder and be pink, with medicine and the food of making various reducing blood-fat.
The production of monascus thalline powder of the present invention is divided into bacterial classification production and fermentation two portions:
One, bacterial classification production:
1, the purebred preparation in inclined-plane:
1., the preparation of slant medium: 100 milliliters of worts, agar 3 grams are transferred PH4.2-4.5, heating agar dissolves fully that to put into the inclined-plane after the sterilization in 1 kilogram/30 minutes of the good tampon of back packing test tube plug stand-by;
2., the purebred preparation in inclined-plane: will make slant medium under aseptic condition, insert red monascus purebred 32 ℃ ± 1 cultivate 10-15 days stand-by.
2, the preparation of shake-flask seed
1., the preparation of one-level shake-flask seed
The preparation of a, seed culture medium: rice meal 5%, peptone 1.5%, corn steep liquor 1%, maltose 1%, amino acid 0.1%, potassium primary phosphate 0.25%, sal epsom 0.1%, divide after the abundant gelatinization of PH4.2-5 rice meal to install in 500 milliliters of triangular flasks, 80 milliliters of liquid amounts, the sterilization postcooling was stand-by in 1 kilogram/30 minutes.
The preparation of b, one-level shake-flask seed: will prepare 500 milliliters (the triangular flask nutrient solution inserts purebred under aseptic condition, filled in tampon and be placed on 32 ℃ ± 1 constant temperature shaking table, 300 rev/mins of revolutions, cultivate 24 hours stand-by.
2., the preparation of secondary shake-flask seed
The preparation of a, seed culture medium: rice meal 5%, peptone 1.5%, corn steep liquor 1%, amino acid 0.1%, maltose 1%, potassium primary phosphate 0.25%, sal epsom 0.1%, be divided in 2000 milliliters of bottles after the abundant gelatinization of the big powder rice of PH4.2-5, liquid amount is 300 milliliters, and the sterilization postcooling was stand-by in 1 kilogram/30 minutes.
The preparation of b, secondary shake-flask seed will have been cultivated one-level kind liquid and inserted in the substratum of 2000 milliliters of triangular flasks by 6% inoculum size under aseptic condition, filled in tampon, be placed on 32 ℃ ± 1 constant temperature shaking table and cultivate, 300 rev/mins of revolutions, cultivate 24 hours stand-by.
C, will cultivate the secondary shake-flask seed and under aseptic condition, pour in the inoculation bottle stand-by.
Two, fermentation:
1, the cultivation of 50 liters of seeding tanks
1., seed culture medium rice meal 5%, maltose 1%, corn steep liquor 1%, amino acid 0.1%, peptone 1.5%, phosphoric acid ammonia dipotassium 0.2%, sal epsom 0.1% is adjusted pH value 4.2-5;
2., liquid amount 70%, control Intake Quantity be 35 kilograms of substratum, sterilization 1 kilogram/30 minutes;
3., 45 ℃ of inoculation temps, will inoculate by 6% inoculum size, and with pressure difference method access shake-flask seed;
4., tank pressure 0.1MPA stirs 380 rev/mins of revolutions, ventilating ratio is 1: 0.7;
5., culture temperature 32 ℃ ± 1;
6., incubation time is 24 hours.
2,500 liters of cultivations of jar of turning sour;
1., the substratum rice meal 7% that turns sour, maltose 1.5%, corn steep liquor 1.5%, peptone 2%, potassium primary phosphate 0.2%, sal epsom 0.1%, PH4.2-5, sterilization 1 kilogram/30 minutes;
2., liquid amount 70% is 350 kilograms, 45 ℃ of inoculation temps;
3., inoculum size by 6% the inoculation, the pressure difference method inserts fermentor tank with cultured seed;
4., 32 ℃ of culture temperature, air flow is 1: 0.7, control PH5.2-5.5, tank pressure 0.1MPA;
5., incubation time (168 hours) stirring in 7 days revolution is 320 rev/mins;
3, defibrination:
With cultured fermented liquid colloidal mill, grinding 2 fineness is 120 orders.
4, homogeneous:
To grind good monascus fermented liquid through the clarifixator homogeneous, homogeneous, fineness 300 orders behind the homogeneous.
5, drying:
130 ℃ of spray-drier inlet temperatures of the monascus fermented liquid that homogeneous is good, 62 ℃ of temperature outs, spraying drying powder-forming promptly obtains containing lovastatin 1.2% monascus thalline powder.
