CN102643749B - Microorganism low-temperature preservation protective agent and microorganism low-temperature preservation method - Google Patents
Microorganism low-temperature preservation protective agent and microorganism low-temperature preservation method Download PDFInfo
- Publication number
- CN102643749B CN102643749B CN201210132750.5A CN201210132750A CN102643749B CN 102643749 B CN102643749 B CN 102643749B CN 201210132750 A CN201210132750 A CN 201210132750A CN 102643749 B CN102643749 B CN 102643749B
- Authority
- CN
- China
- Prior art keywords
- temperature
- microorganism
- low
- preserved
- microorganism low
- 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.)
- Expired - Fee Related
Links
Landscapes
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The invention provides a microorganism low-temperature preservation protective agent, which contains oligosaccharide, glycerol, mannitol, defatted milk powder and agarose. The invention further provides a microorganism low-temperature preservation method, which comprises mixing the microorganism low-temperature preservation protective agent with water and then with microorganisms to be preserved, and then enabling mixed materials to be placed at preservation temperature; or enabling the microorganisms to be preserved to be mixed with the microorganism low-temperature preservation protective agent containing the water, and then enabling the mixed materials to be placed at the preservation temperature. By means of the technical scheme, the survival rate and thallus activity of the microorganisms after the low-temperature preservation are obviously improved.
Description
Technical field
The present invention relates to bioengineering field, particularly, relate to a kind of microorganism low-temperature and preserve protective material and Cryopreservation.
Background technology
Microbial strains is important Biological resources, and culture presevation is important microorganism element task.Culture presevation utilize exactly a blanking method make bacterial classification dead, do not wane, constant so that research and application.Different strain or different requirement, be suitable for different method for preserving, the method for preserving of general bacterial cultures: slant preservation, whiteruss preservation, lyophil preservation, nitrogen ultra low temperature preserving process, cryogenic freezing preserving process.
Slant preservation method, preservation term is short; Whiteruss preserving process, must uprightly deposit, takes up space large, and not Portable belt; Freeze-drying preservation, nitrogen ultra low temperature preserving process, technical sophistication, with high content of technology, equipment investment is high, and running cost is high, is applicable to professional preservation mechanism, is not suitable for manufacturing enterprise.
Low temperature preservation is that bacterial classification is placed on to a kind of method for preserving carrying out to slow down the physiological activity of cell in cryogenic refrigerator.The method for preserving adopting at present has two kinds: one is that 4 DEG C of refrigerators are directly preserved (short-term); The 2nd,, in bacterium liquid, add a certain proportion of glycerine to mix latter-80 DEG C and preserve (medium-term and long-term).
No matter cryopreservation, be 4 DEG C or-20 DEG C ,-80 DEG C, adds a certain proportion of protective material, can effectively protect microorganism active, improves thalline survival rate.And due to various microbe species and structure difference, its best protection agent prescription in cryopreservation is also different.At present, existing microorganism low-temperature preservation protective material is at least one in glycerine, N.F,USP MANNITOL and skim-milk.
But the results show, preserves under protective material existence at existing microorganism low-temperature, survival rate and the microbial activity of microorganism after cryopreservation is still not high.
Summary of the invention
The object of the invention is to overcome at existing microorganism low-temperature and preserve under protective material existence; still not high defect of the survival rate of microorganism after cryopreservation and microbial activity; provide a kind of microorganism low-temperature to preserve protective material and a kind of microorganism low-temperature preservation guard method, so that survival rate and the microbial activity of microorganism after cryopreservation is improved.
To achieve these goals, on the one hand, the invention provides a kind of microorganism low-temperature and preserve protective material, this microorganism low-temperature is preserved protective material and is contained oligose, glycerine, N.F,USP MANNITOL, skim-milk and agarose.
On the other hand, the present invention also provides a kind of microorganism low-temperature store method, this microorganism low-temperature store method comprises: microorganism low-temperature as above is preserved after protective material mixes with water and mixed with microorganism to be preserved, then mixed material is placed under storage temperature; Or microorganism to be preserved and the microorganism low-temperature that contains water are preserved to protective material and mix, then mixed material is placed under storage temperature.
