CN104893983B - Liquid state fermentation low citrinin, the preparation method of High color values monascorubin and product - Google Patents
Liquid state fermentation low citrinin, the preparation method of High color values monascorubin and product Download PDFInfo
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Abstract
The invention discloses a kind of red yeast rice Monascus YZ301 (Monascus anka Yang YZ301) and its breeding method, microbial preservation number is CGMCC9707；Also disclose the method that low citrinin High color values monascorubin is prepared using the red yeast rice strains liquid fermentation comprising following steps: (1) strain improvement；(2) seed culture, expansion culture；(3) liquid state fermentation produce, further include extraction purification, spray drying and etc.；Wherein strain be by the red yeast rice Monascus YZ201 (Monascus anka Nakazawa et Sato) of Dongguan City Tian Yi biofermentation technique Co., Ltd exploitation after culture, the screening of multiple ultraviolet mutagenesis, separating for several times resulting red yeast rice Monascus YZ301 (Monascus anka Yang YZ301), the invention also discloses the above-mentioned preparation method of the use High color values, the low-citrinin monascus pigment products that are prepared.Strain provided by the invention is excellent, rational technology is stablized, gained production height, stay in grade, and the content of citrinin is 10300~10800u/g no more than 0.2mg/kg (in terms of unit color value 500u/g), monascorubin color value in finished product.
Description
Technical field
The present invention relates to technical field of biological fermentation, in particular to a kind of liquid state fermentation low citrinin, High color values Monascus color
The preparation method and its product of pigment.
Background technique
Red yeast rice category Aspergillaceae fungi monascus parpureus Went, also known as Monascus, are that cultivation and fermentation forms in rice with Monascus.
Traditionally, red yeast rice is widely used in the field of food such as red wine, vinegar.Modern medicine research report thinks, red yeast rice have blood pressure lowering,
The effect of reducing blood lipid, contained monacolin K can prevent to generate cholesterol.Monascorubin is also the traditional product in China, is logical
One of the natural pigment the most successful crossing Production by Microorganism Fermentation and being used widely.Monascorubin is as currently the only
A kind of natural safe edible pigment from microorganism be widely used in food, medicine, daily necessities field.It is red
Bent pigment is the alcohol-soluble pigment of ketone of birdsing of the same feather flock together, wherein there is 6 kinds of ingredients: uranidin (Monascin, monascus yellow pigment), haematochrome
(Pan Hongsu, monascorubin), purpurin (Pan Hong amine, monascorubramine), 6 kinds of constituent structure are close, slightly difference.But it is making
During standby monascorubin, it will usually which generating a kind of has carcinogenic, teratogenesis mycotoxin i.e. citrinin, its genotoxicity
Popularization to from monascus product to the whole world generates serious negative effect.
At present, there are mainly two types of the methods for producing monascorubin, first is that producing red yeast rice by solid fermentation, then again
Pigment is extracted, can also directly be applied；Second is that producing pigment by liquid fermentation.However the resulting Monascus color of solid-fermented technique
The color value of pigment is not high and limits its application.Monascorubin is produced using liquid fermentation method at present, pigment content in product,
And yield, finished product color value etc. be not high, and how to improve the color value of monascorubin in fermentation liquid, reduce citrinin content, improve
Product color value and pigment content are largely limited by the factors such as strain, raw material proportioning, fermentation process, production technology, equipment
Influence, Monascus produce citrinin number, in addition to having outside the Pass with strain, also have close relationship with technological condition for fermentation；
However it is actually rare to the optimizing research in the preparation process of monascorubin in the prior art.
Therefore, research and develop a kind of strain that liquid state fermentation is had excellent performance, breeding method and to prepare low tangerine by liquid state fermentation mould
The method of element, High color values monascorubin, improves the production efficiency and quality of monascorubin, becomes more urgent.
Summary of the invention
In view of the deficiencies of the prior art, the object of the present invention is to provide one kind with Dongguan City day benefit biofermentation technique
Co., Ltd exploitation red yeast rice Monascus YZ201 (Monascus anka Nakazawa et Sato) through culture, ultraviolet mutagenesis,
The red yeast rice Monascus YZ301 (Monascus anka) obtained after separation screening is red as production High color values, low citrinin red yeast rice
The strain of element.
