CN103589647A - Monascus purpureus YH-6 strain, application thereof and esterified monascus prepared from strain - Google Patents
Monascus purpureus YH-6 strain, application thereof and esterified monascus prepared from strain Download PDFInfo
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Abstract
The invention discloses a Monascus purpureus YH-6 strain, an application thereof and an esterified monascus prepared from the strain. The esterified monascus prepared by using the strain can be used for accelerating the synthesis of four esters mainly comprising ethyl hexanoate in yeast liquor, shortening the fermentation period of the yeast liquor and increasing the high-quality product rate of the yeast liquor. The Monascus purpureus YH-6 strain disclosed by the invention is separated from the yeast produced by Yanghe Bewery Joint-Stock Co., Ltd., and is preserved with the preservation number of CGMCC (China general microbiological culture collection center) No.7959 in the CGMCC in July 17, 2013.
Description
technical field:
The present invention relates to microbial technology field, be specifically related in wine brewing field
'smonascus purpureus bacterium YH-6 bacterial strain and uses thereof and the esterified red yeast of being prepared by this bacterial strain.
background technology:
Monascus can produce multiple enzyme in process of growth, as amylase, saccharifying enzyme, proteolytic enzyme, polygalacturonase, Esterified Enzyme etc.Since the eighties in last century, wine brewing bound pair monascus has carried out exploring widely and applied research, has obtained breakthrough progress and good economic benefit.Monascus kind is more, and wherein esterified red yeast is mainly made by smoky gray monascus and monascus purpureus, and monascus lyolipase has the stronger synthetic ability of catalysis ethyl hexanoate, is called as the outer ethyl hexanoate synthetic enzyme of Monascus ruber born of the same parents.Esterified red yeast (biotechnological formulation of monascus), as a kind of biological catalyst, has important effect in aromatic Chinese spirit fermenting process.
Esterified red yeast has obvious advantage aspect raw fragrant as strengthening Daqu bacterial classification producing ester, for improving the formation of the Esters such as ethyl hexanoate, ethyl acetate in white wine, has played irreplaceable effect.Therefore, the use of esterified red yeast, to improving the content of ethyl hexanoate, improves base wine quality percentage successful, remarkable in economical benefits.In addition, esterified red yeast is applied to yellow water and processes, and can reduce cost of sewage disposal, can realize the recycling to liquor-making byproduct resource, makes COD, BOD content in yellow water decline on the original basis 80%, and obtains certain economic benefit.Moreover esterified red yeast provides important technical support to promoting the class of secondary wine.In secondary wine, contain a large amount of organic acids and alcohols, the organic acid of esterified red yeast in can catalysis secondary wine and the esterification building-up reactions of alcohols, promote the formation of ester compound in wine, thereby improve taste and the quality of secondary wine, increases the output of top grade wine.
Although the discovery apart from esterified red yeast has had considerable time, different research institutions and liquor-making enterprise have all been carried out the research of esterified red yeast, but the production application of practical large-scale at home also seldom, simultaneously, existing esterified red yeast exists the problem that esterification yield is low and does not also arrive fine solution, and the present invention will promote large-scale promotion and the application of esterified red yeast.
Summary of the invention
The esterified red yeast that the technical problem that will solve of the present invention is to provide a kind of monascus purpureus YH-6 bacterial strain and uses thereof and is prepared by this bacterial strain, the esterified red yeast of preparing with this bacterial strain can promote to take in bent wine that ethyl hexanoate is the synthetic of main four kinds of esters, shorten the fermentation period of bent wine, improve the quality percentage of bent wine.
Monascus purpureus of the present invention (
monascus purpureus) separated obtaining in the homemade Daqu of YH-6 bacterial strain Shi Cong Yanghe River limited-liability company of brewery, this bacterial strain is in July, 2013 17 China Committee for Culture Collection of Microorganisms common micro-organisms center preservation (address: No. 3, Yard 1, BeiChen xi Road, Chaoyang District, Beijing City, Institute of Microorganism, Academia Sinica, postcode 100101), preserving number: CGMCC No.7959.
The present invention is achieved through the following technical solutions:
Bacterium (Monascus purpureus) YH-6, preserving number is: CGMCC No.7959.
The esterified red yeast of preparing with above-mentioned monascus purpureus bacterium (Monascus purpureus) YH-6.
