CN107746813A - A kind of compound freeze drying protectant of Paula enlightening saccharomycete and its preparation and application method - Google Patents

A kind of compound freeze drying protectant of Paula enlightening saccharomycete and its preparation and application method Download PDF

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Publication number
CN107746813A
CN107746813A CN201710964587.1A CN201710964587A CN107746813A CN 107746813 A CN107746813 A CN 107746813A CN 201710964587 A CN201710964587 A CN 201710964587A CN 107746813 A CN107746813 A CN 107746813A
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saccharomycete
paula
enlightening
freeze drying
drying protectant
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陈合
雷张腾
姚春旭
洪海平
舒国伟
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Shaanxi University of Science and Technology
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Shaanxi University of Science and Technology
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    • 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
    • C12N1/14—Fungi; Culture media therefor
    • C12N1/16—Yeasts; Culture media therefor
    • 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
    • C12N1/04—Preserving or maintaining viable microorganisms

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  • Tropical Medicine & Parasitology (AREA)
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  • General Engineering & Computer Science (AREA)
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Abstract

A kind of compound freeze drying protectant of Paula enlightening saccharomycete and its preparation and application method, ascorbic acid sodium is dissolved in and filtered in sterile saline degerming obtains solution A；By xylan, synanthrin, FOS, sodium acid carbonate, sodium glutamate and lactose be dissolved in distilled water it is sterilized after be cooled to room temperature and obtain solution B；Solution A and solution B are well mixed the compound freeze drying protectant of get Bao Ladi saccharomycete.By Paula enlightening saccharomycete and the Paula compound freeze drying protectant of enlightening saccharomycete with 1：40 60min are balanced after 2 mass ratio is well mixed, is then placed in vacuum freeze drier to freeze and produces Paula enlightening saccharomycete bacterium powder.The compound freeze drying protectant of Paula enlightening saccharomycete of the present invention adds during bacterium powder is prepared Paula enlightening saccharomycete is freeze-dried, and lyophilized survival rate can reach 80.23% 86.26%, and bacterium powder viable count is 1.84 × 1010CFU/g‑2.13×1010CFU/g, solves the key technical problem during exploitation high activity Paula enlightening saccharomycete freezes bacterium powder.

