CN104962483B - A kind of monascus purpureus bacterial strain and application thereof - Google Patents
A kind of monascus purpureus bacterial strain and application thereof Download PDFInfo
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Abstract
The invention discloses a kind of monascus purpureus (Monascus purpureus) and its purposes in food production.The monascus purpureus is deposited in China Committee for Culture Collection of Microorganisms's common micro-organisms center, deposit number are as follows: CGMCC No.9712.It hardly produces citrinin, is that citrinin yield is minimum in known monascus category bacterial strain, and it can produce natural monascorubin, possess higher color value, can be used for preparing in dairy products or other food processing technologies.
Description
Technical field
The present invention relates to microorganism fields, and in particular to a kind of monascus purpureus bacterial strain and application thereof.
Background technique
Monascus (Monascus sp.) is a kind of highly important medicinal fungi, is that China is applied to food processing earliest
In one of fungi.And use scope is extensive, is mainly used for culinary art, production fermented bean curd, wine brewing and treatment disease etc..Using red yeast rice
Red yeast rice Gu made from mould fermentation rice claims red song, with a long history, originating from China, is mainly intensively applied in traditional distiller's yeast, system
The fields such as vinegar, coloring and health care product are the tradition materials of dietotherapeutic.Modern medicine study proves that red yeast rice has reduction gallbladder solid
The effects of alcohol, hypoglycemic and blood pressure lowering.It is produced in addition, monascus can generate a variety of metabolism with physiological activity during the fermentation
Object, such as: monascorubin, citrinin (Monacolin K) substance, γ-aminobutyric acid and a variety of enzymes etc..
Although monascus is longer in the edible history of China and pouplarity is higher, since some monascus species exist
Mycotoxin can be generated in fermentation process --- citrinin (citrinin), and citrinin is to human body toxicity with higher, because
This limits the application range of monascus strain to a certain extent.Food safety is the premise of food production and sale, American-European
Various countries have stringent limitation to the content of the citrinin in imported food especially monascus product, meanwhile, in Chinese people's republicanism
Also define that the content of citrinin in red yeast rice (powder) must not exceed 50 μ g/Kg in state light industry standard QB/T2847-2007.Research
The strain variety of the content and monascus ruber that show citrinin is closely related, therefore, to produce the product of Fermentation Condition of Monascus spp class, needs
Bacterial strain and optimized production process are strictly selected, guarantees food safety.
Summary of the invention
The technical problem to be solved by the present invention is to low for the current content for lacking citrinin, meet respective country mark
Quasi- monascus (Monascus sp.) is low, safe using monascus as the food of raw material to the content for being difficult to prepare citrinin
Deficiency, a kind of monascus purpureus bacterial strain of low-yield citrinin is provided and its is preparing the application in food.The monascus ruber
Strain is that citrinin yield is minimum in existing known monascus category bacterial strain, and can produce natural monascorubin, can
Safely apply to food-processing industry.
Technical solution of the present invention first is that: a kind of monascus purpureus (Monascus purpureus) is deposited in China
Microbiological Culture Collection administration committee common micro-organisms center, deposit number are as follows: CGMCC No.9712.
Separation obtains the monascus purpureus CGMCC No.9712 from the red kojic rice powder in China Hubei Wuhan.Described
Monascus purpureus CGMCC No.9712 can produce natural monascorubin, and citrinin yield is lower, can safely apply to eat
Product processing industry.The bacterial strain is identified, result is monascus purpureus Monascus purpureus, is named as BD-M-4.It should
Bacterial strain is preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center on October 8th, 2014, and receives
Collection is registered on the books number CGMCC No.9712, with Microbiological Characteristics below:
1, morphologic characteristic
It is very fast in MEA cultured on solid medium, it is cultivated 10 days under 25 DEG C of dark conditions, 45~50mm of colony diameter.Gas
Raw mycelia is luxuriant, edge white, and middle part is orange；The bacterium colony back side dissolves in MEA culture medium in dark orange.
Micro- sem observation is shown, largely generates conidium, and conidiophore specialization is unobvious, straight or slightly bent, mitogenetic
Spore is spherical in shape, colourless, and wall is slightly coarse, and base portion is truncate, concatenates, and 5.5-10.4 μm of length.Cleistothecium shallowly arrives brown, it is most not at
It is ripe, have no ascospore.
2, the characteristic that culture is learned
It can be grown under the conditions of 20~42 DEG C, proper growth temperature is 25~32 DEG C, and optimum temperature is 30 DEG C；
Suitable pH is 3.5~6.0, optimal pH 5.5.Suitable growth medium can be PDB fluid nutrient medium, the training of YES liquid
Support base or MEA fluid nutrient medium.
3, physiological property
It is nearly free from citrinin, can produce natural monascorubin, and possesses higher color value.
Technical solution of the present invention second is that: a method of preparing the monascus purpureus CGMCC No.9712, packet
Following step is included, cultivates the monascus purpureus CGMCC No.9712 in the medium.
