CN103695407A - Method for increasing trehalose synthase content of thurmus - Google Patents
Method for increasing trehalose synthase content of thurmus Download PDFInfo
- Publication number
- CN103695407A CN103695407A CN201310729535.8A CN201310729535A CN103695407A CN 103695407 A CN103695407 A CN 103695407A CN 201310729535 A CN201310729535 A CN 201310729535A CN 103695407 A CN103695407 A CN 103695407A
- Authority
- CN
- China
- Prior art keywords
- trep
- liquid
- solution
- ammonium sulfate
- phase
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
- C12N9/90—Isomerases (5.)
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Y—ENZYMES
- C12Y504/00—Intramolecular transferases (5.4)
- C12Y504/99—Intramolecular transferases (5.4) transferring other groups (5.4.99)
- C12Y504/99016—Maltose alpha-D-glucosyltransferase (5.4.99.16)
Landscapes
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Organic Chemistry (AREA)
- Health & Medical Sciences (AREA)
- Zoology (AREA)
- Engineering & Computer Science (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Genetics & Genomics (AREA)
- Wood Science & Technology (AREA)
- General Health & Medical Sciences (AREA)
- General Engineering & Computer Science (AREA)
- Biochemistry (AREA)
- Molecular Biology (AREA)
- Microbiology (AREA)
- Biotechnology (AREA)
- Biomedical Technology (AREA)
- Medicinal Chemistry (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The invention relates to a method for increasing the trehalose synthase content of thurmus. The method comprises the following steps: (1) inoculating a liquid seed of a thurmus strain into a liquid culture medium for culturing to prepare a strain cell culture solution; (2) fermenting the strain cell culture solution in a pressure reactor under the pressure to prepare a fermentation solution; (3) centrifuging the fermentation solution, collecting bacterial cells, breaking the cells to prepare a cell breaking solution, and extracting the cell breaking solution by a two-aqueous-phase extraction system to prepare trehalose synthase. According to the method, the trehalose synthase content of the thurmus cells can be increased, the production cost of the trehalose synthase can be reduced, the trehalose synthase can be continuously produced on a large scale, and the production process of the trehalose synthase is simple and convenient.
Description
Technical field
The present invention relates to a kind of the dwell method of hot bacterium TreP content of raising, belong to technical field of bioengineering.
Background technology
TreP (Trehalose synthase, EC5.4.99.16) is glucuronosyltransferases in a kind of molecule, and α-Isosorbide-5-Nitrae glycosidic link of catalysis maltose is converted into α-1 specifically, and 1 glycosidic link generates trehalose.Trehalose is a kind of irreducibility disaccharide being formed by the condensation of hemiacetal hydroxyl by two glucose molecules; have and under adverse circumstance, protect the microbial film of organism and the specific function of biomacromolecule; can give the ability of the severe environment such as the opposing such as lower animal, plant and microorganism nutritive deficiency, high temperature, high osmotic pressure, dry, toxic substance, thereby have a wide range of applications in fields such as biological products, food, medicine, crop breeding and fine chemistry industries.
Enzymic synthesis method is one of main method of producing trehalose, be to utilize transglycosylation that zymin has in vitro, to act on the substrates such as maltose, sucrose to produce trehalose, the method has higher specificity, gentle, the feature such as production cost is low fast.Production by Enzymes trehalose has multiple biosynthetic pathway, can be by trehalose synthesis such as different enzyme difference catalysis starch, glucose, maltose, oligosaccharides.
Utilize TreP to take the flow process that maltose is substrate trehalose synthesis and only need single step reaction, the productive rate of trehalose is high and be not subject to the impact of substrate maltose concentration, technique is simple, be easy to regulation and control, be the simple approach of Production by Enzymes trehalose, thereby TreP has been widely used in the suitability for industrialized production of trehalose.M. smegmatics (
mycobacterium smegmatis), pimelobacter sp (
pimelobacter sp.), Arthrobacter nicotinovorus (
arthrobacter nicotinovorus), pseudomonas putida (
pseudomonas putida) and some hot bacterium that dwell (
thermus) etc. all found TreP in bacterial strain, but the intracellular TreP content of above-mentioned bacterial strains is low, the activity of enzyme is low, causes the production cost of enzyme higher, therefore how to improve TreP content, to reduce production costs be to be badly in need of at present the technical problem that solves.
