CN1216152C - Method for raising yield of fucose produced from microzyme by using high pressure - Google Patents
Method for raising yield of fucose produced from microzyme by using high pressure Download PDFInfo
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- CN1216152C CN1216152C CN 03130410 CN03130410A CN1216152C CN 1216152 C CN1216152 C CN 1216152C CN 03130410 CN03130410 CN 03130410 CN 03130410 A CN03130410 A CN 03130410A CN 1216152 C CN1216152 C CN 1216152C
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Abstract
The present invention relates to a method for improving the output of yeast trehalose by using high pressure, which belongs to a producing technology for obtaining high-output trehalose. The present invention solves the problems of low production output and high cost of the direct extraction of the trehalose from culture yeast. The core of the technical scheme is the adoption of a high-pressure stimulating condition. The method of the present invention specifically comprises: yeast is used as original strains; high-pressure stimulation is applied after seed culture and enlarging culture; the pressure in a tank is from 0.1 to 1.0Mpa; when the required pressure is achieved, the constant pressure is kept for 2 to 4 hours; a stirring rotary speed is from 50 to 300 r/min; a reaction temperature is from 30 to 35 DEG C. The method has the advantages of simple technology and production cost reduction, and the intracellular content of the trehalose in the yeast can be greatly improved. Compared with a normal-pressure comparison condition, the present invention improves the intracellular content of the trehalose by 20 to 70%. Thus, the present invention provides a new technology for producing the trehalose from the yeast by direct fermentation and realizing industrial production.
Description
Technical field
The invention belongs to a kind of technology of obtaining the high yield trehalose, particularly employing directly utilizes high pressure to improve the production technology of yeast trehalose output.
Background technology
Trehalose (Trehalose, α-D-glucopyranosyl-α-D-glucopyranoside) be by 2 furan nucleus glucose molecules by the hemiacetal hydroxyl with a kind of irreducibility disaccharide of α-1,1 glycosidic link bonded, molecular formula is C
12H
22O
112H
2O.Molecular structural formula is:
The common trehalose of occurring in nature is α, α-type, and other two kinds of optics isomers α, β-type and β, β-type is comparatively rare.Since 1853 are obtained from the ergot of rye by WiggersShi, found afterwards in natural animals and plants and microorganism, extensively to exist, especially fungi such as yeast, mould.Trehalose is the best natural disaccharide of stability, does not make fehling reagent reduction variable color, also with amino acid and protein etc. Maillard reaction does not take place, and its most peculiar functional performance is its non-specific defencive function.A large amount of research evidences show that some species degeneration-resistant patience that severe environment showed to external world is all relevant with the trehalose of existence in its body.Trehalose is at aspects such as food-processing, molecular biology, biological medicine preparation, agriculturals, as resist drying food, the preservation of various tool enzyme, antibody, organoid and biological reagent, the preservation of vaccine, hormone, blood products, drug-loaded liposome and active bacteria formulation and transportation, the exploitation of the new crop varieties that patience such as drought resisting, cold-resistant, salt tolerant is strong.All demonstrate its specific superiority.Promoted the trehalose Research on Process thus more.
Because content of trehalose is higher relatively in the yeast cell, and extract simply, what therefore research at present was more is to extract trehalose from yeast, but that its shortcoming is a yield is low, cost is high.Its reason is because trehalose belongs to intracellular product, synthesize to be subjected to the influence of external environmental condition bigger, so output is lower, and the fermentation level that commercialization is produced is only for O.4～0.6%, so the suitability for industrialized production that still is unrealized.Therefore improve this production method output, to reduce cost be its key that realizes suitability for industrialized production.
The yeast trehalose is a kind of irritability meta-bolites that the yeast thalline generates when externally " badly " variation takes place culture condition.Utilize this characteristics, more to saccharomycetic incentive condition research at present, wish to improve yeast trehalose output by the method for pressurization, realize suitability for industrialized production.But adopt the high voltage stimulus condition, the research of handling yeast acquisition trehalose still belongs to blank.
Summary of the invention
The problem that solves:
At the problems referred to above, the invention solves trehalose production problem that yield is low, cost is high of directly from the yeast thalline, extracting.Provide a kind of technology simple, utilize the external stimulus condition of high pressure as thalline, improve the method for yeast trehalose output greatly.
