CN106755171A - A kind of method for promoting red yeast rice strains liquid fermentation to produce red yeast rice citraurin - Google Patents
A kind of method for promoting red yeast rice strains liquid fermentation to produce red yeast rice citraurin Download PDFInfo
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- CN106755171A CN106755171A CN201710071293.6A CN201710071293A CN106755171A CN 106755171 A CN106755171 A CN 106755171A CN 201710071293 A CN201710071293 A CN 201710071293A CN 106755171 A CN106755171 A CN 106755171A
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Abstract
The invention discloses a kind of method for promoting red yeast rice strains liquid fermentation to produce red yeast rice citraurin, belong to technical field of microbial fermentation.Present invention inoculation Monascus seed culture fluid is in 6 8d that after liquid culture medium, fermented at 30 DEG C；Its fermentation start after 24h in fermentation culture in add surfactant, to promote Monascus synthetic active substance red yeast rice citraurin in growth course.The present invention can improve the biosynthesis ability of red yeast rice strains liquid fermentation red yeast rice citraurin by these effective regulation and control；It is convenient and practical only by relatively low additional input, it is possible to improve the concentration of fermentation synthesis high added value target product, and with great economic benefit.
Description
Technical field
The present invention relates to a kind of method for promoting red yeast rice strains liquid fermentation to produce red yeast rice citraurin, belong to microbial fermentation skill
Art field.
Background technology
At present, microbe fermentation method and plant extraction process are two kinds of main methods for producing natural orange pigment.Plant origin
Natural citraurin such as：Annatto pigment, natural capsicum citraurin, carrot citraurin etc., the pigment of these plant origins exists
The limitation of each side is faced in production process, in one side extractive process will use organic reagent, it is relatively costly, yield and should
With being limited, on the other hand its seasonality is very strong, and its production can cause the utilization rate of equipment low under the influence of season, cause money
Source wastes.Conversely, it is nontoxic using the natural citraurin basic security of Production by Microorganism Fermentation, before very wide development
Scape.
To improve the red yeast rice citraurin level of production, there is document report xuwei et al. to obtain one plant of production by mutagenic and breeding
The production bacterium Monascus puepureus W5S8 of red yeast rice citraurin, the yield of liquid production citraurin is only up to 16.38U/
mL.Conventional liquid fermentation process is used before this laboratory, final citraurin colour value is only 80U/mL, and there is fermentation week
Phase is long, a series of problems, such as colour value relatively low high cost.The requirement of industrialized production is not met.
This laboratory is (special in patent of invention " a kind of biliquid phase fermentation method of coupling situ extracting fermentation red yeast rice citraurin "
Profit number 201410117001.4) in, by the content of free amino acid in carbon nitrogen source component reasonable in design, control culture medium with
And Two Liquid Phases fermentation system, the red yeast rice citraurin of synthesis is enriched to rapidly in fat-soluble solvent (fat-soluble solvent is main
Refer to glycerides), so as to greatly improve the production efficiency of red yeast rice citraurin.But in actual production process, find still
So there is the phenomenon of Product inhibiton, the secretion of para chrome still can be restricted, it is therefore necessary to so that citraurin yield is able to
Further improve.
The content of the invention
The route of synthesis that red yeast rice citraurin is produced in current Monascus fermentation does not still obtain in-depth study, using common
Liquid state fermentation medium component, is extremely difficult to the purpose of high yield monascus citraurin.In the original location on the basis of extractive fermentation, product suppression
The releasing of a part is obtained, while mass transfer and biography oxygen are able to the raising of part, but still having further improve red yeast rice orange
Necessity of plain yield.Method of the present invention by adding surfactant, increases membrane passage, suppresses substrate is released
While increase intracellular mass transfer, oxygen mass transfer coefficient.The method is simple to operate, and extra artificial and equipment is not increased, only leads to
Cross relatively low additional input, it is possible to improve the synthesis concentration of target product, thus with great economic benefit.
The invention provides a kind of method for promoting red yeast rice strains liquid fermentation to produce red yeast rice citraurin, methods described is red
The liquid state fermentation of aspergillus initially or in fermentation process adds surfactant.
In one embodiment, the Monascus are the Monascus that can produce red yeast rice citraurin, can be purple red yeast rice
Bacterium or Monascus ruber.