Above-mentioned monascus purpureus bacterium or esterified red yeast exist
bentpurposes in wine fermentation.
Described esterified red yeast preparation comprises the following steps:
(1) 1 g koji powder is added in the 250 mL triangular flasks that 50 mL enrichment mediums are housed, at 32 ℃, 100 r/ m enrichment culture 96 h, pipette 5 mL nutrient solutions and fill continuous 2 enrichments in the triangular flask of enrichment medium to another.After enrichment 3 times, dilution spread selective screening is dull and stereotyped, chooses colony colour for red, and meanwhile, transparent circle is transparent clear, and the bacterium colony line that transparent circle diameter is larger with colony diameter ratio is separated, and purifying obtains pure bacterial strain; Bacterial strain after purifying is stored in wort culture medium slant to 4 ℃ of Refrigerator stores;
(2), by the bacterial classification of step (1) cryopreservation, with inoculating needle picking, a little is inoculated in 100 r/m in 100 mL liquid state fermentation substratum and cultivates 96 h, and mycelium pellet number is reached capacity;
(3) the inoculum size access solid medium with 5 % by the bacterium liquid of step (2), 32 ℃ of standing cultivations 20 days, make esterified red yeast.
Described enrichment medium, liquid fermentation medium are respectively: malt extract medium, and natural pH, 115 ℃ of sterilizing 20 min, and then add ethanol 5 % and sterilized lactic acid 4 ‰.
Selectivity plate culture medium: in 100 mL malt extract mediums, add purpurum bromocresolis 0.004 g, tributyrin 1.5%, agar 2 %, natural pH, 115 ℃ of sterilizing 20 min.
Described solid medium is bran mass: wheat bran 100 g, lactic acid 0.4 mL, glucose 0.5 g, peptone 1.5 g, KH
2pO
40.1 g, MgSO
4.7H
2o 0.1 g, water 70 mL, add ethanol 5 mL after 115 ℃ of sterilizing 20 min.
Bacterial screening step:
By above-mentioned steps (1), obtain pure bacterial strain 10 strains.Bacterial strain after purifying is stored in wort culture medium slant to 4 ℃ of Refrigerator stores.
The pure bacterial strain of 10 strain obtaining is prepared to esterified red yeast by above-mentioned steps (2) and step (3) respectively, and the esterification power of screening is the highest
purplemonascus strain ，You Sangon Biotech (Shanghai) Co., Ltd. carries out the order-checking of 26s RNA molecule and identifies.Classification And Nomenclature is
purplemonascus ruber (Monascus purpureus) YH-6.
Colonial morphology is: mycelia is tightr, and spreading property of mycelia is not strong, the conglobate bacterium colony of shape.Bacterium colony surface is fine hair shape, and there is a projection centre, has outside radial gauffer, and edge is more neat.Bacterium colony is dry, tarnish, and colony colour is white at first, along with the prolongation of incubation time, bacterium colony finally becomes sorrel.Open flat board, bacterium colony has simple and elegant fragrance.
With
purplemonascus ruber YH-6 is starting strain, production obtains esterified red yeast crude zyme preparation catalysis caproic acid synthesizing ethyl hexanoate and has very strong specificity and selectivity, its the suitableeest catalytic temperature is 35 ℃, the suitableeest catalytic pH is 3.5, the suitableeest catalysis ethanol concn is 28 %, the suitableeest caproic acid concentration is 1 %, and it is applicable to being applied in the production of aromatic Chinese spirit and yellow water esterification liquid.
Accompanying drawing explanation
Fig. 1 is 10 strains that filter out
purplemonascus ruber YH-6 bacterial strain esterification ability is schematic diagram relatively.
Fig. 2 is
purplemonascus ruber YH-6 colonial morphology schematic diagram (left side is that bacterium colony is positive, and the right side is bacterium colony reverse side).
Fig. 3 is
purplethe 26S rRNA phylogeny schematic diagram of monascus ruber YH-6.
Fig. 4 be with
purplemonascus ruber YH-6 is esterified red yeast prepared by starting strain, esterification power and time curve schematic diagram.
Fig. 5 be with
purplemonascus ruber YH-6 is esterified red yeast prepared by starting strain, esterification power and temperature curve schematic diagram.
Fig. 6 be with
purplemonascus ruber YH-6 is esterified red yeast prepared by starting strain, esterification power and pH value curve synoptic diagram.