Research is found stress reaction can to occur when microorganism cells during in external stimuluss such as hunger, high temperature, high osmotic pressurees, improve the activity that in cell, trehalose synthesis enzyme is, thereby increase trehalose output is coerced to resist the external world.At present to the existing certain research of variation of cell intracellular trehalose and TreP under these conditions of part microorganism, as Duan Zuoying (biological processing, 2007.5) when the impact research of carrying out heat shock treatment on trehalose synthase of Pseudomonas putida S 1, adopt the method for thermal stimulus to make the enzyme work of TreP improve 76%; As Chinese patent literature CN 12190715A(application number 03129701.3) a kind of method that glucose produces yeast extracellular-trehalose as matrix two-step fermentation of take disclosed, first adopt the methods such as nitrogen hunger, heat-shocked, osmotic stress, what the generation of induction yeast cell was more stress enzyme-TreP be then to utilize a large amount of synthetic extracellular-trehaloses of TreP system in yeast cell; Ambient pressure is also a kind of incentive condition, the pressure of bio-reactor also has remarkably influenced to the content of specific product in extracellular microbial, as Chinese patent literature CN 1480535A(application number 03130410.9) method of utilizing high pressure to improve yeast trehalose output is disclosed, the method has improved 20～70% compared with intracellular trehalose content under normal pressure collating condition.About the research that utilizes pressure to improve the hot bacterium TreP content of dwelling, also there is no relevant report or patent at present.
Summary of the invention
The present invention is directed to the deficiencies in the prior art, a kind of method that provides raising to dwell hot bacterium TreP content.
The technical solution used in the present invention is:
The dwell method of hot bacterium TreP content of raising, is characterized in that, step is as follows:
(1) from test tube slant, the picking 1-2 ring hot bacteria strain of dwelling is inoculated in liquid seed culture medium, under 65～70 ℃, 100～150r/min, cultivate 12～16h, make the liquid seeds after activation, then by liquid seeds by volume 3%～5% inoculum size be inoculated in liquid nutrient medium, under 60～70 ℃, 150～200r/min, cultivate 20～24h, make strain cell nutrient solution;
(2) strain cell nutrient solution is placed in pressure reactor, at 0.5～2.0MPa pressure bottom fermentation, 1～6h, fermentation condition is 60～70 ℃, 150～200r/min, makes fermented liquid;
(3) fermented liquid is centrifugal, collect somatic cells, smudge cells, makes cytoclasis liquid, then adopts two phase aqueous extraction system to prepare TreP cytoclasis liquid.
The hot bacteria strain of dwelling in described step (1) be thermus aquaticus (
thermus aquaticus) ATCC33923 or Thermus bacterium (
thermus sp.) ATCC 43815, above bacterial strain is all preserved in U.S. representative microbial DSMZ (ATCC), can obtain by the mode of buying.
The component of the liquid seed culture medium in described step (1) is: glucose 10g/L, yeast extract paste 10g/L, peptone 10g/L, NaCl5g/L, pH7.5, distilled water 1000mL.
The component of the liquid nutrient medium in described step (1) is: glucose 20g/L, maltose 5～10g/L, yeast extract paste 5g/L, peptone 5g/L, KH
2pO
42g/L, MgSO
41g/L, pH7.5, distilled water 1000mL.
The step that employing two phase aqueous extraction system in described step (3) is prepared TreP is as follows: in cytoclasis liquid, add the polyoxyethylene glycol that ammonium sulfate and molecular weight are 2000, the mass percent concentration that makes ammonium sulfate in total system is that the mass percent concentration of 8wt%～10wt%, polyoxyethylene glycol is 10wt%～12wt%, mix, standing phase-splitting, phase solution on the polyoxyethylene glycol that makes phase solution under ammonium sulfate and contain TreP; In phase solution on polyoxyethylene glycol, add ammonium sulfate, making the mass percent concentration of ammonium sulfate in total system is 5wt%～6wt%, and the pH value of regulator solution is 7.0～8.0, mixes, standing phase-splitting, makes under the ammonium sulfate that contains TreP phase solution on phase solution and polyoxyethylene glycol; By phase solution under the ammonium sulfate making, adopt ultrafiltration membrance filter concentrated, collect trapped fluid, lyophilize, makes TreP.