The present invention also aims to, provide it is a kind of with red yeast rice Monascus YZ301 (Monascus anka) be strain, it is logical
The liquid-state fermentation technology for crossing optimization prepares the preparation method of low citrinin, High color values monascorubin.
The present invention also aims to provide a kind of liquid state fermentation low citrinin, height being prepared using the above method
Color value monascorubin product.
The technical scheme adopted by the invention to achieve the purpose is as follows:
A kind of red yeast rice Monascus YZ301 (Monascus anka), microbial preservation number is CGMCC9707；In It is deposited in China Committee for Culture Collection of Microorganisms's common micro-organisms center on September 26th, 2014, which is located at Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3.
The breeding method of the red yeast rice Monascus YZ301 (Monascus anka) is by the prebiotic object hair in Dongguan City day
The red yeast rice Monascus YZ201 (Monascus anka Nakazawa et Sato) of ferment Technology Co., Ltd. exploitation is through culture, purple
The red yeast rice Monascus YZ301 (Monascus anka) obtained after outer mutagenesis, separation screening.
Described red yeast rice Monascus YZ301 (Monascus anka) its fungus characteristic are as follows: mycelia: 3~5um of diameter, generally
3um, containing particle, long afterwards, particle reddens color, therefore mycelium laking for culture；Conidiospore: spherical, pyriform, diameter
6~9~12um, it is colourless, contain particle, usual 2~3 chainings；Cleistothecium: spherical, diameter 304~0 (50) um, 30um or less person
Seldom；The full son of ascus: ellipse, colourless 3x 2um；Pigment: according to Ridgway chromatography, color is that red pomegranate is red
(Grenadine Red)；Growth temperature: optimum temperature be 33~35 DEG C, 37 DEG C or more, 30 DEG C or less slow growths, 40 DEG C with
On hardly grow.
Red yeast rice Monascus YZ301 (Monascus anka Yang YZ301) AS 3.2636Wa:25 DEG C, 7d, colony diameter
22um, sunset clouds are red；25d colony diameter 56mm, protuberance is red in scar shape raspberry, and bacterium colony center is split.
MEA culture medium: 25 DEG C, 7d, colony diameter 8mm, cologne earth color；25d colony diameter 20mm, Chu Shise；CYA culture
Base: 25 DEG C, 7d, colony diameter 25rnm, baby pink.
Preferably, the culture is 10~15 in wort agar medium。Be, 28~35 DEG C of temperature cultures 6~8
It, bacterium colony growth of planar, on koji agar, in 33~35 DEG C of cultures, 7 days membranaceous bacterium colonies of cutification, no aerial hyphae, shape
If lava flows out, pomegranate water is red, the eugonic on wort agar.
Preferably, the ultraviolet mutagenesis and screening include: A, ultraviolet mutagenesis: will cultivate 7~9 days red yeast rice strains, warp
High temperature 55~65 DEG C of 2~10min of processing, 0.04~0.07Mpa of high pressure are handled 5~10 hours, are put into and are provided with band bead, nothing
The triangular flask of 30~50ml of bacterium water is placed in shaking table and vibrates 1~2 hour, and filtering takes 10ml spore suspension, is added to band magnetic bar
Culture dish in, under magnetic stirring carry out 2~10min of ultraviolet irradiation, apply plate, wrapped up with black cloth, 32~35 DEG C of Yu Wendu
Lower culture 6~8 days；
B, isolation medium: soluble starch 5%, maltose 3%, peptone 0.1% add appropriate lactic acid, adjust pH
Value 5~6, agar 2%；
C, isolation medium PDA:NaNO33.0g, KH2PO41.0g, MgSO47H2O 0.5g, FeSO47H2O 0.01g, sugarcane
Sugared 30g, tap water 1000ml, agar 15g；
D, slant medium MEA: brewer's wort 20g, glucose 20g, peptone 1.0g, tap water 1000ml, agar 20g.