By technique scheme, survival rate and the microbial activity of microorganism after cryopreservation is significantly improved.Particularly; use microorganism low-temperature provided by the invention to preserve protective material; the survival rate that can make escherichia coli (Escherichia coli) preserve at the temperature of subzero 20 DEG C after half a year still can reach 95%; and to use final concentration as the glycerine of 15 % by weight is as protective material, the survival rate that escherichia coli was preserved after half a year at the temperature of subzero 20 DEG C is only 85%.In addition; use microorganism low-temperature provided by the invention to preserve protective material; the seeding tank cycle that can make brevibacterium flavum preserve at the temperature of 4 DEG C after 7 days is only 15h; and to use final concentration as the glycerine of 15 % by weight is as protective material, the seeding tank cycle that brevibacterium flavum preserved after 7 days at the temperature of 4 DEG C reaches 18h.
In the present invention, term " seeding tank cycle " is defined as 10
9after the preservation of individual viable bacteria body experience, be all inoculated in LB substratum and cultivate to 10
12the needed incubation time of the individual viable bacteria scale of construction.Microbial activity after experience preservation is higher, and the seeding tank cycle is shorter.
Other features and advantages of the present invention are described in detail the embodiment part subsequently.
Embodiment
Below the specific embodiment of the present invention is elaborated.Should be understood that, embodiment described herein only, for description and interpretation the present invention, is not limited to the present invention.
Preserve protective material according to microorganism low-temperature provided by the invention, this microorganism low-temperature is preserved protective material and is contained oligose, glycerine, N.F,USP MANNITOL, skim-milk and agarose.
Wherein, it can be the composition of solid state that this microorganism low-temperature is preserved protective material, also can further add water and be mixed into liquid composition.Typically, this microorganism low-temperature is preserved the composition that protectant storing state is solid state, and use state is liquid composition.
Preserve protective material according to microorganism low-temperature provided by the invention; wherein; the content of oligose, glycerine, N.F,USP MANNITOL, skim-milk and agarose can change in a big way; for example; with respect to the oligose of 100 weight parts, the content of glycerine can be 20-70 weight part, and the content of N.F,USP MANNITOL can be 20-70 weight part; the content of skim-milk can be 100-200 weight part, and the content of agarose can be 0.1-1.0 weight part.
Under preferable case, with respect to the oligose of 100 weight parts, the content of glycerine is 40-70 weight part, and the content of N.F,USP MANNITOL is 40-60 weight part, and the content of skim-milk is 150-200 weight part, and the content of agarose is 0.4-0.8 weight part.
Preserve protective material according to microorganism low-temperature provided by the invention, wherein, for making described microorganism low-temperature preserve the convenient directly use of protective material, under preferable case, described microorganism low-temperature is preserved protective material and is also contained water.
Wherein, with respect to the oligose of 100 weight parts, the content of water can be 800-1500 weight part, is preferably 900-1200 weight part.
Wherein, can preserve more uniformly and stably and use in the situation that containing water in order to make described microorganism low-temperature preserve protective material, under preferable case, the microorganism low-temperature that contains water be preserved to protective material and be heated to 90 DEG C and be cooled to room temperature after above.
Preserve protective material according to microorganism low-temperature provided by the invention, wherein, described oligose is the material being connected and composed by glycosidic link by 2-10 monosaccharide unit; typically; described oligose is disaccharides, and under preferable case, described oligose is at least one in trehalose, sucrose and maltose.Described oligose can be all conventional commercially available product.
Preserve protective material according to microorganism low-temperature provided by the invention, wherein, described agarose is to add described microorganism low-temperature to the form of the agarose of agar and/or purification to preserve in protective material, and in described agar, the content of agarose is at least 70 % by weight.The agarose of described agar and described purification can be all conventional commercially available product.Wherein, glycerine, N.F,USP MANNITOL and skim-milk can be all conventional commercially available product.Described skim-milk refers to the skimmed milk powder that meets GB 5410-1999 standard.Well known to a person skilled in the art to be, should be clean for the above-mentioned commercially available product of microorganism field, for example, can be food grade, chemical pure or analytically pure.
