CN102337313B - Method for preparing trehalose - Google Patents
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- CN102337313B CN102337313B CN 201110314098 CN201110314098A CN102337313B CN 102337313 B CN102337313 B CN 102337313B CN 201110314098 CN201110314098 CN 201110314098 CN 201110314098 A CN201110314098 A CN 201110314098A CN 102337313 B CN102337313 B CN 102337313B
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Abstract
The invention belongs to the fields of fermentation and production of microorganisms, and in particular relates to a method for improving the yield of trehalose in a yeast strain by a dry dehydration method. A method for preparing the trehalose comprises the following steps of: fermenting the yeast strain at the temperature of between 30 and 35 DEG C, performing enlargement culture, collecting thalli, performing extrusion dehydration, and performing forced air drying on the dehydrated thalli at the temperature of between 30 and 95 DEG C until the water content of the thalli is 5 to 25 percent; performing ultramicro crushing on the dried thalli to obtain wall breaking thalli powder; and dissolving the thalli powder in water, uniformly mixing, performing ultrafiltration by using an ultrafiltration membrane, and collecting filtration liquor to obtain a solution in which the trehalose is dissolved. By the method, the water content of the yeast thalli is gradually reduced through forced air drying, the yeast thalli is stimulated to synthesize the trehalose in large quantities under a dry environment, the accumulation amount of the trehalose in the yeast thalli is improved, and the content of the trehalose in a yeast cell can be improved by 85 percent.
Description
Technical field
The invention belongs to the microbial fermentation production field, be specifically related to a kind of method that improves yeast strain trehalose output by the drying and dehydrating method.
Background technology
Trehalose (Trehalose) be a kind of by two glucose molecules by the hemiacetal hydroxyl with α-1,1 glycosidic link bonded irreducibility disaccharide.Molecular formula is C
12H
22O
11, relative molecular weight is 378.33.Its structural formula is as follows:
Since 1853 are obtained from the ergot of rye by WiggersShi, found afterwards that trehalose extensively was present in various bacteria, yeast, fungi, algae, insect and the plant.Trehalose is the best natural disaccharide of stability, does not make film reagent reduction variable color, also with amino acid and protein etc. Maillard reactions does not take place, and the most peculiar be that it has nonspecific provide protection to organism.This non-specific provide protection is mainly reflected in when biomass cells is in that hunger, high temperature, low temperature, freezing, radiation, drying, high osmotic pressure and toxic reagent etc. are various coerces environment, and intracellular trehalose content rises rapidly, thus protection life itself.A large amount of research evidences show, some species degeneration-resistant patience of cashing out of severe environment to external world are all relevant with the trehalose that exists in its body.Wherein the most famous example is exactly the so-called latent biology of giving birth to, this class biology can be under extreme exsiccant condition with body in 99% moisture slough and not dead, and life can be kept the several years and even more than the many decades, they can bring back to life again in case condition suits, its secret is exactly to contain abundant trehalose in their cell, so trehalose is called " sugar of life " again.
Discover that ectogenic trehalose has good non-specific provide protection to organism and biomacromolecule equally.For example, the skin of some medical products (as blood products, blood plasma, blood sphaeroprotein, transfer factor etc.), vaccine, vaccine, antibody, drug-loaded liposome, antiserum(antisera), the required storage of surgical operation, organ etc.) and biological products (bacterial classification, molecular biology enzyme, industrial production with enzyme etc.) need preservation at low temperatures, if yet after adding trehalose, can drying make solid phase prod, then do not need freezing, recover to recover active again behind the moisture, the preservation of medical product and biological products and transportation cost are reduced greatly.In addition, trehalose also is widely used in fields such as food, makeup, agriculturals, and demonstrates its specific superiority.
Preparation methods for trehalose can be divided into following two kinds at present:
1, enzyme transforming process: utilize the extracellular microbial endoenzyme, under given conditions the conversion of substrate trehalose synthesis.For example, utilize trehalose synthase, conversion maltose is trehalose; With starch is raw material, utilizes Oligomeric maltose glycosides base trehalose to generate enzyme and Oligomeric maltose glycosides base trehalose lytic enzyme combined action trehalose synthesis.Enzyme transforming process has advantages such as mild condition, specificity are good, but used enzyme is an intracellular enzyme, needs broken thalline earlier, extracts refining enzyme, the process complexity.