Wherein, the culture medium is the culture medium of this field routine, can grow the monascus purpureus CGMCC
No.9712, preferably PDB fluid nutrient medium, YES fluid nutrient medium or MEA fluid nutrient medium.The PDB liquid
Culture medium is the PDB fluid nutrient medium of this field routine, preferably comprising 6~10g/L potato leaching powder and 20~40g/L
Glucose.The YES fluid nutrient medium is the YES fluid nutrient medium of this field routine, preferably comprising 4~6g/L ferment
Mother's leaching powder, 20~40g/L glucose, 5~7g/L potassium dihydrogen phosphate, 3~5g/L sodium dihydrogen phosphate and 0.5~1.5g/L hydroxide
Ammonium.The MEA fluid nutrient medium is the MEA fluid nutrient medium of this field routine, preferably comprising 20~40g/L malt
Medicinal extract and 2~4g/L soy peptone.The time of the culture be this field routine time, preferably 3~10 days, more preferably
Ground is 5~10 days.The temperature of the culture is the temperature of this field routine, and preferably 20~42 DEG C, be more preferably 25~30
℃.Preferably, the culture is shake culture.The revolving speed of the shake culture is the revolving speed of this field routine, preferably
120~220rpm is more preferably 180~220rpm.The inoculum concentration of the culture is the inoculum concentration of this field routine, preferably
5~10%, it is more preferably 5%, the percentage is percent by volume.
Preferably, further including that monascus purpureus CGMCC No.9712 is inoculated in seed culture medium progress before the culture
The step of seed culture.The time of the seed culture is the time of this field routine, and preferably 5~10 days, be more preferably 6
~8 days, be most preferably 7 days.The temperature of the seed culture be this field routine temperature, preferably 15~35 DEG C, more preferably
Ground is 25~32 DEG C, is most preferably 30 DEG C.The culture medium of the seed culture is the culture medium of this field routine, preferably
PDA solid medium, YES solid medium or MEA solid medium.The PDA solid medium is the PDA of this field routine
Solid medium, preferably comprising 6~10g/L potato leaching powder, 20~40g/L glucose and 20~30g/L agar.Institute
The YES solid medium that YES solid medium is this field routine is stated, preferably comprising 4~6g/L yeast extract, 20~
40g/L glucose, 5~7g/L potassium dihydrogen phosphate, 3~5g/L sodium dihydrogen phosphate, 0.5~1.5g/L ammonium hydroxide and 15~25g/
L agar.The MEA solid medium is the MEA solid medium of this field routine, preferably comprising 20~40g/L wheat
Bud medicinal extract, 2~4g/L soy peptone and 12~20g/L agar.Preferably, the seed culture includes will be by the red of refrigeration
After Aspergillus strain slowly heats up, in the step of being coated with or crossing on seed culture medium.The temperature of the heating is that this field is conventional
Temperature, preferably to 15~25 DEG C.The method of the culture is the method for the culture of this field routine, preferably shaking flask
Culture or fermentation tank culture.
Technical solution provided by the invention third is that: the monascus purpureus CGMCC No.9712 is in food production
Purposes.
The food is the food of this field routine, is containing monascus purpureus CGMCC No.9712 or its metabolite
Can, preferably dairy products are more preferably cheese.
The monascus purpureus CGMCC No.9712 hardly produces citrinin, and possesses higher color value, can be used for preparing
In dairy products or other food processing technologies.
Technical solution provided by the invention fourth is that: it is a kind of to utilize cheese made from Fermentation Condition of Monascus spp, the monascus
For monascus purpureus CGMCC No.9712.
The cheese is monascus cheese, has the distinctive flavour of this kind of cheese and smell, and fermenting aroma is strong, tool
There is a distinctive tender texture of such cheese, shell is good, surface corrugationless, and shell takes on a red color or purple, and inside is in uniform purple
Color or red, the cheese are suitble to the eating habit of Chinese Consumer's.
Technical solution provided by the invention fifth is that: a kind of using red kojic rice powder made from Fermentation Condition of Monascus spp, described is red
Aspergillus is monascus purpureus CGMCC No.9712.
The red kojic rice powder is red to dark violet red, and quality is crisp, no mildew, without obvious macroscopic impurity, is in
Irregular graininess, with the intrinsic Qu Xiang of red yeast rice.
On the basis of common knowledge of the art, above-mentioned each optimum condition, can any combination to get each preferable reality of the present invention
Example.
The reagents and materials used in the present invention are commercially available.
The positive effect of the present invention is that: the monascus purpureus CGMCC No.9712 hardly produces citrinin, is
Citrinin yield is minimum in existing known monascus category bacterial strain.And it can produce natural monascorubin, possess compared with
High color value, in the processing technology that can be used for preparing the food such as dairy products, red kojic rice powder.
Biomaterial preservation information
Monascus purpureus BD-M-4 of the invention is deposited in Chinese microorganism strain preservation management on October 8th, 2014
Committee's common micro-organisms center (CGMCC), preservation address: Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3, postcode:
100101, deposit number are as follows: CGMCC No.9712, culture title are BD-M-4, and classification naming is monascus purpureus Monascus
purpureus。
Detailed description of the invention
Fig. 1 is the case where monascus purpureus BD-M-4 biomass changes with fermentation time.
Fig. 2 be different monascus purpureus bacterial strains ferment 10 days when biomass the case where.
Fig. 3 is the case where different monascus purpureus bacterial strains generates citrinin content when fermenting 10 days.
Fig. 4 is the morphological feature of monascus purpureus BD-M-4.Wherein A indicates monascus purpureus BD-M-4 in wort agar culture
Colonial morphology on base (MEA)；The microscopic features of B~D expression monascus purpureus BD-M-4.
Specific embodiment
The present invention is further illustrated below by the mode of embodiment, but does not therefore limit the present invention to the reality
It applies among a range.In the following examples, the experimental methods for specific conditions are not specified, according to conventional methods and conditions, or according to quotient
The selection of product specification.
