CN110592162A - Construction method of optimal fermentation condition of strain for producing 2' -fucosyllactose - Google Patents
Construction method of optimal fermentation condition of strain for producing 2' -fucosyllactose Download PDFInfo
- Publication number
- CN110592162A CN110592162A CN201910753844.6A CN201910753844A CN110592162A CN 110592162 A CN110592162 A CN 110592162A CN 201910753844 A CN201910753844 A CN 201910753844A CN 110592162 A CN110592162 A CN 110592162A
- Authority
- CN
- China
- Prior art keywords
- optimal
- fermentation
- selecting
- medium
- strain
- 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.)
- Pending
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
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P19/00—Preparation of compounds containing saccharide radicals
- C12P19/12—Disaccharides
Abstract
The invention relates to the technical field of escherichia coli strains, firstly, a single-factor level experiment is adopted, the experiment analysis is carried out on the factors of the fermentation rotating speed, the inoculation amount, the liquid loading amount, the time, the temperature, the nitrogen source, the carbon source, the pH value and the lactose concentration to obtain the optimal single-factor level, then, a representative level combination is selected to carry out an orthogonal experiment and analysis to determine the optimal fermentation condition, the content of the obtained 2 '-fucosyllactose is improved by 2.5 percent after the fermentation, the yield is 3.5g/L, the structure and the property of the obtained 2' -fucosyllactose are the same as those of the oligosaccharide 2 '-fucosyllactose in human milk, and the 2' -fucosyllactose is used as a nutrition enhancer and is one of sources of prebiotic substances and is commonly used for infant formula foods, infant formula foods for larger infants and children and infant formula foods.
Description
Technical Field
The invention relates to the technical field of escherichia coli strains, in particular to a construction method of an optimal fermentation condition of a strain for producing 2' -fucosyllactose.
Background
Breast milk is an important source of nutrition for infants, and among infant nutrition, milk powder is the third most abundant solid nutrient, in which Human Milk Oligosaccharides (HMO) are contained, which play a key role in infant health, and fucosylated HMO, including 2 '-fucosyllactose (2' -FL), 3-fucose (3-FL), lactoneon-tetraose (LNnT), etc., which selectively stimulate the growth of bifidobacteria, and can form analogues of pathogen receptors, thereby protecting infants against infection by intestinal pathogens, such as enteropathogenic escherichia coli, vibrio cholerae, and salmonella. Furthermore, fucosylated HMOs play a key role in inflammation and tumor metastasis like the Lewis antigen sugar chain structure. The ability to recognize and metastasize the structure of tumor cells has attracted considerable pharmaceutical interest in fucosylated HMOs, 2' -FL, where they inhibit bacterial or viral adhesion to epithelial cells due to their potential use in nutrition and health care.
Companies have obtained a method capable of producing 2' -fucosyllactose by genetic engineering methods, but the yield thereof is low, the production thereof is limited, and the cost is increased.
The development of metabolic engineering and fermentation microbiology provides possibility for improving the microorganism to synthesize 2 '-fucose lactose, and researches on the content of 2' -fucose lactose are improved by controlling the fermentation conditions and processes and preventing the contamination of the fermentation process.
Disclosure of Invention
In order to solve the problems in the background art, the invention aims to provide a construction method of the optimal fermentation condition of the strain for producing the 2' -fucosyllactose, which provides possibility for being used in infant formula milk powder in the future and can greatly reduce the production cost.