2, yeast extraction method: because yeast cell can synthesize the trehalose of high level, and yeast culture is simple, and the yeast extraction method is the focus of trehalose process study always.A large amount of trehaloses can be accumulated under the extreme conditions such as yeast oozes at high temperature, height, hunger, broken yeast somatocyte can be passed through, the separation and Extraction trehalose.Yet trehalose is an intracellular product, and the yeast trehalose synthesis is subjected to the influence of external environmental condition bigger, and the content of trehalose of intracellular accumulation also is not very high under the common process condition, yeast broken wall difficulty in addition, this method suitability for industrialized production not as yet at present.Therefore improve the yeast intracellular trehalose content, seeking a kind of easy yeast wall breaking technology is the key that realizes yeast extraction method suitability for industrialized production trehalose.
Studies show that trehalose is that a kind of that the yeast thalline generates when externally environment takes place by abominable the variation stress meta-bolites, utilize this principle, derive many researchs that utilize ambient conditions to stimulate the promotion yeast to produce trehalose at present, as utilize elevated temperature, hungry, methods such as high pressure improve the yeast intracellular trehalose content, for example Chinese patent CN1458278A and Chinese patent CN1216152C, especially among the Chinese patent CN1458278A, its stress the attitude yeast cell cultivation cultivate by nitrogen hunger respectively, heat-shocked is cultivated and the osmotic stress best cultivation obtains required thalline, and accumulates trehalose in a large number.But the research of adopting drying and dehydrating to handle yeast thalline acquisition trehalose still belongs to blank, and in the trehalose leaching process, the problem that breaking yeast cellule membrane is raise difficult questions for discussion also still exists.
Summary of the invention
For this reason, technical problem to be solved by this invention is that yeast extraction method in the prior art produces that trehalose exists that the yeast intracellular trehalose is on the low side, the problem of yeast broken wall difficulty in the leaching process, and then a kind of method that improves yeast cell intracellular trehalose content by external stimulus is provided;
The method for preparing trehalose that a kind of breaking yeast cellule membrane method is simple, extraction ratio of trehalose is higher further is provided.
For solving the problems of the technologies described above, the method for preparing trehalose of the present invention is characterized in that, comprises the steps:
(1) yeast culture and collection: after yeast strain 30-35 ℃ of fermentation and enlarged culturing, collect thalline;
(2) with the thalline extrusion dehydration of collecting, and the thalline after will dewatering is 5-25% in 30-95 ℃ of forced air drying to water ratio;
(3) with dried thalline micronizing in the step (2), obtain the thalline powder;
(4) the thalline powder that obtains is water-soluble and mix, adopt the ultra-filtration membrane ultrafiltration, collect the solution that filtered solution obtains being dissolved with required trehalose.
In the described step (2), described forced air drying step comprises:
(a) controlled temperature 30-35 ℃, per minute ventilation and loft drier volume are than being 0.5-2.5:1, and being dried to the thalline water ratio is 50-60%;
(b) elevated temperature to 35-40 ℃, per minute ventilation and loft drier volume than being 0.5-2.5:1, being dried to the thalline water ratio is 40-50%;
(c) elevated temperature to 40-90 ℃, per minute ventilation and loft drier volume than being 0.5-1.5:1, being dried to the thalline water ratio is 5-25%.
In the described step (4), the molecular weight cut-off of ultra-filtration membrane described in the described ultrafiltration step is 1000-4000Da.
In the described step (4), described thalline powder water-soluble and mix after, also comprise the step that adopts the diatomite prefiltration before the ultra-filtration membrane ultrafiltration step.
In the described step (4), the mass volume ratio of described thalline powder and water is 1:2-10.
In the described step (3), the step that described thalline is pulverized is handled by supper micron mill.
In the described step (1), described yeast strain is that the active dry yeast activation obtains.
In the described step (1), the substratum of described fermentation step is: corn steep liquor 5-15%, peptone 2-7%, glucose 8-15%, MgSO
40.01-0.035%, NaH
2PO
40.1-0.25%, all the other are distilled water, more than are mass percent, initial pH is 6.0-7.2.
In the described step (1), the substratum of described enlarged culturing step is: corn steep liquor 5-15%, peptone 2-7%, glucose 8-15%, MgSO
40.01-0.035%, NaH
2PO
40.1-0.25%, all the other are distilled water, more than are mass percent, initial pH is 6.0-7.2.
The weight content of solid substance is 20-25wt% in the described corn steep liquor.