Monascus GL-1 in effect example is red yeast rice (Monascus sp.), which protects on May 8th, 2013
It ensconces China Committee for Culture Collection of Microorganisms's common micro-organisms center (CGMCC), bacterial strain deposit number: CGMCC
No.7603 is announced in CN103444878A.
Monascus purpureus BD-M-1 in effect example is purplish red bent (Monascus purpureus), the bacterial strain in
It is deposited in China Committee for Culture Collection of Microorganisms's common micro-organisms center (CGMCC), bacterial strain preservation on August 28th, 2013
Number: CGMCC No.8120 is announced in CN104585333A.
The acquisition of 1 monascus purpureus BD-M-4 bacterial strain of embodiment
Purification procedures are as follows:
(1) PDA solid medium: potato leaching powder 6g/L, glucose 20g/L and agar 20g/L is configured.
(2) coating culture: the red kojic rice powder in a small amount of China Hubei Wuhan is taken uniformly to be sprinkled into the PDA solid of step (1) preparation
Media surface.30 DEG C are cultivated 2 days in constant incubator.
(3) after white fluffy mycelia grows, picking a little mycelia scribing line switching on another PDA solid medium after
Continuous culture, is cultivated 1 week.Step (2) and step (3) 3 times are continuously repeated, the monascus purpureus of uniform character is obtained, is named as BD-M-
4。
(4) monascus purpureus BD-M-4 bacterial strain is stored on PDA solid medium and is placed in 4 DEG C of refrigerators.
By monascus purpureus BD-M-4 on October 8th, 2014 in China Committee for Culture Collection of Microorganisms's commonly micro- life
Object center (CGMCC) preservation obtains deposit number are as follows: CGMCC No.9712, culture title are BD-M-4, and classification naming is
Monascus purpureus Monascus purpureus.
The operation of same above-mentioned steps (1)~(4), has also obtained the monascus strain of other uniform characters, is respectively designated as
Monascus BD-A, monascus BD-B, monascus BD-C and monascus GL-A.
The cultural character of 2 monascus purpureus BD-M-4 of embodiment
(1), monascus purpureus BD-M-4 is very fast in MEA cultured on solid medium, cultivates 10 days under 25 DEG C of dark conditions, bacterium
It falls diameter and reaches 45~50mm.Aerial hyphae is luxuriant, edge white, and middle part is orange；The bacterium colony back side dissolves in culture in dark orange
In base.Micro- sem observation shows, monascus purpureus BD-M-4 generates a large amount of conidiums, and conidiophore specialization is unobvious, it is straight or
Slightly bent, conidium is spherical, and colourless, wall is slightly coarse, and base portion is truncate, concatenates, and 5.5-10.4 μm.Cleistothecium shallowly arrives brown, more
Number is immature, has no ascospore, meets the feature (A~D referring to fig. 4) of monascus category.
(2), it is found by culture, BD-M-4 bacterial strain can be grown under the conditions of 20~42 DEG C, proper growth temperature
Degree is 25~32 DEG C, and optimum temperature is 30 DEG C；Suitable pH is 3.5~6.0, optimal pH 5.5.Suitable growth medium can
To be PDB fluid nutrient medium, YES fluid nutrient medium or MEA fluid nutrient medium.Wherein, in MEA fluid nutrient medium and PDB liquid
It on body culture medium, cultivates 10 days under the conditions of 30 DEG C, is continuously cultivated through 15 generations, cultural characteristic, morphological feature etc. become without obvious
Change, the biological shapes of the bacterium are basicly stable.
Wherein, PDB culture medium includes 30g/L glucose and 10g/L potato leaching powder.YES culture medium includes 5g/L yeast
Soak powder, 30g/L glucose, 6g/L potassium dihydrogen phosphate, 4g/L sodium dihydrogen phosphate and 1g/L ammonium hydroxide.MEA culture medium includes
25g/L malt extract and 3g/L soy peptone.
The physiological property of 3 monascus purpureus BD-M-4 of embodiment
The constituent of PDA solid medium are as follows: 6g/L potato leaching powder, 20g/L glucose and 20g/L agar.
The constituent of PDB fluid nutrient medium are as follows: 6g/L potato leaching powder and 20g/L glucose, surplus are water.
The monascus purpureus BD-M-4 that 4 DEG C save aseptically is transferred in PDA solid medium, 30 DEG C of activation trainings
Be inoculated in PDA solid medium after supporting 4 days 30 DEG C activation culture 5 days to get first order seed.
250mL triangular flask be packed into 50mL PDB fluid nutrient medium, access diameter be 1cm first order seed bacteria cake, 30 DEG C
Shaking table, 180r/min revolving speed cultivate 4 days to get second order fermentation seed.
It is packed into 100mL PDB fluid nutrient medium in 500mL triangular flask, second order fermentation kind is accessed with the ratio of 5% (v/v)
Son, 30 DEG C of shaking tables, 180r/min revolving speed are cultivated 10 days.
According to the research method of Shao Wei et al., (Shao Wei, Wang Dong, Tang Ming wait " monascus produces property of protease research " [J]
China's brewing, 2006,5:34-37.), measure mycelia weight (i.e. biomass), indicate its upgrowth situation.It will be seen from figure 1 that
Monascus purpureus BD-M-4 is in 3 days that culture starts, and thallus largely generates, and thallus reaches maximum value 6.5g/L within the 4th day or so, and
And thalli growth is in the stabilization sub stage, then the biomass of thallus is slowly reduced, and meets the growth characteristics of monascus.