In order to solve the technical problems, the technical scheme provided by the invention is as follows: a construction method of the optimal fermentation condition of a strain for producing 2' -fucosyllactose is prepared by the following steps: (1) initial fermentation medium: 10g/L glycerol, 12g/L LK2HPO4, 3g/L KH2PO4, 5g/L (NH4)2SO4, 0.1g/L NaCl, MgSO 4.7H 2O.0.3 g/L CaCl 2.2H 2O.015 g/L, FeSO 4.7H 2O/sodium citrate, 15mL/L (7.5g/LFeSO 4.7H 2O plus 100g/L sodium citrate), 7.5 μ g/L thiamine, 1g/L yeast powder, 1mL/L metallic elements (10g/L MnSO 4. nH2O, 4g/L CoCl2, 2g/L Na2MoO 4. multidot.2H 2 BO O, 2g/L ZnSO 4.7H 2O, 1g/L CuCl 2. multidot.2H 2 BO, 0.5g/L Na2MoO 4. multidot. O, pH 593.593 g/L592 BO, pH 9);
(2) activating strains: collecting a preservation strain E.coli K-12 ZK-2 in a refrigerator at-80 ℃, inoculating the preservation strain into an LB culture medium, and activating overnight;
(3) fermentation: inoculating Escherichia coli E.coli K-12 ZK-2 bacterial liquid into a culture medium, fermenting in a fermentation shake flask, adding 0.1mM isopropyl thiogalactoside and lactose when OD600 value reaches 0.6, and inducing expression;
(4) single factor, multi-level experiment: A. rotational speed
5 rotation speed levels of 140rpm, 160rpm, 180rpm, 200rpm and 220rpm are respectively selected, an initial fermentation culture medium is adopted for carrying out a fermentation experiment, other factors are unchanged, and the yield of the 2' -FL is measured after the fermentation is carried out for 24 hours;
B. amount of inoculation
Respectively selecting 5 inoculation quantity levels of 0.5%, 1%, 1.5%, 2% and 2.5%, performing fermentation experiment by using an initial fermentation culture medium, measuring the OD600 of the initial fermentation culture medium every other hour to measure the growth condition of the initial fermentation culture medium, and measuring the yield of 2' -FL of the initial fermentation culture medium after 24 hours of fermentation;
C. liquid loading amount
Respectively selecting 10%, 20%, 30%, 40% and 50% of 5 liquid loading levels, performing fermentation experiments by adopting an initial fermentation culture medium, selecting the optimal rotation speed condition at the rotation speed, selecting the optimal inoculation quantity, and keeping other factors unchanged. Measuring the OD600 of the fermented soybean every other hour to measure the growth condition of the fermented soybean, and measuring the 2' -FL yield of the fermented soybean after 24 hours of fermentation;
D. pH value
Adjusting the pH value of the culture medium to 3.0, 4.5, 6.0, 7.5 and 9.05 pH values by NaOH and HCI respectively, performing fermentation experiments by adopting an initial fermentation culture medium, selecting the optimal rotating speed condition by rotating speed, selecting the optimal inoculation amount by the inoculation amount, selecting the optimal liquid loading amount by the liquid loading amount, and keeping other factors unchanged. Measuring the OD600 of the strain at intervals of one hour in an initial growth stage to measure the growth condition of the strain, and then measuring the yield of 2' -FL of the strain after fermenting for 24 hours in a product generation stage;
E. time of fermentation
Selecting the optimal rotation speed condition, selecting the optimal inoculation amount, selecting the optimal liquid loading amount, selecting the optimal pH value as the pH value, and sampling every 10, 20, 30, 40 and 50 hours to measure the 2' -FL yield, wherein other factors are unchanged;
F. carbon source
Selecting three carbon sources of glucose, sucrose and glycerol, fermenting respectively according to the addition amount of 10g/L, selecting the medium-optimal rotation speed condition at the rotation speed, selecting the medium-optimal inoculation amount at the inoculation amount, selecting the medium-optimal liquid loading amount at the liquid loading amount, selecting the medium-optimal pH value at the pH value, and keeping other factors unchanged. Measuring the OD600 of the fermented soybean every other hour to measure the growth condition of the fermented soybean, and measuring the yield of 2' -FL after the optimal fermentation time;
G. nitrogen source