Beneficial effect
(1) the hot mycetocyte of dwelling that the present invention uses contains TreP, and this endonuclease capable specifically catalysis maltose is converted into trehalose and catalysis productive rate is high, is the zymin of tool application prospect during trehalose enzyme process is produced.
(2) method of the present invention is to utilize pressure as a kind of external stimulus factor, and the TreP content in the hot mycetocyte of dwelling under pressure increases, enzymic activity raises, thereby has reduced the production cost of TreP.
(3) method of the present invention can realize serialization, the mass-producing that TreP is produced, and the production technique of enzyme is easy.
Embodiment
Below in conjunction with embodiment, technical scheme of the present invention is specifically described or is described further, object is that methods of this invention will be better understood, but protection scope of the present invention is not limited to following embodiment.
The pressure reactor adopting in the present invention is the stainless steel compressive reaction device that a set of pressure with whipping appts can be measured and control, and its volume is 3000mL, and sample can inject with syringe.Use high pure air is pressure medium, with the speed of 0.05MPa/min, boosts, and pressurize, is depressurized to normal pressure with the speed of 0.05MPa/min.In operating process, the tank pressure value that the force value of measuring is reactor.
In the present invention, the enzyme activity determination step of TreP is as follows:
Get cell culture fluid centrifugal 20min under 5000r/min, collect thalline, use distilled water wash thalline three times, make wet thallus.Get that 1.0g wet thallus joins the 0.03mol/L of 19.6mL, in the phosphoric acid buffer of pH7.2, then add the toluene of 0.4mL, at 35 ℃, process 2h, make cell treatment solution.Get cell treatment solution centrifugal 15min under 4 ℃, 12000r/min, collect supernatant liquor, supernatant liquor, through the filtering with microporous membrane of 0.45 μ m, is collected filtered liquid, makes crude enzyme liquid.Get in the 10wt% maltose solution that 10.0mL crude enzyme liquid joins 10.0mL, under 50 ℃, 150r/min, reaction 10h, reaction solution is incubated 10min again in boiling water bath.Extract reaction solution centrifugal 20min under 5000r/min, collect supernatant liquor.Get supernatant liquor, adopt high performance liquid chromatograph (purchased from Shanghai Tian Pu Analytical Instrument Co., Ltd) to measure the content of trehalose.
The enzyme activity unit of TreP is defined as: under above-mentioned reaction conditions, the enzyme amount that conversion per hour generates lmg trehalose is 1U.
Raw material sources: toluene, ammonium sulfate are all purchased from Tianjin Zhi Yuan chemical reagent company limited, and polyoxyethylene glycol is purchased from Dow Chemical company.
Embodiment 1
The dwell method of hot bacterium TreP content of raising, step is as follows:
(1) from test tube slant, the picking 2 ring hot bacteria strain of dwelling is inoculated in liquid seed culture medium, under 70 ℃, 150r/min, cultivate 16h, make the liquid seeds after activation, then by liquid seeds by volume 5% inoculum size be inoculated in liquid nutrient medium, under 70 ℃, 200r/min, cultivate 24h, make strain cell nutrient solution;
(2) strain cell nutrient solution step (1) being made is placed in pressure reactor, and at 1.5MPa pressure bottom fermentation 4h, fermentation condition is 70 ℃, 200r/min, makes fermented liquid;
(3) fermented liquid 1000mL step (2) being made centrifugal 20min under 5000r/min, collect somatic cells, somatic cells is resuspended in deionized water, make the suspension that cell mass concentration is 30wt%, suspension is cooled to 10 ℃ of left and right, the APV-2000-1 type high-pressure cell crusher that adopts German APV company to produce carries out three cytoclasis, and crusher working pressure is 60MPa, makes the cytoclasis solution 216mL that contains TreP; In cytoclasis liquid, add the polyoxyethylene glycol that ammonium sulfate and molecular weight are 2000, the mass percent concentration that makes ammonium sulfate in total system is that the mass percent concentration of 10wt%, polyoxyethylene glycol is 11wt%, mix, standing phase-splitting, phase solution on the polyoxyethylene glycol that makes phase solution under ammonium sulfate and contain TreP; In phase solution on polyoxyethylene glycol, add ammonium sulfate, making the mass percent concentration of ammonium sulfate in total system is 5wt%, and the pH value of regulator solution is 7.0, mixes, standing phase-splitting, makes under the ammonium sulfate that contains TreP phase solution on phase solution and polyoxyethylene glycol; By phase solution under the ammonium sulfate making, employing molecular weight cut-off is that the ultrafiltration membrance filter of 20000 Dal is concentrated, collects trapped fluid, and lyophilize, makes TreP 932mg.
