CN106434401B - A kind of preparation method of yeast strain and ergot yeast powder rich in ergosterol - Google Patents
A kind of preparation method of yeast strain and ergot yeast powder rich in ergosterol Download PDFInfo
- Publication number
- CN106434401B CN106434401B CN201610918070.4A CN201610918070A CN106434401B CN 106434401 B CN106434401 B CN 106434401B CN 201610918070 A CN201610918070 A CN 201610918070A CN 106434401 B CN106434401 B CN 106434401B
- Authority
- CN
- China
- Prior art keywords
- yeast
- lotion
- ergot
- preparation
- culture
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/645—Fungi ; Processes using fungi
- C12R2001/85—Saccharomyces
- C12R2001/865—Saccharomyces cerevisiae
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/14—Fungi; Culture media therefor
- C12N1/16—Yeasts; Culture media therefor
- C12N1/18—Baker's yeast; Brewer's yeast
- C12N1/185—Saccharomyces isolates
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/14—Fungi; Culture media therefor
- C12N1/16—Yeasts; Culture media therefor
- C12N1/18—Baker's yeast; Brewer's yeast
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Mycology (AREA)
- Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Organic Chemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Genetics & Genomics (AREA)
- Biotechnology (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- Microbiology (AREA)
- General Engineering & Computer Science (AREA)
- Tropical Medicine & Parasitology (AREA)
- Medicinal Chemistry (AREA)
- Biomedical Technology (AREA)
- Virology (AREA)
- Botany (AREA)
- Coloring Foods And Improving Nutritive Qualities (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Cosmetics (AREA)
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
Abstract
The invention belongs to fermentation engineering fields, and in particular to a kind of preparation method of yeast strain and ergot yeast powder rich in ergosterol.Yeast strain provided by the invention rich in ergosterol has Quantitative Determination of Ergosterol high, fermentation efficiency height and the strong advantage of fermentability.The ergot yeast flakes dry matter and Quantitative Determination of Ergosterol being prepared using the yeast strain provided by the invention rich in ergosterol are high, its dry matter is greater than 1.26g/100ml, Determination of ergosterol is greater than 1.62%, simultaneously, ergot yeast powder simple process is prepared using the yeast strain provided by the invention rich in ergosterol, at low cost, the industry chemical conversion for being conducive to the ergot yeast powder produces.
Description
Technical field
The invention belongs to fermentation engineering fields, and in particular to a kind of yeast strain and ergot ferment rich in ergosterol
The preparation method of female powder.
Background technique
Entitled 24 Beta-methyl cholesterol -5,7 alkene of ergosterol chemistry, 3 beta-hydroxies, molecular formula C28H44O, molecular weight are
396.66, it is a kind of important medicine chemical material and pesticide material.Ergosterol can be used for " cortisone ", " hormone progesterone "
The production of equal drugs can also firmly produce brassin lactones and auximone.Meanwhile ergosterol can also enhance human body
The ability resisted the disease just obtains vitamin D after ultraviolet light suitable radiation2, vitamin D2It must be metabolized adjusting human body and animal
On have critical function, have and promote calcium, the absorption of minerals such as phosphorus and other effects, in addition ergosterol also have it is apparent it is antibacterial,
Antitumor old effect,
Ergosterol is usually isolated from microorganism, due to Quantitative Determination of Ergosterol difference in different microorganisms cell
It is larger, the basic skills that superior strain is strain excellent breeding is screened from existing bacterial strain.It is all kinds of for Quantitative Determination of Ergosterol
Difference is very big between microorganism, and Quantitative Determination of Ergosterol is higher in yeast and certain filamentous fungis (such as mould and aspergillus), especially with
Saccharomyces cerevisiae is height, and therefore, saccharomyces cerevisiae, which becomes, extracts the main research object of ergosterol.
Patent document CN1123838A discloses a kind of manufacturing method rich in ergosterin yeast on June 5th, 1996,
The manufacturing method is using " E " as strain, culture medium using molasses as carbon source, while also comprising suitable urea, ammonium sulfate,
Phosphoric acid etc. carrys out fermented and cultured saccharomycete, and Quantitative Determination of Ergosterol can reach 1.4-1.6% in resulting yeast cells, but above-mentioned wheat
Angle sterol yield is lower, is unfavorable for the industrialized production of the ergosterol.
Patent document CN105176851A discloses a kind of candida tropicalis rich in sterol on December 23rd, 2015
Cultural method, the cultural method first prepare with free fatty acid or fat for sole carbon source culture medium, will be prepared
Culture medium carries out high-temperature sterilization, and after cooling, inoculation candida tropicalis carries out ventilation aerobic fermentation, and after fermentation, centrifugation is received
Collect thallus, that is, obtains the Candida tropicalis cells for being rich in sterol, sitosterol and campesterol contain in yeast cells obtained
Amount can reach 6.23%-7.12%, but its content ergosterol is extremely low.