NH4NO3, (NH4)2SO4, NH4Cl, urea, peptone and yeast powder 6 nitrogen sources are fermented respectively according to the addition amount of 5g/L, the rotation speed is selected from the medium optimal rotation speed condition, the inoculation amount is selected from the medium optimal inoculation amount, the liquid loading amount is selected from the medium optimal liquid loading amount, the pH value is selected from the medium optimal pH value, the carbon source is selected from the medium optimal carbon source, other factors are unchanged, the OD600 is measured every one hour to measure the growth condition, and the 2' -FL yield is measured after the optimal fermentation time;
H. temperature of
Selecting 5 temperature gradients of 16 ℃, 22 ℃, 28 ℃, 34 ℃ and 40 ℃, selecting the medium-optimal rotating speed condition at the rotating speed, selecting the medium-optimal inoculation amount at the inoculation amount, selecting the medium-optimal liquid loading amount at the liquid loading amount, selecting the medium-optimal pH value at the pH value, selecting the medium-optimal carbon source at the carbon source, selecting the medium-optimal nitrogen source at the nitrogen source, keeping other factors unchanged, sampling after the optimal fermentation time, and measuring the yield of 2' -FL;
I. lactose concentration
Respectively selecting 5 temperature gradients of 1g/L, 2g/L, 3g/L, 4g/L and 5g/L, selecting the condition of the rotation speed to be the optimal rotation speed, selecting the inoculation amount to be the optimal inoculation amount, selecting the liquid loading amount to be the optimal liquid loading amount, selecting the pH value to be the optimal pH value, selecting the carbon source to be the optimal carbon source, selecting the nitrogen source to be the optimal nitrogen source, selecting the optimal temperature to be the temperature, keeping other factors unchanged, sampling after the optimal fermentation time, and measuring the yield of 2' -FL.
(5) Multifactor, multilevel experiments: selecting four factors with larger influence factors, namely a nitrogen source, time B, temperature C and lactose concentration D, designing three levels 1, 2 and 3, performing a group of orthogonal tests, and finally determining the optimal component.
As a modification, the liquid content of the culture medium in the fermentation of the E.coli K-12 ZK-2 strain in the step (1) is controlled to be 10%.
As an improvement, the pH value of the culture medium of the E.coli K-12 ZK-2 strain in the step (1) is controlled to be 7.2 during fermentation.
As an improvement, the temperature for activating the E.coli K-12 ZK-2 strain in the step (2) is controlled within 37 ℃.
As an improvement, the temperature of the E.coli K-12 ZK-2 strain in the step (3) is controlled within 37 ℃ before the DO600 reaches 0.6.
As an improvement, the zymophyte liquid in the step (3) is inoculated into a culture medium according to the inoculation amount of 1%.
As an improvement, when the escherichia coli strain e.coli K-12 ZK-2 in the step (3) is fermented, the final concentration of lactose to be added is set at 2g/L, and the production of 2' -fucosyllactose can be realized.
As an improvement, the fermentation time of the Escherichia coli strain E.coli K-12 ZK-2 in the step (3) is controlled within 24 hours.
As an improvement, when the Escherichia coli strain E.coli K-12 ZK-2 in the step (3) is fermented, the rotation speed is adjusted to 120 rpm.
As an improvement, when the escherichia coli strain E.coli K-12 ZK-2 in the step (4) is fermented, the initial fermentation medium and the culture conditions are consistent with those in the step (3).
The invention has the beneficial effects that: the 2' -fucosyllactose is prepared by adopting the escherichia coli strain E.coli K-12 ZK-2, and the shake flask yield is increased from 1g/L to 3.5g/L through the optimization of fermentation conditions, so that the possibility is provided for being used in infant formula milk powder in the future, and the production cost can be greatly reduced.
Drawings
FIG. 1 is an experimental grouping of the multifactor, multi-level experiment of the present invention.
Detailed Description
The invention is illustrated below by means of specific examples, without being restricted thereto.