The TreP vigor of dwelling in hot mycetocyte after testing, is 82.5U/g wet cell.
The hot bacteria strain of dwelling in described step (1) be thermus aquaticus (
thermus aquaticus) ATCC33923, this bacterial classification is purchased from U.S. representative microbial DSMZ (ATCC).
The component of the liquid seed culture medium in described step (1) is: glucose 10g/L, yeast extract paste 10g/L, peptone 10g/L, NaCl5g/L, pH7.5, distilled water 1000mL.
The component of the liquid nutrient medium in described step (1) is: glucose 20g/L, maltose 10g/L, yeast extract paste 5g/L, peptone 5g/L, KH
2pO
42g/L, MgSO
41g/L, pH7.5, distilled water 1000mL.
Embodiment 2
The dwell method of hot bacterium TreP content of raising, step is as follows:
(1) from test tube slant, the picking 1 ring hot bacteria strain of dwelling is inoculated in liquid seed culture medium, under 65 ℃, 100r/min, cultivate 12h, make the liquid seeds after activation, then by liquid seeds by volume 3% inoculum size be inoculated in liquid nutrient medium, under 60 ℃, 150r/min, cultivate 20h, make strain cell nutrient solution;
(2) strain cell nutrient solution step (1) being made is placed in pressure reactor, and at 2.0MPa pressure bottom fermentation 1h, fermentation condition is 60 ℃, 150r/min, makes fermented liquid;
(3) fermented liquid 1000mL step (2) being made centrifugal 20min under 5000r/min, collect somatic cells, somatic cells is resuspended in deionized water, make the suspension that cell mass concentration is 35wt%, suspension is cooled to 10 ℃ of left and right, the APV-2000-1 type high-pressure cell crusher that adopts German APV company to produce carries out three cytoclasis, and crusher working pressure is 60MPa, makes the cytoclasis solution 198mL that contains TreP; In cytoclasis liquid, add the polyoxyethylene glycol that ammonium sulfate and molecular weight are 2000, the mass percent concentration that makes ammonium sulfate in total system is that the mass percent concentration of 8wt%, polyoxyethylene glycol is 12wt%, mix, standing phase-splitting, phase solution on the polyoxyethylene glycol that makes phase solution under ammonium sulfate and contain TreP; In phase solution on polyoxyethylene glycol, add ammonium sulfate, making the mass percent concentration of ammonium sulfate in total system is 5wt%, and the pH value of regulator solution is 8.0, mixes, standing phase-splitting, makes under the ammonium sulfate that contains TreP phase solution on phase solution and polyoxyethylene glycol.By phase solution under the ammonium sulfate making, employing molecular weight cut-off is that the ultrafiltration membrance filter of 20000 Dal is concentrated, collects trapped fluid, and lyophilize, makes TreP 663mg.
The TreP vigor of dwelling in hot mycetocyte after testing, is 54.1U/g wet cell.
The component of the liquid seed culture medium in described step (1) is: glucose 10g/L, yeast extract paste 10g/L, peptone 10g/L, NaCl5g/L, pH7.5, distilled water 1000mL.
The component of the liquid nutrient medium in described step (2) is: glucose 20g/L, maltose 5g/L, yeast extract paste 5g/L, peptone 5g/L, KH
2pO
42g/L, MgSO
41g/L, pH7.5, distilled water 1000mL.
