CN1141375C - Se-enriched high-biomass yeast and its preparing process - Google Patents
Se-enriched high-biomass yeast and its preparing process Download PDFInfo
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- CN1141375C CN1141375C CNB011006005A CN01100600A CN1141375C CN 1141375 C CN1141375 C CN 1141375C CN B011006005 A CNB011006005 A CN B011006005A CN 01100600 A CN01100600 A CN 01100600A CN 1141375 C CN1141375 C CN 1141375C
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- NBIIXXVUZAFLBC-UHFFFAOYSA-N Phosphoric acid Chemical compound OP(O)(O)=O NBIIXXVUZAFLBC-UHFFFAOYSA-N 0.000 claims description 6
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Abstract
The present invention relates to a selenium-enriched high-biomass yeast and a production method thereof. The selenium-enriched high-biomass yeast is prepared by the method that sieving is carried out so as to obtain yeast strains with high biomass, and yeast strains with strong selenium-resistance capacity and a high cytoselenium content; then, a crossbreeding technique is adopted to breed selenium-enriched yeast strains with high biomass and a high selenium content, and the selenium-enriched high-biomass yeast is obtained by means of fermentation cultivation in a fermentation medium and under optimized fermentation conditions. Cells of every gram of dry selenium-enriched high-biomass yeast contain 500 to 1500 microgrammes of organic selenium. The selenium-enriched high-biomass yeast produced by the production method has the advantages of security, no toxicity, less investment, obvious effect and high effectiveness. With the advantages of strong practicability of the production method and simple operation, the selenium-enriched high-biomass yeast is easily produced in a mass or is generally popularized and applied with the meagre means at hand.
Description
Technical field
The present invention relates to the fermentation industry technical field, refer in particular to a kind of yeast rich in selenium and production method thereof that contains high-biomass.
Background technology
As everyone knows, selenium is the intravital a kind of essential trace element of humans and animals, it is the important component of organism glutathion inside peralcohol, many-sided biological function is arranged, wherein topmost biological function is to remove the too much active oxygen radical that produces in the organism, thereby has and improve organism immunizing power, anti-ageing, anticancer effect.In addition, environment arseniasis is a worldwide problem, and it is harmful to the health of humans and animals that its harm not only shows as recent arsenic, and even more serious is to cause that human body malignization at a specified future date changes---canceration, sudden change and distortion.According to arsenic-selenium medicine principle of toxinicide each other, selenium has and suppresses and anticancer change, anti-mutation and anti-distored biological function in addition, and the yeast that is rich in selenium acts on will bring into play significantly aspect prevention and the treatment endemic arsenic poisoning.In addition, yeast cell also contains 14 kinds of essential trace elements of rich in protein, nucleic acid, VITAMIN and body, healthy and helpful to body.
In recent years, although the existing production that is called " yeast rich in selenium ", these productions exist obvious defects and deficiency:
The first is not carried out any screening and breeding to barms, uses a kind of barms at random, owing to the resistance of different barmses to selenium has than big-difference, thus also very big to the absorption and conversion capability difference of plasma selenium.
It two is plasma selenium concentration, the culture condition of the substratum of production fermentation culture yeast rich in selenium not to be optimized, though therefore the yeast of cultivating is called " yeast rich in selenium ", but this being called,, the selenium content of " yeast rich in selenium " cell was lower, and the biomass of yeast cell is also lower.
Producing the method be called " yeast rich in selenium " cell in the prior art has three kinds substantially, and first kind is to adopt mixed processing method, and a certain amount of selenate or selenite and yeast cell are mixed.Its major defect is: it does not transform into organoselenium with inorganic selenium, the what is called that this selenate or selenite and yeast cell mix " yeast rich in selenium " can make humans and animals very low to its specific absorption, the biological action that produces is relatively poor, is not real yeast rich in selenium.
Second kind is to adopt simple adsorption method, bread yeast on the market or cerevisiae be suspended in the aqueous solution that contains selenate or selenite handle certain hour, make plasma selenium be adsorbed in yeast cell surface, its major defect is: this processing still can not transform into organoselenium with inorganic selenium, the yeast cell of this absorption selenate or selenite and above-mentioned first method do not have the difference of essence, can make the human or animal very low to its specific absorption, the biological action that produces is relatively poor, is not real yeast rich in selenium equally.