The present invention also provides a kind of microorganism low-temperature store method, this microorganism low-temperature store method comprises: microorganism low-temperature as above is preserved after protective material mixes with water and mixed with microorganism to be preserved, then mixed material is placed under storage temperature; Or microorganism to be preserved and the microorganism low-temperature that contains water are preserved to protective material and mix, then mixed material is placed under storage temperature.
Wherein, the method for described mixing does not have special requirement, can be conventional piping and druming and/or vibration.Wherein, described microorganism can be for cultivating the culture obtaining, the microbe that also can obtain for this culture centrifugation.
According to microorganism low-temperature store method provided by the invention, wherein, described microorganism low-temperature is preserved protective material and can in a big way, be changed with respect to the consumption of a certain amount of microorganism, with respect to 10
9individual microorganism, it can be 1-5 gram that described microorganism low-temperature is preserved protectant consumption, is preferably 1-3 gram.
According to microorganism low-temperature store method provided by the invention, wherein, described storage temperature can be the storage temperature that the microorganism low-temperature of this area routine is preserved, and typically, described storage temperature can be the arbitrary temp between subzero 260 DEG C to 5 DEG C.Typically, can be conventional various device for the equipment of microorganism low-temperature store method, refrigerator, the temperature setting that refrigerator, the temperature setting that refrigerator, the temperature setting that such as temperature setting is set to 4 DEG C of left and right is set to subzero 20 DEG C of left and right is set to subzero 80 DEG C of left and right be set to the refrigerator of subzero 160 DEG C of left and right or liquid nitrogen and preserves tank (subzero 196 DEG C below) etc.Usually, storage temperature is lower, and the effect of preservation is better.In order to reduce the cost of preserving equipment described in installation and operation, under preferable case, described storage temperature is the arbitrary temp between 0 DEG C to 5 DEG C, or described storage temperature is the arbitrary temp between subzero 30 DEG C to subzero 15 DEG C.
According to microorganism low-temperature store method provided by the invention, wherein, as the particularly preferred embodiment of one, described storage temperature is the arbitrary temp between subzero 30 DEG C to subzero 15 DEG C, and, before the arbitrary temp mixed material being placed between subzero 30 DEG C to subzero 15 DEG C, first mixed material is placed in to 1-5h under the arbitrary temp between 0 DEG C to 5 DEG C, more preferably 2-4h.Under this preferred implementation, survival rate and the microbial activity of microorganism after cryopreservation is further enhanced.
According to microorganism low-temperature store method provided by the invention, wherein, this microorganism low-temperature store method is applicable to conventional various microorganisms, the particularly conventional various microorganisms of field of fermentation engineering, typically, described microorganism is at least one in escherichia coli (Latin formal name used at school is Escherichia coli), brevibacterium flavum (Latin formal name used at school is Brevibacterium flavum) and corynebacterium glutamicum (Latin formal name used at school is Corynebacterium glutamicum).
Below will describe the present invention by embodiment.In following examples, oligose, glycerine, N.F,USP MANNITOL, skim-milk and agarose are the commercially available product purchased from traditional Chinese medicines group company.
Preparation Example 1
According to the weight part shown in table 1, oligose, glycerine, N.F,USP MANNITOL, skim-milk and agarose are mixed, the microorganism low-temperature obtaining respectively under storing state is preserved protective material 1-3.
Table 1 (weight part)
Preparation Example 2
Microorganism low-temperature under the storing state that Preparation Example 1 is obtained preserve protective material 1-3 add water mixes and be heated to 95 DEG C after naturally cool to room temperature, obtain the microorganism low-temperature preservation protective material 1-3 under use state.Wherein, preserve the oligose of every 100 weight parts in protective material 1-3 with respect to the microorganism low-temperature under storing state, the consumption of water is 1000 weight parts.