Lower in view of the Quantitative Determination of Ergosterol generated currently with saccharomycete, strain fermentation ability is unstable, not can guarantee
The production problems of ergosterol.Therefore, the problem of content for improving the cell ergosterol of barms is current urgent need to resolve.
Summary of the invention
In order to overcome the shortcomings in the prior art, the purpose of the present invention is to provide a kind of saccharomycete rich in ergosterol
The preparation method of strain and ergot yeast powder, to solve drawbacks described above.
The present invention provides a kind of yeast strains rich in ergosterol, and it is micro- to be deposited in Guangdong Province on August 22nd, 2016
Biological inoculum collection, systematic name are as follows: saccharomyces cerevisiae (Saccharomyces cerevisiae), deposit number are as follows:
GDMCC No:60064, preservation address: 5 building, the building of compound the 59th of Xianlie Middle Road, Guangzhou City 100, Guangdong Microbes Inst.
In addition, the present invention also provides a kind of preparation methods of ergot yeast powder, comprising the following steps:
The preparation of S1 acidifying solution: taking drinking water to mix with molasses, then with being steam heated to 100-120 DEG C, adjusts sugar hammer
Degree is 20-30Bx, staticly settles 4-8h, filters lime-ash, takes supernatant, obtain acidifying solution；
The preparation of S2 nutrient solution: by 8-12 parts of ammonium sulfate, 8-12 parts of ammonium dihydrogen phosphate, 2-4 parts of phosphoric acid, 2-4 parts of magnesium sulfate,
2-4 parts and urea 4-6 parts of potassium sulfate are uniformly mixed, and 20-50 parts of drinking water dissolutions are added, obtain nutrient solution；Preferably, the nutrition
Liquid is made 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 urea；
S3 actication of culture: by saccharomyces cerevisiae provided by the invention (Saccharomyces cerevisiae) be inoculated in containing
In the slant medium of the agar of the brewer's wort and 1-3% of 4-8Bx, under natural ph, the constant temperature that temperature is 28-32 DEG C
20-26h is cultivated, is activated；Saccharomycete after activation is inoculated in molasses and 0.5- containing 4-8% by 10% culture transferring amount
It is 3.5-5.5 in pH value, shaking table shakes under conditions of temperature is 28-32 DEG C in the basal fermentation medium of 2% Yeast diffusion juice
Swing culture 20-26h；Then bacteria suspension being made in the yeast cells of shake culture maturation, is placed in culture dish, magnetic agitation separates,
Saccharomycete after separation is subjected to basal fermentation culture again, obtains barms；Wherein brewer's wort, agar, molasses and ferment in culture
The percentage (%) of female diffusion juice is percent by volume；
S4 seed culture: the obtained acidifying solution of step S1 and drinking water are added to first class seed pot by the volume ratio of 5:3
It is interior, it is subsequently added into the defoaming agent of the 0.01-0.03% of above-mentioned acidifying solution and drinking water total volume and above-mentioned acidifying solution and drink is added
The nutrient solution obtained with the step S2 of the 0.3-0.6% of water total volume, stirs evenly, and is then heated to 100-120 DEG C, heat preservation
After 12-18min, it is cooled to 28-33 DEG C, obtains base fluid；It is inoculated in base fluid after the barms that step S3 is obtained are expanded culture,
Ventilating fermentation culture 10-12h under conditions of pH value is 8-10 turns in the big tank equipped with base fluid after detecting standard up to standard, in pH
Value is ventilating fermentation culture 22-26h under conditions of 8-10, and the lotion after separation is placed in refrigerated cylinder and saves, obtains ferment by separation
Breast milk liquid；
S5 fermented and cultured: the yeast lotion that step S4 is obtained is put into the big tank equipped with base fluid, is 8-10's in pH value
Under the conditions of ventilating fermentation culture 18-22h, obtain fermentation liquid；
S6 separation: the fermentation liquid that step S5 is obtained is added drinking water and carries out level-one separation, obtains I lotion；Add into I lotion
Enter drinking water and carry out the second-order separation, obtains II lotion；Purified water is added into II lotion and carries out three-level separation, obtains III lotion；
S7 is dry: the III lotion 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 being 0.3-0.4MP, dry, milling to get.
Further, the weight ratio sugar of the acidifying solution of the step S1 is 20-25%, pH value 3.6-5.0, and brix is
27-36Bx。
Further, the pH value in the nutrient solution of the step S2 is 1.2-2.5, brix 12.4-22.6.
Further, the brix of base fluid is 13-19Bx in the first class seed pot of the step S4, the first class seed pot
Examination criteria are as follows: bud ratio >=10% plants liquid capacity >=8000L, wet yeast >=30g/L.
Further, the pH adjusting agent of the step S4 is that sodium hydroxide and sodium carbonate are prepared and formed, the sodium hydroxide
Weight ratio with sodium carbonate is 1:3-6, and the sodium hydroxide additive amount is the 2% of unit volume of water.
