CN104498528A - Monascus fermentation method for low-yield citrinin and high-yield monascorubin - Google Patents
Monascus fermentation method for low-yield citrinin and high-yield monascorubin Download PDFInfo
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Abstract
The invention provides a monascus fermentation method for low-yield citrinin and high-yield monascorubin. The monascus fermentation method comprises seed culture of monascus and liquid-state fermentation, wherein a carbon source in the liquid-state fermentation culture medium is cassava flour, and phosphate is added into the culture medium; the addition amount of the cassava flour in the fermentation culture medium is 1-6%; the phosphate is diammonium hydrogen phosphate, disodium hydrogen phosphate and dipotassium phosphate. The technical method disclosed by the invention can be used for achieving the purpose of lowering and even restraining the production of citrinin, and also can be used for greatly improving the yield of pigments.
Description
Technical field
The invention belongs to technical field of microbial fermentation.More specifically, the present invention relates to a kind of Fermentation Condition of Monascus spp method, and utilize the method can reduce Citrinin output significantly to improve pigment production simultaneously.
Background technology
Monascus is the microorganism that uniquely can be used in large-scale industrial production food dye up to now in the world, and the Red kojic rice of Fermentation Condition of Monascus spp has had the medicine-food two-purpose history of more than one thousand years in China.In recent years, along with people are to the extensive concern of food-safety problem, the security of synthetic colour was under suspicion, and exploitation and the use of natural pigment also more and more come into one's own.From plant, extract the method for natural pigment at present, be the main source of natural pigment.It is short that microbe fermentation method has the time, do not affect, is easy to the advantage of control by natural climate.The monascorubin that Fermentation Condition of Monascus spp produces has that security is high, to ph stability is good, thermotolerance is high, to features such as protein tint permanence are strong, and monascorubin also has certain biocidal property, and therefore it is more and more subject to the favor of food-processing industry.At present, monascorubin has been successfully applied in the commodity such as meat product, biscuit, bread, makeup as tinting material.Along with going deep into of red starter research, other functions of pigment are also found gradually, and its application prospect is by boundless.
Citrinin is a kind of mycotoxins, and experimentation on animals proves that Citrinin can cause the kidney enlargements such as mouse, tubular ectasia and epithelial cell necrosis also to have carcinogenic, mutagenesis simultaneously.Citrinin is separated from the culture filtered liquid of Penicillium citrinum (Penicillium citrinnum) obtaining the earliest, find in Fermentation Condition of Monascus spp product also containing Citrinin in the research process of monascus, and confirm that the synthesis of Citrinin and monascorubin starts from same approach and has common metabolic precursor thereof material through experiment, therefore producing can with the generation of Citrinin in monascorubin process.
Permitted Jiangxi honor etc. and carry out the analysis of product Citrinin to 35 strain monascuses used in food fermentation industry, result shows: 35 strain monascuses all can produce Citrinin in rice medium.Li Fengqin etc. detect 114 parts of red colouring agent for food, also used as a Chinese medicine samples by Citrinin pollution condition for understanding China red koji goods, analyze and find: in 114 increment product, 68 parts detect Citrinin, and content is 0.18-1739.23mg/kg; Monascus product kind difference is also different by Citrinin pollution level, monascorubin powder containing citrinin content the highest and in 27 increment product 25 parts containing Citrinin; Next be in red kojic rice powder 19 increment product 12 parts detect Citrinin, most high-content is 855.67mg/kg; Though all contain Citrinin in functional red yeast rice content all lower than 6mg/kg.
Fermentation Condition of Monascus spp produces the present situation that on the phenomenon of Citrinin and market, red rice products pollutes by Citrinin, bring great impact to the application of monascorubin, therefore ensureing that the generation reducing or suppress Citrinin while Fermentation Condition of Monascus spp produces High color values monascorubin is the problem that we will urgently solve.Current research adopt more mutagenesis or genetic engineering means carry out strain improvement reach fundamentally suppress Citrinin produce object, another control Citrinin produce common method be then training systern.