The physiological property of 4 monascus purpureus BD-M-4 of embodiment
The constituent of PDA solid medium are as follows: 10g/L potato leaching powder, 30g/L glucose and 30g/L agar.
The constituent of PDB fluid nutrient medium are as follows: 10g/L potato leaching powder and 30g/L glucose, surplus are water.
The monascus purpureus BD-M-4 that 4 DEG C save aseptically is transferred in PDA solid medium, 30 DEG C of activation trainings
Be inoculated in PDA solid medium after supporting 4 days 30 DEG C activation culture 5 days to get first order seed.
250mL triangular flask be packed into 50mL PDB fluid nutrient medium, access diameter be 1cm first order seed bacteria cake, 30 DEG C
Shaking table, 180r/min revolving speed cultivate 4 days to get second order fermentation seed.
It is packed into 100mL PDB fluid nutrient medium in 500mL triangular flask, second order fermentation kind is accessed with the ratio of 10% (v/v)
Son, 30 DEG C of shaking tables, 180r/min revolving speed are cultivated 10 days.
According to the measuring method of the honor of Jiangxi perhaps, (detection of Jiangxi honor red yeast rice citrinin and the Jiangnan ferment control technology [D] are big perhaps
Doctoral thesis, 2004.), it is measured using Agilent HPLC 1260, as a result as shown in Figure 3.As seen from Figure 3,
Monascus BD-M-4 production citrinin amount is almost nil, and toxin producing is few, is suitable for preparing cheese.
Illustrated by the data of embodiment 2~4, the monascus purpureus CGMCC No.9712 has microbiology below special
Property:
1, morphologic characteristic
It is very fast in MEA cultured on solid medium, it is cultivated 10 days under 25 DEG C of dark conditions, colony diameter reaches 45~
50mm.Aerial hyphae is luxuriant, edge white, and middle part is orange；The bacterium colony back side dissolves in MEA solid medium in dark orange.It produces
Raw a large amount of conidiums, conidiophore specialization is unobvious, straight or slightly bent, and conidium is spherical, and colourless, wall is slightly coarse, base
Portion is truncate, concatenates, and 5.5-10.4 μm.Cleistothecium shallowly arrives brown, most immature, has no ascospore.
2, the characteristic that culture is learned
It can be grown under the conditions of 20~42 DEG C, proper growth temperature is 25~32 DEG C, and optimum temperature is 30 DEG C；
Suitable pH is 3.5~6.0, optimal pH 5.5.Suitable growth medium can be PDB fluid nutrient medium, the training of YES liquid
Support base or MEA fluid nutrient medium.
3, physiological property
Citrinin is hardly produced, can produce natural monascorubin, and possess higher color value.
The rDNA sequence of 5 monascus purpureus BD-M-4 of embodiment is analyzed
Monascus purpureus BD-M-4 is subjected to rRNA gene sequencing (being sequenced by Institute of Microorganism, Academia Sinica), sequencing knot
For fruit as shown in SEQ No.1 in sequence table, which contains the 18S rRNA segment of monascus purpureus BD-M-4, ITS1,5.8S
The complete sequence and 28S region sequence segment of rRNA, ITS2.By the sequence and document and public database DDBJ, EMBL, GenBank
Equal comparisons discovery, homology 99%.Therefore, monascus purpureus BD-M-4 belongs to monascus purpureus (Monascus purpureus).
6 monascus purpureus BD-M-4 of embodiment prepares red kojic rice powder
Ingredient: long-grained nonglutinous rice 10kg, glucose 40g, potato leaching powder 12g and agar 20g.
The constituent of PDA solid medium are as follows: 6g/L potato leaching powder, 20g/L glucose and 20g/L agar.
The constituent of PDB fluid nutrient medium are as follows: 6g/L potato leaching powder and 20g/L glucose, surplus are water.
Preparation step:
(1) activation and processing of strain: the monascus purpureus BD-M-4 that 4 DEG C save aseptically is transferred and is consolidated in PDA
In body culture medium, 30 DEG C be inoculated in after activation culture 4 days in PDA solid medium 30 DEG C activation culture 5 days to get level-one kind
Son.
250mL triangular flask be packed into 50mL PDB fluid nutrient medium, access diameter be 1cm first order seed bacteria cake, 30 DEG C
Shaking table, 180r/min revolving speed cultivate 4 days to get second order fermentation seed.
It is packed into 100mL PDB fluid nutrient medium in 500mL triangular flask, second order fermentation kind is accessed with the ratio of 10% (v/v)
Son, 30 DEG C of shaking tables, 180r/min revolving speed are cultivated 10 days, its pH value are adjusted to 3.0, obtains the strain of processing.
(2) long-grained nonglutinous rice is impregnated 12 hours in clear water, makes feed moisture content 50%, obtain soaked long-grained nonglutinous rice；
(3) soaked long-grained nonglutinous rice is put into high-pressure steam sterilizing pan, 121 DEG C of steamed rice 20min obtain steamed long-grained nonglutinous rice meal, make Xian
Rice maturation is without the white heart, and uniformity is without agglomeration；
(4) steamed long-grained nonglutinous rice meal is poured out, spreading for cooling is simultaneously sieved, and is inoculated in 38 DEG C.Every 10kg long-grained nonglutinous rice is inoculated with 0.04g step
(1) strain of resulting processing, is mixed thoroughly, is put into constant temperature and humidity incubator, 30 DEG C of constant temperature, the culture of constant relative humidity 80% 6
It, during which turned over bent and moisturizing, rate of water make-up 10% every 4 hours, and the percentage is mass percent, when culture was to the 5th day
Stop moisturizing.Drying is taken out to grind to get red kojic rice powder.