Fig. 7 be with
purplemonascus ruber YH-6 is esterified red yeast prepared by starting strain, esterification power and ethanol concn curve synoptic diagram.
Fig. 8 be with
purplemonascus ruber YH-6 is esterified red yeast prepared by starting strain, esterification power and caproic acid concentration curve schematic diagram.
embodiment:
The chemical reagent such as tributyrin, caproic acid, acetic acid, lactic acid, butyric acid, ethanol are analytical pure, purchased from Chemical Reagent Co., Ltd., Sinopharm Group.Wheat bran is by buying on the market of farm produce.Koji powder is the self-control of Yanghe River limited-liability company of brewery.
Prepare following substratum standby
Enrichment medium, liquid fermentation medium are respectively: malt extract medium, and natural pH, 115 ℃ of sterilizing 20 min, and then add ethanol 5 % and sterilized lactic acid 4 ‰.Be volume percent.
Selectivity plate culture medium: in 100 mL malt extract mediums, add purpurum bromocresolis 0.004 g, tributyrin 1.5%, agar 2 %, natural pH, 115 ℃ of sterilizing 20 min.Be volume percent.
Bran mass: wheat bran 100 g, lactic acid 0.4 mL, glucose 0.5 g, peptone 1.5 g, KH
2pO
40.1 g, MgSO
4.7H
2o 0.1 g, water 70 mL, add ethanol 2.5 mL after 115 ℃ of sterilizing 20 min.
Bacterial screening step:
(1) 1 g koji powder is added in the 250 mL triangular flasks that 50 mL enrichment mediums are housed, at 32 ℃, 100 r/ m enrichment culture 96 h, pipette 5 mL nutrient solutions and fill continuous 2 enrichments in the triangular flask of enrichment medium to another.After enrichment 3 times, dilution spread selective screening is dull and stereotyped, chooses colony colour for red, and meanwhile, transparent circle is transparent clear, and the bacterium colony line that transparent circle diameter is larger with colony diameter ratio is separated, and purifying is until obtain pure bacterial strain (obtaining altogether 10 strains).Bacterial strain after purifying is stored in wort culture medium slant to 4 ℃ of Refrigerator stores.
The pure bacterial strain of 10 strain obtaining is prepared respectively to esterified red yeast, and preparation process is as follows:
(2) by cryopreservation bacterial classification, with inoculating needle picking, a little is inoculated in 100 r/m in 100 mL liquid state fermentation substratum and cultivates 96 h, and mycelium pellet number is reached capacity.
(3) the inoculum size access bran mass with 5 % by the bacterium liquid of step (2), 32 ℃ of standing cultivations 20 days, make esterified red yeast wheat bran.
Obtain 10 kinds of esterified red yeast wheat brans, its esterification power is screened, step is as follows:
(4) take step (3) the esterified red yeast wheat bran of the 5 g dry medium amounts that are equivalent to, add 100 mL containing 1 % caproic acid, in the esterification system of 20 % ethanol (all the other are water), 32 ℃ of esterification 100 h, add water 50 mL distillations, meet distillate 100 mL, finally with the ethyl hexanoate content forming in gas chromatographic detection reaction system.Esterification power is defined as in every 100 mL distillates containing the how many milligrams of ethyl hexanoate.The highest monascus purpureus bacterial strain of screening esterification power.The esterified red yeast wheat bran that wherein No. 6 bacterial strains are made, esterification power the highest (180 ㎎ ∕ 100 mL), as shown in Figure 1.This bacterial strain is identified through Sangon Biotech (Shanghai) Co., Ltd., by its called after
purplemonascus ruber YH-6.This bacterial strain is in July, 2013 17 China Committee for Culture Collection of Microorganisms common micro-organisms center preservation (address: No. 3, Yard 1, BeiChen xi Road, Chaoyang District, Beijing City, Institute of Microorganism, Academia Sinica, postcode 100101), preserving number: CGMCC No.7959
.
purplemonascus ruber YH-6 colonial morphology: mycelia is tightr, spreading property of mycelia is not strong, the conglobate bacterium colony of shape.Bacterium colony surface is fine hair shape, and there is a projection centre, has outside radial gauffer, and edge is (left side is that bacterium colony is positive, and the right side is bacterium colony reverse side) more as shown in Figure 2.Bacterium colony is dry, tarnish, and colony colour is white at first, along with the prolongation of incubation time, bacterium colony finally becomes sorrel.Open flat board, bacterium colony has simple and elegant fragrance.
purplemonascus ruber YH-6 ，You Sangon Biotech (Shanghai) Co., Ltd. carries out the order-checking of 26s RNA molecule and identifies.
