CN101818183B - Method for improving trehalose content of yeast cell - Google Patents
Method for improving trehalose content of yeast cell Download PDFInfo
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- CN101818183B CN101818183B CN 201010146889 CN201010146889A CN101818183B CN 101818183 B CN101818183 B CN 101818183B CN 201010146889 CN201010146889 CN 201010146889 CN 201010146889 A CN201010146889 A CN 201010146889A CN 101818183 B CN101818183 B CN 101818183B
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Abstract
The invention relates to a method for improving trehalose content of a yeast cell, which comprises the following steps of: centrifuging yeast fermentation culture solution to obtain yeast milk with cell concentration of 13 to 17 percent, adding aqueous solution of maltose, sodium chloride and calcium chloride into the yeast milk, culturing the mixture for 2 to 3 hours under the condition that the temperature is 35 DEG C, the pH value is 5.2 and the ventilation rate of 1.0 to 1.2vvm, centrifuging the cultured mixture, and collecting yeast paste, wherein the trehalose content of the yeast reaches over 16 percent. The method can improve the trehalose in the yeast cell to over 16 percent from 8 percent in shorter time; and compared with the prior art, the method has the advantages of shortening the fermentation time and saving fermentation equipment and power consumption.
Description
Technical field
The present invention relates to a kind of method that improves the yeast cell trehalose, particularly a kind of method that improves the rusk trehalose content of yeast cell, belong to fermentation engineering and microbial technology field.
Background technology
Active dry yeast is to be made through the granulating and drying dehydration by fresh yeast or title yeast cake, and the activity of active dry yeast and period of storage are to estimate the important indicator of yeast quality.And the content of these indexs and yeast cell intracellular trehalose is closely related.
Trehalose (Trehalose) has another name called lentinan, ALPHA-D-glycopyranosyl-ALPHA-D-glycopyranoside, and its molecular formula is C
12H
22O
112H
2O, and chemical name α-D-glucopyranosyl α-D-glucopyranoside (α-D-glycopyranosyl-α-D-glycopyranoside), be the irreducibility disaccharide that is formed by the condensation of hemiacetal hydroxyl by two glucose molecules.Trehalose extensively is present in lower plant, algae, bacterium, filamentous fungus, yeast, insect.
some species is severe environment to external world, as arid, high temperature, dehydration, freezing, high osmotic pressure and toxicant etc., the trehalose that exists in the degeneration-resistant tolerance that shows and these species bodies is closely related, has unique biological characteristics that is different from other carbohydrate, can protect biological cell protein under above-mentioned environment, fat, carbohydrate, the components such as nucleic acid are without prejudice, can protect DNA to prevent the damage that radioactive rays cause, ectogenic trehalose also can have good nonspecific provide protection to organism and biomacromolecule, thereby this makes it that many purposes be arranged, as make the cell frostproofer in medicine and microbiology, the stablizer of diagnostic reagent and biological product, the effective ingredient of makeup, food preservatives, cultivate drought-resistant crops etc. on agricultural.
trehalose is as a kind of stress metabolite of yeast, can give the microorganism opposing nutritive deficiencies such as yeast, high temperature, low temperature, dry, high osmotic pressure, the ability of the severe environment such as toxic substance, it has the microbial film of protection organism under harsh and unforgiving environments, liposome, protein, the special efficacy of the structure and functions such as nucleic acid, it makes and keeps moistening in cell, prevent the loss of various nutrients in cell, thereby allow these biologies be in the anhydrobiosis state, as when producing active dry yeast, the dry yeast of content of trehalose below 11% (in solid substance), its active preservation period is short, be difficult for storing, content of trehalose during to 14% left and right, can extend 4-6 month 12% between its preservation period, and when content of trehalose surpassed 16%, its preservation period can reach more than 24 months.