Obtained red kojic rice powder is red to dark violet red, and quality is crisp, no mildew, the obvious macroscopic impurity of nothing,
In irregular graininess, with the intrinsic Qu Xiang of red yeast rice.
7 monascus purpureus BD-M-4 of embodiment prepares cheese
Ingredient: fresh cow milk 100L (being purchased from bright milk industry), MA14 (being purchased from Danisco (China) Co., Ltd product) hair
Ferment agent 3g, calf stomach renin (being purchased from Chr. Hansen A/S) 1g, salt 0.75kg, glucose 40g and potato leaching powder 6g.
(1) after taking 100L fresh cow milk filtering and impurity removing matter, 72 DEG C of sterilizing 30s are subsequently cooled to 30 DEG C, must handle cream.To institute
State inoculating starter MA14, inoculum concentration 3g/100L in processing cream, under 30 DEG C of constant temperature culture to pH6.5 when 1g calf stomach is added
Renin, curdled milk 30min obtain curdled milk；
(2) step (1) resulting curd cutting is stirred into 10min discharging whey at bulk；After whey to be discharged into curdled milk
1.5% salt is added, the percentage is the mass percent that the salt accounts for the curdled milk, enters rectangular mould after mixing evenly
Tool；
(3) curdled milk enters mold forming and obtains ziega and periodically overturn, and frequency is to overturn 1 time every 2 hours, continues 24 hours,
Whey is discharged further；
(4) Monascus fermentation broth that thickness is about 1mm is uniformly sprayed on the ziega surface obtained toward step (3), be packed at
Ripe container, constant incubator is mature, the temperature of the maturation are as follows: and 25 DEG C of initial stage, relative humidity 90%, 10 DEG C of the middle and later periods；It is described
The mature time are as follows: initial stage 2 weeks, the middle and later periods 2 weeks.
The Monascus fermentation broth is made by following methods: monascus purpureus BD-M-4 is inoculated in PDB fluid nutrient medium
30 DEG C are cultivated 2 days, the formula of the PDB fluid nutrient medium are as follows: glucose 40g/L, potato leaching powder 6g/L, surplus are water.
Obtained monascus cheese has the distinctive flavour of monascus cheese and smell, and fermenting aroma is strong, has red
The distinctive tender texture of aspergillus cheese, shell is good, surface corrugationless, and shell takes on a red color or purple, and inside is in uniform purple
Or it is red, it is suitble to the eating habit of Chinese Consumer's.
8 monascus purpureus BD-M-4 of embodiment prepares cheese
Ingredient: fresh sheep cream 100L (being purchased from bright milk industry), MA14 (being purchased from Danisco (China) Co., Ltd product) hair
Ferment agent 3g, microbial rennet (being purchased from Chr. Hansen A/S) 1g, salt 0.5kg, glucose 40g and potato leaching powder 6g.
(1) after taking the fresh sheep cream filtering and impurity removing matter of 100L, 72 DEG C of sterilizing 15s are subsequently cooled to 28 DEG C, must handle cream.To institute
State inoculating starter MA14, inoculum concentration 3g/100L in processing cream, under 28 DEG C of constant temperature culture to pH6.5 when 1g microorganism is added
Renin, curdled milk 30min obtain curdled milk；
(2) step (1) resulting curd cutting is stirred into 10min discharging whey at bulk；After whey to be discharged into curdled milk
1% salt is added, the percentage is the mass percent that the salt accounts for the curdled milk, enters square dies after mixing evenly；
(3) curdled milk enters mold forming and obtains ziega and periodically overturn, and frequency is to overturn 1 time every 2 hours, continues 24 hours,
Whey is discharged further；
(4) Monascus fermentation broth that thickness is about 1.5mm is uniformly sprayed toward the ziega surface that step (3) obtain, be packed into
Mature container, constant incubator is mature, the temperature of the maturation are as follows: and 20 DEG C of initial stage, relative humidity 95%, 8 DEG C of the middle and later periods；It is described
The mature time are as follows: initial stage 2 weeks, the middle and later periods 2 weeks.
The Monascus fermentation broth is made by following methods: monascus purpureus BD-M-4 is inoculated in PDB fluid nutrient medium
30 DEG C are cultivated 2 days, the formula of the PDB fluid nutrient medium are as follows: the formula of the monascus culture medium are as follows: glucose 40g/L, horse
Bell potato soaks powder 6g/L, and surplus is water.
9 monascus purpureus BD-M-4 of embodiment prepares cheese
Ingredient: fresh horse cream 100L (being purchased from bright milk industry), MA14 (being purchased from Danisco (China) Co., Ltd product) hair
Ferment agent 3g, calf stomach renin (being purchased from Chr. Hansen A/S) 1g, salt 1.5kg, glucose 40g and potato leaching powder 6g.