Test implementation example 1
By intestinal bacteria (Escherichia coli, bacterial strain number is 10586, purchased from ATCC) in LB substratum (10 grams of Tryptoness, 5 grams of yeast extracts and 10 grams of sodium-chlor are added water be settled to 1L after autoclaving obtain the LB substratum of 1L), at 37 DEG C, cultivate to 10
9the cell concentration of individual intestinal bacteria/mL, obtains culture.Using this culture as the microorganism for the treatment of cryopreservation.
The above-mentioned culture of 1mL (is contained to 10
9individual intestinal bacteria) microorganism low-temperature under the use state that obtains with the Preparation Example 2 of 1mL respectively preserves protective material 1-3 and mixes; at 4 DEG C, after 2h, go at subzero 20 DEG C cryopreservation half a year; then ((leaf is of heap of stone, Yang Xuemin for reference literature to be at room temperature coated with colony counting method with dilution; microorganism detection technology; Chemical Industry Press, 2009 the 1st edition) in method carry out) record colibacillary survival rate and be respectively 98%, 96% and 97%.
Test implementation example 2
By intestinal bacteria (Escherichia coli, bacterial strain number is 10586, purchased from ATCC) in LB substratum (10 grams of Tryptoness, 5 grams of yeast extracts and 10 grams of sodium-chlor are added water be settled to 1L after autoclaving obtain the LB substratum of 1L), at 37 DEG C, cultivate to 10
9the cell concentration of individual intestinal bacteria/mL, obtains culture.Using this culture as the microorganism for the treatment of cryopreservation.
The above-mentioned culture of 1mL (is contained to 10
9individual intestinal bacteria) microorganism low-temperature under the use state that obtains with the Preparation Example 2 of 1mL respectively preserves protective material 1-3 and mixes; directly be placed at subzero 20 DEG C cryopreservation half a year; then ((leaf is of heap of stone, Yang Xuemin for reference literature to be at room temperature coated with colony counting method with dilution; microorganism detection technology; Chemical Industry Press, 2009 the 1st edition) in method carry out) record colibacillary survival rate and be respectively 95%, 94% and 93%.
Comparative example 1
, obtain microorganism low-temperature and preserve protective material 4-6 each mixing of materials according to the weight part shown in table 2.
Table 2 (weight part)
The intestinal bacteria identical with test implementation example 1, in LB substratum (10 grams of Tryptoness, 5 grams of yeast extracts and 10 grams of sodium-chlor are added water be settled to 1L after autoclaving obtain the LB substratum of 1L), are cultivated to 10 at 37 DEG C
9the cell concentration of/mL, obtains culture.Using this culture as the microorganism for the treatment of cryopreservation.
The above-mentioned culture of 1mL (is contained to 10
9individual intestinal bacteria) mix with the mentioned microorganism cryopreservation protective material 4-6 of 1mL respectively; at 4 DEG C, after 2h, go at subzero 20 DEG C cryopreservation half a year, then at room temperature use the method identical with test implementation example 1 to record colibacillary survival rate and be respectively 90%, 89% and 88%.
By the comparison of test implementation example 1, test implementation example 2 and comparative example 1, can see and appear technical scheme of the present invention, the survival rate of microorganism after cryopreservation is significantly improved; And before preferably mixed material being placed in to the arbitrary temp between subzero 30 DEG C to subzero 15 DEG C, first, by placing in the situation of 1-5h under the arbitrary temp between 0 DEG C to 5 DEG C of mixed materials, can further improve the survival rate of microorganism after cryopreservation.
Test implementation example 3
By brevibacterium flavum (Brevibacterium flavum, article number 1551C0003000002018, purchased from microbial collection center, Sichuan Province) minimum medium (compound method be 10 grams of Tryptoness, 5 grams of yeast extracts and 10 grams of sodium-chlor add water be settled to 1L after autoclaving obtain the LB substratum of 1L) in, at 35 DEG C, cultivate to 10
9the cell concentration of/mL, obtains culture.Using this culture as the microorganism for the treatment of cryopreservation.
