CN106434401A - Yeast strain enriched in ergosterol and method for preparing ergot yeast powder - Google Patents
Yeast strain enriched in ergosterol and method for preparing ergot yeast powder Download PDFInfo
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Abstract
The invention belongs to the technical field of fermentation engineering and particularly relates to a yeast strain enriched in ergosterol and a method for preparing ergot yeast powder. The yeast strain enriched in ergosterol provided by the invention has the advantages of high ergosterol content, high fermentation efficiency and high fermentative capability. The ergot yeast powder prepared by using the yeast strain enriched in ergosterol provided by the invention is high in contents of dry matters and ergosterol, wherein the content of the dry matters is higher than 1.26g/100ml and the content of ergosterol is higher than 1.62%. The ergot yeast powder prepared by using the yeast strain enriched in ergosterol provided by the invention is simple in process and low in cost, and industrial production of the ergot yeast powder is facilitated.
Description
Technical field
The invention belongs to fermentation engineering field is and in particular to a kind of yeast strain rich in ergosterol and Ergota ferment
The preparation method of female powder.
Background technology
Entitled 24 Beta-methyl cholesterol -5 of ergosterol chemistry, 7 alkene, 3 beta-hydroxies, molecular formula is C28H44O, molecular weight is
396.66, it is a kind of important medicine chemical material and pesticide material.Ergosterol can be used for " cortisone ", " hormone progesterone "
Deng the production of medicine, can also firmly produce brassin lactones and auximone.Meanwhile, ergosterol can also strengthen human body
The ability of resist the disease, just obtains vitamin D after ultraviolet suitable radiation2, vitamin D2Obtain metabolism in regulation human body and animal
On have critical function, there is the effects such as promotion calcium, absorption of mineral such as phosphorus, in addition ergosterol also have significantly antibacterial,
The old effect of antitumor,
Ergosterol is usually to separate to obtain, due to Quantitative Determination of Ergosterol difference in different microorganisms cell from microorganism
Larger, from existing bacterial strain, screening superior strain is the basic skills of strain excellent selection-breeding.For Quantitative Determination of Ergosterol, all kinds of
Between microorganism, difference is very big, and in yeast and some filamentous fungis (as penicillium sp and aspergillosis etc.), Quantitative Determination of Ergosterol is higher, especially with
Saccharomyces cerevisiae is height, and therefore, saccharomyces cerevisiae becomes the main object of study of extraction ergosterol.
Patent documentation CN1123838A discloses a kind of manufacture method rich in ergosterin yeast on June 5th, 1996,
Described manufacture method be using " E " as strain, culture medium with molasses as carbon source, also comprise simultaneously appropriate carbamide, ammonium sulfate,
Phosphoric acid etc. carrys out fermentation culture yeast, and in the yeast cells of gained, Quantitative Determination of Ergosterol can reach 1.4-1.6%, but above-mentioned wheat
Angle sterol yield is relatively low, is unfavorable for the industrialized production of this ergosterol.
Patent documentation CN105176851A discloses a kind of candida tropicalises rich in sterol on December 23rd, 2015
Cultural method, described cultural method first prepares the culture medium with free fatty or fat as sole carbon source, by prepare
Culture medium carries out high temperature sterilize, and after cooling, inoculation candida tropicalises carry out aerobic fermentation of ventilating, and after fermentation ends, centrifugation is received
Collection thalline, that is, obtain the Candida tropicalis cells rich in sterol, in the yeast cells being obtained, sitosterol and campesterol contain
Amount can reach 6.23%-7.12%, but its content ergosterol is extremely low.
In view of currently with yeast produce Quantitative Determination of Ergosterol relatively low, strain fermentation ability unstable it is impossible to ensure
The production problems of ergosterol.Therefore, the content improving the cell ergosterol of barmses is the problem of current urgent need to resolve.
Content of the invention
In order to overcome deficiency of the prior art, it is an object of the invention to provide a kind of yeast rich in ergosterol
Strain and the preparation method of Ergota yeast powder, to solve drawbacks described above.
The invention provides a kind of yeast strain rich in ergosterol, it is deposited in Guangdong Province within 22nd in August in 2016 micro-
Biological inoculum collection, systematic name is：Saccharomyces cerevisiae (Saccharomyces cerevisiae), preserving number is：
GDMCC No：60064, preservation address：5th floor, Xianlie Middle Road, Guangzhou City 100 No. 59 building of compound, Guangdong Microbes Inst.