Active dry yeast is to be made through the granulating and drying dehydration by fresh yeast (or claiming yeast cake), and the activity of dry yeast represents with fermenting power usually.Generally, the 3.5g fresh yeast could produce the 1g active dry yeast, but with regard to active (fermenting power), the 1g active dry yeast only is equivalent to the 2-3g fresh yeast: after rehydration, check is surveyed viable cell and is generally 80-90%.Fresh yeast is made active dry yeast, and after the storage and the reconstitution process before use of certain hour, certain loss of activity is arranged.
The content of the trehalose in yeast has a great impact activity and the storage of active dry yeast.Determined to a certain extent activity and the long keeping ability of high activity dried yeast, the amount of trehalose increases, and is conducive to like this stability of cell in dry and storage process.Higher content of trehalose is conducive to extend the prolongation of the storage period of active dry yeast.And the height of the content of phosphorus has certain impact for the activity of Trehalose Content and cell nitrogen content level and enzyme.The content of the trehalose of general active dry yeast reaches 14% left and right of dry-matter, and therefore, culturing yeast technique must be considered the accumulation degree of trehalose.The yeast trehalose synthesis need to be through two stages, and the one, in the amount reproduction stage of yeast cell, this stage yeast body intracellular trehalose content is lower, but the thalline of capacity can be provided; The 2nd, the accumulation stage of trehalose is mainly by controlling ambient conditions, it to be accumulated.
Summary of the invention
The present invention is directed to the deficiencies in the prior art, propose a kind of method that improves trehalose content of yeast cell, can realize in the short period of time that yeast cell intracellular trehalose content reaches more than 16%.
The term explanation:
The vvm of unit of ventilation refers to the volume ratio of the amount of air capacity that per minute passes into and fermented liquid.
Cell concn: refer in fermented liquid or the mass concentration of yeast saccharomyces cerevisiae in yeast-lactic.
" bread yeast " mentioned in the present invention is the product trade(brand)name of being produced by the yeast saccharomyces cerevisiae bacterial classification.
Technical scheme of the present invention is as follows:
a kind of method that improves trehalose content of yeast cell, the yeast-lactic that the yeast fermentation medium centrifugal is separated the cell concn that obtains 13-17%, add maltose in yeast-lactic, the aqueous solution of sodium-chlor and calcium chloride, the mass volume ratio of yeast-lactic and water is 1: 1, maltose, the consumption of sodium-chlor and calcium chloride accounts for yeast-lactic total mass per-cent and is respectively 2.4-3.0% maltose, 3.2-4.0% sodium-chlor and 1.0% calcium chloride, at 35 ℃, pH5.2, 1.0-1.2vvm cultivated 2-3 hour under ventilation condition, centrifugal collection yeast slurry, measure content of trehalose 16-17% in yeast, with the yeast cell dry weight basis.Described mass volume ratio, unit are g/mL or kg/L.Described yeast is yeast saccharomyces cerevisiae.
In method of the present invention, the accumulation volume that can improve trehalose is cultivated in the interpolation of maltose, and this is because yeast cell contains the maltose isomerase and maltose can be converted into trehalose at short notice.
In the method for the invention described above, take the yeast fermentation nutrient solution as starting raw material, can be by prior art about the preparation of yeast fermentation nutrient solution.The invention provides the preparation method of following yeast fermentation nutrient solution, as of the present invention one of preferred:
1. substratum
(1) seed culture medium: glucose 20g, KH
2PO
41g, (NH
4)
2SO
45g, MgSO
47H
2O 0.2g, ZnSO
47H
2O 0.05g, pH5.4, distilled water 1000mL.
(2) fermention medium
1. basic medium ferments
Corn steep liquor (containing solid substance 28wt%) 20%, glucose 1%, nutrient salt solution 5%, water 74% is volume ratio.Wherein, nutrient salt solution: KH
2PO
413.0g, (NH
4)
2SO
494.16g, MgSO
47H
2O 1.4g, ZnSO
47H
2O0.28g adds water and is made into 1000mL.
2. fermenting process stream adds component
Sugar soln: glucose is made into the solution of 250g/L.