In one embodiment, the surfactant is Span series, Tween series, Triton X-100, PEG
Series or gleditsia sinensis.The surfactant plays the role of to release Product inhibiton and improves dissolved oxygen, can greatly improve red yeast rice
Synthesis capability and yield of the bacterium to citraurin.
In one embodiment, the surfactant is in Span-80, Tween-80, Triton X-100, PEG
Any one.
In one embodiment, the addition of the surfactant is 0.1g/L-15g/L.
In one embodiment, the addition of the surfactant is 1g/L-6g/L.
In one embodiment, the addition opportunity of the surfactant for liquid state fermentation culture start after 0~96h
It is interior.
In one embodiment, the addition opportunity of the surfactant for liquid state fermentation culture start after 0~24h
It is interior.
In one embodiment, it is described fermentation be rotating speed be 180r/min, under the conditions of 30 DEG C of temperature, fermented and cultured
8d。
In one embodiment, methods described is that inoculation Monascus seed liquor is sent out in liquid state fermentation culture medium
Ferment；After liquid state fermentation culture starts surfactant is added in 0~96h.
In one embodiment, the liquid state fermentation culture medium is using containing nutritional ingredient needed for red yeast rice bacteria growing
Minimal medium.
In one embodiment, the liquid state fermentation culture medium is sent out by the double liquid phase extraction of water phase and extraction phase composition
Ferment system, the water is mutually the minimal medium containing nutritional ingredient needed for red yeast rice bacteria growing, and the extraction phase contains glyceride
Class material.
In one embodiment, the minimal medium contains cornstarch 20-80g/L, analysis for soybean powder 0.05-10g/L,
Ammonium sulfate 3-10g/L, NaNO32-6g/L, yeast extract 0.05-5g/L, corn starch 0.05-10g/L, zinc sulfate 0.1-0.5g/
L, magnesium sulfate 0.5-1.5g/L, potassium dihydrogen phosphate 0.5-1.5g/L.
In one embodiment, the glycerides in the extraction phase are improved the work of dissolved oxygen as extractant
With, can will be attached in zymotic fluid and the pigment extraction of Monascus ball surface is enriched in extractant phase, increase substantially red yeast rice
The concentration and yield of liquid state fermentation synthesis citraurin product.
In one embodiment, the glycerides are triglyceride materials, and consumption volume accounts for basic culture
The 10-40% of matrix product.The triglyceride material can be tricaprylin, three isooctyl acid glyceride, vegetable oil or
Its mixture.
In one embodiment, the extraction phase is 10 parts of soybean oils, 8 parts of corn oils, 1 part of tricaprylin, 0.3
The mixing of part two glycerol caprylates, 0.2 part of glycerol caprylate.
In one embodiment, the glycerides remove triglyceride material, while also including diglyceride
Class material, monoglyceride class material or its mixture.
Advantages of the present invention and effect：
The method for improving red yeast rice citraurin yield of the invention, economical and efficient, and the structure of pigment will not be changed, it is a kind of
Preferable production method, the feasibility in industrial mass production is higher.
Specific embodiment
The assay method of citraurin colour value：
Colour value is a kind of method for expressing of pigment concentration, and size is equal to extension rate and is inhaled at it with the pigment solution after dilution
The product of corresponding OD values at peak is received, unit is U/mL.
With absolute ethyl alcohol dilute suitable multiple (OD values are controlled between 0.2-0.8) to solution to be measured, and with anhydrous second
The absorbance of alcohol carries out full wavelength scanner for blank in the range of 200-600nm, determines the OD of peak value between 450-480nm
Value.