The third is a culturing yeast cell in containing the substratum of plasma selenium, but barms is not carried out any screening and breeding, use a kind of barms at random, its major defect is: owing to the resistance of different barmses to selenium has than big-difference, thus widely different to the absorption and conversion capability of plasma selenium.In addition, this technology is not optimized plasma selenium concentration, the culture condition of substratum, therefore adopt in the yeast of this technology cultivation, organic selenium content is lower, and organoselenium 500 micrograms are following, the also low economic benefit of cellular biomass is also very low thereby every gram dry yeast cell contains.
Summary of the invention
The object of the present invention is to provide a kind of yeast rich in selenium and production method thereof that contains high-biomass, utilize modern high-new biotechnology, physio-biochemical characteristics according to different barmses, by screening, obtain high barms and the high barms of cell selenium content of biomass, the yeast rich in selenium bacterial classification that adopts crossbreeding technology to select then to contain high-biomass and high selenium content, reaching the fermentation condition bottom fermentation in the substratum of optimizing cultivates, by suitable production technique, acquisition contains the yeast rich in selenium product of high-biomass, and the dried yeast rich in selenium cell of every gram contains organoselenium and reaches the 500-1500 microgram.
The object of the present invention is achieved like this: a kind of yeast rich in selenium that contains high-biomass, it is characterized in that: it is by screening, make the high barms of biomass and anti-selenium ability reaches the high barms of cell selenium content by force, adopt crossbreeding technology then, select the yeast rich in selenium bacterial classification that contains high-biomass and high selenium content, fermentation culture obtains to contain the yeast rich in selenium of high-biomass and high selenium content in the substratum of optimizing, the dried yeast rich in selenium cell of every gram contains organoselenium and reaches the 500-1500 microgram, classification called after: yeast saccharomyces cerevisiae (yeast rich in selenium) Saccharomyces cerevisiae, preserving number is: CGMCC NO.05 39, preservation day is: January 15 calendar year 2001.
The present invention also provides a kind of production method that contains the yeast rich in selenium of high-biomass, and it is characterized in that: it comprises the steps:
1), preparation contains the yeast rich in selenium bacterial classification of high-biomass: obtain to contain the yeast rich in selenium excellent species of high-biomass and high selenium content by the means of molecular breeding, its method that adopts is:
1. according to the physio-biochemical characteristics of different barmses, in containing the substratum of different selenium concentrations, screen the high barms of biomass respectively and anti-selenium ability reaches the high barms of cell selenium content by force;
2. the high barms of biomass and anti-selenium ability are reached by force that the high barms of cell selenium content is given birth to spore, monoploid separates, the monoploid mutant strain that has different mating types and different genetic markers through mutagenesis screening;
3. the biomass that is separated to is high yeast monoploid mutant strain and anti-selenium ability reach the high yeast monoploid mutant strain of cell selenium content by force and hybridize;
4. at the diploid hybrid strain excellent of selecting to screen on the substratum high-biomass and high selenium content, i.e. yeast rich in selenium bacterial classification;
2), fermentation culture contains the yeast rich in selenium of high-biomass and high selenium content:
(1), fermentative medium formula:
The wort of 8%～20% total reducing sugar amount of selection includes the selenate of 200～400 mcg/ml or the selenite of 400～600 mcg/ml, and the pH of substratum is 4.5-7.0;
(2), fermentation culture conditions: the culture condition that fermentation culture contains the yeast rich in selenium of high-biomass and high selenium content is: the inoculum size of seed liquor is 5-30%, under 25-32 ℃ condition, stir culture 16-30 hour;
3), produce the yeast rich in selenium that contains high-biomass and high selenium content: its production technique comprises: slant strains → liquid spawn → level liquid seed culture → secondary liquid seeds is cultivated → three grades of liquid seeds cultivation → ferment tank cultivation → collection yeast cell and drying → pulverizing and packing, and its each production technique is as follows:
(1), slant strains: the yeast rich in selenium bacterial classification is inoculated in that to contain total sugar content be that 8-20%, selenate concentration are 200～400 mcg/ml or use selenite, its concentration is on the wort solid inclined-plane of 400～600 mcg/ml,, put into 4 ℃ of refrigerators and preserve after 24-72 hour 25-32 ℃ of cultivation;
(2), liquid spawn: after the yeast rich in selenium bacterial classification activation of preserving, connecing a garland cells is that 8-20%, selenate concentration are 200～400 mcg/ml or selenite in 150 milliliters of total sugar contents are housed, its concentration is in the triangular flask of malt extract medium of 400～600 mcg/ml, in 25-32 ℃ of stir culture 14-20 hour, be liquid spawn;
(3), level liquid seed culture: it is that 8-20%, selenate concentration are 200～400 mcg/ml or selenite that liquid spawn is equipped with 150 milliliters of total sugar contents by 5～20% 30 of inoculum size accesses, its concentration is in the triangular flask of malt extract medium of 400～600 mcg/ml, at 25-32 ℃ of stir culture 14-20 hour, be the level liquid inoculum;
(4), the secondary liquid seeds is cultivated: it is that 8-20%, selenate concentration are 200～400 mcg/ml or use selenite that the first order seed nutrient solution is equipped with 45 liters of total sugar contents by 5～20% inoculum size access, its concentration is the Xiao Kashi fermentor tank of the malt extract medium of 400～600 mcg/ml, at 25-32 ℃ of stir culture 14-20 hour, be secondary liquid seeds culture;
(5), three grades of liquid seeds are cultivated: it is that 8-20%, selenate concentration are 200～600 mcg/ml or use selenite that the secondary seed nutrient solution is equipped with 450 liters of total sugar contents by 10～30% inoculum size access, its concentration is the Da Kashi fermentor tank of the malt extract medium of 400～600 mcg/ml, at 25-32 ℃ of stir culture 14-20 hour, be three grades of liquid seeds cultures;
(6), ferment tank is cultivated: it is that 8-20%, selenate concentration are 200～400 mcg/ml or use selenite that three grades of seed culture fluids are equipped with 2250 liters of total sugar contents by 20～40% inoculum size access, its concentration was the fermentor tank of the malt extract medium of 400～600 mcg/ml, at 25-32 ℃ of stir culture 14-24 hour;
(7), collect yeast cell and drying: adopt sheet frame squeezing or centrifugal collection yeast rich in selenium cell.Air-dry under 45～85 ℃ of conditions, moisture is less than 10%;
(8), pulverize: with pulverizer with air-dry yeast rich in selenium cell pulverization;
(9), packing: use air-locked packaging bags, be the yeast rich in selenium of the selenium-enriched content of high-biomass, the dried yeast rich in selenium cell of every gram contains organoselenium and reaches the 500-1500 microgram, classification called after: yeast saccharomyces cerevisiae (yeast rich in selenium) Saccharomyces cerevisiae, preserving number is: CGMCC NO.0539, preservation day is: January 15 calendar year 2001.
This fermentative medium formula and fermentation culture conditions are: the total reducing sugar amount is 12% hydrolysis sugar, 0.8% corn hydrolyzed solution, 0.5% ammonium sulfate, the selenite of the selenate of 300 mcg/ml or 500 mcg/ml, the pH of substratum is 5.0, inoculum size is 15%, under 28-30 ℃ condition, and stir culture 30 hours.This fermentative medium formula and fermentation culture conditions are: the molasses of total reducing sugar amount 15%, 1.0% corn hydrolyzed solution, 0.5% ammonium sulfate, 0.1% phosphoric acid, the selenite of the selenate of 350 mcg/ml or 450 mcg/ml, the pH of substratum was 6.0, and inoculum size is 20%, 26-28 ℃ of stir culture 24 hours.This production technique that contains the yeast rich in selenium of high-biomass and high selenium content comprises the cultivation of level Four seed liquor.This production technique that contains the yeast rich in selenium of high-biomass and high selenium content comprises the variation of doing suitable increase or minimizing technology with actual production conditions.
Major advantage of the present invention is as follows:
1, the yeast rich in selenium bacterial classification of the present invention's use, not only has high-biomass, and the organic selenium content of cell is higher, and its organic selenium content contains the 500-1500 microgram up to every gram stem cell, and the organic selenium content of the product that prior art is produced is that every gram stem cell contains below organoselenium 500 micrograms.