A method of low citrinin, High color values monascorubin are prepared using aforementioned red yeast rice strains liquid fermentation comprising
Following steps:
(1) strain improvement: using the red yeast rice Monascus YZ301 (Monascus anka) isolated by ultraviolet mutagenesis
As strain, microbial preservation number is CGMCC9707；
(2) seed culture, expansion culture: the red yeast rice Monascus YZ301 that step (1) breeding obtains is sequentially ingressed into equipped with kind
It is cultivated respectively in the I and II seeding tank of sub- culture medium 6~9 hours, it is small to be inoculated into expansion culture 75~85 in fermentor later
When, obtain liquid seed culture fluid；
(3) liquid state fermentation produces:
(31) use capacity for 30m3Large capacity fermentor, be packed into liquid fermentation medium；
(32) the resulting liquid seed culture fluid of inoculation step (2), inoculum concentration be 5~10%, access secondary seed tank in
It is cultivated 6~9 hours at 32~35 DEG C of temperature, fermentation liquid seeds culture solution is made；
(33) the resulting fermentation of step (32) is accessed in fermentor with liquid seeds culture solution after, sets fermentation condition
Carry out liquid state fermentation afterwards to get low citrinin, High color values monascorubin fermentation liquid.
Wherein, in the step (2) further include: by red yeast rice Monascus YZ301 merging equipped with sterile water, bead three
It is smashed 1~2 hour in the bottle of angle, accesses triangular flask by 5~10% inoculum concentrations, cultivate 22~28 at 32~35 DEG C of temperature through shaking flask
Hour；The raw material of seed culture medium in the step (2) forms and weight proportion are as follows: starch 4~6%, NaNO30.1~
0.3%, MgSO40.1~0.3%, KH2PO40.1~0.3%, corn pulp 1~5%.
It further include that the liquid fermentation for accounting for 3/4 volume of fermentation tank is packed into first in large capacity fermentor in the step (31)
Culture medium, sterilize 30min in the case where 121~125 DEG C of tank temperature, tank press 0.1~0.15Mpa, cools to 30~35 DEG C；Described
In step (31) liquid fermentation medium raw material composition and weight proportion are as follows: rice flour 8~15%, soybean 0.1~0.5%,
NaNO30.1%~0.3%, corn pulp 1~5%；Further include with lactic acid adjust fermentation medium initial pH value be 3.0~4.5.
Described step (33) fermentation condition sets as follows: temperature is initial 0~34h heat preservation of fermenting in 30~35 DEG C, hair
It is 33~36 DEG C that ferment, which continues 35~85h adjustment temperature,；Ventilation quantity is that the initial 0~25h of fermentation is set as 300~500Lmin, fermentation
Middle and later periods, 26~85h was adjusted to 550~700Lmin；Entire fermentation process at the uniform velocity stirs always, and revolving speed is 200~400r/
Min, tank pressure are 0.04~0.07Mpa；The incubation time that continuously ferments carries out 75~85h altogether.
It is 3.0~4.5 that the step (33), which further includes with lactic acid adjusting fermentation medium initial pH value, temperature 32~36
℃；Feed supplement, feed supplement amount of the total volume 5~10% are carried out after fermentation lasts arrival in 35~45 hours fermentation animated period；Described
Feed supplement is rice flour 10~20%, soybean 0.1~0.5%, with newborn acid for adjusting pH value 3.0~4.5.
The preparation method of liquid state fermentation low citrinin above-mentioned, High color values monascorubin, further includes following steps:
(4) extract, purify: low citrinin, the High color values monascorubin fermentation liquid warp that the step (33) is obtained are de-
After water, the mycelium pH that will be left behind is the extraction of 7.0~9.0,60~80v/v alcohol, then secondary filters pressing, will extract filtrate through film
(i.e. nanofiltration), vacuum concentration (20~35Be brix) are filtered, the concentrate of low citrinin, High color values monascorubin is obtained；
(5) it is spray-dried: step (4) resulting concentrate being spray-dried at 100~210 DEG C, low tangerine is made
Mycin, High color values monascorubin powder product.
During fermented and cultured, its physical and chemical index of regular sampling analysis simultaneously carries out appropriate Additional nutrient solution i.e. feed supplement, can
It is improved fermentation level and output capacity, the feed supplement time is preferable in ferment middle and animated period.
For detecting the fermentation liquid, monascorubin using ultraviolet spectral photometer in aforesaid fermentation incubation
The absorbance of finished product, absorption peak wavelength are 495~505nm.
During the fermentation, the content of the citrinin of the monascorubin finished product is detected, citrinin contains in finished product
Amount is not more than 0.2mg/kg (in terms of unit color value 500u/g).
Low citrinin, the High color values monascorubin being prepared using preparation method above-mentioned, citrinin in finished product
Content no more than 0.2mg/kg (in terms of unit color value 500u/g), monascorubin color value be 10300~10800u/g.