(1) after taking the fresh horse cream filtering and impurity removing matter of 100L, 72 DEG C of sterilizing 30s are subsequently cooled to 32 DEG C, must handle cream.To institute
State inoculating starter MA14, inoculum concentration 3g/100L in processing cream, under 32 DEG C of constant temperature culture to pH6.5 when 1g calf stomach is added
Renin, curdled milk 30min obtain curdled milk；
(2) step (1) resulting curd cutting is stirred into 10min discharging whey at bulk；After whey to be discharged into curdled milk
1.25% salt is added, the percentage is the mass percent that the salt accounts for the curdled milk, enters rectangular mould after mixing evenly
Tool；
(3) curdled milk enters mold forming and obtains ziega and periodically overturn, and frequency is to overturn 1 time every 2 hours, continues 24 hours,
Whey is discharged further；
(4) Monascus fermentation broth that thickness is about 1mm is uniformly sprayed on the ziega surface obtained toward step (3), be packed at
Ripe container, constant incubator is mature, the temperature of the maturation are as follows: and 22 DEG C of initial stage, relative humidity 85%, 12 DEG C of the middle and later periods；It is described
The mature time are as follows: initial stage 2 weeks, the middle and later periods 2 weeks.
The Monascus fermentation broth is made by following methods: monascus purpureus BD-M-4 is inoculated in PDB fluid nutrient medium
30 DEG C are cultivated 2 days, the formula of the PDB fluid nutrient medium are as follows: glucose 40g/L, potato leaching powder 6g/L, surplus are water.
10 monascus purpureus BD-M-4 of embodiment prepares cheese
Ingredient: fresh camel cream 100L (being purchased from bright milk industry), MA14 (being purchased from Danisco (China) Co., Ltd product) hair
Ferment agent 3g, calf stomach renin (being purchased from Chr. Hansen A/S) 1g, salt 0.5kg, glucose 40g and potato leaching powder 6g.
(1) after taking the newborn filtering and impurity removing matter of the fresh camel of 100L, 72 DEG C of sterilizing 30s are subsequently cooled to 30 DEG C, must handle cream.To institute
State inoculating starter MA14, inoculum concentration 3g/100L in processing cream, under 30 DEG C of constant temperature culture to pH6.5 when 1g calf stomach is added
Renin, curdled milk 30min obtain curdled milk；
(2) step (1) resulting curd cutting is stirred into 10min discharging whey at bulk；After whey to be discharged into curdled milk
1% salt is added, the percentage is the mass percent that the salt accounts for the curdled milk, enters square dies after mixing evenly；
(3) curdled milk enters mold forming and obtains ziega and periodically overturn, and frequency is to overturn 1 time every 2 hours, continues 24 hours,
Whey is discharged further；
(4) Monascus fermentation broth that thickness is about 1mm is uniformly sprayed on the ziega surface obtained toward step (3), be packed at
Ripe container, constant incubator is mature, the temperature of the maturation are as follows: and 25 DEG C of initial stage, relative humidity 90%, 10 DEG C of the middle and later periods；It is described
The mature time are as follows: initial stage 2 weeks, the middle and later periods 2 weeks.
The Monascus fermentation broth is made by following methods: monascus purpureus BD-M-4 is inoculated in PDB fluid nutrient medium
30 DEG C are cultivated 2 days, the formula of the PDB fluid nutrient medium are as follows: glucose 40g/L, potato leaching powder 6g/L, surplus are water.
11 monascus purpureus BD-M-4 of embodiment fermentation
Seed culture: the monascus purpureus BD-M-4 that 4 DEG C save is to slowly warm up to 15 DEG C, after aseptically will heat up
Bacterial strain is crossed on PDA solid medium, 30 DEG C activation culture 4 days, obtain seed liquor.
PDA solid medium are as follows: 6g/L potato leaching powder, 20g/L glucose and 20g/L agar.
Fermented and cultured: being inoculated in PDB fluid nutrient medium for seed liquor with 5%, 42 DEG C shake culture 3 days, revolving speed is
220rpm, the percentage are percent by volume.
PDB fluid nutrient medium are as follows: 6g/L potato leaching powder and 20g/L glucose, surplus are water.
12 monascus purpureus BD-M-4 of embodiment fermentation
Seed culture: the monascus purpureus BD-M-4 that 4 DEG C save is to slowly warm up to 25 DEG C, after aseptically will heat up
Bacterial strain is crossed on YES solid medium, 30 DEG C activation culture 7 days, obtain seed liquor.
YES solid medium are as follows: 5g/L yeast extract, 30g/L glucose, 6g/L potassium dihydrogen phosphate, 4g/L biphosphate
Sodium, 1g/L ammonium hydroxide and 20g/L agar.
Fermented and cultured: being inoculated in YES fluid nutrient medium for seed liquor with 5%, 20 DEG C shake culture 10 days, revolving speed is
120rpm, the percentage are percent by volume.
YES fluid nutrient medium are as follows: 5g/L yeast extract, 30g/L glucose, 6g/L potassium dihydrogen phosphate, 4g/L biphosphate
Sodium and 1g/L ammonium hydroxide, surplus are water.
13 monascus purpureus BD-M-4 of embodiment fermentation
Seed culture: the monascus purpureus BD-M-4 that 4 DEG C save is to slowly warm up to 15 DEG C, after aseptically will heat up
Bacterial strain is crossed on YES solid medium, 25 DEG C activation culture 8 days, obtain seed liquor.
YES solid medium are as follows: 4g/L yeast extract, 20g/L glucose, 5g/L potassium dihydrogen phosphate, 3g/L biphosphate
Sodium, 0.5g/L ammonium hydroxide and 15g/L agar.
Fermented and cultured: being inoculated in YES fluid nutrient medium for seed liquor with 10%, 25 DEG C shake culture 5 days, revolving speed is
220rpm, the percentage are percent by volume.