2, the fermentation substrate that uses of method of the present invention is common wort or makes fermentation substrate with amylum hydrolysate of the sugar or molasses, only needs to regulate its total sugar content, adds selenate or selenite, and method is simple, and is with low cost.
3, method of the present invention does not have particular requirement to fermentation equipment and condition, and the equipment and the condition of general fermentation plant all can be produced, and the production investment is less.
4, the yeast rich in selenium product safety of method production of the present invention is nontoxic, less investment, instant effect, high efficiency.
5, method of the present invention is practical, and is easy and simple to handle, is easy to generally promote.
6, press the biomass height of the yeast rich in selenium product of method production of the present invention, the organic selenium content height of yeast cell, quality is good, and the dried yeast rich in selenium cell of every gram contains organoselenium and reaches the 500-1500 microgram.
Further specify below in conjunction with preferred embodiment and accompanying drawing.
Description of drawings
Fig. 1 is a technological process of production synoptic diagram of the present invention.
Embodiment
Embodiment 1
Consult Fig. 1, the production method that contains the yeast rich in selenium of high-biomass of the present invention comprises the steps:
1), preparation contains the yeast rich in selenium bacterial classification of high-biomass: because occurring in nature does not have biomass and the simultaneously all high barms of cell selenium content, can only could obtain to contain the yeast rich in selenium excellent species of high-biomass and high selenium content by the means of molecular breeding.The method that the present invention adopts is:
1. according to the physio-biochemical characteristics of different barmses, in containing the substratum of different selenium concentrations, screen the high barms of biomass respectively and anti-selenium ability reaches the high barms of cell selenium content by force.
2. the high barms of biomass and anti-selenium ability are reached by force that the high barms of cell selenium content is given birth to spore, monoploid separates, the monoploid mutant strain that has different mating types and different genetic markers through mutagenesis screening.
3. high yeast monoploid mutant strain and the anti-selenium ability of biomass that is separated to reached the high yeast monoploid mutant strain cross-breeding of cell selenium content by force.
4. at the diploid hybrid strain excellent of selecting to screen on the substratum high-biomass and high selenium content, i.e. yeast rich in selenium bacterial classification.
2), cultivate the yeast rich in selenium that contains high-biomass and high selenium content in the fermentation culture conditions top fermentation of fermention medium and optimization:
(1), fermentative medium formula:
The wort of 8%～20% total reducing sugar amount of selection includes the selenate of 200～400 mcg/ml or the selenite of 400～600 mcg/ml, and the pH of substratum is 4.5-7.0.
(2), the fermentation culture conditions of You Huaing:
The culture condition that fermentative production contains the yeast rich in selenium of high-biomass and high selenium content is: the inoculum size of yeast rich in selenium bacterial classification is 5-30%, under 25-32 ℃ of condition, and stir culture 16-30 hour;
3), produce the yeast rich in selenium product that contains high-biomass and high selenium content: its production technique comprises:
Slant strains → liquid spawn → level liquid seed culture → secondary liquid seeds is cultivated the yeast rich in selenium of the selenium-enriched content of → three grades of liquid seeds cultivation → ferment tank cultivation → collection yeast cell and drying → pulverizing and packing high-biomass.Further specify as follows to each production technique below:
(1), slant strains: the yeast rich in selenium bacterial classification is inoculated in that to contain total sugar content be that 8-20%, selenate concentration are that 200～400 mcg/ml (or are used selenite, its concentration is 400～600 mcg/ml) wort solid inclined-plane on, under 25-32 ℃ condition, cultivate after 24-72 hour, put into 4 ℃ of refrigerators and preserve.
(2), liquid spawn: after the yeast rich in selenium bacterial classification activation of preserving, connecing a garland cells is that 8-20%, selenate concentration are that 200～400 mcg/ml (or are used selenite in 150 milliliters of total sugar contents are housed, its concentration is 400～600 mcg/ml) the triangular flask of malt extract medium in, under 25-32 ℃ condition, stir culture 14-20 hour, be liquid spawn.
(3), level liquid seed culture: it is that 8-20%, selenate concentration are that 400～600 mcg/ml (or are used selenite that liquid spawn is equipped with 150 milliliters of total sugar contents by 5～20% inoculum size access, its concentration is 400～600 mcg/ml) the triangular flask of malt extract medium in, under 25-32 ℃ condition, stir culture 14-20 hour, be the level liquid inoculum.