Example one
Coli K-12 ZK-2 fermentation optimization preparation method: inoculating the fresh culture of Escherichia coli E.coli K-12 ZK-2 strain into fermentation medium, and culturing at 34 deg.C for 24 hr.
The culture medium comprises the following components in percentage by weight: 10g/L glycerol, 12g/L K HPO4, 3g/L KH2 5964, 5g/L peptone, 0.1g/L NaCl, MgSO4 & 7H 2O.3 g/L CaCl2 & 2H 2O.015 g/L, FeSO4 & 7H 2O/sodium citrate 15mL/L (7.5g/LFeSO4 & 7H2O plus 100g/L sodium citrate) 7.5. mu.g/L thiamine, 1g/L yeast powder, 1mL/L metallic elements (10g/L MnSO4 & nH2O, 4g/L CoCl2, 2g/L Na2MoO4 & 2H2O, 2g/L O4 & 7H2O, 1g/L LCuCl2 & 2H2O, 0. 3/L BO 3.9, pH 9.7 BO 2. multidot.7).
The inoculation amount of the bacterial liquid is 2.5 percent of the volume of the culture medium, the liquid loading amount is 10 percent, the initial pH value is 7.2, the bacterial liquid and the liquid are fully and uniformly mixed and then are placed into a fermentation shake flask, the rotation speed is adjusted to 190rpm under the conditions of 37 ℃ and 220rpm when the OD600 value reaches 0.6, 0.1mM IPTG is added to induce expression at 34 ℃ and 10g/L lactose is added to induce expression for 5 d.
The above description is only for the preferred embodiment of the present invention, but the scope of the present invention is not limited thereto, and any person skilled in the art should be able to cover the technical scope of the present invention, the technical solutions and the inventive concepts of the present invention with equivalent or modified alternatives and modifications within the technical scope of the present invention.
Claims (10)
1. A construction method of the optimal fermentation condition of a strain for producing 2' -fucosyllactose is characterized by comprising the following steps: (1) initial fermentation medium: 10g/L glycerol, 12g/L K2HPO4, 3g/L KH2PO4, 5g/L (NH4)2SO4, 0.1g/L NaCl, MgSO4 & 7H2O0.3g/L CaCl 2.2 H2O0.015g/L, FeSO4 & 7H 2O/sodium citrate, 15mL/L (7.5g/LFeSO4 & 7H2O plus 100g/L sodium citrate), 7.5 μ g/L thiamine, 1g/L yeast powder, 1mL/L metallic element (10g/L MnSO4 & nH2O, 4g/L CoCl2, 2g/LNa 2MoO4 & 2H2O, 2g/L pHO 4 & 7H2O, 1g/L LCuCl2 & 2H2O, 670.5 g/L BO 3. 3.);
(2) activating strains: collecting a preservation strain E.coli K-12 ZK-2 in a refrigerator at-80 ℃, inoculating the preservation strain into an LB culture medium, and activating overnight;
(3) fermentation: inoculating Escherichia coli E.coli K-12 ZK-2 bacterial liquid into a culture medium, fermenting in a fermentation shake flask, adding 0.1mM isopropyl thiogalactoside and lactose when OD600 value reaches 0.6, and inducing expression;
(4) single factor, multi-level experiment: A. rotational speed
5 rotation speed levels of 140rpm, 160rpm, 180rpm, 200rpm and 220rpm are respectively selected, an initial fermentation culture medium is adopted for carrying out a fermentation experiment, other factors are unchanged, and the yield of the 2' -FL is measured after the fermentation is carried out for 24 hours;
B. amount of inoculation
Respectively selecting 5 inoculation quantity levels of 0.5%, 1%, 1.5%, 2% and 2.5%, performing fermentation experiment by using an initial fermentation culture medium, measuring the OD600 of the initial fermentation culture medium every other hour to measure the growth condition of the initial fermentation culture medium, and measuring the yield of 2' -FL of the initial fermentation culture medium after 24 hours of fermentation;
C. liquid loading amount
Respectively selecting 10%, 20%, 30%, 40% and 50% of 5 liquid loading levels, performing fermentation experiments by adopting an initial fermentation culture medium, selecting the optimal rotation speed condition at the rotation speed, selecting the optimal inoculation quantity, keeping other factors unchanged, measuring the OD600 of the strain every other hour to measure the growth condition of the strain, and measuring the yield of 2' -FL of the strain after fermenting for 24 hours;