Denomination of invention is method and the goods that monascus yellow pigment is prepared in monascus mutant strain and submerged fermentation thereof, application number: 201010279425.2, invention provides monascus mutant strain Monascus purpureusyang TY622, and adopt this bacterial strain submerged fermentation to prepare the method for monascus yellow pigment pigment, it comprises strain improvement, seed culture, bulk fermentation, monascus yellow pigment pigment extract, wherein seed culture, fermentation all adopts with rice is the substratum of carbon source, multistage cultivation is carried out, secondary fermentation in Large Copacity fermentor tank; Technological condition for fermentation: temperature 32 ~ 36 DEG C, mixing speed is 200 ~ 400r/min; Ventilation is 400 ~ 450Lmin in earlier stage, and the middle and later periods is adjusted to 500 ~ 600Lmin.Present invention also offers a kind of goods adopting aforesaid method to prepare.Preparation method provided by the invention, technique is simple, and be easy to suitability for industrialized production, in the fermented liquid obtained, monascus yellow pigment pigment content is high, citrinin content is low.
A kind of production method of monascus liquid state fermentation song, application number: a kind of production method of monascus liquid state fermentation song of 201310034337.X. disclosure of the invention, what the method related to set out monascus strain L high yield pigment and Citrinin, by China typical culture collection center preservation, deposit number is: CCTCC M2013039.The liquid state fermentation utilizing this monascus strain L to produce under the technology of the present invention condition is bent, its look valency level is 110 ~ 120U/mL, fermentation period is 5 ~ 7d, saccharifying enzymic activity is 630 ~ 740U/mL, citrinin content is only 2 ~ 3mg/L simultaneously, promulgate in 1999 lower than Japanese health ministry, citrinin content limit index 0.2mg/kg to monascorubin in " the Japanese food additives standard " of revision in 2003, there is not the hidden danger in safety in this liquid state fermentation song, can be applicable to red rice yellow wine and brewage system.
Make a general survey of the patent utilizing Fermentation Condition of Monascus spp to produce monascorubin, the overwhelming majority still adopts rice fermentation, do not relate in research in the past or patent and adopt Tapioca Starch to do substrate and add the report that phosphoric acid salt fermentation monascus investigates pigment and Citrinin correlation with yield, the present invention does Fermentation Condition of Monascus spp substratum main component with Tapioca Starch and phosphoric acid salt, realizes the object reducing Citrinin output or suppress Citrinin to produce.
Summary of the invention
The invention provides a kind of Fermentation Condition of Monascus spp method of simple and effective low-yield citrinin high yield monascus pigment, the method take monascus parpureus Went as starting strain, fermention medium does substrate with Tapioca Starch and adds phosphoric acid salt and carries out Fermentation Condition of Monascus spp, and the present invention can significantly reduce Citrinin output and significantly improve pigment production.
The technological method that the present invention realizes object is as follows:
A Fermentation Condition of Monascus spp method for low-yield citrinin high yield monascus pigment, comprises seed culture and the liquid state fermentation of monascus, and the carbon source in described liquid state fermentation substratum is Tapioca Starch, is added with phosphoric acid salt in substratum.
Tapioca Starch addition in described fermention medium is 1-6%.
Phosphoric acid salt is Secondary ammonium phosphate, Sodium phosphate dibasic, dipotassium hydrogen phosphate;
The addition of described Secondary ammonium phosphate is 0.001-0.1%.
The optimal set of Fermentation Condition of Monascus spp substratum becomes 2% Tapioca Starch, 1% peptone, 2% glucose, 0.05% Secondary ammonium phosphate, pH 5.8.
Preferred Fermentation Condition of Monascus spp substratum consist of 2% Tapioca Starch, 1% peptone, 2% glucose, 0.001% Secondary ammonium phosphate, pH 5.8.
Described Fermentation Condition of Monascus spp condition is: by 4% inoculation seed liquor, 160r/min 35 DEG C cultivates 3-6d.
Described monascus seed liquor culture condition is 1% sucrose, 0.3% peptone, 0.05% dipotassium hydrogen phosphate, 0.05% magnesium sulfate, pH 5.7,121 DEG C of sterilizing 20min, and by 2% inoculating spores suspension, 160r/min 35 DEG C cultivates 1d.