In addition, present invention also offers a kind of preparation method of Ergota yeast powder, comprising the following steps：
The preparation of S1 acidifying solution：Take drinking water to mix with molasses, then with being steam heated to 100-120 DEG C, adjust sugar hammer
Spend for 20-30Bx, staticly settle 4-8h, filter lime-ash, take supernatant, obtain acidifying solution；
The preparation of S2 nutritional solution：By ammonium sulfate 8-12 part, ammonium dihydrogen phosphate 8-12 part, phosphoric acid 2-4 part, magnesium sulfate 2-4 part,
Potassium sulfate 2-4 part and carbamide 4-6 part mix homogeneously, add 20-50 part to drink water dissolution, obtain nutritional solution；Preferably, described nutrition
Liquid is made up of 10 parts of ammonium sulfate, 10 parts of ammonium dihydrogen phosphate, 3 parts of phosphoric acid, 3 parts of magnesium sulfate, 3 parts of potassium sulfate and 5 parts of carbamide；
S3 actication of culture：The saccharomyces cerevisiae (Saccharomyces cerevisiae) that the present invention is provided be inoculated in containing
In the slant medium of the agar of the beerwort of 4-8Bx and 1-3%, in natural ph, temperature is under 28-32 DEG C of constant temperature
Culture 20-26h, is activated；Yeast after activation is inoculated in the molasses containing 4-8% and 0.5- by 10% subcultivation amount
In the basal fermentation medium of 2% Yeast diffusion juice, it is 3.5-5.5 in pH value, under conditions of temperature is 28-32 DEG C, shaking table shakes
Swing culture 20-26h；Then the ripe yeast cells of concussion and cultivate are made bacteria suspension, are placed in culture dish, magnetic agitation, separate,
Yeast after separation is carried out basal fermentation culture again, obtains barmses；Beerwort, agar, molasses and ferment in wherein cultivating
The percentage ratio (%) of female diffusion juice is percent by volume；
S4 seed culture：The acidifying solution that step S1 is obtained and drinking water are by 5:3 volume ratio is added to first class seed pot
Interior, it is subsequently added into the defoamer of 0.01-0.03% of above-mentioned acidifying solution and drinking water cumulative volume and add above-mentioned acidifying solution and drink
The nutritional solution being obtained with step S2 of the 0.3-0.6% of water cumulative volume, stirs, and is then heated to 100-120 DEG C, insulation
After 12-18min, it is cooled to 28-33 DEG C, obtain base fluid；It is inoculated in base fluid after the barmses amplification culture that step S3 is obtained,
Ventilating fermentation culture 10-12h under conditions of pH value is for 8-10, turns in the big tank equipped with base fluid, in pH after detection standard up to standard
It is worth and cultivates 22-26h for ventilating fermentation under conditions of 8-10, separate, the emulsion after separation is positioned in refrigerated cylinder and preserves, obtain ferment
Breast milk liquid；
S5 fermentation culture：The yeast emulsion that step S4 is obtained is put into equipped with the big tank of base fluid, in pH value for 8-10's
Under the conditions of ventilating fermentation culture 18-22h, obtain fermentation liquid；
S6 separates：The fermentation liquid that step S5 is obtained adds drinking water to carry out flash trapping stage, obtains I emulsion；Add toward in I emulsion
Enter drinking water and carry out the second-order separation, obtain II emulsion；Add purified water to carry out three-level toward in II emulsion to separate, obtain III emulsion；
S7 is dried：The III emulsion that step S6 is obtained is with 1.2-4.8m3The flow velocity of/h is 70-100 DEG C in temperature, steam
Pressure carries out yeasts non-viable, sterilizing under conditions of 0.3-0.4MP, is dried, pulverizing, obtains final product.
Further, the weight of the acidifying solution of described step S1 is 20-25% than sugar, and pH value is 3.6-5.0, and brix is
27-36Bx.
Further, the pH value in the nutritional solution of described step S2 is 1.2-2.5, and brix is 12.4-22.6.
Further, in the first class seed pot of described step S4, the brix of base fluid is 13-19Bx, described first class seed pot
Examination criteria is：Bud ratio >=10%, plants liquid capacity >=8000L, wet yeast >=30g/L.
Further, the pH adjusting agent of described step S4 is that sodium hydroxide prepares composition, described sodium hydroxide with sodium carbonate
With the weight of sodium carbonate than for 1:3-6, described sodium hydroxide addition is the 2% of unit volume of water.