D. pH value
Adjusting the pH value of the culture medium to 3.0, 4.5, 6.0, 7.5 and 9.05 pH values by NaOH and HCI respectively, performing fermentation experiments by adopting an initial fermentation culture medium, selecting the optimal rotating speed condition by rotating speed, selecting the optimal inoculation amount by the inoculation amount, selecting the optimal liquid loading amount by the liquid loading amount, and keeping other factors unchanged. Measuring the OD600 of the strain at intervals of one hour in an initial growth stage to measure the growth condition of the strain, and then measuring the yield of 2' -FL of the strain after fermenting for 24 hours in a product generation stage;
E. time of fermentation
Selecting the optimal rotation speed condition, selecting the optimal inoculation amount, selecting the optimal liquid loading amount, selecting the optimal pH value as the pH value, and sampling every 10, 20, 30, 40 and 50 hours to measure the 2' -FL yield, wherein other factors are unchanged;
F. carbon source
Selecting three carbon sources of glucose, sucrose and glycerol, fermenting respectively according to the addition amount of 10g/L, selecting the medium-optimal rotating speed condition at the rotating speed, selecting the medium-optimal inoculation amount, selecting the medium-optimal liquid loading amount, selecting the medium-optimal pH value at the pH value, keeping other factors unchanged, measuring the OD600 of the medium-optimal liquid loading amount every other hour to measure the growth condition of the medium-optimal liquid loading amount, and measuring the yield of 2' -FL of the medium-optimal liquid loading amount after the optimal fermentation time;
G. nitrogen source
NH4NO3, (NH4)2SO4, NH4Cl, urea, peptone and yeast powder 6 nitrogen sources are fermented respectively according to the addition amount of 5g/L, the rotation speed selects the medium optimal rotation speed condition, the inoculation amount selects the medium optimal inoculation amount, the liquid loading amount selects the medium optimal liquid loading amount, the pH value selects the medium optimal pH value, the carbon source selects the medium optimal carbon source, other factors are unchanged, the OD600 is measured every one hour to measure the growth condition, and the 2' -FL yield is measured after the optimal fermentation time;
H. temperature of
Selecting 5 temperature gradients of 16 ℃, 22 ℃, 28 ℃, 34 ℃ and 40 ℃, selecting the medium-optimal rotating speed condition at the rotating speed, selecting the medium-optimal inoculation amount at the inoculation amount, selecting the medium-optimal liquid loading amount at the liquid loading amount, selecting the medium-optimal pH value at the pH value, selecting the medium-optimal carbon source at the carbon source, selecting the medium-optimal nitrogen source at the nitrogen source, keeping other factors unchanged, sampling after the optimal fermentation time, and measuring the yield of 2' -FL;
I. lactose concentration
Respectively selecting 5 temperature gradients of 1g/L, 2g/L, 3g/L, 4g/L and 5g/L, selecting the medium-optimal rotating speed condition by rotating speed, selecting the medium-optimal inoculation amount by inoculation amount, selecting the medium-optimal liquid filling amount by liquid filling amount, selecting the medium-optimal pH value by pH value, selecting the medium-optimal carbon source by carbon source, selecting the medium-optimal nitrogen source by nitrogen source, selecting the optimal temperature by temperature, and keeping other factors unchanged. After the optimal fermentation time, samples were taken and the 2' -FL yield was determined.
(5) Multifactor, multilevel experiments: selecting four factors with larger influence factors, namely a nitrogen source, time B, temperature C and lactose concentration D, designing three levels 1, 2 and 3, performing a group of orthogonal tests, and finally determining the optimal component.