YES fluid nutrient medium are as follows: 4g/L yeast extract, 20g/L glucose, 5g/L potassium dihydrogen phosphate, 3g/L biphosphate
Sodium and 0.5g/L ammonium hydroxide, surplus are water.
14 monascus purpureus BD-M-4 of embodiment fermentation
Seed culture: the monascus purpureus BD-M-4 that 4 DEG C save is to slowly warm up to 15 DEG C, after aseptically will heat up
Bacterial strain is crossed on YES solid medium, 25 DEG C activation culture 8 days, obtain seed liquor.
YES solid medium are as follows: 6g/L yeast extract, 40g/L glucose, 7g/L potassium dihydrogen phosphate, 5g/L biphosphate
Sodium, 1.5g/L ammonium hydroxide and 25g/L agar.
Fermented and cultured: being inoculated in YES fluid nutrient medium for seed liquor with 10%, 30 DEG C shake culture 5 days, revolving speed is
180rpm, the percentage are percent by volume.
YES fluid nutrient medium are as follows: 6g/L yeast extract, 40g/L glucose, 7g/L potassium dihydrogen phosphate, 5g/L biphosphate
Sodium and 1.5g/L ammonium hydroxide, surplus are water.
15 monascus purpureus BD-M-4 of embodiment fermentation
Seed culture: the monascus purpureus BD-M-4 that 4 DEG C save is to slowly warm up to 20 DEG C, after aseptically will heat up
Bacterial strain is crossed on MEA solid medium, 15 DEG C activation culture 10 days, obtain seed liquor.
MEA solid medium are as follows: 25g/L malt extract, 3g/L soy peptone and 15g/L agar.
Fermented and cultured: being inoculated in MEA fluid nutrient medium for seed liquor with 5%, 30 DEG C shake culture 10 days, revolving speed is
180rpm, the percentage are percent by volume.
MEA fluid nutrient medium are as follows: 25g/L malt extract and 3g/L soy peptone, surplus are water.
16 monascus purpureus BD-M-4 of embodiment fermentation
Seed culture: the monascus purpureus BD-M-4 that 4 DEG C save is to slowly warm up to 20 DEG C, after aseptically will heat up
Bacterial strain is crossed on MEA solid medium, 15 DEG C activation culture 10 days, obtain seed liquor.
MEA solid medium are as follows: 20g/L malt extract, 2g/L soy peptone and 12g/L agar.
Fermented and cultured: being inoculated in MEA fluid nutrient medium for seed liquor with 8%, 42 DEG C shake culture 3 days, revolving speed is
120rpm, the percentage are percent by volume.
MEA fluid nutrient medium are as follows: 20g/L malt extract and 2g/L soy peptone, surplus are water.
17 monascus purpureus BD-M-4 of embodiment fermentation
Seed culture: the monascus purpureus BD-M-4 that 4 DEG C save is to slowly warm up to 20 DEG C, after aseptically will heat up
Bacterial strain is crossed on MEA solid medium, 15 DEG C activation culture 10 days, obtain seed liquor.
MEA solid medium are as follows: 40g/L malt extract, 4g/L soy peptone and 20g/L agar.
Fermented and cultured: being inoculated in MEA fluid nutrient medium for seed liquor with 10%, 42 DEG C shake culture 3 days, revolving speed is
150rpm, the percentage are percent by volume.
MEA fluid nutrient medium are as follows: 40g/L malt extract and 4g/L soy peptone, surplus are water.
Effect example 1
By purplish red song BD-M-4, monascus BD-A, monascus BD-B, monascus BD-C, monascus GL-A, monascus GL-1
The step of being activated, cultivated according to the step of embodiment 3 with monascus purpureus BD-M-1 measures mycelia weight, as a result such as Fig. 2 and
Shown in table 1.
The result of Fig. 2 illustrates that the biomass of other bacterial strains is compared compared with, the maximum biomass of monascus strain BD-M-4
It is larger, there is significant difference with other bacterial strains, there is apparent growth vigor under 30 DEG C of environment, it is dry particularly suitable for preparing
Junket.
The different monascus purpureus bacterial strain of table 1 is when fermenting 10 days the case where biomass
| Strain name | Biomass (g/L) |
| Purplish red song BD-M-4 | 6.5 |
| Monascus BD-A | 1.5 |
| Monascus BD-B | 1.8 |
| Monascus BD-C | 3 |
| Monascus GL-A | 1.3 |
| Monascus GL-1 | 1.6 |
| Monascus purpureus BD-M-1 | 3.2 |
Effect example 2
By purplish red song BD-M-4, monascus BD-A, monascus BD-B, monascus BD-C, monascus GL-A, monascus GL-1
The step of being activated, cultivated according to the step of embodiment 4 with monascus purpureus BD-M-1 measures citrinin content, as a result such as Fig. 3
With shown in table 2.
The different monascus purpureus bacterial strain of table 2 is when fermenting 10 days the case where citrinin content
| Strain name | Citrinin content (μ g/L) |
| Purplish red song BD-M-4 | 10 |
| Monascus BD-A | 100 |
| Monascus BD-B | 500 |
| Monascus BD-C | 1200 |
| Monascus GL-A | 20 |
| Monascus GL-1 | 15 |
| Monascus purpureus BD-M-1 | 15 |
Table 2 illustrates that the citrinin content that purplish red song BD-M-4 fermentation generates is currently used in the monascus of food preparation
Minimum, relative to other monascuses, the citrinin that purplish red song BD-M-4 fermentation generates is very little, can be widely used in
In the processing technology for preparing the food such as dairy products, red kojic rice powder.