Utilize PCR sequencing technologies, obtain to be identified
purple587 bp 26S rRNA gene D1/D2 conserved sequences of monascus ruber YH-6 bacterial strain, result is as follows:
GAAAAGAAACCAACCGGGATTGCCTCAGTAACGGCGAGTGAAGCGGCAAGAGCTCAAATTTGAAAGCTGGCCCCTCCGGGGTCCGCGTTGTAATTTGCAGAGGATGCTTCGGGCTCAGCCCCCGTCTAAGTGCCCTGGAACGGGCCGTCGGAGAGGGTGAGAATCCCGTCTGGGACGGGGTGCCTGGGTCCATGTGAAGCTCCTTCGACGAGTCGAGTTGTTTGGGAATGCAGCTCTAAATGGGTGGTAAATTTCATCTAAAGCTAAATACTGGCCGGAGACCGATAGCGCACAAGTAGAGTGATCGAAAGATGAAAAGCACTTTGAAAAGAGAGTTAAACAGCACGTGAAATTGTTGAAAGGGAAGCGCTTGCGATCAGACTCGCCTGCGGGGTTCAGCCGGCATTCGTGCCGGTGTACTTCCCCGTGGGCGGGCCAGCGTCGGTTCGGGTGGCCGGTCAAAGGCCCCGGGAATGTGTCGCCCTCCGGGGCGTCTTATAGCCCGGGGTGCCATGCGGCCTACCTGGACCGAGGAACGCGCTTCGGCTCGGACGCTGGCGTAATGGTCGTAAGCGACCCGTCTT
According to 26S rRNA gene order conservative property, carry out Phylogenetic Analysis.BLAST by NCBI compares, and obtains the higher known taxon of homology, then with phylip software, these sequence construct phylogenetic trees is shown in to accompanying drawing 3.
From phylogenetic tree Fig. 3, the 26S rRNA sequence of the known monascus purpureus YH-6 bacterial strain screening and monascus purpureus (
monascus purpureus) evolutionary distance nearest.
The tolerance of monascus purpureus YH-6 bacterial strain to acid and alcohol:
By YH-6 bacterial strain respectively picking to the malt extract medium that contains 2 ‰, 4 ‰, 6 ‰, 8 ‰, 10 ‰ and 12 ‰ lactic acid, cultivate, result shows when lactic acid concn is over 8 ‰ time, without mycelial growth.By YH-6 bacterial strain respectively picking to the malt extract medium that contains 2 %, 4 %, 6 %, 8 %, 10 % and 12 % alcohol, cultivate, result shows when ethanol concn is during over 10 %, without mycelial growth.Therefore,, when lactic acid and alcohol are used separately, YH-6 can stand 8 ‰ lactic acid and the alcohol of 10 %.
Esterified red yeast wheat bran esterification power and time curve are as shown in Figure 4.
As can be seen from Figure 4, when esterified red yeast YH-6 cultivates the 7th day in bran mass, the esterification power of wheat bran is relatively the highest.While surpassing 7 days, the esterification power of wheat bran can keep relative stability.The incubation time of determining esterified red yeast is 7 days.The same above-mentioned steps of esterification system and culture condition (4).
Esterified red yeast YH-6 wheat bran is measured respectively the esterification power of several acid:
According to esterification power measuring method (above-mentioned steps (4)), in catalyst system, reaction substrate is respectively caproic acid, acetic acid, butyric acid or lactic acid, other condition remains unchanged, and measures respectively the generation content of esterified red yeast YH-6 catalysis ethyl hexanoate, ethyl acetate, ethyl butyrate and ethyl lactate in system.It the results are shown in Table 1.