2. The method for constructing the optimal fermentation condition of the strain for producing 2' -fucosyllactose as claimed in claim 1, wherein the liquid content of the culture medium in the fermentation of the E.coli K-12 ZK-2 strain of step (1) is controlled to 10%.
3. The method for constructing the optimal fermentation condition of the strain for producing 2' -fucosyllactose as claimed in claim 1, wherein the pH value of the culture medium of the E.coli K-12 ZK-2 strain of step (1) is controlled to be 7.2 during fermentation.
4. The method for constructing the optimal fermentation condition of the strain for producing 2' -fucosyllactose as claimed in claim 1, wherein the temperature for activating the E.coli K-12 ZK-2 strain in step (2) is controlled within 37 ℃.
5. The method for constructing the optimal fermentation condition of the strain for producing 2' -fucosyllactose as claimed in claim 1, wherein the temperature of the E.coli K-12 ZK-2 strain in the step (3) is controlled within 37 ℃ before the DO600 reaches 0.6.
6. The method for constructing the optimal fermentation condition of the strain for producing 2' -fucosyllactose as claimed in claim 1, wherein the fermentation broth of step (3) is inoculated into the culture medium in an amount of 1%.
7. The method for constructing the optimal fermentation conditions of the strain for producing 2 '-fucosyllactose as claimed in claim 1, wherein the final concentration of lactose to be added is set at 2g/L during the fermentation of the E.coli strain E.coli K-12 ZK-2 in step (3), so as to realize the production of 2' -fucosyllactose.
8. The method for constructing the optimal fermentation conditions of the strain for producing 2' -fucosyllactose as claimed in claim 1, wherein the fermentation time of the E.coli strain E.coli K-12 ZK-2 in step (3) is controlled within 24 hours.
9. The method for constructing the optimal fermentation conditions for the strain producing 2' -fucosyllactose as claimed in claim 1, wherein the rotation speed is adjusted to 120rpm when the E.coli strain E.coli K-12 ZK-2 in step (3) is fermented.
10. The method for constructing the optimal fermentation conditions of the strain for producing 2' -fucosyllactose as claimed in claim 1, wherein the initial fermentation medium and the culture conditions of the strain E.coli K-12 ZK-2 in step (4) are the same as those in step (3).
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910753844.6A CN110592162A (en) | 2019-08-15 | 2019-08-15 | Construction method of optimal fermentation condition of strain for producing 2' -fucosyllactose |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910753844.6A CN110592162A (en) | 2019-08-15 | 2019-08-15 | Construction method of optimal fermentation condition of strain for producing 2' -fucosyllactose |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN110592162A true CN110592162A (en) | 2019-12-20 |
Family
ID=68854349
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201910753844.6A Pending CN110592162A (en) | 2019-08-15 | 2019-08-15 | Construction method of optimal fermentation condition of strain for producing 2' -fucosyllactose |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN110592162A (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2022122721A1 (en) * | 2020-12-11 | 2022-06-16 | Dsm Ip Assets B.V. | Improved prebiotic formulations |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2016095924A1 (en) * | 2014-12-16 | 2016-06-23 | Glycom A/S | Separation of 2'-fl from a fermentation broth |
| CN106190937A (en) * | 2016-07-18 | 2016-12-07 | 南开大学 | A kind of method building recombination bacillus coli biosynthesis 2 ' rock algae lactose |
| CN109609422A (en) * | 2018-11-20 | 2019-04-12 | 安徽天凯生物科技有限公司 | One kind can produce the construction method of 2 '-rock algae base lactose bacterial strains |
-
2019
- 2019-08-15 CN CN201910753844.6A patent/CN110592162A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2016095924A1 (en) * | 2014-12-16 | 2016-06-23 | Glycom A/S | Separation of 2'-fl from a fermentation broth |