Described step (4) also comprises afterwards with gained aqueous trehalose crystalline step (5): it is 70-90% that the filtered solution that ultrafiltration in the step (4) is obtained is evaporated to trehalose concentration, and crystallization, separates, is drying to obtain required trehalose.
The dry weight ratio that accounts for thalline with content of trehalose among the present invention is represented the condition of production of trehalose, and concrete trehalose measuring method is: high performance liquid chromatography, see trehalose national standard (GB/T 23529-2009).
Technique scheme of the present invention compared with prior art has the following advantages:
1, the present invention reduces moisture content in the yeast thalline gradually by forced air drying, stimulate yeast thalline a large amount of trehalose synthesis under dry environment, promote the intravital trehalose accumulation volume of yeast, under top condition, content of trehalose can account for 26% of yeast dry weight, and common high temperature, starvation conditions stimulate yeast, and the accumulation volume of trehalose is about 14% of dry cell weight in the thalline, and the inventive method can make trehalose content of yeast cell improve 85%;
2, because the thalline after the forced air drying of the present invention presents the stem cell state, therefore can collect the back adopts supper micron mill to destroy the cell walls of yeast thalline, this is dry milling, with traditional wet crushing (cell solution, adopt ultrasonic disruption method, pressure difference method, organic solvent method, smash to pieces, fracturing cell walls such as grinding) compare, have that technology is simple, easy to operate, energy consumption is little, advantage such as be easy to accomplish scale production;
3, described forced air drying step adopts the method for temperature-controlled drying step by step, the two steps temperature control air seasoning by 30-35 ℃ of temperature control respectively and 35-40 ℃ earlier, realize that thalline accumulates trehalose in a large number, 40-90 ℃ of air seasoning then, further with the thalline drying, for follow-up dry type fragmentation is prepared, also further promote the accumulation volume of trehalose simultaneously;
4, select active dry yeast activation culture and collect thalline for use, active dry yeast cell itself promptly is dry lack of water bacterial strain, and the output of its body intracellular trehalose will be higher than common yeast strain, helps a large amount of trehalose synthesis;
5, adopting corn steep liquor is the major nitrogen source of bacterial strain activation and enlarged culturing, saves cost.
Description of drawings
For the easier quilt of content of the present invention is clearly understood, below according to a particular embodiment of the invention and in conjunction with the accompanying drawings, the present invention is further detailed explanation, wherein
Fig. 1 is a trehalose preparation technology flow process of the present invention.
Embodiment
According to following embodiment, the present invention may be better understood.Yet, those skilled in the art will readily understand that the described concrete processing condition of embodiment, material proportion and result thereof only are used to illustrate the present invention, and should also can not limit the present invention described in claims.
As shown in Figure 1, the described method for preparing trehalose of various embodiments of the present invention will be carried out according to process flow steps shown in it.
Embodiment 1
Seed culture medium and fermention medium that present embodiment is selected for use are: corn steep liquor (containing solid substance 25%) 10%, peptone 7.0%, glucose 15%, MgSO
40.01%, NaH
2PO
40.1%, all the other are distilled water, more than are mass percent, and initial pH is 6.0.
The described method for preparing trehalose comprises:
(1) strain inclined plane that obtains after the active dry yeast activation of preserving is scraped and is got two ring bacterium, is inoculated into 1 L that the above-mentioned seed culture medium of 100 ml is housed and shakes in the bottle, and 33 ℃, 220 rpm are cultivated 20 h, and are standby as bacterial classification.After 121 ℃ of 7.5 L fermentor tanks sterilization, 20 min of 4.5 L fermention mediums are housed, above-mentioned bacterial classification is seeded to 4.5 L ferment tanks by 10% inoculum size.In the fermenting process, control pH 5.0 ± 0.1 by auto-feeding 5 mol/L NaOH and HCI, rotating speed 300 rpm, air flow is 1.5:1 with the fermentating liquid volume ratio, 33 ℃ of constant temperature culture 20 h, thalline (dry weight) content is 120 g/L, and centrifugal 10 min of fermented liquid 3000 rpm that collect are collected thalline.
(2) the thalline water ratio is about about 80% in the centrifugal back bacterium mud, and after the squeezer extruding, the water ratio of thalline is about 65% in the bacterium mud, then thalline is placed air dry oven to carry out forced air drying.