Further, the pH adjusting agent of the step S4 is that sodium hydroxide and sodium carbonate are prepared and formed, the sodium oxide molybdena and
The weight ratio of sodium carbonate is 1:4, and the sodium hydroxide additive amount is the 2% of unit volume of water.
Further, wet yeast number >=85g/L in the step S4 yeast lotion, alcoholic strength are 0.1-0.9% (V/V).
Further, the concentration of I lotion in the step S6 is 7-10Bx, and the concentration of II lotion is 7-13Bx,
The concentration of III lotion is 10-15Bx.
Further, the step S7 is dried using double drum type drying machine, and the double drum type drying machine is done
Dry condition are as follows: revolving speed 6-9r/min, steam pressure 0.3-0.4MPa.
Yeast diffusion juice provided by the invention the preparation method comprises the following steps: weigh the food malt of 2kg, be put into powder in pulverizer
Broken 5-10min takes out and 5L drinking water is added, 4-6h is stirred under the conditions of 60-80 DEG C, filters, goes supernatant under the conditions of 100 DEG C
Boil 10-20min, be cooled to room temperature, adjust pH to 3.0-3.5 to get.
Nutrient solution is in addition to being added ammonium sulfate, phosphoric acid, magnesium sulfate, urine in the preparation method of ergot yeast powder provided by the invention
Outside plain basic element, be also especially added with ammonium dihydrogen phosphate and potassium sulfate, ammonium dihydrogen phosphate can stable pH value very well, sulfuric acid
The thickness of yeast cell wall then can be improved in potassium, improves the nutrient absorption ability of barms, prevents the loss of ergosterol.Together
When, saccharomycete seed growth phase pays attention to increasing air quantity, so that yeast is obtained sufficient oxygen, in conjunction with provided by the invention
The characteristic of saccharomyces cerevisiae (Saccharomyces cerevisiae), regulates ammonium sulfate in nutrient solution, phosphoric acid, magnesium sulfate, urine
The ratio of plain basic element reaches the source N, the source P and C element and coordinates, breed saccharomyces cerevisiae sufficiently, saccharomyces cerevisiae is promoted to synthesize wheat
Angle sterol.
Matched in the preparation method of ergot yeast powder provided by the invention using seed growth phase sodium hydroxide and sodium carbonate
The lye of system can promote ergosterol that can largely synthesize as pH adjusting agent.Meanwhile after seed growth phase will separate
Yeast carry out freezen protective and can increase the cell wall flexibility of strain, keep the nutriment of the inside.
Yeast strain provided by the invention rich in ergosterol has Quantitative Determination of Ergosterol height, and fermentation efficiency is high and ferments
The dry matter of the strong advantage of ability, the ergot yeast powder being prepared using yeast strain provided by the invention is greater than 1.26g/
100ml, Determination of ergosterol are greater than 1.83%, compared with commercially available yeast strain, the dry matter for the ergot yeast powder being prepared
19.4% is improved, Quantitative Determination of Ergosterol improves 42.4%, is one plant of saccharomycete ideal at production ergosterol.
Compared with prior art, technical solution provided by the invention has the advantage that
(1) yeast strain provided by the invention rich in ergosterol has Quantitative Determination of Ergosterol high, fermentation efficiency height and
The strong advantage of fermentability；
(2) the preparation method simple process and low cost of ergot yeast powder provided by the invention is conducive to the ergot yeast powder
Industry chemical conversion produce.
Yeast strain provided by the invention rich in ergosterol was deposited in Guangdong Province microorganism on August 22nd, 2016
Culture Collection Center, systematic name are as follows: saccharomyces cerevisiae (Saccharomyces cerevisiae), deposit number are as follows: GDMCC
No:60064, preservation address: 5 building, the building of compound the 59th of Xianlie Middle Road, Guangzhou City 100, Guangdong Microbes Inst.
Specific embodiment
The following describes the present invention further through the description of specific embodiments, but it is to limit of the invention that this, which is not,
System, those skilled in the art's basic thought according to the present invention can make various modifications or improvements, but without departing from this
The basic thought of invention, is all within the scope of the present invention.