Compared with the prior art, the invention has the following advantages:
1, red yeast rice Monascus YZ301 (Monascus anka) provided by the invention, microbial preservation number are
CGMCC9707 is that inventor obtains excellent species, integrative biology after culture, ultraviolet mutagenesis, separating for several times screening
Can be excellent, it is most suitable for liquid-state fermentation technology, breeding performance stabilization, high yield pigment, which can be resistant to during ultraviolet mutagenesis
High temperature (35~36.5 DEG C), high pressure (0.06~0.08Mpa), as increased inoculum concentration and keeping lower in fermentation liquid incubation
PH value can be such that fermentation level stablizes, and can control the content of red yeast rice time metabolin citrinin in fermentation process, and it is red to improve fermentation liquid
The content (i.e. High color values) and yield of bent pigment, it is highly-safe, stability is good.
2, the present invention uses the reasonable control of strain, process conditions and each step specially cultivated, to the Monascus of selection
Mutagenic fungi carries out liquid state fermentation, obtains the monascorubin fermentation liquid of high-content, resulting red after handling using abstraction process
Bent haematochrome, citrinin content is very low, highly-safe, stability is good, monascorubin color value is high, and the present invention is by preferred strain
After seed culture, it before liquid state fermentation, additionally uses and expands culture step by step, so that the strain content at fermentation initial stage is increased,
It improves the activity of fermentation phase microorganism, shorten fermentation period, be in a large amount of microorganisms within the fermentation phase and grow the vigorous stage,
Vigorous metabolism is kept, is conducive to improve pigment production.
3, the present invention is by extraction, purifying process, effectively by high molecular weight protein, small molecular organic acid, sugar and natural colour
Element separation, removes part citrinin, significantly improves the purity and stability of monascorubin product.
4, the present invention during fermented and cultured, simultaneously suitably mend in fermentation animated period by its physical and chemical index of regular sampling analysis
Material, is improved fermentation level and output capacity.
5, the present invention uses liquid state fermentation technology, and raw material is environmentally protective, pollution-free, safety, is formulated rationally, and preparation process is held
It is easy to control, rational technology is stable, high-efficient, makes that stable product quality, effective component are high, harmful components are low, meet food quality
Safety standard is, it can be achieved that large-scale industrial production.
6, to be up to 10300~10800u/g, tangerine mould for monascorubin color value in finished product made from preparation method of the invention
Cellulose content is not more than 0.2mg/kg (in terms of unit color value 500u/g).
Above-mentioned is the general introduction of inventive technique scheme, and below in conjunction with specific embodiment, the present invention will be further described.
Specific embodiment:
The red yeast rice Monascus YZ301 (Monascus anka) provided in this embodiment of embodiment 1, microbial preservation number
For CGMCC9707；The breeding method of the red yeast rice Monascus YZ301 (Monascus anka) 1 is by Dongguan City day benefit
The red yeast rice Monascus YZ201 (Monascus anka Nakazawa et Sato) of biofermentation technique Co., Ltd exploitation is through training
The red yeast rice Monascus supported, obtained after the screening of ultraviolet mutagenesis, separating for several times, number YZ301 (Monascus anka).
The culture is in wort agar medium 10~15。Be, 28~35 DEG C of temperature cultivate 6~8 days, bacterium colony is raw
Length is flat, on koji agar, in 33~35 DEG C of cultures, 7 days membranaceous bacterium colonies of cutification, no aerial hyphae, shape such as lava flow
Out, pomegranate water is red, the eugonic on wort agar；
Preferably, the ultraviolet mutagenesis and screening include: A, ultraviolet mutagenesis: will cultivate 7~9 days red yeast rice strains, warp
High temperature 55~65 DEG C of 2~10min of processing, 0.04~0.07Mpa of high pressure are handled 5~10 hours, are put into and are provided with band bead, nothing
The 500ml triangular flask of 80~100ml of bacterium water, is placed in shaking table and vibrates 1~2 hour, and filtering takes 10ml spore suspension, is added to band
In the culture dish of magnetic bar, 2~10min of ultraviolet irradiation is carried out under magnetic stirring, is applied plate, is wrapped up with black cloth, Yu Wendu 32
It is cultivated 6~8 days at~35 DEG C；
B, isolation medium: soluble starch 5%, maltose 3%, peptone 0.1% add appropriate lactic acid, adjust pH
Value 5~6, agar 2%；
C, isolation medium PDA:NaNO33.0g, KH2PO41.0g, MgSO47H2O 0.5g, FeSO47H2O 0.01g, sugarcane
Sugared 30g, tap water 1000ml, agar 15g；
D, slant medium MEA: brewer's wort 20g, glucose 20g, peptone 1.0g, tap water 1000ml, agar 20g.