Embodiment 3
Improve as described in Example 1 the method for the hot bacterium TreP content of dwelling, difference is:
In step (2), strain cell nutrient solution is placed in pressure reactor, at 0.5MPa pressure bottom fermentation 6h, makes fermented liquid.
With this understanding, make TreP 862mg.
The TreP vigor of dwelling in hot mycetocyte after testing, is 78.1U/g wet cell.
Embodiment 4
Improve as described in Example 2 the method for the hot bacterium TreP content of dwelling, difference is:
In step (2), strain cell nutrient solution is placed in pressure reactor, at 1.0MPa pressure bottom fermentation 2h, makes fermented liquid.
With this understanding, make TreP 824mg.
The TreP vigor of dwelling in hot mycetocyte after testing, is 73.2U/g wet cell.
Embodiment 5
Improve as described in Example 1 the method for the hot bacterium TreP content of dwelling, difference is:
In step (1), the hot bacteria strain of dwelling be Thermus bacterium (
thermus sp.) ATCC 43815, this bacterial classification is purchased from U.S. representative microbial DSMZ (ATCC).
With this understanding, make TreP 918mg.
The TreP vigor of dwelling in hot mycetocyte after testing, is 84.2U/g wet cell.
Embodiment 6
Improve as described in Example 2 the method for the hot bacterium TreP content of dwelling, difference is:
In step (1), the hot bacteria strain of dwelling be Thermus bacterium (
thermus sp.) ATCC 43815, this bacterial classification is purchased from U.S. representative microbial DSMZ (ATCC).
With this understanding, make TreP 657mg.
The TreP vigor of dwelling in hot mycetocyte after testing, is 52.2U/g wet cell.
Comparative example 1
Improve as described in Example 1 the method for the hot bacterium TreP content of dwelling, difference is:
In step (2), strain cell nutrient solution is placed in pressure reactor, at normal pressure bottom fermentation 4h, fermentation condition is 70 ℃, 200r/min, makes fermented liquid.
With this understanding, make TreP 506mg.
The TreP vigor of dwelling in hot mycetocyte after testing, is 43.3U/g wet cell.
Comparative example 2
Improve as described in Example 6 the method for the hot bacterium TreP content of dwelling, difference is:
In step (2), get strain cell nutrient solution and be placed in pressure reactor, at normal pressure bottom fermentation 1h, fermentation condition is 60 ℃, 150r/min, makes fermented liquid.
With this understanding, make TreP 502mg.
The TreP vigor of dwelling in hot mycetocyte after testing, is 41.7U/g wet cell.
In above-described embodiment and comparative example, when the hot bacteria strain of dwelling under stress ferments, its intracellular TreP content increases, and enzyme activity also raises.In embodiment 1 and comparative example 1, the hot bacteria strain of dwelling is at 1.5MPa pressure bottom fermentation 4h than the normal pressure 4h that ferments, and its intracellular TreP content increases by 84.19%, enzyme activity raises 90.53%; In embodiment 6 and comparative example 2, the hot bacteria strain of dwelling is at 2.0MPa pressure bottom fermentation 1h than the normal pressure 1h that ferments, and its intracellular TreP content increases by 30.88%, enzyme activity raises 25.18%.Show thus, pressure can obviously improve the content of the TreP in the hot mycetocyte of dwelling.
Claims (5)
1. the raising method for hot bacterium TreP content of dwelling, is characterized in that, step is as follows:
(1) from test tube slant, the picking 1-2 ring hot bacteria strain of dwelling is inoculated in liquid seed culture medium, under 65～70 ℃, 100～150r/min, cultivate 12～16h, make the liquid seeds after activation, then by liquid seeds by volume 3%～5% inoculum size be inoculated in liquid nutrient medium, under 60～70 ℃, 150～200r/min, cultivate 20～24h, make strain cell nutrient solution;
(2) strain cell nutrient solution is placed in pressure reactor, at 0.5～2.0MPa pressure bottom fermentation, 1～6h, fermentation condition is 60～70 ℃, 150～200r/min, makes fermented liquid;
(3) fermented liquid is centrifugal, collect somatic cells, smudge cells, makes cytoclasis liquid, then adopts two phase aqueous extraction system to prepare TreP cytoclasis liquid.