Further, the pH adjusting agent of described step S4 is that sodium hydroxide and sodium carbonate prepare composition, described sodium oxide and
The weight of sodium carbonate is than for 1:4, described sodium hydroxide addition is the 2% of unit volume of water.
Further, wet yeast number >=85g/L in described step S4 yeast emulsion, alcoholic strength is 0.1-0.9% (V/V).
Further, the concentration of I emulsion in described step S6 is 7-10Bx, and the concentration of described II emulsion is 7-13Bx,
The concentration of described III emulsion is 10-15Bx.
Further, described step S7 is dried using double drum type drying machine, described double drum type drying machine dry
Dry condition is：Rotating speed is 6-9r/min, and steam pressure is 0.3-0.4MPa.
The preparation method of Yeast diffusion juice that the present invention provides is：Weigh the food wheat tooth of 2kg, put into powder in pulverizer
Broken 5-10min, takes out and adds 5L drinking water, stirs 4-6h under the conditions of 60-80 DEG C, filters, removes supernatant under the conditions of 100 DEG C
Boil 10-20min, be cooled to room temperature, adjust pH to 3.0-3.5, obtain final product.
In the preparation method of Ergota yeast powder that the present invention provides, nutritional solution is except adding ammonium sulfate, phosphoric acid, magnesium sulfate, urine
Outside plain basic element, also it has been especially added with ammonium dihydrogen phosphate and potassium sulfate, the pH value that ammonium dihydrogen phosphate can be stable very well, sulphuric acid
Potassium then can improve the thickness of yeast cell wall, improve the alimentation ability of barmses, prevent the loss of ergosterol.With
When, yeast seed growth phase, note increasing air quantity, enable yeast to obtain sufficient oxygen, provide in conjunction with the present invention
The characteristic of saccharomyces cerevisiae (Saccharomyces cerevisiae), regulates ammonium sulfate in nutritional solution, phosphoric acid, magnesium sulfate, urine
The ratio of plain basic element, reaches N source, P source and C element and coordinates, so that saccharomyces cerevisiae is fully bred, and promotes saccharomyces cerevisiae synthesis wheat
Angle sterol.
Joined using seed growth phase sodium hydroxide and sodium carbonate in the preparation method of Ergota yeast powder that the present invention provides
The alkali liquor of system can promote ergosterol can synthesize in a large number as pH adjusting agent.Meanwhile, after seed growth phase will separate
Yeast carry out freezen protective and can increase the cell wall pliability of strain, so that the nutrient substance of the inside is kept.
The yeast strain rich in ergosterol that the present invention provides has Quantitative Determination of Ergosterol height, and fermentation efficiency is high and ferments
The strong advantage of ability, the dry of the Ergota yeast powder being prepared using the yeast strain that the present invention provides is more than 1.26g/
100ml, Determination of ergosterol is more than 1.83%, compared with commercially available yeast strain, the dry of the Ergota yeast powder preparing
Improve 19.4%, Quantitative Determination of Ergosterol improves 42.4%, be one plant and become to produce the ideal yeast of ergosterol.
Compared with prior art, the technical scheme that the present invention provides has the advantage that：
(1) yeast strain rich in ergosterol that the present invention provides to have a Quantitative Determination of Ergosterol high, fermentation efficiency high and
The strong advantage of fermentability；
(2) the preparation method process is simple of the Ergota yeast powder that the present invention provides, low cost, are conducive to this Ergota yeast powder
Industry chemical conversion produce.
The yeast strain rich in ergosterol that the present invention provides, is deposited in Guangdong Province's microorganism on 22nd in August in 2016
DSMZ, systematic name is：Saccharomyces cerevisiae (Saccharomyces cerevisiae), preserving number is：GDMCC
No：60064, preservation address：5th floor, Xianlie Middle Road, Guangzhou City 100 No. 59 building of compound, Guangdong Microbes Inst.
Specific embodiment
Below by way of the description of specific embodiment, the invention will be further described, but this is not the limit to the present invention
System, according to the basic thought of the present invention, various modifications may be made or improves for those skilled in the art, but without departing from this
The basic thought of invention, all within the scope of the present invention.