Described forced air drying comprises: (a) at 35 ℃ of insulation 5 h, and with the per minute air quantity: loft drier volume (down with) is the ratio forced air drying of 2.5:1, and moisture contains rate and reduces to 52.5% in the thalline; (b) then controlled temperature is incubated 5 h for 40 ℃, and ventilation is 0.5:1, and thalline moisture contains rate and reduces to 41%; (c) 90 ℃ of drying 3.5 h of temperature control then, ventilation is 0.5:1, obtains the dry yeast thalline, water ratio is about 5.5%(W/W), this moment, content of trehalose can reach the 26%(W/W of dry cell weight).
(3) the above-mentioned dry yeast thalline that obtains is pulverized 30 min at supper micron mill, stocking volume is 30%, and the bacterial cell disruption rate reaches 95%.
(4) then broken wall thalline powder and water are prepared into the aqueous solution by the 1:7 dissolving, evenly stir 1 h.Above-mentioned solution is adopted the diatomite prefiltration, remove large granular impurities such as bacterial chip, adopt the organic ultrafiltration membrance filter of rolling then, the ultra-filtration membrane molecular weight cut-off is 1000Da, service temperature is 35 ℃, and pressure is 6 bar, removes impurity such as high molecular weight protein, polysaccharide, collect and see through liquid, the trehalose rate of recovery can arrive 88.5%.
(5) with under 70 ℃ of conditions of rotatory evaporator above-mentioned aqueous trehalose being concentrated in vacuo to 70%, put the crystallizer crystallization, at first solution is at 70 ℃ of insulation 5 min, press 2% of content of trehalose and add crystal seed, slowly cooling drops to 5 ℃, centrifugation then with temperature from 70 ℃ in 5 h, drying obtains the trehalose product, and purity is 98% relatively.
Embodiment 2
Seed culture medium and fermention medium that present embodiment is selected for use are: corn steep liquor (containing solid substance 20%) 15%, peptone 2%, glucose 8%, MgSO
40.035%, NaH
2PO
40.25%, all the other are distilled water, more than are mass percent, and initial pH is 7.2.
The described method for preparing trehalose comprises:
(1) strain inclined plane that obtains after the active dry yeast activation of preserving is scraped and is got two ring bacterium, is inoculated into 1 L that 100 ml seed culture mediums are housed and shakes in the bottle, and 30 ℃, 150 rpm are cultivated 15 h, as bacterial classification.After 121 ℃ of sterilizations of 7.5 L fermentor tanks, 20 min of 4.5 L fermention mediums are housed, bacterial classification is pressed in 15% the inoculum size inoculation fermentation jar.In the fermenting process, control pH 6.5 by auto-feeding 5 mol/L NaOH and HCI, rotating speed 250 rpm, air flow is 1.5:1 with the fermentating liquid volume ratio, 30 ℃ of constant temperature culture 28 h, thalline (dry weight) content is 95 g/L, filtering fermentation liquor is collected thalline.
(2) the thalline water ratio is about about 83% in the bacterium mud of filtration back, and after the squeezer extruding, the water ratio of thalline is about 65% in the bacterium mud, then thalline is placed air dry oven to carry out forced air drying.
Described forced air drying comprises: (a) at 33 ℃ of insulation 5 h, and with the per minute air quantity: the loft drier volume is the ratio forced air drying of 0.5:1, and moisture contains rate and reduces to about 55.5% in the thalline; (b) then control 37 ℃ of insulation 5 h, ventilation is 2.5:1, and moisture contains rate and reduces to 45% in the thalline; (c) then at 70 ℃ of insulation 5 h, ventilation is 1:1, obtains the dry yeast thalline, and it is 13%(W/W that moisture contains rate), this moment, content of trehalose can reach the 22%(W/W of dry cell weight).
(3) the above-mentioned dry yeast thalline that obtains is pulverized 30 min at supper micron mill, stocking volume is 20%, and the bacterial cell disruption rate reaches 97%.
(4) then broken wall thalline powder and water are pressed the 1:10 dissolving, be prepared into the aqueous solution, stir 1 h.Above-mentioned solution is adopted the diatomite prefiltration, remove large granular impurities such as bacterial chip, adopt the organic ultrafiltration membrance filter of rolling then, the ultra-filtration membrane molecular weight cut-off is 2500Da, service temperature is 35 ℃, and pressure is 4 bar, removes impurity such as high molecular weight protein, polysaccharide, collect and see through liquid, the trehalose rate of recovery can arrive 90.5%.