The esterification power of table 1 esterified red yeast wheat bran YH-6 to several acid
| Organic acid ethyl ester | Ethyl hexanoate | Ethyl acetate | Ethyl butyrate | Ethyl lactate |
| Content (mg/100 mL) | 180.13 | 19.86 | 80.94 | 44.07 |
Esterified red yeast wheat bran is measured the esterification power of mixing acid:
With the mixing acid of each 1 mL of caproic acid, acetic acid, butyric acid and lactic acid, as substrate (the same above-mentioned steps of other condition (4)), measure the generation content of esterified red yeast YH-6 wheat bran catalysis ethyl hexanoate, ethyl acetate, ethyl butyrate and ethyl lactate in system.It the results are shown in Table 2.
The catalytic force of table 2 esterified red yeast YH-6 wheat bran to mixing acid
| Organic acid ethyl ester | Ethyl hexanoate | Ethyl acetate | Ethyl butyrate | Ethyl lactate |
| Content (mg/100 mL) | 78.85 | 14.02 | 48.98 | 11.16 |
As can be seen from Table 2, in nitration mixture, esterified red yeast YH-6 wheat bran is best to the catalytic selectivity of caproic acid, and the growing amount of ethyl hexanoate is the highest, and other three kinds of esters are all fewer.This illustrates when this esterified red yeast wheat bran is applied in the production of aroma daqu liquor and yellow water esterification liquid, in various acid, mix in the substrate existing, selectivity to caproic acid is fairly good, can improve in a large number the content that represents aromatic Chinese spirit Main Fragrance ethyl hexanoate, has sizable utility value.
The mensuration of the best esterification temperature of esterified red yeast YH-6 wheat bran:
According to esterification power measuring method (above-mentioned steps 4), in catalyst system, other condition remains unchanged, and only investigates at different temperature the generation content of esterified red yeast YH-6 Esterified Enzyme catalysis ethyl hexanoate in system.The results are shown in accompanying drawing 5.
As shown in Figure 5, the suitableeest catalytic temperature of esterified red yeast YH-6 wheat bran Esterified Enzyme is 35 ℃, and within the scope of 25 ℃～35 ℃, the esterification ability of esterified red yeast strengthens with the rising of temperature.Its major cause is that enzyme does not have deactivation in this temperature range, and along with the raising of temperature, the activity of enzyme constantly improves.When temperature surpasses more than 35 ℃, can there is protein denaturation in Esterified Enzyme, and the esterification ability of esterified red yeast weakens gradually with the rising of temperature.
The mensuration of the best esterification pH of esterified red yeast YH-6 wheat bran:
According to esterification power measuring method (above-mentioned steps 4), temperature is selected 35 ℃, and in catalyst system, other condition remains unchanged, and investigates under different pH the generation content of esterified red yeast YH-6 Esterified Enzyme catalysis ethyl hexanoate in system.The results are shown in accompanying drawing 6.As shown in Figure 6, the suitable pH of esterified red yeast YH-6 is 3.5.
The mensuration of the best ethanol concn of esterified red yeast YH-6 wheat bran:
According to esterification power measuring method (above-mentioned steps 4), temperature is selected 35 ℃, and pH selects 3.5, and in catalyst system, other condition remains unchanged, and investigates under different ethanol concns the generation content of esterified red yeast YH-6 Esterified Enzyme catalysis ethyl hexanoate in system.The results are shown in accompanying drawing 7.As can be seen from Figure 7, the suitableeest catalysis ethanol concn of esterified red yeast YH-6 Esterified Enzyme is 28 %, and the esterification ability of ethanol concn esterified red yeast when lower than 28 % strengthens with the rising of ethanol concn; And at 28 % when above, the esterification ability of esterified red yeast weakens with the rising of ethanol concn.
The mensuration of the best caproic acid concentration of esterified red yeast YH-6 wheat bran:
According to esterification power measuring method, temperature is selected 35 ℃, and pH selects 3.5, ethanol concn is selected 28 %, in catalyst system, other condition remains unchanged, and investigates under different caproic acid concentration the generation content of esterified red yeast YH-6 Esterified Enzyme catalysis ethyl hexanoate in system.The results are shown in accompanying drawing 8.As can be seen from Figure 8, the suitableeest catalysis caproic acid concentration of esterified red yeast YH-6 Esterified Enzyme is 1 %, and the esterification ability of caproic acid concentration esterified red yeast when lower than 1 % strengthens with the rising of caproic acid concentration; And at 1 % when above, the esterification ability of esterified red yeast weakens with the rising of ethanol concn.In actual production, note controlling the acidity of wine unstrained spirits, acidity is excessive or too small, all can affect the efficiency of esterification.