Embodiment 1, a kind of preparation method of Ergota yeast powder
The preparation of S1 acidifying solution：Drinking water is taken to mix with molasses, then with being steam heated to 100 DEG C, adjusting sugared brix is
20Bx, staticly settles 4h, filters lime-ash, takes supernatant, obtains acidifying solution, and the weight of described acidifying solution is 20% than sugar, pH value
For 3.6, brix is 27Bx；
The preparation of S2 nutritional solution：By 8 parts of ammonium sulfate, 8 parts of ammonium dihydrogen phosphate, 2 parts of phosphoric acid, 2 parts of magnesium sulfate, 2 parts of potassium sulfate
With 4 parts of mix homogeneously of carbamide, add 20 parts and drink water dissolution, obtain nutritional solution, in described nutritional solution, pH value is 1.2, brix is
12.4；
S3 actication of culture：Saccharomyces cerevisiae (Saccharomyces cerevisiae) is inoculated in the beerwort containing 4Bx
In the slant medium of 1% agar, in natural ph, temperature is to cultivate 26h under 28 DEG C of constant temperature, is activated；
By activation after yeast by 10% subcultivation amount be inoculated in containing 4% molasses and 0.5% Yeast diffusion juice basal fermentation
In culture medium, it is 3.5 in pH value, temperature is shaking table concussion and cultivate 26h under conditions of 28 DEG C；Then by the ferment of concussion and cultivate maturation
Blast cell makes bacteria suspension, is placed in culture dish, magnetic agitation, separates, the yeast after separation is carried out basal fermentation culture again,
Obtain barmses；
S4 seed culture：The acidifying solution that step S1 is obtained and drinking water are by 5:3 volume ratio is added to first class seed pot
Interior, it is subsequently added into the defoamer of above-mentioned acidifying solution and the 0.01% of drinking water cumulative volume and add above-mentioned acidifying solution total with drinking water
The nutritional solution that step S2 of the 0.3% of volume obtains, stirs, and is then heated to 100 DEG C, after insulation 18min, is cooled to 28
DEG C, obtain base fluid；It is inoculated in base fluid after the barmses amplification culture that step S3 is obtained, divulge information under conditions of pH value is for 8
Fermentation culture 10h, turns in the big tank equipped with base fluid, ventilating fermentation culture under conditions of pH value is for 8 after detection standard up to standard
22h, separates, and the emulsion after separation is positioned in refrigerated cylinder and preserves, obtain yeast emulsion, the hammer of base fluid in described first class seed pot
Spend for 13Bx, the examination criteria of described first class seed pot is：Bud ratio >=10%, plants liquid capacity >=8000L, wet yeast >=30g/
L, described pH adjusting agent is that sodium hydroxide and sodium carbonate prepare composition, and the weight of described sodium hydroxide and sodium carbonate ratio is for 1:3, institute
State 2%, the wet yeast number >=85g/L in described yeast emulsion that sodium hydroxide addition is unit volume of water, alcoholic strength is 0.2%
(V/V)；
S5 fermentation culture：The yeast emulsion that step S4 is obtained is put into equipped with the big tank of base fluid, the condition being 8 in pH value
Lower ventilating fermentation cultivates 22h, obtains fermentation liquid；
S6 separates：The fermentation liquid that step S5 is obtained adds drinking water to carry out flash trapping stage, obtains I emulsion；Add toward in I emulsion
Enter drinking water and carry out the second-order separation, obtain II emulsion；Add purified water to carry out three-level toward in II emulsion to separate, obtain III emulsion, described
The concentration of I emulsion is 7Bx, and the concentration of described II emulsion is 7Bx, and the concentration of described III emulsion is 10Bx；
S7 is dried：The III emulsion that step S6 is obtained is with 1.2m3The flow velocity of/h is 70 DEG C in temperature, and steam pressure is
Carry out yeasts non-viable, sterilizing under conditions of 0.3MP, be dried using double drum type drying machine, described double drum type drying machine
Drying condition be：Rotating speed is 6r/min, and steam pressure is 0.3MPa, and pulverizing obtains final product.