Technical scheme:
The present invention has studied the influence for the tired intracellular trehalose of yeast volume of different pressure, temperature of reaction, treatment time and mixing speed; Having proposed to improve the production technology of yeast intracellular trehalose output, is starting strain with the yeast promptly, obtains bacterial culture fluid through enlarged culturing, then utilizes high-pressure reaction vessel, as incentive condition, improves the output of yeast intracellular trehalose with high pressure.
Concrete grammar is as follows:
Spawn culture: the inclined-plane barms is transferred in the Erlenmeyer flask that seed culture medium is housed 30 ℃ of constant temperature culture 16h.
Enlarged culturing: add the yeast culture base of sterilization in advance in Erlenmeyer flask or ventilated type fermentor tank, the inoculum size by 10% inserts the yeast starter nutrient solution.Placing temperature then is on 30 ℃ the shaking table or fermentor tank, carries out enlarged culturing, and incubation time is 16～24h.
Autoclaving: the yeast culture liquid of enlarged culturing is placed in the high-pressure reaction vessel, and high-pressure stimulates then, stirs simultaneously, and temperature of reaction is 30～35 ℃.Related pressure all is tank pressure later on
Need to prove that high-pressure stimulates, tank pressure is 0.1～1.0Mpa (in the present patent application file, related pressure value all is the tank pressure value); After reaching required pressure, constant voltage keeps 2～4h; In the autoclaving process, changing pressure must at the uniform velocity carry out; When stirring, the mixing speed of yeast culture liquid is 50～300r/min.This method improves yeast intracellular trehalose content 20～70%.
Beneficial effect:
The present invention has explored a kind of method of new raising trehalose output, promptly utilizes the high voltage stimulus condition, yeast is produced trehalose carried out new trial; Remedied the blank of the research; Solved when utilizing the yeast extraction method to produce trehalose at present, the output of existence is crossed low problem.By improving yeast thalline intracellular trehalose output significantly, reduced production cost.This method technology is simple, has improved than intracellular trehalose content under the normal pressure collating condition to be about 20～70%; For directly utilizing yeast to produce trehalose, provide a kind of new production process.
Embodiment
Embodiment 1:
Get 1～2 ring transition of inclined-plane barms in the Erlenmeyer flask that seed culture medium is housed, 30 ℃ of constant temperature culture 16h.Add the yeast culture base of sterilization in advance in fermentor tank, the inoculum size by 10% inserts cultured yeast starter nutrient solution.Placing temperature then is 30 ℃, carries out enlarged culturing 16h in the fermentor tank of ventilation 1.5vvm.The yeast culture liquid of enlarged culturing is placed autoclave, and the pressure that raises then is to 0.1MPa, and constant voltage keeps 2h to handle, and mixing speed is 120r/min, and temperature of reaction is 30 ℃.In the autoclaving process, changing pressure must at the uniform velocity carry out.With this understanding, improved under the trehalose rate ratio normal pressure collating condition and be about 20%.
Embodiment 2:
In the present embodiment, the seed culture temperature is 25 ℃, and the reaction pressure in the high-pressure reaction vessel is 0.5Mpa, and other is with embodiment 1.With this understanding, improved more than 20% under the trehalose rate ratio normal pressure collating condition.
Embodiment 3:
In the present embodiment, the enlarged culturing temperature is 25 ℃, constant temperature culture 16h, and the reaction pressure in the high-pressure reaction vessel is 0.75Mpa, other is with embodiment 1.With this understanding, improved under the trehalose rate ratio normal pressure collating condition and be about 25%.
Embodiment 4:
Reaction pressure in the present embodiment high-pressure reaction vessel is 1.0Mpa, and other is with embodiment 1.With this understanding, improved about 30% under the trehalose rate ratio normal pressure collating condition.
Embodiment 5:
Reaction pressure in the present embodiment high-pressure reaction vessel is 0.5Mpa, and the constant voltage treatment time is 3h, and temperature of reaction is 33 ℃, and other is with embodiment 1.With this understanding, improved under the trehalose rate ratio normal pressure collating condition and be about 35%.
Embodiment 6:
Reaction pressure in the present embodiment high-pressure reaction vessel is 0.75Mpa, and the constant voltage treatment time is 3h, and temperature of reaction is 34 ℃, and other is with embodiment 1.With this understanding, improved under the trehalose rate ratio normal pressure collating condition and be about 50%.