The above-mentioned culture of 1mL (is contained to 10
9individual brevibacterium flavum) microorganism low-temperature under the use state that obtains with the Preparation Example 2 of 1mL respectively preserves protective material 1-3 and mixes; cryopreservation 7 days at 4 DEG C; then be all inoculated in the LB substratum (compound method be 10 grams of Tryptoness, 5 grams of yeast extracts and 10 grams of sodium-chlor add water be settled to autoclaving after 1L and obtain the LB substratum of 1L) of 1L, to cultivate 10
12the needed incubation time of the individual viable bacteria scale of construction is the seeding tank cycle, records the seeding tank cycle to be respectively 15h, 15.5h and 14.5h.
Comparative example 2
By the brevibacterium flavum identical with test implementation example 3 LB substratum (compound method be 10 grams of Tryptoness, 5 grams of yeast extracts and 10 grams of sodium-chlor add water be settled to 1L after autoclaving obtain the LB substratum of 1L) in, at 35 DEG C, cultivate to 10
9the cell concentration of/mL, obtains culture.Using this culture as the microorganism for the treatment of cryopreservation.
The above-mentioned culture of 1mL (is contained to 10
7individual Methionin produces bacterium) microorganism low-temperature that obtains with the comparative example 1 of 1mL respectively preserves protective material 4-6 and mixes; cryopreservation 7 days at 4 DEG C, then uses the method identical with test implementation example 3 to record the seeding tank cycle and is respectively 18h, 18.5h and 17.5h.
By the comparison of test implementation example 3 and comparative example 2, can see and appear technical scheme of the present invention, the microbial activity of microorganism after cryopreservation is significantly improved.
More than describe the preferred embodiment of the present invention in detail; but the present invention is not limited to the detail in above-mentioned embodiment, within the scope of technical conceive of the present invention; can carry out multiple simple variant to technical scheme of the present invention, these simple variant all belong to protection scope of the present invention.
It should be noted that in addition, each concrete technical characterictic described in above-mentioned embodiment, in reconcilable situation, can combine by any suitable mode, for fear of unnecessary repetition, the present invention is to the explanation no longer separately of various possible array modes.
In addition, also can carry out arbitrary combination between various embodiment of the present invention, as long as it is without prejudice to thought of the present invention, it should be considered as content disclosed in this invention equally.
Claims (10)
1. microorganism low-temperature is preserved a protective material, and this microorganism low-temperature is preserved protective material and contained oligose, glycerine, N.F,USP MANNITOL, skim-milk and agarose; Wherein, with respect to the oligose of 100 weight parts, the content of glycerine is 40-70 weight part, and the content of N.F,USP MANNITOL is 40-60 weight part, and the content of skim-milk is 150-200 weight part, and the content of agarose is 0.4-0.8 weight part.
2. microorganism low-temperature according to claim 1 is preserved protective material, and wherein, described oligose is at least one in trehalose, sucrose and maltose.
3. microorganism low-temperature according to claim 1 is preserved protective material; wherein; described agarose is to add described microorganism low-temperature to the form of the agarose of agar and/or purification to preserve in protective material, and in described agar, the content of agarose is at least 70 % by weight.
4. preserve protective material according to the microorganism low-temperature described in any one in claim 1-3, wherein, described microorganism low-temperature is preserved protective material and is also contained water, and with respect to the oligose of 100 weight parts, the content of water is 800-1500 weight part.
5. microorganism low-temperature according to claim 4 is preserved protective material, and wherein, with respect to the oligose of 100 weight parts, the content of water is 900-1200 weight part.
6. a microorganism low-temperature store method, this microorganism low-temperature store method comprises: the microorganism low-temperature described in any one in claim 1-3 is preserved after protective material mixes with water and mixed with microorganism to be preserved, then mixed material is placed under storage temperature; Or microorganism to be preserved is preserved to protective material with the microorganism low-temperature described in claim 4 or 5 and mix, then mixed material is placed under storage temperature; Wherein, described microorganism is escherichia coli (Escherichia coli) and/or brevibacterium flavum (Brevibacterium flavum).