Embodiment 2, a kind of preparation method of Ergota yeast powder
The preparation of S1 acidifying solution：Drinking water is taken to mix with molasses, then with being steam heated to 110 DEG C, adjusting sugared brix is
25Bx, staticly settles 6h, filters lime-ash, takes supernatant, obtains acidifying solution, and the weight of described acidifying solution is 22% than sugar, pH value
For 4.2, brix is 30Bx；
The preparation of S2 nutritional solution：By 10 parts of ammonium sulfate, 10 parts of ammonium dihydrogen phosphate, 3 parts of phosphoric acid, 3 parts of magnesium sulfate, potassium sulfate 3
Part and 5 parts of mix homogeneously of carbamide, add 50 parts and drink water dissolution, obtain nutritional solution, and the pH value in described nutritional solution is 2.0, brix
For 18.4；
S3 actication of culture：Saccharomyces cerevisiae (Saccharomyces cerevisiae) is inoculated in the beerwort containing 6Bx
In the slant medium of 2% agar, in natural ph, temperature is to cultivate 24h under 30 DEG C of constant temperature, is activated；
By activation after yeast by 10% subcultivation amount be inoculated in containing 6% molasses and 1% Yeast diffusion juice basal fermentation training
In foster base, it is 4.5 in pH value, temperature is shaking table concussion and cultivate 24h under conditions of 30 DEG C；Then by the yeast of concussion and cultivate maturation
Cell makes bacteria suspension, is placed in culture dish, magnetic agitation, separates, the yeast after separation is carried out basal fermentation culture again, obtains
Barmses；
S4 seed culture：The acidifying solution that step S1 is obtained and drinking water are by 5:3 volume ratio is added to first class seed pot
Interior, it is subsequently added into the defoamer of above-mentioned acidifying solution and the 0.02% of drinking water cumulative volume and add above-mentioned acidifying solution total with drinking water
The nutritional solution that step S2 of the 0.5% of volume obtains, stirs, and is then heated to 110 DEG C, after insulation 16min, is cooled to 30
DEG C, obtain base fluid；It is inoculated in base fluid after the barmses amplification culture that step S3 is obtained, divulge information under conditions of pH value is for 9
Fermentation culture 11h, turns in the big tank equipped with base fluid, ventilating fermentation culture under conditions of pH value is for 9 after detection standard up to standard
24h, separates, and the emulsion after separation is positioned in refrigerated cylinder and preserves, obtain yeast emulsion, the hammer of base fluid in described first class seed pot
Spend for 15Bx, the examination criteria of described first class seed pot is：Bud ratio >=10%, plants liquid capacity >=8000L, wet yeast >=30g/
L, described pH adjusting agent is that sodium hydroxide and sodium carbonate prepare composition, and the weight of described sodium hydroxide and sodium carbonate ratio is for 1:4, institute
State 2%, the wet yeast number >=85g/L in described yeast emulsion that sodium hydroxide addition is unit volume of water, alcoholic strength is 0.4%
(V/V)；
S5 fermentation culture：The yeast emulsion that step S4 is obtained is put into equipped with the big tank of base fluid, the condition being 9 in pH value
Lower ventilating fermentation cultivates 20h, obtains fermentation liquid；
S6 separates：The fermentation liquid that step S5 is obtained adds drinking water to carry out flash trapping stage, obtains I emulsion；Add toward in I emulsion
Enter drinking water and carry out the second-order separation, obtain II emulsion；Add purified water to carry out three-level toward in II emulsion to separate, obtain III emulsion, described
The concentration of I emulsion in step S6 is 8Bx, and the concentration of described II emulsion is 10Bx, and the concentration of described III emulsion is 12Bx；
S7 is dried：The III emulsion that step S6 is obtained is with 2.8m3The flow velocity of/h is 80 DEG C in temperature, and steam pressure is
Carry out yeasts non-viable, sterilizing under conditions of 0.35MP, it is dried using double drum type drying machine, described double drum type is dried
The drying condition of machine is：Rotating speed is 8r/min, and steam pressure is 0.35MPa, and pulverizing obtains final product.