Citraurin colour value=OD460nm× total extension rate (U/mL)
Embodiment 1：
Selection purple Monascus CH2, CICC numbering is that 40225, CICC numberings are purple that 40937, CICC numberings are 5042
Color Monascus, and CGMCC numberings are that 3.4701, CGMCC numberings are that 3.4652, CGMCC numberings are that 3.465, CGMCC numberings are
3.4649 Monascus ruber is shaking flask strain, its fermentation minimal medium (g/L)：Cornstarch 60, ammonium sulfate 4, sodium nitrate
2, magnesium sulfate 0.5, potassium dihydrogen phosphate 2, dipotassium hydrogen phosphate 2, calcium chloride 0.1；
Monascus liquid seeds in exponential phase are accessed equipped with 50mL fermentation bases with the inoculum concentration of volume ratio 10%
It is 180r/min in rotating speed in the 500mL triangular flasks of basal culture medium, under the conditions of 30 DEG C of temperature, shake flask fermentation culture 8d.After end
Measure this eight plants of bacterium CH2, CICC 40225, CICC 40937, CICC 5042, CGMCC 3.4701, CGMCC 3.4652,
CGMCC 3.465, the colour value of the red yeast rice citraurin of CGMCC 3.4649 be respectively 75U/mL, 64U/mL, 44U/mL, 42U/mL,
30U/mL、62U/mL、26U/mL、35U/mL。
Embodiment 2：
Selection purple Monascus CH2, CICC numbering is that 40225, CICC numberings are purple that 40937, CICC numberings are 5042
Color Monascus, and CGMCC numberings are that 3.4701, CGMCC numberings are that 3.4652, CGMCC numberings are that 3.465, CGMCC numberings are
3.4649 Monascus ruber is shaking flask strain, its fermentation minimal medium (g/L)：Cornstarch 60, ammonium sulfate 4, sodium nitrate
2, magnesium sulfate 0.5, potassium dihydrogen phosphate 2, dipotassium hydrogen phosphate 2, calcium chloride 0.1；
Monascus liquid seeds in exponential phase are accessed equipped with 50mL fermentation bases with the inoculum concentration of volume ratio 10%
In the 500mL triangular flasks of basal culture medium, while span-80, Tween-80, Triton of 2g/L is added when fermenting initial respectively
X-100, PEG are 180r/min in rotating speed, are 180r/min in rotating speed, under the conditions of 30 DEG C of temperature, shake flask fermentation culture 8d.Terminate
Measure the colour value of red yeast rice citraurin respectively afterwards, as a result as shown in table 1.
The different surfaces activating agent of table 1 produces the influence of red yeast rice citraurin to bacterial strain
Embodiment 3：
Selection purple Monascus CH2, CICC numbering is that 40225, CICC numberings are purple that 40937, CICC numberings are 5042
Color Monascus are shaking flask strain, its fermentation minimal medium (g/L)：Cornstarch 60, ammonium sulfate 4, sodium nitrate 2, magnesium sulfate
0.5, potassium dihydrogen phosphate 2, dipotassium hydrogen phosphate 2, calcium chloride 0.1；
Monascus liquid seeds in exponential phase are accessed equipped with 50mL fermentation bases with the inoculum concentration of volume ratio 10%
In the 500mL triangular flasks of basal culture medium, (extraction phase is 10 parts of soybean oils, 8 parts of corns to add glycerolipid situ extracting phase 20mL
Oil, 1 part of tricaprylin, 0.3 part of two glycerol caprylate, the mixing of 0.2 part of glycerol caprylate), it is 180r/min in rotating speed,
Under the conditions of 30 DEG C of temperature, shake flask fermentation culture 8d.Colour value 330U/mL, 280U/mL, the 260U/ of red yeast rice citraurin are measured after end
mL、250U/mL。
Embodiment 4：
Selection Monascus CH2 is shaking flask strain, its fermentation minimal medium (g/L)：Cornstarch 60, ammonium sulfate 4, nitric acid
Sodium 2, magnesium sulfate 0.5, potassium dihydrogen phosphate 2, dipotassium hydrogen phosphate 2, calcium chloride 0.1；
Monascus liquid seeds in exponential phase are accessed equipped with 50mL fermentation bases with the inoculum concentration of volume ratio 10%
In the 500mL triangular flasks of basal culture medium, glycerolipid situ extracting phase 20mL (with embodiment 3) is added, while initial in fermentation
Tween-80, span-80, Triton X-100, PEG of Shi Tianjia 1g/L, is 180r/min, 30 DEG C of conditions of temperature in rotating speed
Under, shake flask fermentation culture 8d.Measure the colour value of red yeast rice citraurin after end, respectively 450U/mL, 360U/mL, 374U/mL,
348U/mL.Compared with Example 3, it has been respectively increased 36.4%, 9.1%, 13.3%, 5.5%.