Advantage of the present invention and positively effect are:
Add phosphate radical fermentation monascus using Tapioca Starch as fermention medium main component, not only reduce the application of grain rice and reduce the production cost of monascorubin, simultaneously few the and successful of phosphate adding quantity, method simple application is strong.
Reach according to technological method of the present invention the object that reduction even suppresses Citrinin to produce, largely improve the output of pigment simultaneously.
Embodiment
Below in conjunction with embodiment, the present invention is further described.The following examples are illustrative, are not restrictive, can not limit protection scope of the present invention with this.
In the present invention, all per-cent is all weight percentage, and all equipment and raw material all can obtain from market or the industry is conventional.Method below in embodiment, if no special instructions, is the ordinary method of this area.
Embodiment 1
When the present embodiment illustrates and there is phosphate radical in Fermentation Condition of Monascus spp substratum, on the impact that the output of monascorubin and Citrinin causes.
Monascus seed culture: 1% sucrose, 0.3% peptone, 0.05% dipotassium hydrogen phosphate, 0.05% magnesium sulfate, hydrochloric acid is adjusted to pH 5.7,121 DEG C of sterilizing 20min, spore suspension to above-mentioned culture medium inoculated 2% from wash-out wort agar inclined-plane, is placed in and cultivates 1d in 160r/min 35 DEG C.
Fermentation Condition of Monascus spp is cultivated: 2% Tapioca Starch, 1% peptone, and 2% glucose, is adjusted to pH5.8 with phosphoric acid, hydrochloric acid respectively, 121 DEG C of sterilizing 20min, and to the seed liquor that above-mentioned fermention medium inoculation 4% has been cultivated, ventilate 35 DEG C of fermentation 5d.Result is as shown in table 1.
Table 1 phosphate radical is on the impact of tunning
As can be seen from Table 1, identical Fermentation Condition of Monascus spp condition, fermention medium can largely reduce Citrinin output with phosphorus acid for adjusting pH, improves monascorubin output simultaneously.There is phosphoric acid in this experiment fermention medium and achieve obvious technique effect, is the result that contriver surprisingly obtains.
Embodiment 2
The present embodiment illustrates and add different sorts phosphoric acid salt in Fermentation Condition of Monascus spp substratum, on the impact that the output of monascorubin and Citrinin causes.
Monascus seed culture method is with embodiment 1.
Fermentation Condition of Monascus spp cultural method adds Sodium phosphate dibasic, dipotassium hydrogen phosphate or the Secondary ammonium phosphate of 0.05% on the basis of embodiment 1 in fermention medium, and regulates medium pH to avoid the interference of phosphate radical with hydrochloric acid, and result is as shown in table 2.
Table 2 different phosphate hydrochlorate is on the impact of Fermentation Condition of Monascus spp
As can be seen from Table 2, not add phosphatic experiment for contrast, different sorts phosphoric acid salt is added in fermention medium, in tunning, citrinin content is very low maybe can not detect, and the output of monascorubin all increases, wherein when adding Secondary ammonium phosphate, the fermentation results of monascus is optimum.
Embodiment 3
The present embodiment illustrates the impact that Secondary ammonium phosphate in Fermentation Condition of Monascus spp substratum adds the output of concentration on monascorubin and Citrinin and causes.
Monascus seed culture method is with embodiment 1.
Fermentation Condition of Monascus spp cultural method adds the Secondary ammonium phosphate of 0.025%, 0.05%, 0.075% and 0.10% respectively on the basis of embodiment 1 in Fermentation Condition of Monascus spp substratum, and regulating medium pH to avoid the interference of phosphate radical with hydrochloric acid, result is as shown in table 3.
Table 3 different concns Secondary ammonium phosphate is on the impact of Fermentation Condition of Monascus spp
As can be seen from Table 3, not add the experiment of Secondary ammonium phosphate for contrast, in fermention medium, add different concns Secondary ammonium phosphate, in tunning, all do not detect Citrinin, and the output of pigment all significantly improves, when being wherein 0.05% with addition, effect is best.