Embodiment 3, a kind of preparation method of Ergota yeast powder
The preparation of S1 acidifying solution：Drinking water is taken to mix with molasses, then with being steam heated to 120 DEG C, adjusting sugared brix is
30Bx, staticly settles 8h, filters lime-ash, takes supernatant, obtains acidifying solution, and the weight of described acidifying solution is 25% than sugar, pH value
For 5.0, brix is 36Bx；
The preparation of S2 nutritional solution：By 12 parts of ammonium sulfate, 12 parts of ammonium dihydrogen phosphate, phosphatase 24 part, 4 parts of magnesium sulfate, potassium sulfate 4
Part and 6 parts of mix homogeneously of carbamide, add 60 parts and drink water dissolution, obtain nutritional solution, and the pH value in described nutritional solution is 2.5, brix
For 22.6；
S3 actication of culture：Saccharomyces cerevisiae (Saccharomyces cerevisiae) is inoculated in the beerwort containing 8Bx
In the slant medium of 3% agar, in natural ph, temperature is to cultivate 20h under 32 DEG C of constant temperature, is activated；
By activation after yeast by 10% subcultivation amount be inoculated in containing 8% molasses and 2% Yeast diffusion juice basal fermentation training
In foster base, it is 5.5 in pH value, temperature is shaking table concussion and cultivate 20h under conditions of 32 DEG C；Then by the yeast of concussion and cultivate maturation
Cell makes bacteria suspension, is placed in culture dish, magnetic agitation, separates, the yeast after separation is carried out basal fermentation culture again, obtains
Barmses；
S4 seed culture：The acidifying solution that step S1 is obtained and drinking water are by 5:3 volume ratio is added to first class seed pot
Interior, it is subsequently added into the defoamer of above-mentioned acidifying solution and the 0.03% of drinking water cumulative volume and add above-mentioned acidifying solution total with drinking water
The nutritional solution that step S2 of the 0.6% of volume obtains, stirs, and is then heated to 120 DEG C, after insulation 12min, is cooled to 33
DEG C, obtain base fluid；It is inoculated in base fluid after the barmses amplification culture that step S3 is obtained, divulge information under conditions of pH value is for 10
Fermentation culture 12h, turns in the big tank equipped with base fluid, ventilating fermentation culture under conditions of pH value is for 10 after detection standard up to standard
26h, separates, and the emulsion after separation is positioned in refrigerated cylinder and preserves, obtain yeast emulsion, the hammer of base fluid in described first class seed pot
Spend for 19Bx, the examination criteria of described first class seed pot is：Bud ratio >=10%, plants liquid capacity >=8000L, wet yeast >=30g/
L, described pH adjusting agent is that sodium hydroxide and sodium carbonate prepare composition, and the weight of described sodium hydroxide and sodium carbonate ratio is for 1:6, institute
State 2%, the wet yeast number >=85g/L in described yeast emulsion that sodium hydroxide addition is unit volume of water, alcoholic strength is 0.9%
(V/V)；
S5 fermentation culture：The yeast emulsion that step S4 is obtained is put into equipped with the big tank of base fluid, the bar being 10 in pH value
Under part, ventilating fermentation culture 18h, obtains fermentation liquid；
S6 separates：The fermentation liquid that step S5 is obtained adds drinking water to carry out flash trapping stage, obtains I emulsion；Add toward in I emulsion
Enter drinking water and carry out the second-order separation, obtain II emulsion；Add purified water to carry out three-level toward in II emulsion to separate, obtain III emulsion, described
The concentration of I emulsion in step S6 is 10Bx, and the concentration of described II emulsion is 13Bx, and the concentration of described III emulsion is 15Bx；
S7 is dried：The III emulsion that step S6 is obtained is with 4.8m3The flow velocity of/h is 100 DEG C in temperature, and steam pressure is
Carry out yeasts non-viable, sterilizing under conditions of 0.4MP, be dried using double drum type drying machine, described double drum type drying machine
Drying condition be：Rotating speed is 9r/min, and steam pressure is 0.4MPa, and pulverizing obtains final product.
Comparative example 1, a kind of preparation method of Ergota yeast powder
The difference of preparation method is：(genotype is the commercially available ferment of FY4 to adopt commercially available yeast strain in described step S4
Mother strains) be inoculated in the beerwort containing 6Bx and 2% the slant medium of agar in, in natural ph, temperature is 30 DEG C
Cultivate 24h under constant temperature, activated, remaining step such as embodiment 2 is similar to.
Comparative example 2, a kind of preparation method of Ergota yeast powder
The difference of preparation method is：In described step S2 nutritional solution by 10 parts of ammonium sulfate, 3 parts of phosphoric acid, 3 parts of magnesium sulfate,
13 parts of potassium sulfate and 5 parts of mix homogeneously compositions of carbamide, do not add ammonium dihydrogen phosphate, increase the parts by weight of potassium sulfate, remaining
Step such as embodiment 2 is similar to.
Comparative example 3, a kind of preparation method of Ergota yeast powder
The difference of preparation method is：In described step S4, the pH value of ventilating fermentation culture is adjusted to 7, and remaining step is strictly according to the facts
Apply example 2 to be similar to.