A method of low citrinin, High color values monascorubin are prepared using aforementioned red yeast rice strains liquid fermentation comprising
Following steps:
(1) strain improvement: using the red yeast rice Monascus YZ301 (Monascus anka) isolated by ultraviolet mutagenesis
As strain, deposit number CGMCC9707；
(2) seed culture, expansion culture: the red yeast rice Monascus YZ301 that step (1) breeding obtains is sequentially ingressed into equipped with kind
It is cultivated respectively in the I and II seeding tank of sub- culture medium 6~9 hours, it is small to be inoculated into expansion culture 75~85 in fermentor later
When, obtain liquid seed culture fluid；
(3) liquid state fermentation produces:
(31) use capacity for 30m3Large capacity fermentor, be packed into liquid fermentation medium；
(32) the resulting liquid seed culture fluid of inoculation step (2), inoculum concentration be 5~10%, access secondary seed tank in
It is cultivated 6~9 hours at 32~35 DEG C of temperature, fermentation liquid seeds culture solution is made；
(33) the resulting fermentation of step (32) is accessed in fermentor with liquid seeds culture solution after, sets fermentation condition
Carry out liquid state fermentation afterwards to get low citrinin, High color values monascorubin fermentation liquid.
Wherein, in the step (2) further include: by red yeast rice Monascus YZ301 merging equipped with sterile water, bead three
It is smashed 1~2 hour in the bottle of angle, accesses triangular flask by 5~10% inoculum concentrations, cultivate 22~28 at 32~35 DEG C of temperature through shaking flask
Hour；The raw material of seed culture medium in the step (2) forms and weight proportion are as follows: starch 4~6%, NaNO30.1~
0.3%, MgSO40.1~0.3%, KH2PO40.1~0.3%, corn pulp 1~5%.
It further include that the liquid fermentation for accounting for 3/4 volume of fermentation tank is packed into first in large capacity fermentor in the step (31)
Culture medium, sterilize 30min in the case where 121~125 DEG C of tank temperature, tank press 0.1~0.15Mpa, cools to 30~35 DEG C；Described
In step (31) liquid fermentation medium raw material composition and weight proportion are as follows: rice flour 8~15%, soybean 0.1~0.5%,
NaNO30.1%~0.3%, corn pulp 1~5%；Further include with lactic acid adjust fermentation medium initial pH value be 3.0~4.5.
Described step (33) fermentation condition sets as follows: temperature is initial 0~34h heat preservation of fermenting in 30~35 DEG C, hair
It is 33~36 DEG C that ferment, which continues 35~85h adjustment temperature,；Ventilation quantity is that the initial 0~25h of fermentation is set as 300~500Lmin, fermentation
Middle and later periods, 26~85h was adjusted to 550~700Lmin；Entire fermentation process at the uniform velocity stirs always, and revolving speed is 200~400r/
Min, tank pressure are 0.04~0.07Mpa；The incubation time that continuously ferments carries out 75~85h altogether.
It is 3.0~4.5 that the step (33), which further includes with lactic acid adjusting fermentation medium initial pH value, temperature 32~36
℃；Feed supplement, feed supplement amount of the total volume 5~10% are carried out after fermentation lasts arrival in 35~45 hours fermentation animated period；Described
Feed supplement is rice flour 10~20%, soybean 0.1~0.5%, newborn acid for adjusting pH value are 3.0~4.5.
The preparation method of liquid state fermentation low citrinin above-mentioned, High color values monascorubin, further includes following steps:
(4) extract, purify: the low citrinin High color values monascorubin fermentation liquid that the step (33) is obtained is through being dehydrated
Afterwards, the mycelium pH that will be left behind is that 7.0~9.0,60~80v/v alcohol extracts, secondary filters pressing, extracts filtrate again through film mistake
(nanofiltration), vacuum concentration (20~35Be brix) are filtered, the concentrate of low citrinin, High color values monascorubin is obtained；
(5) it is spray-dried: step (4) resulting concentrate being spray-dried at 100~210 DEG C, low tangerine is made
Mycin, High color values monascorubin powder product.