Nutrient salt solution: KH
2PO
413.0g, (NH
4)
2SO
494.16g, MgSO
47H
2O 1.4g, ZnSO
47H
2O0.28g adds water and is made into 1000mL.
The Na of alkali lye: 100g/L
2CO
3Solution.
2. seed culture
Barms is available from Institute of Microorganism, Academia Sinica, title: yeast saccharomyces cerevisiae (Saccharomyces cerevisiae), and code name: AS 2.146, AS 2.502, AS 2.503 or AS 2.504.With the yeast slant strains, in the 250mL triangular flask of access dress 20mL seed culture medium, after 30 ℃ of 200r/min shake-flask culture 24h, change the 500mL triangular flask that the 80mL seed culture medium is housed over to, 30 ℃ of 200r/min shake-flask culture 16h obtain seed culture fluid, and are standby.
3. fermentation culture
Carry out real tank sterilization with being equipped with in the 50L fermentor tank of 25L fermentation basic medium, sterilising conditions is 120 ℃, 30min.When then being cooled to 32 ℃, access seed culture fluid 3L, regulate pH5.2, control 30 ℃ of culture temperature, stream sugared soln and nutrient salt solution, the control total fermentation time is 18-21h, nutrient salt solution 3-5 before fermentation ends added in individual hour, the stream dosage of sugar soln is according to the yeast growth situation, makes that in fermentation system, glucose content is controlled at below 0.5% always.In fermenting process, control pH5.2, mixing speed is controlled at 200r/min.Other processing condition are preferably as follows:
Culture temperature is controlled: 30 ℃ of fermentation time 1-8h culture temperature, and 32 ℃ of 9-15h culture temperature, 16h is to 35 ℃ of fermentation ends culture temperature.
Air flow: fermentation (front, mid-term) time 1-15h control the sterile air volume with fermentating liquid volume than per minute at 2.5vvm, in the Trehalose Content stage of fermentation (later stage) time 16h to fermentation ends, adjust valve, air flow is dropped to 1.2vvm.
Preferably, in above method fermentation culture step, control the sugar soln dosage and make glucose content 0.30-0.50% volume ratio in fermentation system; PH adjusting agent is 10wt%Na
2CO
3Solution.
The yeast fermentation nutrient solution that above method makes, cell concn 4.5-5.5%, wherein content of trehalose is 8-10% after measured.Further be used as the raw material of the inventive method.
Content of trehalose measuring method in yeast
In yeast cell, contain mannosans and insoluble dextran, can react with anthrone reagent.Therefore, selecting suitable extraction agent to control the interference that these materials measure trehalose is the key that in yeast, trehalose is measured.Yeast extracts with the cold trichoroacetic acid(TCA) of 0.5mol/L, only has trehalose to exist in gained solution.Trichoroacetic acid(TCA) extracting solution and 0.2% sulfuric acid anthrone solution are mixed boiling water bath reaction 2min with the ratio of 1: 4, and in the light absorption value of 590nm place measured reaction liquid.The content that calculates trehalose in extracting solution is the trehalose amount in yeast.
Adopt paper chromatography to separate elution method: during with paper chromatography quantitative analysis trehalose, two samples putting on the filter paper bar are same sample, and the point sample amount is controlled at 40～100 μ g, and one of them point is reference point, and another is the quantitative measurment point.With the filter paper bar after point sample at the developping agent propyl carbinol: acetone: water: acetic acid=10: 3: 6: launch in 2 (v/v), air-dry, and it is cut off from the centre.Half spraying colour developing with reference point, to determine the accurate location of trehalose spot, fragment is cut and be cut into to the part that on second half filter paper bar, spot is corresponding therewith, put into elutriant 0.1mol/LHCl room temperature wash-out 16h, supernatant liquor wherein content of trehalose of sulfuric acid-anthrone colorimetric method for determining after wash-out.
Method of the present invention is as follows by technical characterstic and the excellent results that yeast culture accumulates trehalose:
1. temperature: 30 ℃ of earlier fermentation (1-8h) culture temperature, mid-term, (9-15h) culture temperature was 32 ℃, 35 ℃ of later stage (16h is to fermentation ends) culture temperature.