(4), the secondary liquid seeds is cultivated: it is that 8-20%, selenate concentration are that 200～400 mcg/ml (or are used selenite that the first order seed nutrient solution is equipped with 45 liters of total sugar contents by 5～20% inoculum size access, its concentration is 400～600 mcg/ml) the Xiao Kashi fermentor tank of malt extract medium, under 25-32 ℃ condition, stir culture 14-20 hour, be secondary liquid seeds culture.
(5), three grades of liquid seeds are cultivated: it is that 8-20%, selenate concentration are that 200～400 mcg/ml (or are used selenite that the secondary seed nutrient solution is equipped with 450 liters of total sugar contents by 10～30% inoculum size access, its concentration is 400～600 mcg/ml) the Da Kashi fermentor tank of malt extract medium, under 25-32 ℃ condition, stir culture 14-20 hour, be three grades of liquid seeds cultures.
(6), ferment tank is cultivated: it is that 8-20%, selenate concentration are that 200～400 mcg/ml (or are used selenite that three grades of seed culture fluids are equipped with 2250 liters of total sugar contents by 20～40% inoculum size access, its concentration is 400～600 mcg/ml) the fermentor tank of malt extract medium, under 25-32 ℃ condition, stir culture 14-24 hour.
(7), collect yeast cell and drying: adopt sheet frame squeezing or centrifugal collection yeast rich in selenium cell.Air-dry under 45～85 ℃ of conditions, moisture is less than 10%.
(8), pulverize: with pulverizer with air-dry yeast rich in selenium cell pulverization.
(9), packing: use air-locked packaging bags, be the yeast rich in selenium of the selenium-enriched content of high-biomass, the dried yeast rich in selenium cell of every gram contains organoselenium and reaches the 500-1500 microgram, classification called after: yeast saccharomyces cerevisiae (yeast rich in selenium) Saccharomyces cerevisiae, preserving number is: CGMCC NO.0539, preservation day is: January 15 calendar year 2001.
Production method of the present invention can be according to the fermentation equipment and the condition of producer, and the adjustment that above technology is done suitably to increase or reduce changes.
Embodiment 2
Production method of the present invention can be selected different fermentative medium formulas, culture condition and contain high-biomass and the suitable adjustment change that increases or reduce of the production technique of the yeast rich in selenium of high selenium content do according to the different material of different areas.
Present embodiment is that selection is a substratum with the amylum hydrolysate of the sugar, its fermentative medium formula and culture condition are: the total reducing sugar amount is 12% amylum hydrolysate of the sugar, 0.8% corn hydrolyzed solution, 0.5% ammonium sulfate, the selenite of the selenate of 350 mcg/ml or 550 mcg/ml, the pH of substratum are 5.0, and the inoculum size of seed liquor is 15%, under 28-30 ℃ condition, stir culture 30 hours.
Its production technique comprises: slant strains → liquid spawn → level liquid seed culture → secondary liquid seeds is cultivated the yeast rich in selenium of → three grades of liquid seeds cultivation → level Four liquid seeds cultivation → ferment tank cultivation → collection yeast cell and drying → pulverizing and the high selenium content of packing high-biomass.The dried yeast rich in selenium cell of every gram contains organoselenium and reaches 1000 micrograms.
The production technique of present embodiment increases the level Four liquid seeds cultivates, and its condition is identical substantially with three grades of liquid seeds cultivations, and other processing condition are substantially the same manner as Example 1, so do not repeat.
Embodiment 3
Present embodiment is that selection is a substratum with sucrose molasses or beet sirup, its fermentative medium formula and culture condition are: the total reducing sugar amount is 15% sucrose molasses or beet sirup, 1.0% corn hydrolyzed solution, 0.8% ammonium sulfate, 0.1% phosphoric acid, the selenite of the selenate of 300 mcg/ml or 500 mcg/ml, the pH of substratum is 6.0, and the inoculum size of seed liquor is 20%, 26-28 ℃ of stir culture 28 hours.