The manufacture method of lovastatin-containing red koji mold body powder of the present invention is that the production of present China contains the exclusive production method of lovastatin red colouring agent for food, also used as a Chinese medicine, produce red colouring agent for food, also used as a Chinese medicine thalline powder and contain lovastatin 1.2%, this method production contains lovastatin monascus body powder and all oneself controls sealing production, do not pollute, impurity is few, easy to control, labour intensity is little, lovastatin content height.The protective foods of the reducing blood-fat of making is effective.
Embodiment:
The specific implementation method of the production of monascus thalline powder provided by the invention is divided into five parts:
1, the purebred cultivation in monascus inclined-plane.
2, the cultivation of I and II shake-flask seed.
3, the cultivation of monascus seeding tank.
4, the cultivation of monascus fermentation;
5, defibrination, even matter are finished product to spraying drying powder-forming---contain the monascus body powder of 1.2% lovastatin.
Below we describe the present invention in detail by specific measures for implementation:
1, the purebred cultivation in inclined-plane:
1. the preparation of slant medium:
With 100 milliliters of worts, agar 3 gram mixes, and pH value is aligned 3.5, and heating fully dissolves agar to put into the inclined-plane after the sterilization in 1 kilogram/30 minutes of the good tampon of back packing test tube plug stand-by;
2. the purebred preparation in inclined-plane:
With the above-mentioned slant medium that makes under aseptic condition, insert red monascus purebred 32 ℃ ± 1 cultivate 15 days stand-by.
2, the preparation of shake-flask seed:
1. the preparation of one-level shake-flask seed
The preparation of a, seed culture medium:
Prepare this seed culture medium according to following composition and ratio and method:
Rice meal 5%, peptone 1.5%, corn steep liquor 1%, maltose 1%, amino acid 0.1%, potassium primary phosphate 0.25%, sal epsom 0.1% is after the abundant gelatinization of the rice meal of pH value 4.5, divide to install in 500 milliliters of triangular flasks, liquid amount is 80 milliliters, and the sterilization postcooling was stand-by in 1 kilogram/30 minutes;
The preparation of b, one-level shake-flask seed:
In 500 milliliters of triangular flasks, aseptic condition inserts purebred down with the nutrient solution for preparing, filled in tampon to be placed on 32 ℃ ± 1 constant temperature shaking table, 300 rev/mins of revolutions, cultivate 24 hours stand-by.
2., the preparation of secondary shake-flask seed:
The preparation of a, seed culture medium:
Prepare this seed culture medium according to following composition and ratio and method:
Rice meal 5%, peptone 1.5%, corn steep liquor 1%, amino acid 0.1%, maltose 1%, potassium primary phosphate 0.25%, sal epsom 0.1%, be divided in 2000 milliliters of triangular flasks after the abundant gelatinization of big powder rice with pH value 4.5, liquid amount is 300 milliliters, and the sterilization postcooling was stand-by in 1 kilogram/30 minutes;
The preparation of b, secondary shake-flask seed:
Under aseptic condition, will cultivate one-level kind liquid and insert in the substratum of 2000 milliliters of triangular flasks, fill in tampon, and be placed on 32 ℃ ± 1 constant temperature shaking table and cultivate by 6% inoculum size, 300 rev/mins of revolutions, cultivate 24 hours stand-by;
C, will cultivate the secondary shake-flask seed and under aseptic condition, pour in the inoculation bottle, stand-by;
Fermenting process:
1, the cultivation of 50 liters of seeding tanks:
1., the prescription of seed culture medium:
Rice meal 5%, maltose 1%, corn steep liquor 1%, amino acid 0.1%, peptone 1.5%, phosphoric acid ammonia dipotassium 0.2%, sal epsom 0.1% is adjusted into 4.5 with pH value;
Concrete working method is:
2., liquid amount 70%, the 35 kilograms of substratum of packing into; Sterilization 1 kilogram/30 minutes;
3., inoculation temp is controlled at about 45 ℃, inoculate by 6% inoculum size, and insert shake-flask seed with conventional pressure difference method;
4., tank pressure is controlled at about 0.1MPA, stirs simultaneously, stirs 380 rev/mins of revolutions, ventilating ratio is 1: 0.7;
5., culture temperature is controlled at about 32 ℃;
6., incubation time is 24 hours;
2,500 liters of cultivations of jar of turning sour:
1., the turn sour prescription of substratum is:
Rice meal 7%, maltose 1.5%, corn steep liquor 1.5%, peptone 2%, potassium primary phosphate 0.2%, sal epsom 0.1% is adjusted pH value 4.5, sterilization 1 kilogram/30 minutes;
2., liquid amount 70%, approximately be controlled at 350 kilograms, 45 ℃ of inoculation temps;
3., the pressure difference method inserts fermentor tank with cultured seed, the seed of seed tank culture is inoculated in the ratio of inoculum size 6%;
4., 32 ℃ of culture temperature, air flow is 1: 0.7, about control PH5.2-5.5, tank pressure is controlled at about 0.1MPA;
5., incubation time 7 days (168 hours), stirring revolution is 320 rev/mins.