7. microorganism low-temperature store method according to claim 6, wherein, preserves protective material by microorganism to be preserved with the microorganism low-temperature described in claim 4 or 5 and mixes, with respect to 10
9individual microorganism, it is 1mL that described microorganism low-temperature is preserved protectant consumption.
8. according to the microorganism low-temperature store method described in claim 6 or 7, wherein, described storage temperature is the arbitrary temp between 0 DEG C to 5 DEG C, or described storage temperature is the arbitrary temp between subzero 20 DEG C to subzero 15 DEG C.
9. microorganism low-temperature store method according to claim 8, wherein, described storage temperature is the arbitrary temp between subzero 20 DEG C to subzero 15 DEG C, and, before the arbitrary temp mixed material being placed between subzero 20 DEG C to subzero 15 DEG C, first mixed material is placed to 1-5h under the arbitrary temp between 0 DEG C to 5 DEG C.
10. microorganism low-temperature store method according to claim 9 wherein, before the arbitrary temp mixed material being placed between subzero 20 DEG C to subzero 15 DEG C, is first placed 2-4h by mixed material under the arbitrary temp between 0 DEG C to 5 DEG C.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201210132750.5A CN102643749B (en) | 2012-04-28 | 2012-04-28 | Microorganism low-temperature preservation protective agent and microorganism low-temperature preservation method |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201210132750.5A CN102643749B (en) | 2012-04-28 | 2012-04-28 | Microorganism low-temperature preservation protective agent and microorganism low-temperature preservation method |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN102643749A CN102643749A (en) | 2012-08-22 |
| CN102643749B true CN102643749B (en) | 2014-07-16 |
Family
ID=46656827
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201210132750.5A Expired - Fee Related CN102643749B (en) | 2012-04-28 | 2012-04-28 | Microorganism low-temperature preservation protective agent and microorganism low-temperature preservation method |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN102643749B (en) |
Families Citing this family (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103146580B (en) * | 2013-03-07 | 2015-02-25 | 西北民族大学 | Anaerobic ultralow temperature strain preserving agent |
| CN105238693B (en) * | 2015-10-28 | 2021-06-22 | 上海市农业科学院 | Conventional low-temperature preservation method for volvariella volvacea strain |
| CN106867905A (en) * | 2015-12-11 | 2017-06-20 | 深圳华大基因研究院 | Preservative fluid and recombination bacillus coli fermentation process in high density for culture presevation |
| CN108359613B (en) * | 2018-05-14 | 2022-04-29 | 安发（福建）生物科技有限公司 | Cordyceps sinensis antifreeze agent and ultralow-temperature preservation and recovery method based on antifreeze agent |
| CN109628311A (en) * | 2018-12-28 | 2019-04-16 | 美霖佳生物医学科技(苏州)有限公司 | Freeze drying protectant, freeze-dried powder, capsule and preparation method thereof for protecting enterobacteriaceae |
| CN109749972B (en) * | 2019-03-15 | 2022-06-03 | 南京工业大学 | Fungus agent spray drying protective agent and application thereof |
| CN114214246A (en) * | 2021-12-27 | 2022-03-22 | 贵州百灵企业集团制药股份有限公司 | Preparation method of countable strain with long shelf life at low temperature |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101264062A (en) * | 2007-03-12 | 2008-09-17 | 袁红杰 | Production technology for preparing freeze-drying genetic engineering bacterium competence cell and protective agent formula |
| CN101486986A (en) * | 2009-02-12 | 2009-07-22 | 上海谱莱生物技术有限公司 | Preparation of freeze-dried Lactobacillus acidophilus powder |
Family Cites Families (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1305705A (en) * | 2001-01-08 | 2001-08-01 | 卢杲 | Method for permanently preserving all species genes and biological samples in the world |