Embodiment 1, a kind of preparation method of ergot yeast powder
The preparation of S1 acidifying solution: taking drinking water to mix with molasses, and 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 ratio sugar of the acidifying solution is 20%, pH value
It is 3.6, brix 27Bx；
The preparation of S2 nutrient 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
It is uniformly mixed with 4 parts of urea, 20 parts of drinking water dissolutions is added, obtain nutrient solution, pH value is 1.2 in the nutrient solution, and brix is
12.4；
S3 actication of culture: saccharomyces cerevisiae (Saccharomyces cerevisiae) is inoculated in the brewer's wort containing 4Bx
In the slant medium of 1% agar, 26h is cultivated under natural ph, the constant temperature that temperature is 28 DEG C, is activated；
Saccharomycete after activation is inoculated in the basal fermentation containing 4% molasses and 0.5% Yeast diffusion juice by 10% culture transferring amount
It is 3.5 in pH value, temperature is shaking table shake culture 26h under conditions of 28 DEG C in culture medium；Then by the ferment of shake culture maturation
Bacteria suspension is made in mother cell, is placed in culture dish, magnetic agitation, and the saccharomycete after separation is carried out basal fermentation culture by separation again,
Obtain barms；
S4 seed culture: the obtained acidifying solution of step S1 and drinking water are added to first class seed pot by the volume ratio of 5:3
Interior, the defoaming agent and the above-mentioned acidifying solution of addition and drinking water for being subsequently added into above-mentioned acidifying solution with the 0.01% of drinking water total volume are total
The nutrient solution that the step S2 of the 0.3% of volume is obtained, stirs evenly, and is then heated to 100 DEG C, after keeping the temperature 18min, is cooled to 28
DEG C, obtain base fluid；It is inoculated in base fluid after the barms that step S3 is obtained are expanded culture, divulges information under conditions of pH value is 8
Fermented and cultured 10h turns in the big tank equipped with base fluid, ventilating fermentation culture under conditions of pH value is 8 after detecting standard up to standard
22h, separation, the lotion after separation is placed in refrigerated cylinder and is saved, yeast lotion is obtained, the hammer of base fluid in the first class seed pot
Degree is 13Bx, the examination criteria of the first class seed pot are as follows: bud ratio >=10% plants liquid capacity >=8000L, wet yeast >=30g/
L, the pH adjusting agent are that sodium hydroxide and sodium carbonate are prepared and formed, and the weight ratio of the sodium hydroxide and sodium carbonate is 1:3, institute
State 2% that sodium hydroxide additive amount is unit volume of water, wet yeast number >=85g/L in the yeast lotion, alcoholic strength 0.2%
(V/V)；
S5 fermented and cultured: the yeast lotion that step S4 is obtained is put into the big tank equipped with base fluid, the condition for being 8 in pH value
Lower ventilating fermentation culture 22h, obtains fermentation liquid；
S6 separation: the fermentation liquid that step S5 is obtained is added drinking water and carries out level-one separation, obtains I lotion；Add into I lotion
Enter drinking water and carry out the second-order separation, obtains II lotion；Purified water is added into II lotion and carries out three-level separation, obtains III lotion, it is described
The concentration of I lotion is 7Bx, and the concentration of II lotion is 7Bx, and the concentration of III lotion is 10Bx；
S7 is dry: the III lotion that step S6 is obtained is with 1.2m3The flow velocity of/h is 70 DEG C in temperature, and steam pressure is
Yeasts non-viable, sterilizing are carried out under conditions of 0.3MP, are dried using double drum type drying machine, the double drum type drying machine
Drying condition are as follows: revolving speed 6r/min, steam pressure 0.3MPa, milling to get.
Embodiment 2, a kind of preparation method of ergot yeast powder
The preparation of S1 acidifying solution: taking drinking water to mix with molasses, and 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 ratio sugar of the acidifying solution is 22%, pH value
It is 4.2, brix 30Bx；
The preparation of S2 nutrient 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 is uniformly mixed with 5 parts of urea, and 50 parts of drinking water dissolutions are added, obtain nutrient solution, the pH value in the nutrient solution is 2.0, brix
It is 18.4；
S3 actication of culture: saccharomyces cerevisiae (Saccharomyces cerevisiae) is inoculated in the brewer's wort containing 6Bx
In the slant medium of 2% agar, cultivates for 24 hours, activated under natural ph, the constant temperature that temperature is 30 DEG C；
Saccharomycete after activation is inoculated in by 10% culture transferring amount and is trained containing the basal fermentation of 6% molasses and 1% Yeast diffusion juice
Support base in, pH value be 4.5, temperature be 30 DEG C under conditions of shaking table shake culture for 24 hours；Then by the yeast of shake culture maturation
Bacteria suspension is made in cell, is placed in culture dish, magnetic agitation, and the saccharomycete after separation is carried out basal fermentation culture again, obtained by separation
Barms；
S4 seed culture: the obtained acidifying solution of step S1 and drinking water are added to first class seed pot by the volume ratio of 5:3
Interior, the defoaming agent and the above-mentioned acidifying solution of addition and drinking water for being subsequently added into above-mentioned acidifying solution with the 0.02% of drinking water total volume are total
The nutrient solution that the step S2 of the 0.5% of volume is obtained, stirs evenly, and is then heated to 110 DEG C, after keeping the temperature 16min, is cooled to 30
DEG C, obtain base fluid；It is inoculated in base fluid after the barms that step S3 is obtained are expanded culture, divulges information under conditions of pH value is 9