Its production technique comprises: liquid spawn → level liquid seed culture → secondary liquid seeds is cultivated the yeast rich in selenium of → three grades of liquid seeds cultivation → level Four liquid seeds cultivation → Pyatyi liquid seeds cultivation → ferment tank cultivation → collection yeast cell and drying → pulverizing and the high selenium content of packing high-biomass.The dried yeast rich in selenium cell of every gram contains organoselenium and reaches 1200 micrograms.
The production technique of present embodiment increases the Pyatyi liquid seeds cultivates, and its condition is identical substantially with three grades of liquid seeds cultivations, and other processing condition are substantially the same manner as Example 2, so do not repeat.
In sum, method of the present invention is to make full use of microbial technique, physio-biochemical characteristics according to different barmses, by screening, obtain high barms and the anti-selenium ability of biomass and reach the high barms of cell selenium content by force, the yeast rich in selenium bacterial classification that adopts crossbreeding technology to select then to contain high-biomass and high selenium content, fermentation culture in the fermention medium of optimizing, acquisition contains the yeast rich in selenium of high-biomass and high selenium content, and it is low that this yeast rich in selenium that contains high-biomass and high selenium content has a cost, raw material is extensive, easy and simple to handle, fermentation period is short, characteristics such as the biomass height of yeast cell and organic selenium content height.Adopt production yeast rich in selenium method of the present invention, be fit to large-scale production and the production of short run on the basis of the existing conditions no matter how simple and crude, have actual application prospect widely, and have important social benefit and huge economic benefit.
Claims (6)
1, a kind of yeast rich in selenium that contains high-biomass, it is characterized in that: it is by screening, obtain high barms and the anti-selenium ability of biomass and reach the high barms of cell selenium content by force, adopt crossbreeding technology then, select the yeast rich in selenium bacterial classification that contains high-biomass and high selenium content, under the fermentation condition of fermention medium and optimization, fermentation culture production contains the yeast rich in selenium of high-biomass, and the dried yeast rich in selenium cell of every gram contains organoselenium and reaches the 500-1500 microgram; The classification called after: yeast saccharomyces cerevisiae (Saccharomycescerevisiae), preserving number is: CGMCC NO.0539.
2, a kind of production method that contains the yeast rich in selenium of high-biomass, it is characterized in that: it comprises the steps:
1), preparation contains the yeast rich in selenium bacterial classification of high-biomass: obtain to contain the yeast rich in selenium excellent species of high-biomass and high selenium content by the means of molecular breeding, its method that adopts is:
1. according to the physio-biochemical characteristics of different barmses, in containing the substratum of different selenium concentrations, screen the high barms of biomass respectively and anti-selenium ability reaches the high barms of cell selenium content by force;
2. the high barms of biomass and anti-selenium ability are reached by force that the high barms of cell selenium content is given birth to spore, monoploid separates, the monoploid mutant strain that has different mating types and different genetic markers through mutagenesis screening;
3. the biomass that is separated to is high yeast monoploid mutant strain and anti-selenium ability reach the high yeast monoploid mutant strain of cell selenium content by force and hybridize;
4. at the diploid hybrid strain excellent of selecting to screen on the substratum high-biomass and high selenium content, i.e. yeast rich in selenium bacterial classification;
2), fermentation culture contains the yeast rich in selenium of high-biomass and high selenium content:
(1), fermentative medium formula: select the wort of 8%～20% total reducing sugar amount, contain the selenate of 200～400 mcg/ml or the selenite of 400～600 mcg/ml, the pH of substratum is 4.5-7.0;
(2), fermentation culture conditions: the culture condition that fermentative production contains the yeast rich in selenium of high-biomass and high selenium content is: the seed liquor inoculum size is 5-30%, under 25-32 ℃ condition, stir culture 16-30 hour;