(5) with under 75 ℃ of conditions of rotatory evaporator above-mentioned aqueous trehalose being concentrated in vacuo to 80%, put the crystallizer crystallization, at first solution is at 75 ℃ of insulation 5 min, press 3% of content of trehalose and add crystal seed, slowly cooling drops to 10 ℃, centrifugation then with temperature from 75 ℃ in 3 h, drying obtains the trehalose product, and purity is 98.5% relatively.
Embodiment 3
Seed culture medium and fermention medium that present embodiment is selected for use are: corn steep liquor (containing solid substance 23%) 5%, peptone 5.0%, glucose 11%, MgSO
40.02%, NaH
2PO
40.2%, all the other are distilled water, more than are mass percent, and initial pH is 6.5.
The described method for preparing trehalose comprises:
(1) strain inclined plane that obtains after the active dry yeast activation of preserving is scraped and is got two ring bacterium, is inoculated into 1 L that 100 ml seed culture mediums are housed and shakes in the bottle, and 35 ℃, 180 rpm are cultivated 25 h, and are standby as bacterial classification.121 ℃ of 7.5 L fermentor tanks sterilization, 20 min of 4.5 L fermention mediums are housed, then bacterial classification are inoculated in the fermentor tank by 8% inoculum size.In the fermenting process, control pH 6.0 by auto-feeding 5 mol/L NaOH and HCI, rotating speed 250 rpm, air flow is 1.0:1 with the fermentating liquid volume ratio, 35 ℃ of constant temperature culture 24 h, thalline (dry weight) content is 105.5 g/L, centrifugal 10 min of fermented liquid 5000 rpm collect thalline.
(2) the thalline water ratio is about about 80% in the centrifugal back bacterium mud, and after the squeezer extruding, the water ratio of thalline is about 65% in the bacterium mud, then thalline is placed air dry oven to carry out forced air drying.
Described forced air drying comprises: (a) at 30 ℃ of insulation 5 h, ventilation is a 1.5:1(per minute air quantity: the loft drier volume), moisture contains rate and reduces to 57.5% in the thalline; (b) then be warming up to 35 ℃ of insulation 5 h, ventilation is 1.5:1, and moisture contains rate and reduces to 40.5% in the thalline; (c) then at 40 ℃ of insulation 8 h, ventilation is 1.5:1, obtains the dry yeast thalline, and it is 23%(W/W that moisture contains rate), this moment, content of trehalose can reach the 24%(W/W of dry cell weight).
(3) the dry yeast thalline is pulverized 30 min at supper micron mill, and stocking volume is 25%, and the bacterial cell disruption rate reaches 96%.
(4) then broken wall thalline powder and water are pressed the 1:2 dissolving, be prepared into the aqueous solution, evenly stir 1 h.Above-mentioned solution is adopted the diatomite prefiltration, remove large granular impurities such as bacterial chip, adopt the organic ultrafiltration membrance filter of rolling then, the ultra-filtration membrane molecular weight cut-off is 4000Da, service temperature is 35 ℃, and pressure is 2.5bar, removes impurity such as high molecular weight protein, polysaccharide, collect and see through liquid, the trehalose rate of recovery can arrive 91.5%.
(5) with under 80 ℃ of conditions of rotatory evaporator above-mentioned aqueous trehalose being concentrated in vacuo to 90%, place the crystallizer crystallization, at first solution is at 80 ℃ of insulation 5 min, press 2.5% of content of trehalose and add crystal seed, slowly cooling drops to 15 ℃, centrifugation then with temperature from 80 ℃ in 4 h, drying obtains the trehalose product, and purity is 97.5% relatively.
Embodiment 4
Carry out the activation and the enlarged culturing of bacterial classification according to the prescription of seed culture medium described in the embodiment 1 and fermentation culture, its difference is, present embodiment is selected for use and scraped the fresh yeast strain of getting from the inclined-plane flat board and cultivate and be used to prepare trehalose.
And forced air drying treatment process that it is follow-up and extraction process condition, all identical with method and the processing condition described in the embodiment 1, obtain dried dry yeast thalline, the content that detects trehalose can reach the 20.4%(W/W of dry cell weight).
Embodiment 5
Carry out the enlarged culturing of active dry yeast activated spawn according to the prescription of seed culture medium described in the embodiment 1 and fermentation culture, be used to prepare the bacterial strain of trehalose.