2. pH: in the culturing process of yeast, stream adds 10%Na
2CO
3Solution is regulated pH5.2.
3. dissolved oxygen: in the culturing process of yeast, in the situation that air flow is constant, improve the oxygen transfer rate that stir speed (S.S.) can effectively improve fermentor tank, thereby guarantee that Growth of Cells and the required dissolved oxygen of metabolism supply with.In the inventive method, mixing speed is controlled at 200r/min, and air flow 1-15h is controlled at 2.5vvm, is the Trehalose Content stage at fermentation time 16h to fermentation ends, and air flow is dropped to 1.2vvm.
4. after yeast culture obtains a large amount of cells, obtain the yeast-lactic of the concentration of 13-17% through centrifugation, add maltose in yeast-lactic, and add sodium-chlor, calcium chloride, cultivated 2-3 hour under 35 ℃ of ventilation conditions, to realize the high density accumulation of trehalose in yeast.The present invention can within a short period of time, and the trehalose in yeast cell is brought up to more than 16% from 8%.Existing general technique has been dwindled fermentation time.
Because raw material of the present invention is yeast-lactic, barm cell concentration is 13-17%, and the concentration 4.5-5.5% of yeast in yeast fermentation broth, thereby can save fermentation equipment and power consumption.
Description of drawings
Fig. 1 the inventive method embodiment 1 yeast-lactic is in the situation that salt adding, add the maltose Trehalose Content;
Fig. 2 content of trehalose in sulfuric acid-anthrone colorimetric method for determining embodiment 1 yeast slurry: during with paper chromatography quantitative analysis trehalose, two samples putting on the filter paper bar are same sample, the point sample amount is controlled at 40～100 μ g, and one of them point is reference point, and another is the quantitative measurment point.
Embodiment
Below in conjunction with embodiment, technical solution of the present invention is further elaborated, but institute of the present invention protection domain is not limited only to this.In embodiment, bacterial classification used and substratum are:
Yeast saccharomyces cerevisiae (Saccharomyces cerevisiae) bacterial classification is available from Institute of Microorganism, Academia Sinica, and DSMZ of Institute of Microorganism, Academia Sinica bacterial classification catalogue code name: AS 2.146; AS 2.502; AS 2.503; AS 2.504.
Seed culture medium: glucose 20g, KH
2PO
41g, (NH
4)
2SO
45g, MgSO
47H
2O 0.2g, ZnSO
47H
2O 0.05g, pH5.4, distilled water 1000mL.
Fermention medium: comprise that fermentation basic medium and fermenting process stream add component, as previously mentioned.
Barms: yeast saccharomyces cerevisiae (Saccharomyces cerevisiae) AS 2.146.With the yeast slant strains, in the 250mL triangular flask of access dress 20mL seed culture medium, after 30 ℃ of 200r/min shake-flask culture 24h, change the 500mL triangular flask that the 80mL seed culture medium is housed over to, 30 ℃ of 200r/min shake-flask culture 16h obtain seed culture fluid, and are standby.
Above-mentioned seed culture fluid 3L is accessed in the 25L fermentation basic medium of having sterilized (fermentor tank volume 50L), and culture condition is as follows.1. temperature: at the 1-8h of fermentation in the time, 30 ℃ of culture temperature, within the time of fermentation 9-15h, 32 ℃ of culture temperature, at fermentation 16-20h in the time, 35 ℃ of culture temperature.2. pH: in the culturing process of yeast, stream adds 10%Na
2CO
3Solution is regulated pH5.2.3. dissolved oxygen: mixing speed is controlled at 200r/min, and air flow 1-16h is controlled at 2.5vvm, in the time, by valve, air flow is dropped to 1.2vvm at fermentation 16-20h.Stream sugared soln and nutrient salt solution, nutrient salt solution stream adds total amount 2.5L, 16h adds at fermentation time, sugar soln carries out stream according to the growing state of ethanol feedback and yeast and adds, make that in fermented liquid, glucose content is controlled at 0.40-0.45% (volume ratio) always, the 20h fermentation ends, yeast cell 4.78% (dry weight) in the gained fermented liquid, content of trehalose 8.2%.