Fermented and cultured 11h turns in the big tank equipped with base fluid, ventilating fermentation culture under conditions of pH value is 9 after detecting standard up to standard
For 24 hours, it separates, the lotion after separation is placed in refrigerated cylinder and is saved, yeast lotion is obtained, the hammer of base fluid in the first class seed pot
Degree is 15Bx, the examination criteria of the first class seed pot are as follows: bud ratio >=10% plants liquid capacity >=8000L, wet yeast >=30g/
L, the pH adjusting agent are that sodium hydroxide and sodium carbonate are prepared and formed, and the weight ratio of the sodium hydroxide and sodium carbonate is 1:4, institute
State 2% that sodium hydroxide additive amount is unit volume of water, wet yeast number >=85g/L in the yeast lotion, alcoholic strength 0.4%
(V/V)；
S5 fermented and cultured: the yeast lotion that step S4 is obtained is put into the big tank equipped with base fluid, the condition for being 9 in pH value
Lower ventilating fermentation culture 20h, obtains fermentation liquid；
S6 separation: the fermentation liquid that step S5 is obtained is added drinking water and carries out level-one separation, obtains I lotion；Add into I lotion
Enter drinking water and carry out the second-order separation, obtains II lotion；Purified water is added into II lotion and carries out three-level separation, obtains III lotion, it is described
The concentration of I lotion in step S6 is 8Bx, and the concentration of II lotion is 10Bx, and the concentration of III lotion is 12Bx；
S7 is dry: the III lotion that step S6 is obtained is with 2.8m3The flow velocity of/h is 80 DEG C in temperature, and steam pressure is
Yeasts non-viable, sterilizing are carried out under conditions of 0.35MP, it is dried using double drum type drying machine, the double drum type is dry
The drying condition of machine are as follows: revolving speed 8r/min, steam pressure 0.35MPa, milling to get.
Embodiment 3, a kind of preparation method of ergot yeast powder
The preparation of S1 acidifying solution: taking drinking water to mix with molasses, and 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 ratio sugar of the acidifying solution is 25%, pH value
It is 5.0, brix 36Bx；
The preparation of S2 nutrient 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 is uniformly mixed with 6 parts of urea, and 60 parts of drinking water dissolutions are added, obtain nutrient solution, the pH value in the nutrient solution is 2.5, brix
It is 22.6；
S3 actication of culture: saccharomyces cerevisiae (Saccharomyces cerevisiae) is inoculated in the brewer's wort containing 8Bx
In the slant medium of 3% agar, 20h is cultivated under natural ph, the constant temperature that temperature is 32 DEG C, is activated；
Saccharomycete after activation is inoculated in by 10% culture transferring amount and is trained containing the basal fermentation of 8% molasses and 2% Yeast diffusion juice
It supports in base, is 5.5 in pH value, temperature is shaking table shake culture 20h under conditions of 32 DEG C；Then by the yeast of shake culture maturation
Bacteria suspension is made in cell, is placed in culture dish, magnetic agitation, and the saccharomycete after separation is carried out basal fermentation culture again, obtained by separation
Barms；
S4 seed culture: the obtained acidifying solution of step S1 and drinking water are added to first class seed pot by the volume ratio of 5:3
Interior, the defoaming agent and the above-mentioned acidifying solution of addition and drinking water for being subsequently added into above-mentioned acidifying solution with the 0.03% of drinking water total volume are total
The nutrient solution that the step S2 of the 0.6% of volume is obtained, stirs evenly, and is then heated to 120 DEG C, after keeping the temperature 12min, is cooled to 33
DEG C, obtain base fluid；It is inoculated in base fluid after the barms that step S3 is obtained are expanded culture, divulges information under conditions of pH value is 10
Fermented and cultured 12h turns in the big tank equipped with base fluid, ventilating fermentation culture under conditions of pH value is 10 after detecting standard up to standard
26h, separation, the lotion after separation is placed in refrigerated cylinder and is saved, yeast lotion is obtained, the hammer of base fluid in the first class seed pot
Degree is 19Bx, the examination criteria of the first class seed pot are as follows: bud ratio >=10% plants liquid capacity >=8000L, wet yeast >=30g/
L, the pH adjusting agent are that sodium hydroxide and sodium carbonate are prepared and formed, and the weight ratio of the sodium hydroxide and sodium carbonate is 1:6, institute
State 2% that sodium hydroxide additive amount is unit volume of water, wet yeast number >=85g/L in the yeast lotion, alcoholic strength 0.9%
(V/V)；
S5 fermented and cultured: the yeast lotion that step S4 is obtained is put into the big tank equipped with base fluid, the item for being 10 in pH value
Ventilating fermentation culture 18h, obtains fermentation liquid under part；
S6 separation: the fermentation liquid that step S5 is obtained is added drinking water and carries out level-one separation, obtains I lotion；Add into I lotion
Enter drinking water and carry out the second-order separation, obtains II lotion；Purified water is added into II lotion and carries out three-level separation, obtains III lotion, it is described
The concentration of I lotion in step S6 is 10Bx, and the concentration of II lotion is 13Bx, and the concentration of III lotion is 15Bx；
S7 is dry: the III lotion that step S6 is obtained is with 4.8m3The flow velocity of/h is 100 DEG C in temperature, and steam pressure is
Yeasts non-viable, sterilizing are carried out under conditions of 0.4MP, are dried using double drum type drying machine, the double drum type drying machine
Drying condition are as follows: revolving speed 9r/min, steam pressure 0.4MPa, milling to get.