2. the method for claim 1, is characterized in that, the hot bacteria strain of dwelling in described step (1) be thermus aquaticus (
thermos aquatics) ATCC33923 or Thermus bacterium (
thermos sp.) ATCC 43815.
3. the method for claim 1, is characterized in that, the component of the liquid seed culture medium in described step (1) is: glucose 10g/L, yeast extract paste 10g/L, peptone 10g/L, NaCl5g/L, pH7.5, distilled water 1000mL.
4. the method for claim 1, is characterized in that, the component of the liquid nutrient medium in described step (1) is: glucose 20g/L, maltose 5～10g/L, yeast extract paste 5g/L, peptone 5g/L, KH
2pO
42g/L, MgSO
41g/L, pH7.5, distilled water 1000mL.
5. the method for claim 1, it is characterized in that, the step that employing two phase aqueous extraction system in described step (3) is prepared TreP is as follows: in cytoclasis liquid, add the polyoxyethylene glycol that ammonium sulfate and molecular weight are 2000, the mass percent concentration that makes ammonium sulfate in total system is that the mass percent concentration of 8wt%～10wt%, polyoxyethylene glycol is 10wt%～12wt%, mix, standing phase-splitting, phase solution on the polyoxyethylene glycol that makes phase solution under ammonium sulfate and contain TreP; In phase solution on polyoxyethylene glycol, add ammonium sulfate, making the mass percent concentration of ammonium sulfate in total system is 5wt%～6wt%, and the pH value of regulator solution is 7.0～8.0, mixes, standing phase-splitting, makes under the ammonium sulfate that contains TreP phase solution on phase solution and polyoxyethylene glycol; By phase solution under the ammonium sulfate making, adopt ultrafiltration membrance filter concentrated, collect trapped fluid, lyophilize, makes TreP.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310729535.8A CN103695407B (en) | 2013-12-26 | 2013-12-26 | A kind of raising is dwelt the method for hot bacterium TreP content |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310729535.8A CN103695407B (en) | 2013-12-26 | 2013-12-26 | A kind of raising is dwelt the method for hot bacterium TreP content |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN103695407A true CN103695407A (en) | 2014-04-02 |
| CN103695407B CN103695407B (en) | 2016-01-20 |
Family
ID=50357071
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201310729535.8A Expired - Fee Related CN103695407B (en) | 2013-12-26 | 2013-12-26 | A kind of raising is dwelt the method for hot bacterium TreP content |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN103695407B (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104073479A (en) * | 2014-06-27 | 2014-10-01 | 齐鲁工业大学 | Method for purifying alliinase by double-water-phase separation |
| CN114525230A (en) * | 2022-03-31 | 2022-05-24 | 杭州优玛达生物科技有限公司 | Fermentation culture and fermentation method of thermophilic thermus using fermentation culture |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1216152C (en) * | 2003-07-18 | 2005-08-24 | 天津科技大学 | Method for raising yield of fucose produced from microzyme by using high pressure |
| CN103194432A (en) * | 2013-04-15 | 2013-07-10 | 山东天力药业有限公司 | Novel maltooligosyl trehalose synthase, gene of synthase, recombinant expression vector containing gene, and recombinant bacterium, and preparation of synthase |
-
2013
- 2013-12-26 CN CN201310729535.8A patent/CN103695407B/en not_active Expired - Fee Related
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1216152C (en) * | 2003-07-18 | 2005-08-24 | 天津科技大学 | Method for raising yield of fucose produced from microzyme by using high pressure |
| CN103194432A (en) * | 2013-04-15 | 2013-07-10 | 山东天力药业有限公司 | Novel maltooligosyl trehalose synthase, gene of synthase, recombinant expression vector containing gene, and recombinant bacterium, and preparation of synthase |
Non-Patent Citations (4)
| Title |
|---|
| YONGSHENG DONG 等: "Effects of high pressure on the accumulation of trehalose and glutathione in the Saccharomyces cerevisiae cells", 《BIOCHEMICAL ENGINEERING JOURNAL》, vol. 37, 31 December 2007 (2007-12-31) * |