With above-mentioned fermented liquid, with large capacity whizzer, at 3500r/min, under the condition of time 10min, centrifugal collection yeast-lactic.Abandon the centrifuge tube supernatant liquor, take out yeast-lactic.In 1000mL distilled water, add sodium-chlor 40g, calcium chloride 10g, maltose 30g, dissolving fully, 100 ℃ of sterilizations with water-bath temperature control to 30 ℃, are added in the 1000g yeast-lactic, are placed in the 10L fermentor tank.Control 35 ℃ of culture temperature, use 10wt%Na
2CO
3Solution is regulated pH5.2; 1.20vvm under ventilation condition, incubation time 3 hours, centrifugal collection yeast slurry, in mensuration yeast cell dry weight, content of trehalose is 16.88%.
Barms: yeast saccharomyces cerevisiae (Saccharomyces cerevisiae) AS 2.502.The seed culture fluid preparation is with embodiment 1.
With the yeast slant strains, in the 250mL triangular flask of access dress 20mL seed culture medium, after 30 ℃ of 200r/min shake-flask culture 24h, change the 500mL triangular flask that the 80mL seed culture medium is housed over to, 30 ℃ of 200r/min shake-flask culture 16h obtain seed culture fluid, and are standby.
Above-mentioned seed culture fluid 3L is accessed in the 25L fermentation basic medium of having sterilized (fermentor tank volume 50L), and culture condition is as follows.1. temperature: at the 1-8h of fermentation in the time, 30 ℃ of culture temperature, within the time of fermentation 9-15h, 32 ℃ of culture temperature, at fermentation 16-19h in the time, 35 ℃ of culture temperature.2. pH: in the culturing process of yeast, stream adds 10%Na
2CO
3Solution is regulated pH5.2.3. dissolved oxygen: mixing speed is controlled at 200r/min, and air flow 1-16h is controlled at 2.5vvm, in the time, by valve, air flow is dropped to 1.2vvm at fermentation 16-20h.Stream sugared soln and nutrient salt solution, nutrient salt solution stream adds total amount 2.5L, 16h adds at fermentation time, sugar soln carries out stream according to the growing state of ethanol feedback and yeast and adds, make that in fermented liquid, glucose content is controlled at 0.40-0.48% (volume ratio) always, the 19h fermentation ends, yeast cell 4.56% (dry weight) in the gained fermented liquid, content of trehalose 9.0%.
With above-mentioned fermented liquid, in the centrifugal 10min of 3500r/min, collect yeast-lactic.Abandon the centrifuge tube supernatant liquor, take out yeast-lactic.In 1000mL distilled water, add sodium-chlor 32g, calcium chloride 10g, maltose 24g, dissolving fully, 100 ℃ of sterilizations with water-bath temperature control to 30 ℃, are added in the 1000g yeast-lactic, are placed in the 10L fermentor tank.Control 35 ℃ of culture temperature, use 10%Na
2CO
3Solution is regulated pH5.2; 1.20vvm under ventilation condition, incubation time 3 hours, centrifugal collection yeast slurry, in mensuration yeast cell dry weight, content of trehalose is 16.00%.
Embodiment 3
Barms: yeast saccharomyces cerevisiae (Saccharomyces cerevisiae) AS 2.503.The seed culture fluid preparation is with embodiment 1.
With the yeast slant strains, in the 250mL triangular flask of access dress 20mL seed culture medium, after 30 ℃ of 200r/min shake-flask culture 24h, change the 500mL triangular flask that the 80mL seed culture medium is housed over to, 30 ℃ of 200r/min shake-flask culture 16h obtain seed culture fluid, and are standby.