Comparative example 1, a kind of preparation method of ergot yeast powder
The difference of preparation method is: commercially available yeast strain is used in the step S4, and (genotype is the commercially available ferment of FY4
Mother strains) be inoculated in the brewer's wort containing 6Bx and 2% agar slant medium in, in natural ph, temperature is 30 DEG C
It cultivates for 24 hours, is activated under constant temperature, remaining step such as embodiment 2 is similar.
Comparative example 2, a kind of preparation method of ergot yeast powder
The difference of preparation method is: in the step S2 nutrient solution by 10 parts of ammonium sulfate, 3 parts of phosphoric acid, 3 parts of magnesium sulfate,
13 parts and 5 parts of urea of potassium sulfate are uniformly mixed composition, do not add ammonium dihydrogen phosphate, increase the parts by weight of potassium sulfate, remaining
Step such as embodiment 2 is similar.
Comparative example 3, a kind of preparation method of ergot yeast powder
The difference of preparation method is: the pH value of ventilating fermentation culture is adjusted to 7 in the step S4, remaining step is strictly according to the facts
It is similar to apply example 2.
The quality testing test of test example one, ergot yeast powder
1, test material: ergot yeast powder prepared by embodiment 1, embodiment 2, comparative example 1, comparative example 2 and comparative example 3.
2, test method:
Ergot yeast powder prepared by Example 1, embodiment 2, comparative example 1, comparative example 2 and comparative example 3 carries out dry matter
It is measured with Determination of ergosterol, the dry matter of the ergot yeast powder is measured using weight method, the ergot yeast powder
Ergosterol is measured using high performance liquid chromatograph.
4, test result:
Test result is as shown in table 1.
The dry matter of 1 ergot yeast powder of table 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 1,1.26g/ is greater than using the dry matter of 1-3 of the embodiment of the present invention ergot yeast powder being prepared
100ml, Determination of ergosterol are greater than 1.83%, and the dry matter for the obtained ergot yeast powder that wherein prepared by embodiment 2 is
1.34g/100ml Determination of ergosterol 1.96% is most preferred embodiment.Compared with commercially available yeast strain, the wheat that is prepared
The dry matter of angle yeast powder improves 19.4%, and Quantitative Determination of Ergosterol improves 42.4%, and the wheat that comparative example 1-3 is prepared
The dry matter and Determination of ergosterol of angle yeast powder are relatively low, illustrate saccharomyces cerevisiae (Saccharomyces provided by the invention
Cerevisiae) Quantitative Determination of Ergosterol is high, and fermentation efficiency height and fermentability are strong.
Claims (10)
1. a kind of yeast strain rich in ergosterol was deposited in Guangdong Province's Microbiological Culture Collection on August 22nd, 2016
The heart, deposit number are as follows: GDMCC No:60064, preservation address: 5 building, the building of compound the 59th of Xianlie Middle Road, Guangzhou City 100, Guangdong Province is micro-
Biological study institute.
2. a kind of preparation method of ergot yeast powder, which comprises the following steps:
The preparation of S1 acidifying solution: taking drinking water to mix with molasses, and 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 nutrient solution: by 8-12 parts of ammonium sulfate, 8-12 parts of ammonium dihydrogen phosphate, 2-4 parts of phosphoric acid, 2-4 parts of magnesium sulfate, sulfuric acid
2-4 parts and urea 4-6 parts of potassium are uniformly mixed, and 20-50 parts of drinking water dissolutions are added, obtain nutrient solution；
S3 actication of culture: yeast strain described in claim 1 is inoculated in the brewer's wort containing 4-8 Bx and the agar of 1-3%
Slant medium in, in natural pH, cultivate 20-26h under the constant temperature that temperature is 28-32 DEG C, activated；After activating
Saccharomycete the basal fermentation mediums of the molasses containing 4-8% and the Yeast diffusion juice of 0.5-2% is inoculated in by 10% culture transferring amount
In, it is 3.5-5.5 in pH value, temperature is shaking table shake culture 20-26h under conditions of 28-32 DEG C；Then by shake culture maturation
Yeast cells bacteria suspension is made, be placed in culture dish, the saccharomycete after separation is carried out basal fermentation by magnetic agitation, separation again
Culture, obtains barms；
S4 seed culture: the obtained acidifying solution of step S1 and drinking water are added in first class seed pot by the volume ratio of 5:3, connect
Be added above-mentioned acidifying solution and the 0.01-0.03% of drinking water total volume defoaming agent and the above-mentioned acidifying solution of addition and drinking water it is total
The nutrient solution that the step S2 of the 0.3-0.6% of volume is obtained, stirs evenly, and is then heated to 100-120 DEG C, keeps the temperature 12-18min
Afterwards, it is cooled to 28-33 DEG C, obtains base fluid；It is inoculated in base fluid after the barms that step S3 is obtained are expanded culture, is in pH value
Ventilating fermentation culture 10-12h under conditions of 8-10 turns in the big tank equipped with base fluid after detecting standard up to standard, is 8-10 in pH value
Under conditions of ventilating fermentation culture 22-26h, separation, the lotion after separation is placed in refrigerated cylinder and is saved, yeast lotion is obtained；
S5 fermented and cultured: the yeast lotion that step S4 is obtained is put into the big tank equipped with base fluid, in the condition that pH value is 8-10
Lower ventilating fermentation culture 18-22h, obtains fermentation liquid；
S6 separation: the fermentation liquid that step S5 is obtained is added drinking water and carries out level-one separation, obtains I lotion；Drink is added into I lotion
The second-order separation is carried out with water, obtains II lotion；Purified water is added into II lotion and carries out three-level separation, obtains III lotion；
S7 is dry: the III lotion that step S6 is obtained is with 1.2-4.8 m3The flow velocity of/h is 70-100 DEG C in temperature, steam pressure
It is dry to carry out yeasts non-viable, sterilizing under conditions of 0.3-0.4 MP, milling to get.