The difference of the processing condition described in itself and the embodiment 1 only is in the forced air drying step, control drying temperature is 60 ℃ always, be dried to water ratio and be lower than 6%, obtain dried yeast thalline, this moment, content of trehalose can reach the 6.5%(W/W of dry cell weight).
Obviously, the foregoing description only is for example clearly is described, and is not the qualification to embodiment.For those of ordinary skill in the field, can also make other changes in different forms on the basis of the above description.Here need not also can't give exhaustive to all embodiments.And conspicuous variation of being extended out thus or change still are among the protection domain of the invention.
Claims (9)
1. a method for preparing trehalose is characterized in that, comprises the steps:
(1) yeast culture and collection: yeast strain after 30-35 ℃ of fermentation and enlarged culturing, is collected thalline;
(2) with the thalline extrusion dehydration of collecting, and the thalline after will dewatering passes through 30-35 ℃ of (a) controlled temperature, the per minute ventilation is 0.5-2.5:1 with loft drier volume ratio, and being dried to the thalline water ratio is 50-60%; (b) elevated temperature to 35-40 ℃, per minute ventilation and loft drier volume than being 0.5-2.5:1, being dried to the thalline water ratio is 40-50%; (c) elevated temperature to 40-90 ℃, per minute ventilation and loft drier volume than being 0.5-1.5:1, being dried to the thalline water ratio is 5-25%;
(3) with dried thalline micronizing in the step (2), obtain broken wall thalline powder;
(4) the thalline powder that obtains is water-soluble and mix, adopt the ultra-filtration membrane ultrafiltration, collect the solution that filtered solution obtains being dissolved with required trehalose.
2. the method for preparing trehalose according to claim 1 is characterized in that:
In the described step (4), the molecular weight cut-off of ultra-filtration membrane described in the described ultrafiltration step is 1000-4000 Da.
3. the method for preparing trehalose according to claim 2 is characterized in that:
In the described step (4), described thalline powder water-soluble and mix after, also comprise the step that adopts the diatomite prefiltration before the ultra-filtration membrane ultrafiltration step.
4. the method for preparing trehalose according to claim 3 is characterized in that:
In the described step (4), the mass volume ratio of described thalline powder and water is 1:2-10.
5. the method for preparing trehalose according to claim 4 is characterized in that:
In the described step (1), described yeast strain is that the active dry yeast activation obtains.
6. the method for preparing trehalose according to claim 5 is characterized in that:
In the described step (1), the substratum of described fermentation step is: corn steep liquor 5-15%, peptone 2-7%, glucose 8-15%, MgSO
40.01-0.035%, NaH
2PO
40.1-0.25%, all the other are distilled water, more than are mass percent, initial pH is 6.0-7.2.
7. the method for preparing trehalose according to claim 6 is characterized in that:
In the described step (1), the substratum of described enlarged culturing step is: corn steep liquor 5-15%, peptone 2-7%, glucose 8-15%, MgSO
40.01-0.035%, NaH
2PO
40.1-0.25%, all the other are distilled water, more than are mass percent, initial pH is 6.0-7.2.
8. the method for preparing trehalose according to claim 7 is characterized in that:
The content of solid substance is 20-25wt% in the described corn steep liquor.
9. according to the arbitrary described method for preparing trehalose of claim 1-8, it is characterized in that:
Described step (4) also comprises afterwards with gained aqueous trehalose crystalline step (5): it is 70-90% that the filtered solution that ultrafiltration in the step (4) is obtained is evaporated to trehalose concentration, and crystallization, separates, is drying to obtain required trehalose.
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| CN104829275B (en) * | 2015-03-25 | 2017-11-17 | 冉令波 | Production process of liquid seaweed fertilizer and liquid seaweed fertilizer |
| CN105925642B (en) * | 2016-06-14 | 2019-08-13 | 湖南汇升生物科技有限公司 | With the method for microbe fermentation method industrialized production trehalose |
| CN106317131B (en) * | 2016-08-24 | 2019-04-16 | 山东福洋生物科技有限公司 | A kind of method for crystallising of trehalose |
| 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 |
| CN114438072B (en) * | 2022-04-08 | 2022-05-31 | 山东天力药业有限公司 | Production method of trehalose |
| CN117887784A (en) * | 2024-03-13 | 2024-04-16 | 山东天力药业有限公司 | Method for preparing trehalose from starch |
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