Embodiment 7:
Reaction pressure in the present embodiment high-pressure reaction vessel is 1.0Mpa, and the constant voltage treatment time is 3h, and temperature of reaction is 35 ℃, and other is with embodiment 1.With this understanding, improved about 70% under the trehalose rate ratio normal pressure collating condition.
Embodiment 8:
Reaction pressure in the present embodiment high-pressure reaction vessel is 1.0Mpa, and the constant voltage treatment time is 4h, and temperature of reaction is 35 ℃, and other is with embodiment 1.With this understanding, improved under the trehalose rate ratio normal pressure collating condition and be about 65%.
Embodiment 9:
Reaction pressure in the present embodiment high-pressure reaction vessel is 0.75Mpa, and the constant voltage treatment time is 3h, and temperature of reaction is 34 ℃, and during autoclaving, mixing speed is 50r/min, and other is with embodiment 1.With this understanding, improved under the rate ratio normal pressure collating condition of trehalose and be about 35%.
Embodiment 10:
Reaction pressure in the present embodiment high-pressure reaction vessel is 1.0Mpa, and the constant voltage treatment time is 3h, and temperature of reaction is 34 ℃, and during autoclaving, mixing speed is 50r/min, and other is with embodiment 1.With this understanding, improved about 40% under the rate ratio normal pressure collating condition of trehalose.
Embodiment 11:
Reaction pressure in the present embodiment high-pressure reaction vessel is 1.0Mpa, and the constant voltage treatment time is 3h, and temperature of reaction is 34 ℃, and during autoclaving, mixing speed is 300r/min, and other is with embodiment 1.With this understanding, improved about 30% under the rate ratio normal pressure collating condition of trehalose.
Claims (3)
1. utilize high pressure to improve the method for yeast trehalose output, it is characterized in that with the yeast being starting strain, obtain nutrient solution, bacterial culture fluid is placed under the condition of high voltage through enlarged culturing, as incentive condition, improve the output of yeast intracellular trehalose with high pressure;
Concrete grammar is as follows:
Seed culture: get the inclined-plane barms and be transferred in the Erlenmeyer flask that seed culture medium is housed, 25～30 ℃ of constant temperature culture 16h;
Enlarged culturing: in Erlenmeyer flask or ventilation fermentation tank, add the yeast culture base of sterilization in advance, inoculum size by 10% inserts the yeast starter nutrient solution, placing temperature then is on 25～30 ℃ the shaking table or fermentor tank, carries out enlarged culturing, and incubation time is 16～24h;
Autoclaving: the yeast culture liquid of enlarged culturing is placed in the high-pressure reaction vessel, high-pressure, promptly tank pressure is that 0.1～1.0Mpa stimulates, and in the autoclaving process, changing pressure must at the uniform velocity carry out, and stirs simultaneously, and temperature of reaction is 30～35 ℃.
2. according to the described method of utilizing high pressure to improve yeast trehalose output of claim 1, it is characterized in that high-pressure stimulates, after reaching required pressure, constant voltage keeps 2～4h.
3. according to the described method of utilizing condition of high voltage to improve yeast trehalose output of claim 1, when it is characterized in that autoclaving, mixing speed is 50～300r/min.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 03130410 CN1216152C (en) | 2003-07-18 | 2003-07-18 | Method for raising yield of fucose produced from microzyme by using high pressure |
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| CN 03130410 CN1216152C (en) | 2003-07-18 | 2003-07-18 | Method for raising yield of fucose produced from microzyme by using high pressure |
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| CN1216152C true CN1216152C (en) | 2005-08-24 |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103695407A (en) * | 2013-12-26 | 2014-04-02 | 齐鲁工业大学 | Method for increasing trehalose synthase content of thurmus |
Families Citing this family (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105695349B (en) * | 2016-04-15 | 2019-04-05 | 湖北工业大学 | A kind of method of phosphate starvation culture and improvement yeast cells intracellular trehalose |
| CN112458131A (en) * | 2020-12-22 | 2021-03-09 | 吉林奥谷生物科技有限公司 | Method for preparing trehalose by using waste beer yeast |
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2003
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103695407A (en) * | 2013-12-26 | 2014-04-02 | 齐鲁工业大学 | Method for increasing trehalose synthase content of thurmus |
| CN103695407B (en) * | 2013-12-26 | 2016-01-20 | 齐鲁工业大学 | A kind of raising is dwelt the method for hot bacterium TreP content |
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