Test example one, the quality testing test of Ergota yeast powder
1st, test material：Embodiment 1, embodiment 2, the Ergota yeast powder of comparative example 1, comparative example 2 and comparative example 3 preparation.
2nd, test method：
Example 1, embodiment 2, the Ergota yeast powder of comparative example 1, comparative example 2 and comparative example 3 preparation carry out dry
Measure with Determination of ergosterol, the dry of described Ergota yeast powder is measured using weight method, described Ergota yeast powder
Ergosterol is measured using high performance liquid chromatograph.
4th, result of the test：
Result of the test is as shown in table 1.
The dry of table 1 Ergota yeast powder and the content of ergosterol
| Group | Biomass (g/100ml) | Determination of ergosterol % |
| Embodiment 1 | 1.26 | 1.83 |
| Embodiment 2 | 1.34 | 1.96 |
| Embodiment 3 | 1.28 | 1.87 |
| Comparative example 1 | 1.08 | 1.32 |
| Comparative example 2 | 1.18 | 1.64 |
| Comparative example 3 | 1.16 | 1.60 |
As shown in Table 1, the dry of the Ergota yeast powder being prepared using embodiment of the present invention 1-3 is more than 1.26g/
100ml, Determination of ergosterol is more than 1.83%, and the dry of the wherein Ergota yeast powder obtaining of embodiment 2 preparation is
1.34g/100ml, Determination of ergosterol is 1.96%, is most preferred embodiment.Compared with commercially available yeast strain, the wheat for preparing
The dry of angle yeast powder improves 19.4%, and Quantitative Determination of Ergosterol improves 42.4%, and the wheat that comparative example 1-3 prepares
The dry of angle yeast powder and Determination of ergosterol are relatively low, and the saccharomyces cerevisiae (Saccharomyces that the present invention provides is described
Cerevisiae) Quantitative Determination of Ergosterol is high, and fermentation efficiency is high and fermentability is strong.
Claims (10)
1. a kind of yeast strain rich in ergosterol, is deposited in Guangdong Province's Microbiological Culture Collection in August in 2016 on the 22nd
The heart, preserving number is：GDMCC No：60064, preservation address：5th floor, Xianlie Middle Road, Guangzhou City 100 No. 59 building of compound, Guangdong Province is micro-
Biological study institute.
2. a kind of preparation method of Ergota yeast powder is it is characterised in that comprise the following steps：
The preparation of S1 acidifying solution：Drinking water is taken to mix with molasses, then with being steam heated to 100-120 DEG C, adjusting sugared brix is
20-30Bx, staticly settles 4-8h, filters lime-ash, takes supernatant, obtain acidifying solution；
The preparation of S2 nutritional solution：By ammonium sulfate 8-12 part, ammonium dihydrogen phosphate 8-12 part, phosphoric acid 2-4 part, magnesium sulfate 2-4 part, sulphuric acid
Potassium 2-4 part and carbamide 4-6 part mix homogeneously, add 20-50 part to drink water dissolution, obtain nutritional solution；
S3 actication of culture：Yeast strain described in claim 1 is inoculated in the agar of the beerwort containing 4-8Bx and 1-3%
Slant medium in, in natural pH, temperature is to cultivate 20-26h under 28-32 DEG C of constant temperature, is activated；After activating
Yeast be inoculated in by 10% subcultivation amount the molasses containing 4-8% and 0.5-2% Yeast diffusion juice basal fermentation training
In foster base, it is 3.5-5.5 in pH value, temperature is shaking table concussion and cultivate 20-26h under conditions of 28-32 DEG C；Then by concussion and cultivate
Ripe yeast cells make bacteria suspension, are placed in culture dish, magnetic agitation, separate, the yeast after separation is carried out basis again
Fermentation culture, obtains barmses；
S4 seed culture：The acidifying solution that step S1 is obtained and drinking water are by 5:3 volume ratio is added in first class seed pot, connects
And add above-mentioned acidifying solution total with drinking water with the defoamer of the 0.01-0.03% of drinking water cumulative volume and the above-mentioned acidifying solution of addition
The nutritional solution that step S2 of the 0.3-0.6% of volume obtains, stirs, and is then heated to 100-120 DEG C, is incubated 12-18min
Afterwards, it is cooled to 28-33 DEG C, obtain base fluid；It is inoculated in base fluid after the barmses amplification culture that step S3 is obtained, in pH value be
Ventilating fermentation culture 10-12h under conditions of 8-10, turns in the big tank equipped with base fluid after detection standard up to standard, is 8-10 in pH value
Under conditions of ventilating fermentation culture 22-26h, separate, by the emulsion after separation be positioned in refrigerated cylinder preserve, obtain yeast emulsion；
S5 fermentation culture：The yeast emulsion that step S4 is obtained is put into equipped with the big tank of base fluid, in the condition for 8-10 for the pH value
Lower ventilating fermentation cultivates 18-22h, obtains fermentation liquid；
S6 separates：The fermentation liquid that step S5 is obtained adds drinking water to carry out flash trapping stage, obtains I emulsion；Drink is added toward in I emulsion
Carry out the second-order separation with water, obtain II emulsion；Add purified water to carry out three-level toward in II emulsion to separate, obtain III emulsion；
S7 is dried：The III emulsion that step S6 is obtained is with 1.2-4.8m3The flow velocity of/h is 70-100 DEG C in temperature, and steam pressure is
Carry out yeasts non-viable, sterilizing under conditions of 0.3-0.4MP, be dried, pulverizing, obtain final product.