| CN106190937A (en) * | 2016-07-18 | 2016-12-07 | 南开大学 | A kind of method building recombination bacillus coli biosynthesis 2 ' rock algae lactose |
| CN109609422A (en) * | 2018-11-20 | 2019-04-12 | 安徽天凯生物科技有限公司 | One kind can produce the construction method of 2 '-rock algae base lactose bacterial strains |
Non-Patent Citations (2)
| Title |
|---|
| 王华: "《定点突变β-环糊精葡萄糖基转移酶及其突变菌株发酵条件优化》", 30 November 2016, 黑龙江大学出版社 * |
| 胡兰兰等: "重组大肠杆菌E. coli BL21/pET28a(+)-mle产苹果酸乳酸酶发酵条件的优化", 《中国酿造》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2022122721A1 (en) * | 2020-12-11 | 2022-06-16 | Dsm Ip Assets B.V. | Improved prebiotic formulations |
| CN116615203A (en) * | 2020-12-11 | 2023-08-18 | 帝斯曼知识产权资产管理有限公司 | Improved prebiotic formulation |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Zhao et al. | Isolation and characterization of a gamma-aminobutyric acid producing strain Lactobacillus buchneri WPZ001 that could efficiently utilize xylose and corncob hydrolysate | |
| CN110734884B (en) | Lactobacillus paracasei low-salt culture medium and culture method | |
| CN101037661A (en) | Pseudoalteromonas and its usage | |
| CN103409480B (en) | Method for producing Pulullans with different molecular weights | |
| CN105018544B (en) | With the carrageenan oligosaccharide and its preparation method and application for promoting sea cucumber growth and degeneration-resistant effect | |
| WO2021082352A1 (en) | Method for synthesizing lacto-n-biose | |
| CN102258059A (en) | Method for preparing ralstonia solanacearum-resistant bacillus subtilis microbial inoculum | |
| CN115141757A (en) | Aureobasidium pullulans strain and method for producing pullulan polysaccharide by fermentation method thereof | |
| CN111820419A (en) | Composition for targeted regulation and control of enteron-bacterium and short-chain fatty acid producing bacterium | |
| CN102994430B (en) | Bacterial cellulose production strain and application thereof | |
| CN109880786A (en) | A method of promoting Bacillus coagulans spore formation | |
| JPS5953834B2 (en) | Bifidobacterium growth promoter | |
| CN110592162A (en) | Construction method of optimal fermentation condition of strain for producing 2' -fucosyllactose | |
| CN112391318B (en) | Preparation method of bifidobacterium longum and selenium-enriched microorganism and preparation method of freeze-dried preparation thereof | |
| CN113046253B (en) | Culture method for improving heat resistance of kluyveromyces marxianus | |
| CN112852891A (en) | Artificial dual-bacterium system for producing mcl-PHA and application thereof | |
| CN112592860A (en) | Bifidobacterium infantis fermentation medium without animal-derived nitrogen source and application thereof | |
| Ramya et al. | Studies on the production and optimization of levan from Bacillus sp | |
| CN100390295C (en) | Microorganism polysaccharide and its preparation method and application | |
| CN115895974A (en) | Lactobacillus plantarum rich in selenium and capable of producing gamma-aminobutyric acid at high yield and application of lactobacillus plantarum | |
| Takagi et al. | Characterization of polysaccharide production of Haemophilus influenzae type b and its relationship to bacterial cell growth | |
| CN101982089B (en) | Micro-ecological preparation for livestock, preparation method and application thereof | |
| CN114573732A (en) | Preparation method and application of xylan zinc complex with probiotic effect | |
| CN108192848A (en) | A kind of Psychrobacter bacterial strain of galactopoiesis carbohydrase and the method that low temperature lactase is prepared using the bacterial strain | |
| Al-Baarri et al. | Glucose and D-Allulose contained medium to support the growth of lactic acid bacteria |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20191220 |
|
| RJ01 | Rejection of invention patent application after publication |