Above-mentioned seed culture fluid 3L is accessed in the 25L fermentation basic medium of having sterilized (fermentor tank volume 50L), and culture condition is as follows.1. temperature: at the 1-8h of fermentation in the time, 30 ℃ of culture temperature, within the time of fermentation 9-15h, 32 ℃ of culture temperature, at fermentation 16-20h in the time, 35 ℃ of culture temperature.2. pH: in the culturing process of yeast, stream adds 10%Na
2CO
3Solution is regulated pH5.2.3. dissolved oxygen: mixing speed is controlled at 200r/min, and air flow 1-16h is controlled at 2.5vvm, in the time, by valve, air flow is dropped to 1.2vvm at fermentation 16-20h.Stream sugared soln and nutrient salt solution, nutrient salt solution stream adds total amount 2.5L, 16h adds at fermentation time, sugar soln carries out stream according to the growing state of ethanol feedback and yeast and adds, make that in fermented liquid, glucose content is controlled at 0.40-0.45% (volume ratio) always, the 20h fermentation ends, yeast cell 5.72% (dry weight) in the gained fermented liquid, content of trehalose 7.72%.
With above-mentioned fermented liquid at the centrifugal 10min of 3500r/min,, centrifugal collection yeast-lactic.Abandon the centrifuge tube supernatant liquor, take out yeast-lactic.In 1000mL distilled water, add sodium-chlor 36g, calcium chloride 10g, maltose 26g, dissolving fully, 100 ℃ of sterilizations with water-bath temperature control to 30 ℃, are added in the 1000g yeast-lactic, are placed in the 10L fermentor tank.Control 35 ℃ of culture temperature, use 10%Na
2CO
3Solution is regulated pH5.2; 1.20vvm under ventilation condition, incubation time 3 hours, centrifugal collection yeast slurry, in mensuration yeast cell dry weight, content of trehalose is 16.01%.
Embodiment 4
Barms: yeast saccharomyces cerevisiae (Saccharomyces cerevisiae) AS 2.504.The seed culture fluid preparation is with embodiment 1.
With the yeast slant strains, in the 250mL triangular flask of access dress 20mL seed culture medium, after 30 ℃ of 200r/min shake-flask culture 24h, change the 500mL triangular flask that the 80mL seed culture medium is housed over to, 30 ℃ of 200r/min shake-flask culture 16h obtain seed culture fluid, and are standby.
Above-mentioned seed culture fluid 3L is accessed in the 25L fermentation basic medium of having sterilized (fermentor tank volume 50L) culture condition such as embodiment 1,20h fermentation ends, yeast cell 5.11% (dry weight) in the gained fermented liquid, content of trehalose 6.2%.
Above-mentioned fermented liquid at the centrifugal 10min of 3500r/min, is collected yeast-lactic.Abandon the centrifuge tube supernatant liquor, take out yeast-lactic.In 1000mL distilled water, add sodium-chlor 38g, calcium chloride 10g, maltose 28g, dissolving fully, 100 ℃ of sterilizations with water-bath temperature control to 30 ℃, are added in the 1000g yeast-lactic, are placed in the 10L fermentor tank.Control 35 ℃ of culture temperature, use 10%Na
2CO
3Solution is regulated pH5.2; 1.20vvm under ventilation condition, incubation time 3 hours, centrifugal collection yeast slurry, in mensuration yeast cell dry weight, content of trehalose is 16.22%.