3, defibrination: with cultured fermented liquid colloidal mill, grind 2 times, fineness is 120 orders.
4, homogeneous: will grind good monascus fermented liquid through the clarifixator homogeneous, homogeneous, fineness 300 orders behind the homogeneous.
5, drying: 130 ℃ of spray-drier inlet temperatures of the monascus fermented liquid that homogeneous is good, 62 ℃ of temperature outs, spraying drying powder-forming promptly obtains containing lovastatin 1.2% monascus thalline powder.
Claims (6)
1, a kind of liquid submerged fermentation method is produced the method for the thalline powder that contains 1.2% lovastatin, it is characterized in that: this method is to adopt a plant height to produce the red monascus mutagenic strain CGMCC1148 of lovastatin, is prepared into the monascus thalline powder that contains 1.2% lovastatin through the purebred cultivation of bacterial classification, shake-flask seed cultivation, seed tank culture, fermentation culture, defibrination homogeneous, spray drying process.
2, production as claimed in claim 1 contains the method for the thalline powder of 1.2% lovastatin, and it is characterized in that: the method for described spawn culture is:
1, the purebred preparation in inclined-plane:
1., the preparation of slant medium:
100 milliliters of worts, agar 3 grams are transferred PH4.2-4.5, and heating agar fully dissolves back packing test tube, has filled in tampon, and it is stand-by to put into the inclined-plane after the sterilization in 1 kilogram/30 minutes;
2., the purebred cultivation of bacterial classification: will make slant medium under aseptic condition, it is purebred to insert red monascus, cultivates 10-15 days 32 ℃ ± 1, stand-by.
3, production as claimed in claim 1 contains the method for the thalline powder of 1.2% lovastatin, it is characterized in that:
The method of described defibrination is: with cultured fermented liquid colloidal mill, grinding 2 fineness is 120 orders.
4, production as claimed in claim 1 contains the method for the thalline powder of 1.2% lovastatin, and it is characterized in that: the method for described homogeneous is: will grind good monascus fermented liquid through the clarifixator homogeneous, homogeneous, fineness 300 orders behind the homogeneous.
5, production as claimed in claim 1 contains the method for the thalline powder of 1.2% lovastatin, it is characterized in that: described exsiccant method is: 130 ℃ of spray-drier inlet temperatures of the monascus fermented liquid that homogeneous is good, 62 ℃ of temperature outs, spraying drying powder-forming promptly obtains containing the monascus thalline powder of lovastatin 1.2%.
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| Application Number | Priority Date | Filing Date | Title |
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| CN 200410038293 CN1242068C (en) | 2004-05-21 | 2004-05-21 | Method for producing somatic powder containing lovastatin by liquid submerged fermentation |
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| CN 200410038293 CN1242068C (en) | 2004-05-21 | 2004-05-21 | Method for producing somatic powder containing lovastatin by liquid submerged fermentation |
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| CN1580272A CN1580272A (en) | 2005-02-16 |
| CN1242068C true CN1242068C (en) | 2006-02-15 |
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Families Citing this family (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102559795A (en) * | 2010-12-20 | 2012-07-11 | 北大方正集团有限公司 | Method for producing lovastatin by fermentation and fermentation medium used by same |
| CN102533894A (en) * | 2011-12-22 | 2012-07-04 | 中山大学 | Method for maintaining stable content of Monacolin K and its acid structure in monascus |
| CN105054021A (en) * | 2015-08-04 | 2015-11-18 | 江苏大学 | Monascus purpureus transformed corn bran fermentation liquid preparation method and application thereof |
| CN105695527A (en) * | 2016-04-27 | 2016-06-22 | 山东鲁抗医药股份有限公司 | Culture medium for lovastatin fermentation and supplementing method for fermentation process |
| CN108865897A (en) * | 2018-06-29 | 2018-11-23 | 夏放军 | A kind of high yield type monascus strain culture medium and preparation method thereof |
| CN110592155B (en) * | 2019-09-12 | 2021-07-06 | 北京工商大学 | Application of arginine in improving monacolin K production by monascus purpureus |
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