During fermented and cultured, its physical and chemical index of regular sampling analysis simultaneously carries out appropriate Additional nutrient solution i.e. feed supplement, can
It is improved fermentation level and output capacity, the feed supplement time is preferable in ferment middle and animated period.
For detecting the fermentation liquid, monascorubin using ultraviolet spectral photometer in aforesaid fermentation incubation
The absorbance of finished product, absorption peak wavelength are 495~505nm.
During the fermentation, the content of the citrinin of the monascorubin finished product is detected, citrinin contains in finished product
Amount is not more than 0.2mg/kg (in terms of unit color value 500u/g).
Low citrinin, the High color values monascorubin being prepared using preparation method above-mentioned, citrinin in finished product
Content no more than 0.2mg/kg (in terms of unit color value 500u/g), monascorubin color value be 10300~10800u/g.
In the following, providing four kinds according to the embodiment above and having representative specific embodiment.
The red yeast rice Monascus YZ301 (Monascus anka) provided in this embodiment of embodiment 2, and use the red yeast rice bacterium solution
State fermentation prepares the method and product of low citrinin High color values monascorubin, and substantially the same manner as Example 1, difference exists
In:
It is a kind of that low citrinin, High color values red yeast rice are prepared using red yeast rice Monascus YZ301 (Monascus anka) liquid state fermentation
The method of haematochrome, the difference of specific steps are as follows:
(2) seed culture, expansion culture: red yeast rice Monascus YZ301 merging is equipped in the triangular flask of sterile water, bead
It smashes 2 hours, accesses triangular flask by 10% inoculum concentration, cultivated 28 hours at 35 DEG C of temperature through shaking flask, is sequentially ingressed into equipped with kind
It is cultivated respectively in the I and II seeding tank of sub- culture medium 6 hours, is inoculated into fermentor later and expands culture 85 hours, obtained
Liquid seed culture fluid；
The raw material of the seed culture medium forms and weight proportion are as follows: starch 4%, NaNO30.2%, MgSO40.2%,
KH2PO40.2%, corn pulp 1%.
(3) liquid state fermentation produces:
(31) use capacity for 30m3Large capacity fermentor, first in large capacity fermentor be packed into account for 3/4 body of fermentation tank
Long-pending liquid fermentation medium, sterilize 30min at 121 DEG C of tank temperature, tank pressure 0.1Mpa, cools to 30 DEG C；
The raw material of the liquid fermentation medium forms and weight proportion are as follows: rice flour 8%, soybean 0.2%,
NaNO30.1%, corn pulp 1%；Further include with lactic acid adjust fermentation medium initial pH value be 3.0~4.5.
(32) the resulting liquid seed culture fluid of inoculation step (2), inoculum concentration 5% access secondary seed tank in temperature
It is cultivated 9 hours at 32 DEG C, fermentation liquid seeds culture solution is made；
(33) the resulting fermentation of step (32) is accessed in fermentor with liquid seeds culture solution after, obtain low citrinin,
High color values monascorubin fermentation liquid；The fermentation condition wherein set are as follows: 1. temperature controls: 30~35 DEG C of initial temperature, 35 is small
When after be adjusted to 33~36 DEG C；2. speed of agitator: 220~400r/min；3. ventilation quantity: 350~500 (Lmin) are initially,
After 35 hours, it is then adjusted to 550~700/ (Lmin)；4. tank pressure: 0.04~0.07MPa.
It is 3.0~4.5 that the step (33), which further includes with lactic acid adjusting fermentation medium initial pH value, temperature 32~36
℃；Feed supplement is carried out after fermentation lasts 35~45 hours, feed supplement amount of the total volume 5~10% was finished through fermentation in 75~85 hours,
Low citrinin, the high fermentation liquid containing monascorubin can be obtained；The feed supplement be rice flour 10~20%, soybean 0.1~
0.5%, newborn acid for adjusting pH value is 3.0~4.5.
(4) it extracts, purify: by obtained low citrinin, High color values monascorubin fermentation liquid after being dehydrated, will be left behind
Mycelium pH is that 7.0~9.0,60~80v/v alcohol extracts, through secondary filters pressing, extract again filtrate through film filtering (nanofiltration),
It is concentrated in vacuo (20~35Be brix), obtains the concentrate of low citrinin, High color values monascorubin；
(5) it is spray-dried: resulting concentrate is spray-dried at 100~210 DEG C, the high color of low citrinin is made
Valence monascorubin powder product.