| CN102140423B (en) * | 2010-12-30 | 2012-11-21 | 广东环凯微生物科技有限公司 | Protective agent for quantitatively preserving microbial strains |
-
2012
- 2012-04-28 CN CN201210132750.5A patent/CN102643749B/en not_active Expired - Fee Related
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101264062A (en) * | 2007-03-12 | 2008-09-17 | 袁红杰 | Production technology for preparing freeze-drying genetic engineering bacterium competence cell and protective agent formula |
| CN101486986A (en) * | 2009-02-12 | 2009-07-22 | 上海谱莱生物技术有限公司 | Preparation of freeze-dried Lactobacillus acidophilus powder |
Non-Patent Citations (4)
| Title |
|---|
| 保护剂在乳酸菌冻干过程中的应用;徐致远等;《乳业科学与技术》;20061231;第156-157页 * |
| 双歧杆菌冻干菌粉制备过程中保护剂的研究;李德斌 等;《中国乳品工业》;20111231;第39卷(第11期);第32-34页 * |
| 徐致远等.保护剂在乳酸菌冻干过程中的应用.《乳业科学与技术》.2006,第156-157页. |
| 李德斌 等.双歧杆菌冻干菌粉制备过程中保护剂的研究.《中国乳品工业》.2011,第39卷(第11期),第32-34页. |
Also Published As
| Publication number | Publication date |
|---|---|
| CN102643749A (en) | 2012-08-22 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN102643749B (en) | Microorganism low-temperature preservation protective agent and microorganism low-temperature preservation method | |
| Zheng et al. | The effect of cultivar, wilting and storage period on fermentation and the clostridial community of alfalfa silage | |
| CN108570421A (en) | One lactobacillus plantarum strain and its ensilage fermentation agent | |
| CN107312732B (en) | Probiotic feed additive | |
| CN102524518B (en) | Method for producing antibacterial peptide by using brevibacillus laterosporu | |
| CN109439577A (en) | One plant of broad-spectrum antiseptic bacillus amyloliquefaciens and its application | |
| CN103421715B (en) | Lactobacillus rhamnosus and application thereof | |
| CN101548716A (en) | A porcine lactobacillu plantarurn freeze-dry preparation and its preparation method | |
| CN107217020B (en) | Culture medium suitable for lactobacillus acidophilus and preparation method thereof | |
| CN105602875A (en) | Preparation method of lactic acid bacteria freeze-dried powder suitable for storage at room temperature | |
| CN105861333A (en) | Eurotium cristatum LS1 strain | |
| CN107177522A (en) | One plant height activity forage plant lactobacillus and its cultural method and application | |
| CN101933620A (en) | Probiotic powder and preparation method | |
| CN108117992A (en) | A kind of preservation under room temperature method of denitrifying bacterium | |
| CN110295126A (en) | A kind of mixing probiotics preparation and its preparation process | |
| CN101606552A (en) | Freezing direct vat inoculation of a kind of bifidobacterium deep and composite frozen protective agent thereof | |
| CN105341425A (en) | Pig feed containing bacteriostatic microbial ecological agent and application thereof | |
| CN1162536C (en) | Preservation method for live bacterial preparation | |
| CN102286411B (en) | Lactobacillus plantarum and application thereof in fermenting cabbage wrapper leaf | |
| CN108570422A (en) | One plant of lactobacillus buchneri bacterial strain and its ensilage fermentation agent | |
| Qu et al. | Protective effect of crop by-products on Lactobacillus gasseri H87 during freeze-drying and storage | |
| CN101124317A (en) | Method of modifying the viability of a lyophilized microorganism by treating the growth medium thereof with gases | |
| CN102409012A (en) | Klebsiella pneumonia being separated from Termite gut | |
| CN109897800A (en) | The strong enterococcus A8-1 of one plant of selenium-rich and its application | |
| US11479749B2 (en) | Production and cryopreservation of high concentration inocula |
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: 20140716 Termination date: 20180428 |
|
| CF01 | Termination of patent right due to non-payment of annual fee |