Monascus is in saprophytic property fungi, and it has stronger capacity of decomposition to biomass.This mainly gives the credit in Natural growth process, and monascus just can produce multiple decomposition enzyme, as amylase, saccharifying enzyme, proteolytic enzyme, Esterified Enzyme etc.If can be used in conjunction with traditional Daqu, can increase substantially saccharogenic power, fermenting power, liquefaction power and the esterification power of Daqu.The homemade bag Bao Qu of esterified red yeast YH-6 wheat bran ，Yu Yanghe River limited-liability company of brewery obtaining with embodiment 1, and carry out production performance comparison with the good red colouring agent for food, also used as a Chinese medicine that becomes to produce in Wuhan of outsourcing.The results are shown in Table 3.
Table 3 esterified red yeast YH-6 wheat bran and bag Bao Qu, become the comparison of red colouring agent for food, also used as a Chinese medicine production performance with good
| ? | YH-6 | Bag Bao Qu | Good one-tenth red colouring agent for food, also used as a Chinese medicine |
| Moisture (%) | 48.4 | 13.06 | 7.96 |
| Acidity (mmoL/10g is bent) | 1.21 | 1.01 | 0.81 |
| Fermenting power (g CO 2The bent .48 hr of/10g) | 0.72 | 0.25 | 2.14 |
| Saccharogenic power (mg C 6H 12O 6The bent .hr of/g) | 635 | 520 | 580 |
| Liquefaction power (the bent .hr of g starch/g) | 1.45 | 1.84 | 1.57 |
| Aspartic protease (U/g is bent) | 58.104 | 58.65 | 57.26 |
| Esterification power (mg/100 mL) | 183.16 | 16.66 | 137.23 |
| Ester decomposing force (mg/100 mL) | 22.32 | 11.94 | 15.25 |
As seen from table, esterified red yeast YH-6 with bag Bao Qu compare, except liquefaction power and, all indexs are all better than Bao Qu.Illustrate that esterified red yeast can be used in combination with bag Bao Qu completely, improves the service efficiency of bag Bao Qu.
Esterified red yeast YH-6 wheat bran is compared with the red colouring agent for food, also used as a Chinese medicine that the good one-tenth in Wuhan is produced, and esterification power is far away higher than good one-tenth red colouring agent for food, also used as a Chinese medicine.In addition, a little less than the red colouring agent for food, also used as a Chinese medicine that the ester decomposing force of esterified red yeast YH-6 wheat bran is also produced than good one-tenth.Esterified red yeast prepared by the YH-6 bacterial strain that the present invention filters out, better than good one-tenth red colouring agent for food, also used as a Chinese medicine, be more suitable for producing in wine brewing.The fermenting power of good one-tenth red colouring agent for food, also used as a Chinese medicine is relatively high, and major cause is to have added a large amount of yeast in good one-tenth red colouring agent for food, also used as a Chinese medicine.
The esterified red yeast YH-6 wheat bran that embodiment 1 obtains, is applied to the processing of liquor-making byproduct with the good red colouring agent for food, also used as a Chinese medicine that becomes to produce in Wuhan of outsourcing.After secondary wine, yellow water and the wine tail that Yanghe River limited-liability company of brewery is produced mixes in the ratio of 2:1:1, add again caproic acid 2.5 %, then add respectively esterified red yeast YH-6 wheat bran 5 %(weight and volume percent), red colouring agent for food, also used as a Chinese medicine 5 %(weight and the volume percent that the good one-tenth in Wuhan is produced), under 35 ℃ of conditions, esterification is 30 days.The results are shown in Table 4.
Table 4 esterified red yeast YH-6 wheat bran becomes red colouring agent for food, also used as a Chinese medicine to liquor-making byproduct esterification Performance Ratio with good
| ? | Esterified red yeast YH-6 wheat bran | Good one-tenth red colouring agent for food, also used as a Chinese medicine |
| Esterification power (mg/100 mL) | 1569.53 | 1287.73 |
As seen from table, the esterification ability successful of esterified red yeast YH-6 to liquor-making byproduct, far above the good one-tenth red colouring agent for food, also used as a Chinese medicine in Wuhan.
Claims (7)
1. monascus purpureus bacterium (Monascus purpureus) YH-6, preserving number is: CGMCC No.7959.