| 吕斌 等: "双水相萃取技术及其在生物、食品工业中的应用", 《食品与发酵工业》, vol. 27, no. 6, 31 December 2001 (2001-12-31), pages 70 - 74 * |
| 胡耀辉 等: "水生栖热菌FL-03海藻糖合酶的分离纯化及特性研究", 《吉林农业大学学报》, vol. 28, no. 1, 31 December 2006 (2006-12-31) * |
| 范志华: "压力对酵母菌及其产海藻糖的影响", 《中国优秀硕士学位论文全文数据库（工程科技Ⅰ辑）》, no. 4, 15 December 2004 (2004-12-15) * |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104073479A (en) * | 2014-06-27 | 2014-10-01 | 齐鲁工业大学 | Method for purifying alliinase by double-water-phase separation |
| CN114525230A (en) * | 2022-03-31 | 2022-05-24 | 杭州优玛达生物科技有限公司 | Fermentation culture and fermentation method of thermophilic thermus using fermentation culture |
| CN114525230B (en) * | 2022-03-31 | 2024-04-09 | 杭州优玛达生物科技有限公司 | Fermentation culture and fermentation method of thermophilic thermus strain using fermentation culture |
Also Published As
| Publication number | Publication date |
|---|---|
| CN103695407B (en) | 2016-01-20 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN101935623B (en) | Agrobacterium ZX09, water-soluble beta-glucan prepared from Agrobacterium ZX09 and preparation method thereof and application on reducing blood sugar | |
| CN102220270B (en) | Screening method for producing chondroitin sulfate bacterial strain and application of bacterial strain fermentation method in production of chondroitin sulfate | |
| US11279961B2 (en) | Aspergillus oryzae BLCY-006 strain and application thereof in preparation of galactooligosaccharide | |
| CN102533605B (en) | Strain capable of producing levansucrase and method for producing levan by using levansucrase | |
| CN103468624B (en) | Genetic engineering bacteria used for high efficient production of mycose | |
| CN102586151B (en) | High-yield polysaccharide strain and method for producing polysaccharide by utilizing strain through fermentation | |
| CN111944730B (en) | Lactobacillus paracasei capable of efficiently utilizing jerusalem artichoke powder and application thereof | |
| CN102226159A (en) | Strain of Enterobacter cloacae and its application in the preparation of 2,3-butylene glycol | |
| CN101899407B (en) | Screening and application of bacillus licheniformis MEL09 with high 3-hydroxy butanone yield | |
| CN100558884C (en) | A kind of acid-producing Klebsiella bacterium and application thereof | |
| CN103695407B (en) | A kind of raising is dwelt the method for hot bacterium TreP content | |
| CN105567779A (en) | Fermentation method of high-yield and low-molecular-weight thermal gel | |
| CN106995789B (en) | Resveratrol zymocyte and application thereof | |
| CN105820988B (en) | One bacillus and its application in hot fermentation production 3-hydroxy-2-butanone and 2,3- butanediol | |
| CN104651284A (en) | Sphingomonas sp. T-3 and method for producing biological polysaccharide and poly-beta-hydroxy-butanoic acid by co-fermentation | |
| CN105176859A (en) | Strain MQO-153 for production of arginine deiminase | |
| CN104059857A (en) | Aspergillus and application of aspergillus in fructosyltransferase preparing | |
| CN109161569B (en) | Method for producing bacterial cellulose by fermenting tobacco extracts through microorganism combination | |
| CN103468606B (en) | Klebsiella oxytoca and application thereof in allitol production | |
| CN102363757A (en) | Screening method for 2-keto-D-gluconic acid high-yield bacterial strain, and fermentation method of such bacterial strain | |
| CN105950482B (en) | One plant of bacterial strain for producing inulinase and its application | |
| CN102399734B (en) | Thermophilic anaerobic strain and application thereof | |
| CN103173398B (en) | Short bacillus and method for preparing trehalose by virtue of fermentation | |
| CN105969811B (en) | The method for preparing pyrogallic acid using lactobacillus plantarum fermentation gallic acid | |
| CN102417888A (en) | Clostridium acetobutylicum for producing butanol by utilizing manihot as raw materials and application thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20160120 Termination date: 20171226 |
|
| CF01 | Termination of patent right due to non-payment of annual fee |