Claims (1)
1. method that improves trehalose content of yeast cell, the yeast-lactic that the yeast fermentation medium centrifugal is separated the cell concn that obtains 13-17%, add maltose in yeast-lactic, the aqueous solution of sodium-chlor and calcium chloride, the mass volume ratio of yeast-lactic and water is 1: 1, maltose, the consumption of sodium-chlor and calcium chloride accounts for yeast-lactic total mass per-cent and is respectively 2.4-3.0% maltose, 3.2-4.0% sodium-chlor and 1.0% calcium chloride, at 35 ℃, pH5.2, 1.0-1.2vvm cultivated 2-3 hour under ventilation condition, centrifugal collection yeast slurry, measure content of trehalose 16-17% in yeast, with the yeast cell dry weight basis,
Described yeast is yeast saccharomyces cerevisiae;
Described mass volume ratio, unit are g/mL or kg/L;
The preparation of described yeast fermentation nutrient solution, step is as follows:
(1) substratum
Seed culture medium: glucose 20g, KH
2PO
41g, (NH
4)
2SO
45g, MgSO
47H
2O 0.2g, ZnSO
47H
2O 0.05g, pH5.4, distilled water 1000mL;
Fermention medium:
1. basic medium ferments
The corn steep liquor 20% that contains solid substance 28wt%, glucose 1%, nutrient salt solution 5%, water 74% is volume ratio; Wherein, the nutrient salt solution formula is KH
2PO
413.0g, (NH
4)
2SO
494.16g, MgSO
47H
2O 1.4g, ZnSO
47H
2O0.28g adds water and is made into 1000mL;
2. fermenting process stream adds component
Sugar soln: glucose is made into the solution of 250g/L; Nutrient salt solution: KH
2PO
413.0g, (NH
4)
2SO
494.16g, MgSO
47H
2O 1.4g, ZnSO
47H
2O 0.28g adds water and is made into 1000mL; The Na of alkali lye: 100g/L
2CO
3Solution;
(2) seed culture
Barms is yeast saccharomyces cerevisiae (Saccharomyces cerevisiae), code name: AS 2.146, and AS 2.502, AS 2.503 or AS 2.504; With the yeast slant strains, in the 250mL triangular flask of access dress 20mL seed culture medium, 30 ℃ of 200r/min shake-flask culture 24h change the 500mL triangular flask that the 80mL seed culture medium is housed over to, and 30 ℃ of 200r/min shake-flask culture 16h obtain seed culture fluid, and are standby;
(3) fermentation culture
The 50L fermentor tank that 25L fermentation basic medium is housed is carried out real tank sterilization, and sterilising conditions is 120 ℃, 30min; When then being cooled to 32 ℃, access seed culture fluid 3L regulates pH5.2, controls 30 ℃ of culture temperature, and stream adds sugar soln and the nutrient salt solution of step (1) described in 2.;
The control total fermentation time is 18-21h, 30 ℃ of fermentation time 1-8h culture temperature, and 32 ℃ of 9-15h culture temperature, 16h is to 35 ℃ of fermentation ends culture temperature; Fermentation time 1-15h control the sterile air volume with fermentating liquid volume than per minute at 2.5vvm, in the Trehalose Content stage of fermentation time 16h to fermentation ends, adjust valve, air flow is dropped to 1.2vvm;
Nutrient salt solution added before fermentation ends in 3-5 hour, and the stream dosage of sugar soln is according to the yeast growth situation, made that in fermentation system, glucose content is controlled at below 0.5% always; In fermenting process, control pH5.2, mixing speed is controlled at 200r/min.
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| CN105861344A (en) * | 2016-05-30 | 2016-08-17 | 湖北工业大学 | Synchronous culture method for improving yeast biomass and intracellular trehalose content |
| CN106399423B (en) * | 2016-09-14 | 2019-11-29 | 中美食品有限公司 | A method of trehalose is prepared under the conditions of environment stress using beer waste yeast |
| CN109868229A (en) * | 2019-03-11 | 2019-06-11 | 江南大学 | A method of keeping yeast activity |
| CN110656080A (en) * | 2019-11-22 | 2020-01-07 | 顾霆 | Directional culture method of yeast cells |
| CN113151367A (en) * | 2021-05-10 | 2021-07-23 | 湖北工业大学 | Fermentation method for synthesizing xylitol de novo by self-protected zygosaccharomyces rouxii |
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2010
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| Yoshida M.production of trehalose from starch by maltose phosphorylase from a strain of plesiommnas.《Starch》.1997,第49卷(第1期),26-29. * |
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