Embodiment 4
The present embodiment illustrates to be made Fermentation Condition of Monascus spp culture medium carbon source with Tapioca Starch and adds the impact of phosphoric acid salt on monascorubin and Citrinin output.
Monascus seed culture method is with embodiment 1.
Fermentation Condition of Monascus spp is cultivated:
Control group: 2% rice meal, 1% peptone, 2% glucose;
Experimental group 1:2% Tapioca Starch, 1% peptone, 2% glucose, 0.001% Secondary ammonium phosphate.
Experimental group 2:2% Tapioca Starch, 1% peptone, 2% glucose, 0.05% Secondary ammonium phosphate.
Substratum hydrochloric acid is adjusted to pH5.8,121 DEG C of sterilizing 20min, and to the seed liquor that above-mentioned fermention medium inoculation 4% has been cultivated, ventilate 35 DEG C of fermentation 5d.Result is as shown in table 4.
Table 4 Tapioca Starch compound phosphoric acid salt culture medium is on the impact of Fermentation Condition of Monascus spp
Rice meal is conventional Fermentation Condition of Monascus spp substrate, as can be seen from Table 4, compares with rice meal fermentation monascus, adopts Tapioca Starch to do substrate and the experimental group of adding Secondary ammonium phosphate can significantly improve the output that pigment production significantly reduces Citrinin simultaneously.Add 0.001% Secondary ammonium phosphate and reach good ferment effect, the combination of this content Secondary ammonium phosphate and cassava can significantly improve the output that pigment production significantly reduces Citrinin simultaneously.
Claims (6)
1. a Fermentation Condition of Monascus spp method for low-yield citrinin high yield monascus pigment, comprises seed culture and the liquid state fermentation of monascus, and the carbon source in described liquid state fermentation substratum is Tapioca Starch, is added with phosphoric acid salt in substratum; Tapioca Starch addition in described fermention medium is 1-6%; Phosphoric acid salt is Secondary ammonium phosphate, Sodium phosphate dibasic, dipotassium hydrogen phosphate.
2. the Fermentation Condition of Monascus spp method of low-yield citrinin high yield monascus pigment according to claim 1, the addition of described Secondary ammonium phosphate is 0.001-0.1%.
3. the Fermentation Condition of Monascus spp method of low-yield citrinin high yield monascus pigment according to claim 1, specific as follows: monascus seed culture: 1% sucrose, 0.3% peptone, 0.05% dipotassium hydrogen phosphate, 0.05% magnesium sulfate, hydrochloric acid is adjusted to pH 5.7,121 DEG C of sterilizing 20min, spore suspension to above-mentioned culture medium inoculated 2% from wash-out wort agar inclined-plane, is placed in 160r/min 35 DEG C and cultivates 1d; Fermentation Condition of Monascus spp is cultivated: 2% Tapioca Starch, 1% peptone, and 2% glucose, is adjusted to pH5.8 with phosphoric acid, 121 DEG C of sterilizing 20min, and to the seed liquor that above-mentioned fermention medium inoculation 4% has been cultivated, ventilate 35 DEG C of fermentation 5d.
4. the Fermentation Condition of Monascus spp method of low-yield citrinin high yield monascus pigment according to claim 1, described Fermentation Condition of Monascus spp substratum consists of: 2% Tapioca Starch, 1% peptone, 2% glucose, 0.05% Secondary ammonium phosphate.
5. the Fermentation Condition of Monascus spp method of low-yield citrinin high yield monascus pigment according to claim 1, described Fermentation Condition of Monascus spp substratum consists of: 2% Tapioca Starch, 1% peptone, 2% glucose, 0.001% Secondary ammonium phosphate, pH 5.8.
6. the Fermentation Condition of Monascus spp substratum of low-yield citrinin high yield monascus pigment, consists of: 2% Tapioca Starch, 1% peptone, 2% glucose, 0.001% Secondary ammonium phosphate, pH 5.8.