Effect example 3
By purplish red song BD-M-4, monascus BD-A, monascus BD-B, monascus BD-C, monascus GL-A, monascus GL-1
With monascus purpureus BD-M-1 with the ratio of 5% (v/v) be inoculated in YES culture medium (including 5g/L yeast extract, 30g/L glucose,
6g/L potassium dihydrogen phosphate, 4g/L sodium dihydrogen phosphate and 1g/L ammonium hydroxide) in, fermentation liquid is cultivated 5 days to obtain at 28 DEG C, according to Jiangxi perhaps
Flourish measuring method (detection of Jiangxi honor red yeast rice citrinin and ferment control technology [D] Southern Yangtze University doctoral thesis perhaps,
2004.) the isometric 70% ethyl alcohol extraction 30min of 10mL fermentation liquid, is taken, filtering, filtrate carries out after suitably diluting, in 190-
Scanning in 700nm wave-length coverage.
The calculation formula of color value: color value (CV)=OD505× extension rate (U/ml).The results are shown in Table 3:
Color value situation of the different monascus purpureus bacterial strain of table 3 when fermenting 5 days
| Strain name | Color value (CV) |
| Purplish red song BD-M-4 | 6 |
| Monascus BD-A | 4 |
| Monascus BD-B | 4 |
| Monascus BD-C | 5 |
| Monascus GL-A | 5 |
| Monascus GL-1 | 4 |
| Monascus purpureus BD-M-1 | 5 |
Table 3 illustrates that the color value that purplish red song BD-M-4 produces monascorubin is high, illustrates that the ability for producing Monascouruarin is strong.
It should be understood that those skilled in the art can make the present invention various after having read above content of the invention
Change or modification, these equivalent forms also fall within the scope of the appended claims of the present application.
Claims (13)
1. a kind of monascus purpureus (Monascus purpureus), which is characterized in that it is deposited in Chinese microorganism strain preservation
Administration committee's common micro-organisms center, deposit number are as follows: CGMCC No.9712.
2. a kind of method for preparing monascus purpureus CGMCC No.9712, which is characterized in that it includes the following steps, is cultivating
Monascus purpureus CGMCC No.9712 is cultivated in base.
3. method according to claim 2, which is characterized in that the culture medium is PDB fluid nutrient medium, the training of YES liquid
Support base or MEA fluid nutrient medium；The PDB fluid nutrient medium includes 6~10g/L potato leaching powder and 20~40g/L grape
Sugar；The YES fluid nutrient medium includes 4~6g/L yeast extract, 20~40g/L glucose, 5~7g/L potassium dihydrogen phosphate, 3
~5g/L sodium dihydrogen phosphate and 0.5~1.5g/L ammonium hydroxide；The MEA fluid nutrient medium is soaked including 20~40g/L malt
Cream and 2~4g/L soy peptone；The time of the culture is 3~10 days；The temperature of the culture is 20~42 DEG C；Described
Culture is shake culture, and the revolving speed of the shake culture is 120~220rpm；And/or the inoculum concentration of the culture be 5~
10%, the percentage is percent by volume.
4. method as claimed in claim 3, which is characterized in that the time of the culture is 5~10 days.
5. method as claimed in claim 3, which is characterized in that the temperature of the culture is 25~30 DEG C.
6. method as claimed in claim 3, which is characterized in that the revolving speed of the shake culture is 180~220rpm.
7. method according to claim 2, which is characterized in that further include by monascus purpureus CGMCC before the culture
No.9712 is inoculated in the step of seed culture medium carries out seed culture.
8. the method for claim 7, which is characterized in that the time of the seed culture is 5~10 days；The seed training
Feeding temperature is 15~35 DEG C；And/or the culture medium of the seed culture be PDA solid medium, YES solid medium or
MEA solid medium.
9. method according to claim 8, which is characterized in that the time of the seed culture is 6~8 days.
10. method according to claim 8, which is characterized in that the temperature of the seed culture is 25~32 DEG C.
11. the method for claim 7, which is characterized in that the seed culture includes the monascus purpureus bacterium that will be refrigerated
The step of strain slowly heats up.
12. method as claimed in claim 11, which is characterized in that the temperature of the heating is 15~25 DEG C.
13. purposes of the monascus purpureus CGMCC No.9712 as described in claim 1 in food production.
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| CN105505789B (en) * | 2015-12-28 | 2019-01-29 | 光明乳业股份有限公司 | One plant of monascus purpureus bacterial strain and its preparing the application in food |
| CN105420123B (en) * | 2015-12-28 | 2019-02-01 | 光明乳业股份有限公司 | One plant of monascus purpureus bacterial strain and its preparing the application in food |
| CN105505790A (en) * | 2015-12-28 | 2016-04-20 | 光明乳业股份有限公司 | Monascus purpureus strain and application thereof to food preparation |
| CN105505791B (en) * | 2015-12-28 | 2019-02-01 | 光明乳业股份有限公司 | One plant of monascus purpureus bacterial strain and its preparing the application in food |
| 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 |
| CN108624511B (en) * | 2018-05-18 | 2020-12-22 | 浙江师范大学 | Monascus mutant strain capable of producing fluorescent substance and application thereof |
| CN111117898B (en) * | 2019-12-31 | 2022-06-28 | 光明乳业股份有限公司 | Monascus purpureus and application thereof |
| CN113679004B (en) * | 2021-09-04 | 2024-01-05 | 南昌市检验检测中心 | Fermented bean curd and preparation method thereof |
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