Embodiment 5：
Selection Monascus CH2 is shaking flask strain, its fermentation minimal medium (g/L)：Cornstarch 60, ammonium sulfate 4, nitric acid
Sodium 2, magnesium sulfate 0.5, potassium dihydrogen phosphate 2, dipotassium hydrogen phosphate 2, calcium chloride 0.1；
Monascus liquid seeds in exponential phase are accessed equipped with 50mL fermentation bases with the inoculum concentration of volume ratio 10%
In the 500mL triangular flasks of basal culture medium, glycerolipid situ extracting phase 20mL (with embodiment 3) is added, while when fermenting initial
The Tween-80 of 4g/L is added, is 180r/min in rotating speed, under the conditions of 30 DEG C of temperature, shake flask fermentation culture 8d.Measured after end
The colour value of red yeast rice citraurin is 530U/mL.
Embodiment 6：
Selection Monascus CH2 is shaking flask strain, its fermentation minimal medium (g/L)：Cornstarch 60, ammonium sulfate 4, nitric acid
Sodium 2, magnesium sulfate 0.5, potassium dihydrogen phosphate 2, dipotassium hydrogen phosphate 2, calcium chloride 0.1；
Monascus liquid seeds in exponential phase are accessed equipped with 50mL fermentation bases with the inoculum concentration of volume ratio 10%
In the 500mL triangular flasks of basal culture medium, glycerolipid situ extracting phase 20mL (with embodiment 3) is added, while when fermenting initial
The Tween-80 of 5g/L is added, is 180r/min in rotating speed, under the conditions of 30 DEG C of temperature, shake flask fermentation culture 8d.Measured after end
The colour value of red yeast rice citraurin is 495U/mL.
Embodiment 7：
Selection Monascus CH2 is shaking flask strain, its fermentation minimal medium (g/L)：Cornstarch 60, ammonium sulfate 4, nitric acid
Sodium 2, magnesium sulfate 0.5, potassium dihydrogen phosphate 2, dipotassium hydrogen phosphate 2, calcium chloride 0.1；
Monascus liquid seeds in exponential phase are accessed equipped with 50mL fermentation bases with the inoculum concentration of volume ratio 10%
In the 500mL triangular flasks of basal culture medium, glycerolipid situ extracting phase 20mL (with embodiment 3) is added, while when fermenting initial
The Tween-80 of 6g/L is added, is 180r/min in rotating speed, under the conditions of 30 DEG C of temperature, shake flask fermentation culture 8d.Measured after end
The colour value of red yeast rice citraurin is 410U/mL.
Embodiment 8：
Selection Monascus CH2 is shaking flask strain, its fermentation minimal medium (g/L)：Cornstarch 60, ammonium sulfate 4, nitric acid
Sodium 2, magnesium sulfate 0.5, potassium dihydrogen phosphate 2, dipotassium hydrogen phosphate 2, calcium chloride 0.1；
Monascus liquid seeds in exponential phase are accessed equipped with 50mL fermentation bases with the inoculum concentration of volume ratio 10%
In the 500mL triangular flasks of basal culture medium, glycerolipid situ extracting phase 20mL (with embodiment 3) is added, while adding during fermentation 24h
Plus the Tween-80 of 4g/L, it is 180r/min in rotating speed, under the conditions of 30 DEG C of temperature, shake flask fermentation culture 8d.Measured after end red
The colour value of bent citraurin is 550U/mL.
Embodiment 9：
Selection Monascus CH2 is shaking flask strain, its fermentation minimal medium (g/L)：Cornstarch 60, ammonium sulfate 4, nitric acid
Sodium 2, magnesium sulfate 0.5, potassium dihydrogen phosphate 2, dipotassium hydrogen phosphate 2, calcium chloride 0.1；
Monascus liquid seeds in exponential phase are accessed equipped with 50mL fermentation bases with the inoculum concentration of volume ratio 10%
In the 500mL triangular flasks of basal culture medium, glycerolipid situ extracting phase 20mL (with embodiment 3) is added, while adding during fermentation 48h
Plus the Tween-80 of 4g/L, it is 180r/min in rotating speed, under the conditions of 30 DEG C of temperature, shake flask fermentation culture 8d.Measured after end red
The colour value of bent citraurin is 500U/mL.