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Cited By (12)
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| CN106399382A (en) * | 2016-10-21 | 2017-02-15 | 福州大学 | Fermentation preparation method of monascus pigment |
| CN107296266A (en) * | 2017-05-20 | 2017-10-27 | 河南福切尔生物科技有限公司 | A kind of jujube ferment and preparation method thereof |
| CN108486164A (en) * | 2018-03-30 | 2018-09-04 | 福建省农业科学院农业工程技术研究所 | The production technology of low citrinin Monascus color |
| CN110184303A (en) * | 2019-06-20 | 2019-08-30 | 中南林业科技大学 | A method of monascorubin is prepared by raw material of yellow serofluid |
| CN110305802A (en) * | 2019-08-22 | 2019-10-08 | 光明乳业股份有限公司 | The culture medium and cultural method of high spore output monascus purpureus are bred in a kind of liquid state fermentation |
| CN110499338A (en) * | 2018-05-17 | 2019-11-26 | 华中农业大学 | A kind of additive that can improve monascorubin yield while reducing citrinin yield |
| CN111394404A (en) * | 2019-05-14 | 2020-07-10 | 江苏臻大天园健康科技有限公司 | Method for producing coenzyme Q10 by using red yeast rice |
| CN112430635A (en) * | 2020-12-01 | 2021-03-02 | 广东肇庆星湖生物科技股份有限公司 | Fermentation method for low-yield citrinin and high-yield water-soluble monascus yellow pigment |
| CN114806894A (en) * | 2022-05-20 | 2022-07-29 | 南昌大学 | Monascus liquid fermentation method and application |
| CN115261423A (en) * | 2022-07-12 | 2022-11-01 | 北京工商大学 | Fermentation method for changing anabolism mode of monascus pigment in monascus purpureus and improving synthetic amount |
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| CN104962483B (en) * | 2015-07-20 | 2019-01-29 | 光明乳业股份有限公司 | A kind of monascus purpureus bacterial strain and application thereof |
| CN105062894A (en) * | 2015-07-20 | 2015-11-18 | 光明乳业股份有限公司 | Monascus purpureus strain and application thereof |
| CN104962483A (en) * | 2015-07-20 | 2015-10-07 | 光明乳业股份有限公司 | Monascus purpureus strain and application thereof |
| CN105062894B (en) * | 2015-07-20 | 2019-01-29 | 光明乳业股份有限公司 | A kind of purple Monascus Strains and its application |
| CN106399382A (en) * | 2016-10-21 | 2017-02-15 | 福州大学 | Fermentation preparation method of monascus pigment |
| CN107296266A (en) * | 2017-05-20 | 2017-10-27 | 河南福切尔生物科技有限公司 | A kind of jujube ferment and preparation method thereof |
| CN108486164A (en) * | 2018-03-30 | 2018-09-04 | 福建省农业科学院农业工程技术研究所 | The production technology of low citrinin Monascus color |
| CN110499338A (en) * | 2018-05-17 | 2019-11-26 | 华中农业大学 | A kind of additive that can improve monascorubin yield while reducing citrinin yield |
| CN111394404A (en) * | 2019-05-14 | 2020-07-10 | 江苏臻大天园健康科技有限公司 | Method for producing coenzyme Q10 by using red yeast rice |
| CN110184303A (en) * | 2019-06-20 | 2019-08-30 | 中南林业科技大学 | A method of monascorubin is prepared by raw material of yellow serofluid |
| CN110305802A (en) * | 2019-08-22 | 2019-10-08 | 光明乳业股份有限公司 | The culture medium and cultural method of high spore output monascus purpureus are bred in a kind of liquid state fermentation |
| CN112430635A (en) * | 2020-12-01 | 2021-03-02 | 广东肇庆星湖生物科技股份有限公司 | Fermentation method for low-yield citrinin and high-yield water-soluble monascus yellow pigment |
| CN114806894A (en) * | 2022-05-20 | 2022-07-29 | 南昌大学 | Monascus liquid fermentation method and application |
| CN115261423A (en) * | 2022-07-12 | 2022-11-01 | 北京工商大学 | Fermentation method for changing anabolism mode of monascus pigment in monascus purpureus and improving synthetic amount |
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