3. the preparation method of ergot yeast powder as claimed in claim 2, which is characterized in that the weight of the acidifying solution of the step S1
Amount is 20-25%, pH value 3.6-5.0, brix 27-36Bx than sugar.
4. the preparation method of ergot yeast powder as claimed in claim 2, which is characterized in that the pH of the nutrient solution of the step S2
Value is 1.2-2.5, brix 12.4-22.6Bx.
5. the preparation method of ergot yeast powder as claimed in claim 2, which is characterized in that the first class seed pot of the step S4
The brix of middle base fluid is 13-19Bx, the examination criteria of the first class seed pot are as follows: bud ratio >=10% plants liquid capacity >=8000L,
Wet yeast >=30g/L.
6. the preparation method of ergot yeast powder as claimed in claim 2, which is characterized in that adjusted in the step S4 using pH
PH value is adjusted to 8-10 by agent, and the pH adjusting agent is that sodium hydroxide and sodium carbonate are prepared and formed, the sodium hydroxide and carbonic acid
The adding proportion of sodium is 1:3-6, and the sodium hydroxide additive amount is the 2% of unit volume of water.
7. the preparation method of ergot yeast powder as claimed in claim 2, which is characterized in that adjusted in the step S4 using pH
PH value is adjusted to 8-10 by agent, and the pH adjusting agent is that sodium hydroxide and sodium carbonate are prepared and formed, the sodium hydroxide and carbonic acid
The adding proportion of sodium is 1:4, and the sodium hydroxide additive amount is the 2% of unit volume of water.
8. the preparation method of ergot yeast powder as claimed in claim 2, which is characterized in that wet in the step S4 yeast lotion
Yeast number >=85g/L, alcoholic strength 0.1-0.9%(V/V).
9. the preparation method of ergot yeast powder as claimed in claim 2, which is characterized in that I lotion in the step S6
Concentration is 7-10Bx, and the concentration of II lotion is 7-13Bx, and the concentration of III lotion is 10-15Bx.
10. the preparation method of ergot yeast powder as claimed in claim 2, which is characterized in that the step S7 uses twin-roll
Formula drying machine is dried, the drying condition of the double drum type drying machine are as follows: revolving speed is 6-9 r/min, and steam pressure is
0.3-0.4MPa。
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610918070.4A CN106434401B (en) | 2016-10-21 | 2016-10-21 | A kind of preparation method of yeast strain and ergot yeast powder rich in ergosterol |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610918070.4A CN106434401B (en) | 2016-10-21 | 2016-10-21 | A kind of preparation method of yeast strain and ergot yeast powder rich in ergosterol |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN106434401A CN106434401A (en) | 2017-02-22 |
| CN106434401B true CN106434401B (en) | 2019-07-19 |
Family
ID=58176522
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201610918070.4A Active CN106434401B (en) | 2016-10-21 | 2016-10-21 | A kind of preparation method of yeast strain and ergot yeast powder rich in ergosterol |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN106434401B (en) |
Families Citing this family (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106901377B (en) * | 2017-03-13 | 2020-12-18 | 广东省科学院生物工程研究所 | Calcium tablet containing ergosterol and preparation method thereof |
| CN111334460A (en) * | 2020-03-30 | 2020-06-26 | 江苏师范大学 | Method for inhibiting temperature tolerance of aspergillus flavus |
| CN113980825B (en) * | 2020-07-27 | 2023-12-12 | 安琪酵母股份有限公司 | Ergosterol-enriched strain and method for co-producing ergosterol and extract thereof |
| CN111849793B (en) * | 2020-08-10 | 2022-06-24 | 常州悦朋科技有限公司 | 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 |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1683517A (en) * | 2004-04-16 | 2005-10-19 | 中国科学院微生物研究所 | A Strain of yeast engineering fungus for producing ergosterol and its selective breeding method and use |
| CN1844407A (en) * | 2005-04-06 | 2006-10-11 | 北京化工大学 | Method for simultaneous production of ergosterol and glutathione by yeast fermentation |
| CN102911883A (en) * | 2012-09-27 | 2013-02-06 | 华南理工大学 | Saccharomyces ergosterol high-producing strain, and breeding method and application for same |