3), produce the yeast rich in selenium that contains high-biomass and high selenium content: its production technique in turn includes the following steps: slant strains, liquid spawn, level liquid seed culture, secondary liquid seeds are cultivated, three grades of liquid seeds cultivations, ferment tanks are cultivated, collected yeast cell and drying, pulverizing and packing, and its each production technique is as follows:
(1), slant strains: the yeast rich in selenium bacterial classification is inoculated in that to contain total sugar content be that 8-20%, selenate concentration are 200～400 mcg/ml or use selenite, its concentration is on the wort solid inclined-plane of 400～600 mcg/ml,, put into 4 ℃ of refrigerators and preserve after 24-72 hour 25-32 ℃ of cultivation;
(2), liquid spawn: after the yeast rich in selenium bacterial classification activation of preserving, connecing a garland cells is that 8-20%, selenate concentration are 200～400 mcg/ml or use selenite in 150 milliliters of total sugar contents are housed, its concentration was in the triangular flask of malt extract medium of 400～800 mcg/ml, was liquid spawn in 25-32 ℃ of stir culture 14-20 hour;
(3), level liquid seed culture: it is that 8-20%, selenate concentration are 200～400 mcg/ml or use selenite that liquid spawn is equipped with 150 milliliters of total sugar contents by 5～20% 30 of inoculum size accesses, its concentration is in the triangular flask of malt extract medium of 400～600 mcg/ml, at 25-32 ℃ of stir culture 14-20 hour, be the level liquid inoculum;
(4), the secondary liquid seeds is cultivated: it is that 8-20%, selenate concentration are 200～400 mcg/ml or use selenite that the first order seed nutrient solution is equipped with 45 liters of total sugar contents by 5～20% inoculum size access, its concentration is the Xiao Kashi fermentor tank of the malt extract medium of 400～600 mcg/ml, at 25-32 ℃ of stir culture 14-20 hour, be secondary liquid seeds culture;
(5), three grades of liquid seeds are cultivated: it is that 8-20%, selenate concentration are 200～400 mcg/ml or use selenite that the secondary seed nutrient solution is equipped with 450 liters of total sugar contents by 10～30% inoculum size access, its concentration is the Da Kashi fermentor tank of the malt extract medium of 400～600 mcg/ml, at 25-32 ℃ of stir culture 14-20 hour, be three grades of liquid seeds cultures;
(6), ferment tank is cultivated: it is that 8-20%, selenate concentration are 200～400 mcg/ml or use selenite that three grades of seed culture fluids are equipped with 2250 liters of total sugar contents by 20～40% inoculum size access, its concentration was the fermentor tank of the malt extract medium of 400～600 mcg/ml, at 25-32 ℃ of stir culture 14-24 hour;
(7), collect yeast cell and drying: adopt sheet frame squeezing or centrifugal collection yeast rich in selenium cell.Air-dry under 45～85 ℃ of conditions, moisture is less than 10%;
(8), pulverize: with pulverizer with air-dry yeast rich in selenium cell pulverization;
(9), packing: use air-locked packaging bags, be the yeast rich in selenium that contains high-biomass, the dried yeast rich in selenium cell of every gram contains organoselenium and reaches the 500-1500 microgram, and the classification called after: yeast saccharomyces cerevisiae (Saccharomyces cerevisiae), preserving number is: CGMCC NO.0539.
3, production method as claimed in claim 2, it is characterized in that: this fermentative medium formula and culture condition are: the total reducing sugar amount is 12% amylum hydrolysate of the sugar, 0.8% corn hydrolyzed solution, 0.5% ammonium sulfate, the selenite of the selenate of 350 mcg/ml or 550 mcg/ml, the pH of substratum are 5.0, and the inoculum size of seed liquor is 15%, under 28-30 ℃ condition, stir culture 30 hours.
4, production method as claimed in claim 2, it is characterized in that: the prescription and the culture condition of this fermention medium are: the total reducing sugar amount is 15% molasses, 1.0% corn hydrolyzed solution, 0.5% ammonium sulfate, 0.1% phosphoric acid, the selenite of the selenate of 300 mcg/ml or 500 mcg/ml, the pH of substratum was 6.0, and the inoculum size of seed liquor is 20%, 26-28 ℃ of stir culture 24 hours.
5, production method as claimed in claim 2 is characterized in that: this production technique that contains the yeast rich in selenium of high-biomass comprises that increasing the level Four liquid seeds cultivates.
6, production method as claimed in claim 5 is characterized in that: this production technique that contains the yeast rich in selenium of high-biomass comprises that increasing the Pyatyi liquid seeds cultivates.
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- 2001-01-17 CN CNB011006005A patent/CN1141375C/en not_active Expired - Lifetime
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