3. the preparation method of Ergota yeast powder as claimed in claim 2 is it is characterised in that the weight of the acidifying solution of described step S1
Amount is 20-25% than sugar, and pH value is 3.6-5.0, and brix is 27-36Bx.
4. the preparation method of Ergota yeast powder as claimed in claim 2 is it is characterised in that in the nutritional solution of described step S2
PH value is 1.2-2.5, and brix is 12.4-22.6.
5. the preparation method of Ergota yeast powder as claimed in claim 2 is it is characterised in that the first class seed pot of described step S4
The brix of middle base fluid is 13-19Bx, and the examination criteria of described first class seed pot is：Bud ratio >=10%, kind liquid capacity >=
8000L, wet yeast >=30g/L.
6. the preparation method of Ergota yeast powder as claimed in claim 2 is it is characterised in that the pH adjusting agent of described step S4 is
Sodium hydroxide and sodium carbonate prepare composition, and the adding proportion of described sodium hydroxide and sodium carbonate is 1:3-6, described sodium hydroxide adds
Dosage is the 2% of unit volume of water.
7. the preparation method of Ergota yeast powder as claimed in claim 2 is it is characterised in that the pH adjusting agent of described step S4 is
Sodium hydroxide and sodium carbonate prepare composition, and the adding proportion of described sodium hydroxide and sodium carbonate is 1:4, described sodium hydroxide adds
Measure 2% for unit volume of water.
8. the preparation method of Ergota yeast powder as claimed in claim 2 is it is characterised in that wet in described step S4 yeast emulsion
Yeast number >=85g/L, alcoholic strength is 0.1-0.9% (V/V).
9. the preparation method of Ergota yeast powder as claimed in claim 2 is it is characterised in that I emulsion in described step S6
Concentration is 7-10Bx, and the concentration of described II emulsion is 7-13Bx, and the concentration of described III emulsion is 10-15Bx.
10. the preparation method of Ergota yeast powder as claimed in claim 2 is it is characterised in that described step S7 uses twin-roll
Formula drying machine is dried, and the drying condition of described double drum type drying machine is：Rotating speed is 6-9r/min, and steam pressure is 0.3-
0.4MPa.
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| CN111849793A (en) * | 2020-08-10 | 2020-10-30 | 南京金浩医药科技有限公司 | Yeast JHpharm5-1 for high-yield ergosterol and mutagenesis method thereof |
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| CN106901377A (en) * | 2017-03-13 | 2017-06-30 | 广东省生物工程研究所（广州甘蔗糖业研究所） | A kind of calcium tablet containing ergosterol and preparation method thereof |
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| CN111849793A (en) * | 2020-08-10 | 2020-10-30 | 南京金浩医药科技有限公司 | Yeast JHpharm5-1 for high-yield ergosterol and mutagenesis method thereof |
| CN114196527A (en) * | 2021-12-17 | 2022-03-18 | 广东五洲药业有限公司 | Automatic control system for yeast fermentation |
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Denomination of invention: Yeast strain rich in ergosterol and preparation method of ergot yeast powder Effective date of registration: 20221229 Granted publication date: 20190719 Pledgee: Guangdong Suixi Rural Commercial Bank Co.,Ltd. Fucheng Sub branch Pledgor: GUANGDONG WUZHOU PHARMACEUTICAL Co.,Ltd. Registration number: Y2022980029279 |
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