| CN104293686A (en) * | 2014-09-06 | 2015-01-21 | 浙江大学 | Saccharomyces cerevisiae ZG27 (MATa/alpha) and application thereof |
-
2016
- 2016-10-21 CN CN201610918070.4A patent/CN106434401B/en active Active
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1683517A (en) * | 2004-04-16 | 2005-10-19 | 中国科学院微生物研究所 | A Strain of yeast engineering fungus for producing ergosterol and its selective breeding method and use |
| CN1844407A (en) * | 2005-04-06 | 2006-10-11 | 北京化工大学 | Method for simultaneous production of ergosterol and glutathione by yeast fermentation |
| CN102911883A (en) * | 2012-09-27 | 2013-02-06 | 华南理工大学 | Saccharomyces ergosterol high-producing strain, and breeding method and application for same |
| CN104293686A (en) * | 2014-09-06 | 2015-01-21 | 浙江大学 | Saccharomyces cerevisiae ZG27 (MATa/alpha) and application thereof |
Non-Patent Citations (2)
| Title |
|---|
| Ergosterol production from molasses by genetically modified Saccharomyces cerevisiae;Xiuping He等;《Appl Microbiol Biotechnol》;20070116;第75卷;摘要，第58页右栏最后一段，第59页左栏 * |
| 酵母菌利用糖蜜发酵产麦角甾醇的工艺条件优化;王绍杰等;《生物工程学报》;20131125;第29卷(第11期);摘要，第1677页第1.1-1.2节 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN106434401A (en) | 2017-02-22 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN106434401B (en) | A kind of preparation method of yeast strain and ergot yeast powder rich in ergosterol | |
| CN102160595B (en) | Preparation process of complex microorganism fermented active feed | |
| CN1970718B (en) | Method for preparation of white spirit by fermentation of waste distiller's grains | |
| CN108504621B (en) | Culture medium for paecilomyces hepiali Cs-4 and preparation method thereof | |
| CN101270329B (en) | Method for preparing high concentration fruit vinegar with liquid state submerged fermentation | |
| CN101703214A (en) | Lucid Ganoderma hypra powder or Lucid Ganoderma tea and double fermentation process | |
| CN103484421B (en) | A kind of chlamydosporic method of liquid fermenting scale up test Gliocladium roseum | |
| CN102630490B (en) | Artificial cultivation method for cordyceps militaris mycelium for increasing cordycepin content | |
| CN102550293B (en) | Method for liquid fermentation cultivation of Agaricus bisporus strain | |
| CN104893983B (en) | Liquid state fermentation low citrinin, the preparation method of High color values monascorubin and product | |
| CN106635934B (en) | Thermophilic lactobacillus and corn soaking method by artificially adding thermophilic lactobacillus | |
| CN102533889B (en) | Method for continuously fermenting lysine | |
| CN102249752A (en) | Microbial fertilizer and preparation method and application thereof | |
| CN103194410A (en) | Paenibacillus mucilaginosus and method for producing compound microorganism bacterium agent by utilizing same | |
| CN105925495B (en) | A kind of high activity powdery Pichia yeast powder and preparation method thereof | |
| CN102851330A (en) | Method for preparing citric acid through fermentation | |
| CN102533570B (en) | Aspergillus niger, application of Aspergillus niger and method for preparing citric acid by fermentation | |
| CN102399702B (en) | Aspergillus niger and application thereof as well as citric acid preparation method through fermentation | |
| CN104087632B (en) | A kind of deep layer liquid fermentation produces the method for Phellinus igniarius (L. ex Fr.) Quel. extracellular polysaccharide | |
| CN102443611B (en) | Production method of citric acid | |
| CN109266578A (en) | Escherichia coli ACThr1032 and its application in production of L-threonine by fermentation | |
| CN104694405A (en) | Bacterial strain for generating low-temperature acid alpha-amylase and industrial fermentation enzyme production method of bacterial strain | |
| CN110387343B (en) | Solid culture medium for fermentation of streptomyces noursensis and preparation method and application thereof | |
| CN108522136A (en) | Black collybia albuminosa culture medium and the preparation method and application thereof | |
| CN103497977A (en) | Method for preparing citric acid by fermentation |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant | ||
| PE01 | Entry into force of the registration of the contract for pledge of patent right | ||
| PE01 | Entry into force of the registration of the contract for pledge of patent right |
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 |