The content of citrinin in fermentation process, periodic detection product detects absorbance, evaluation with ultraviolet spectral photometer
The color value of monascorubin in finished product.
The red yeast rice Monascus YZ301 (Monascus anka) provided in this embodiment of embodiment 3, and use the red yeast rice bacterium solution
State fermentation prepares low citrinin, the method for High color values monascorubin and product, essentially identical with embodiment 1,2, difference
Be in:
In the method for preparing low citrinin, High color values monascorubin, in specific steps (3) liquid state fermentation production: described
The step of (31) in liquid fermentation medium raw material composition and weight proportion adjustment are as follows: rice flour 10%, soybean 0.2%,
NaNO30.1%, corn pulp 3%；Further include with lactic acid adjust fermentation medium initial pH value be 3.0~4.5.The step
(32) the resulting liquid seed culture fluid of inoculation step (2) in, inoculum concentration 6% access secondary seed tank at 32 DEG C of temperature
Culture 6 hours.
Other steps are the same as the essentially identical of embodiment 1 or 2.The content of citrinin in fermentation process, periodic detection product,
Absorbance is detected with ultraviolet spectral photometer, evaluates the color value of monascorubin in finished product.
The red yeast rice Monascus YZ301 (Monascus anka) provided in this embodiment of embodiment 4, and use the red yeast rice bacterium solution
State fermentation prepares low citrinin, the method for High color values monascorubin and product, essentially identical with embodiment 1,2,3, different
Place is:
In the method for preparing low citrinin, High color values monascorubin, specific steps (2) seed culture expands culture: institute
The raw material composition and ratio for the liquid seed culture medium stated: starch 4%, NaNO30.1%, MgSO40.1%, KH2PO40.1%,
Corn pulp 1%；In specific steps (3) liquid state fermentation production, the raw material composition of liquid fermentation medium in the step (31)
And weight proportion adjustment are as follows: rice flour 12%, soybean 0.2%, NaNO30.1%, corn pulp 5%；It further include being adjusted to ferment with lactic acid
Initial pH value of medium is 3.0~4.5.The resulting liquid seed culture fluid of inoculation step (2) in the step (32), inoculation
Amount is 8%, and access secondary seed tank is cultivated 6 hours at 34 DEG C of temperature.The step (33) further include fermentation lasts 35~
Feed supplement, feed supplement amount of the total volume 10% are carried out after 45 hours；The feed supplement is rice flour 10%, soybean 0.1%, lactic acid adjusting
PH value is 3.0~4.5.
Other steps with embodiment 1 or 2 or 3 it is essentially identical.Citrinin contains in fermentation process, periodic detection product
Amount detects absorbance with ultraviolet spectral photometer, evaluates the color value of monascorubin in finished product.
The red yeast rice Monascus YZ301 (Monascus anka) provided in this embodiment of embodiment 5, and use the red yeast rice bacterium solution
State fermentation prepares low citrinin, the method for High color values monascorubin and product, essentially identical with embodiment 1,2,3,4, no
It is with place:
In the method for preparing low citrinin, High color values monascorubin, specific steps (2) seed culture expands culture: institute
The raw material composition and ratio for the liquid seed culture medium stated: starch 4%, NaNO30.1%, MgSO40.1%, KH2PO40.1%,
Corn pulp 1%；In specific steps (3) liquid state fermentation production, the raw material composition of liquid fermentation medium in the step (31)
And weight proportion adjustment are as follows: rice flour 15%, soybean 0.3%, NaNO30.1%, corn pulp 5%；It further include being adjusted to ferment with lactic acid
Initial pH value of medium is 3.0~4.5.The resulting liquid seed culture fluid of inoculation step (2) in the step (32), inoculation
Amount is 10%, and access secondary seed tank is cultivated 8 hours at 35 DEG C of temperature.The step (33) further include fermentation lasts 35~
Feed supplement, feed supplement amount of the total volume 5% are carried out after 45 hours；The feed supplement is rice flour 10%, soybean 0.5%, with lactic acid tune
Saving pH value is 3.0~4.5.