Embodiment 10：
Selection Monascus CH2 is shaking flask strain, its fermentation minimal medium (g/L)：Cornstarch 60, ammonium sulfate 4, nitric acid
Sodium 2, magnesium sulfate 0.5, potassium dihydrogen phosphate 2, dipotassium hydrogen phosphate 2, calcium chloride 0.1；
Monascus liquid seeds in exponential phase are accessed equipped with 50mL fermentation bases with the inoculum concentration of volume ratio 10%
In the 500mL triangular flasks of basal culture medium, glycerolipid situ extracting phase 20mL (with embodiment 3) is added, while adding during fermentation 96h
Plus the Tween-80 of 4g/L, it is 180r/min in rotating speed, under the conditions of 30 DEG C of temperature, shake flask fermentation culture 8d.Measured after end red
The colour value of bent citraurin is 470U/mL.
Although the present invention is disclosed as above with preferred embodiment, it is not limited to the present invention, any to be familiar with this skill
The people of art, without departing from the spirit and scope of the present invention, can do various changes with modification, therefore protection model of the invention
Enclose being defined of being defined by claims.
Claims (10)
1. a kind of method that promotion red yeast rice strains liquid fermentation produces red yeast rice citraurin, methods described is the liquid state fermentation in Monascus
Surfactant is added in initial or fermentation process.
2. method according to claim 1, it is characterised in that the surfactant be Span series, Tween series,
Triton X-100, PEG series or gleditsia sinensis.
3. method according to claim 1, it is characterised in that the surfactant be Span-80, Tween-80,
Any one in Triton X-100, PEG.
4. method according to claim 1, it is characterised in that the Monascus are the Monascus that can produce red yeast rice citraurin
Kind.
5. method according to claim 1, it is characterised in that the addition of the surfactant is 0.1g/L-15g/
L。
6. method according to claim 1, it is characterised in that the addition opportunity of the surfactant is liquid state fermentation training
After foster beginning in 0~96h.
7. method according to claim 1, it is characterised in that the addition opportunity of the surfactant is liquid state fermentation training
After foster beginning in 0~24h.
8. method according to claim 1, it is characterised in that methods described is inoculation Monascus seed liquor in liquid state fermentation
Fermented in culture medium；After liquid state fermentation culture starts surfactant is added in 0~96h；Wherein liquid state fermentation culture
Base is minimal medium.
9. method according to claim 8, it is characterised in that the minimal medium contains cornstarch 20-80g/L,
Analysis for soybean powder 0.05-10g/L, ammonium sulfate 3-10g/L, NaNO32-6g/L, yeast extract 0.05-5g/L, corn starch 0.05-10g/
L, zinc sulfate 0.1-0.5g/L, magnesium sulfate 0.5-1.5g/L, potassium dihydrogen phosphate 0.5-1.5g/L.
10. method according to claim 1, it is characterised in that methods described is that inoculation Monascus seed liquor is sent out in liquid
Fermented in ferment culture medium；After liquid state fermentation culture starts surfactant is added in 0~96h；The liquid state fermentation training
Foster base is by the double liquid phase extraction fermentation system of water phase and extraction phase composition；The water is mutually to contain nutrition needed for red yeast rice bacteria growing
The minimal medium of composition, the extraction phase contains glycerides.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| 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 |
| CN114058648A (en) * | 2021-12-14 | 2022-02-18 | 广东天益生物科技有限公司 | Method for high-yield fermentation of extracellular monascus red pigment by using monascus purpureus |
| CN114350526A (en) * | 2022-01-14 | 2022-04-15 | 北京工商大学 | Method for improving monascus pigment tone |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| 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 |
| CN114058648A (en) * | 2021-12-14 | 2022-02-18 | 广东天益生物科技有限公司 | Method for high-yield fermentation of extracellular monascus red pigment by using monascus purpureus |
| CN114058648B (en) * | 2021-12-14 | 2023-11-21 | 广东天益生物科技有限公司 | Method for fermenting high-yield extracellular monascus red pigment by using monascus purpureus |
| CN114350526A (en) * | 2022-01-14 | 2022-04-15 | 北京工商大学 | Method for improving monascus pigment tone |
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