2. the esterified red yeast of preparing with monascus purpureus bacterium YH-6 claimed in claim 1.
3. monascus purpureus bacterium claimed in claim 1 or the purposes of esterified red yeast claimed in claim 2 in bent wine fermentation.
4. esterified red yeast as claimed in claim 2, is characterized in that described esterified red yeast preparation comprises the following steps:
(1) 1 g koji powder is added in the 250 mL triangular flasks that 50 mL enrichment mediums are housed, at 32 ℃, 100 r/ m enrichment culture 96 h, pipette 5 mL nutrient solutions and fill continuous 2 enrichments in the triangular flask of enrichment medium to another, after enrichment 3 times, dilution spread selective screening is dull and stereotyped, choose colony colour for red, meanwhile, transparent circle is transparent clear, the bacterium colony line that transparent circle diameter is larger with colony diameter ratio is separated, and purifying obtains pure bacterial strain; Bacterial strain after purifying is stored in wort culture medium slant to 4 ℃ of Refrigerator stores;
(2), by the bacterial classification of step (1) cryopreservation, with inoculating needle picking, a little is inoculated in 100 r/m in 100 mL liquid state fermentation substratum and cultivates 96 h, and mycelium pellet number is reached capacity;
(3) the inoculum size access solid medium with 5 % by the bacterium liquid of step (2), 32 ℃ of standing cultivations 20 days, make esterified red yeast.
5. esterified red yeast as claimed in claim 4, is characterized in that: described enrichment medium, liquid fermentation medium are respectively malt extract medium, 115 ℃ of sterilizing 20 min, and then add ethanol 5 % and sterilized lactic acid 4 ‰.
6. esterified red yeast as claimed in claim 4, is characterized in that: selectivity plate culture medium, in 100 mL malt extract mediums, adds purpurum bromocresolis 0.004 g, tributyrin 1.5%, agar 2 %, 115 ℃ of sterilizing 20 min.
7. esterified red yeast as claimed in claim 4, is characterized in that: described solid medium is bran mass, composed of the following components, wheat bran 100 g, lactic acid 0.4 mL, glucose 0.5 g, peptone 1.5 g, KH
2pO
40.1 g, MgSO
4.7H
2o 0.1 g, water 70 mL, add ethanol 2.5 mL after 115 ℃ of sterilizing 20 min.
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| CN108004152A (en) * | 2018-01-23 | 2018-05-08 | 武汉雅仕博科技有限公司 | A kind of Monascus and its application |
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| CN110229753A (en) * | 2019-06-28 | 2019-09-13 | 福建生物工程职业技术学院 | A kind of simple Monascus solid-state separation method |
| CN110699263A (en) * | 2019-10-29 | 2020-01-17 | 浙江工业大学 | Aspergillus niger YH-6 and application thereof in improving content of icaritin in epimedium |
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| CN107418901A (en) * | 2017-08-31 | 2017-12-01 | 麻城佳成生物科技有限公司 | A kind of monascus purpureus bacterium of high-yield glucoamylase and its method for producing red yeast rice |
| CN107418901B (en) * | 2017-08-31 | 2019-11-19 | 湖北佳成生物科技有限公司 | A kind of monascus purpureus bacterium of high-yield glucoamylase and its method for producing red yeast rice |
| CN108004152A (en) * | 2018-01-23 | 2018-05-08 | 武汉雅仕博科技有限公司 | A kind of Monascus and its application |
| CN108004152B (en) * | 2018-01-23 | 2021-11-19 | 武汉雅仕博科技有限公司 | Monascus and application thereof |
| CN109439548A (en) * | 2018-11-15 | 2019-03-08 | 中国科学院成都生物研究所 | A kind of strengthening porcelain and Preparation method and use that can generate tyrosol and hydroxytyrosol |
| CN110229753A (en) * | 2019-06-28 | 2019-09-13 | 福建生物工程职业技术学院 | A kind of simple Monascus solid-state separation method |
| CN110699263A (en) * | 2019-10-29 | 2020-01-17 | 浙江工业大学 | Aspergillus niger YH-6 and application thereof in improving content of icaritin in epimedium |
| CN111304102A (en) * | 2020-04-29 | 2020-06-19 | 河南省宋河酒业股份有限公司 | Method for preparing and applying esterified red yeast rice |
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