Other steps with embodiment 1 or 2 or 3 or 4 it is essentially identical.The citrinin in fermentation process, periodic detection product
Content, with ultraviolet spectral photometer detect absorbance, evaluate finished product in monascorubin color value.
The performance test results of low citrinin, High color values monascorubin product that the present invention obtains are shown in Table 1.
The performance test results of 1 low citrinin of table, High color values monascorubin
Present invention is not limited to the embodiments described above, using raw material same as or similar to the above embodiments of the present invention, work
Skill, and the preparation method and product of obtained low citrinin, High color values monascorubin, within that scope of the present invention.
Claims (4)
1. a kind of method that liquid state fermentation prepares low citrinin, High color values monascorubin, which is characterized in that including walking as follows
It is rapid:
(1) strain is used as using red yeast rice Monascus YZ301 (Monascus anka Yang YZ301), microbial preservation number is
CGMCC9707；
(2) seed culture, expansion culture: the red yeast rice Monascus YZ301 of step (1) is sequentially ingressed into equipped with seed culture medium
It is cultivated respectively in I and II seeding tank 6~9 hours, is inoculated into fermentor later and expands culture 75~85 hours, obtain liquid
Seed culture fluid；
(3) liquid state fermentation produces:
(31) use capacity for 30m3Large capacity fermentor, be packed into liquid fermentation medium；
(32) the resulting liquid seed culture fluid of inoculation step (2), inoculum concentration are to cultivate 6 at 32~35 DEG C of 5~10%, Yu Wendu
~9 hours, fermentation liquid seeds culture solution is made；
(33) the resulting fermentation of step (32) is accessed in large capacity fermentor with liquid seeds culture solution after, setting fermentation item
Liquid state fermentation is carried out after part to get low citrinin, High color values monascorubin fermentation liquid.
2. the method that liquid state fermentation according to claim 1 prepares low citrinin, High color values monascorubin, feature exist
In in the step (2) further include: before seed culture by red yeast rice Monascus YZ301 merging equipped with sterile water, bead three
Processing is smashed 1~2 hour in the bottle of angle, is linked into another triangular flask by 5~10% inoculum concentrations, through shaking flask in temperature 32~35
It is cultivated 22~28 hours at DEG C；The raw material of seed culture medium in the step (2) forms and weight proportion are as follows: and starch 4~
6%, NaNO30.1~0.3%, MgSO40.1~0.3%, KH2PO40.1~0.3%, corn pulp 1~5%.
3. the method that liquid state fermentation according to claim 1 prepares low citrinin, High color values monascorubin, feature exist
In the step (33) fermentation condition sets as follows: temperature is initial 0~34h heat preservation of fermenting in 30~35 DEG C, fermentation lasts
It is 33~36 DEG C that 35~85h, which adjusts temperature,；Ventilation quantity is that the initial 0~25h of fermentation is set as 300~500L/min, fermentation middle and later periods
26~85h is adjusted to 550~700L/min；Entire fermentation process at the uniform velocity stirs always, and revolving speed is 200~400r/min, tank pressure
For 0.04~0.07Mpa.
4. the method that liquid state fermentation according to claim 1 prepares low citrinin, High color values monascorubin, feature exist
In further including following steps:
(4) extract, purify: low citrinin that the step (33) is obtained, High color values monascorubin fermentation liquid after being dehydrated,
The mycelium pH that will be left behind is the extraction of 7.0~9.0,60~80%v/v alcohol, then secondary filters pressing, and extraction filtrate is carried out film
Nanofiltration, vacuum concentration are handled, obtains low citrinin, the concentrate that the brix of High color values monascorubin is 20~35Be；
(5) it is spray-dried: step (4) resulting concentrate is spray-dried at 100~210 DEG C, low citrinin is made
High color values monascorubin powder product.
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| 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 |
| 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 |
| CN110195022B (en) * | 2019-06-28 | 2021-04-27 | 福州大学 | Method for reducing diameter of monascus mycelium pellet and simultaneously improving biomass and pigment content |
| 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 |
| CN112481327B (en) * | 2020-12-01 | 2023-04-14 | 广东肇庆星湖生物科技股份有限公司 | Fermentation regulation and control method of water-soluble monascus yellow pigment |
| 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 |
| CN114540340A (en) * | 2022-04-18 | 2022-05-27 | 安徽农业大学 | Method for improving ultraviolet mutagenesis effect of monascus |
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