Description

A kind of compound freeze drying protectant of Paula enlightening saccharomycete and its preparation and application method
Technical field
The invention belongs to saccharomycete production of articles technical field, is related to a kind of Paula enlightening saccharomycete that improves and freezes survival rate The compound freeze drying protectant of Paula enlightening saccharomycete and its preparation and application method.
Background technology
Paula enlightening saccharomycete (Saccharomyces boulardii) be by French scientist Henri Boulard in A kind of nineteen twenty budding fungus probiotics isolated from the fruit such as Indonesia's lichee, it is a kind of non-pathogenic yeast bacterium, There is no toxic side effect to humans and animals.Paula enlightening Blastocystis is the subspecies of saccharomyces cerevisiae, had heat-resisting in unicellular fungi Property, it can be grown at 37 DEG C.
At present, it is external that the preparation for the treatment of diarrhoea has been developed using Paula enlightening yeast, and be widely used in animal husbandry and Human body is resisted in the medicine of diarrhoea, and it can effectively reduce pathogen and the concentration about toxin, stimulating immune system, improve machine Body immunity.And the country not yet carries out systematic research in terms of the enlightening saccharomycete formulation development of Paula, only a small number of clinics should Report.Therefore the Paula enlightening yeast preparation of exploitation high activity shelf-stable is necessary.
The main method for preparing bacterium powder at present both at home and abroad is spray drying process and vacuum freeze-drying method.Wherein it is spray-dried Technical units product heat consumption is big, and the thermal efficiency of equipment is low, and EAT and outlet temperature are high, cause thalline survival rate It is low.If reducing temperature, easily go out damp powder；Higher is required to the separator of bacterium powder simultaneously, product is lost and is polluted, But the treating capacity of spray drying is big, and cost is low.Compared with spray drying process, Vacuum Freezing ＆ Drying Technology has obvious advantage： Strain is dried at low temperature, and the loss of activity of various composition and enzyme is less；The bacterium powder prepared by Vacuum Freezing ＆ Drying Technology has Have that color and luster, form are excellent, dehydration is thorough, and rehydration is good, the features such as being easy to preservation and transport.But vacuum freeze drying process In, thalline is subjected to freezing and dries the effect of two kinds of fierce factors, causes thalline dead.Influenceing unit bacterium powder viable count and jelly In the factor of dry survival rate, freeze drying protectant is key factor the most complicated, and it not only influences thalline in freeze-drying process Cell survival rate, also influence the cell stability during preservation.
The research to Paula enlightening saccharomycete freeze drying protectant such as Chu Jie finds that the formula of compound freeze drying protectant is：10% The normal saline solution of skimmed milk powder, 8% glycerine.The optimum results table to Paula enlightening saccharomycete freeze drying protectant such as Shao Mingqian Bright, best protection agent prescription is：10% skimmed milk, 10 trehaloses, 5% glycerine, 8% sucrose；All add in above-mentioned two protective agent Skimmed milk is added, the application of the bacterium powder of production such as medicament etc. in non-dairy product is restricted.In latter protective agent Trehalose is with the addition of, the water imbibition of trehalose is very strong, the easy moisture absorption, can cause dead very fast in bacterium powder storage process.
The content of the invention
Paula enlightening saccharomycete, which can be significantly improved, it is an object of the invention to provide one kind freezes survival rate and the work of unit bacterium powder The compound freeze drying protectant of Paula enlightening saccharomycete and its preparation and application method of bacterium number, solve Paula enlightening saccharomycete bacterium powder and are freezing Existing lyophilized survival rate is low during dry, and viable count is few, hygroscopic to become the problems such as damp.
In order to achieve the above object, the present invention and the Paula compound freeze drying protectant of enlightening saccharomycete include：2-4g oligomeric wood Sugar, 5-8g synanthrin, 8-12g FOS, 0.3-0.6g sodium acid carbonate, 4-6g sodium glutamate, 20-23g lactose, 0.7g-1g ascorbic acid sodium and 100mL distilled water.
The preparation method of the compound freeze drying protectant of Paula enlightening saccharomycete of the present invention is as follows：
1) first, take 0.7g-1g ascorbic acid sodium to be dissolved in and filtered in 20mL sterile salines degerming obtain solution A；
2) 2-4g xylo-oligosaccharide, 5-8g synanthrin, 8-12g FOS, 0.3-0.6g bicarbonate secondly, are taken The lactose of sodium, 4-6g sodium glutamate and 20-23g is dissolved in 80mL distilled water, is then cooled to after 100-118 DEG C of sterilizing Room temperature obtains solution B；
3) solution A and solution B finally, are well mixed the compound freeze drying protectant of get Bao Ladi saccharomycete.
The sterile saline of described step 1) uses 0.9% sterile saline.
The filtration sterilization of described step 1) is degerming using 0.22 μm of membrane filtration.
The application process of the compound freeze drying protectant of Paula enlightening saccharomycete of the present invention is：By Paula enlightening saccharomycete and Paula enlightening The compound freeze drying protectant of saccharomycete is with 1：40-60min is balanced after 2 mass ratio is well mixed, is then placed in vacuum freeze drying Freezed in machine and produce Paula enlightening saccharomycete bacterium powder.
The compound freeze drying protectant of Paula enlightening saccharomycete of the present invention is added to Paula enlightening yeast during bacterium powder is prepared Bacterium is freeze-dried, and lyophilized survival rate can reach 80.23%-86.26%, and bacterium powder viable count is 1.84 × 1010CFU/g- 2.13×1010CFU/g, solve the key technical problem during exploitation high activity Paula enlightening saccharomycete freezes bacterium powder, system Standby high activity bacterium powder can be used as additive, leavening to be used to develop Paula enlightening saccharomycete functional food and pharmaceutical preparation, this Invent the Paula enlightening saccharomycete and be free of skimmed milk powder and trehalose, and with the addition of the benefits such as xylo-oligosaccharide, synanthrin and FOS Raw member, solve that Paula enlightening saccharomycete bacterium powder is hygroscopic present in freeze-drying process to become the problems such as damp, while prebiotics adds Add the healthcare function for further increasing bacterium powder.
Embodiment
The present invention is described in further detail with reference to embodiment.
Embodiment 1：
The Paula compound freeze drying protectant of enlightening saccharomycete of the present embodiment includes：2g xylo-oligosaccharide, 5g synanthrin, 8g it is low Fructooligosaccharides, 0.4g sodium acid carbonate, 4g sodium glutamate, 20g lactose, 0.7g ascorbic acid sodium and 100mL distilled water.
Its preparation method is as follows：
1) 0.7g ascorbic acid sodium first, is taken to be dissolved in 20mL, 0.9% sterile saline, then by 0.22 μm of filter Membrane filtration is degerming to obtain solution A；
2) 2g xylo-oligosaccharide, 5g synanthrin, 8g FOS, 0.4g sodium acid carbonate, 4g glutamic acid secondly, are taken Sodium, 20g lactose are dissolved in 80mL distilled water, and being then cooled to room temperature after 100 DEG C of sterilizings obtains solution B；
3) solution A and solution B finally, are well mixed the compound freeze drying protectant of get Bao Ladi saccharomycete.
The application process of the compound freeze drying protectant of Paula enlightening saccharomycete of the present invention is：By Paula enlightening saccharomycete and Paula enlightening The compound freeze drying protectant of saccharomycete is with 1：50min is balanced after 2 mass ratio is well mixed, is then placed in vacuum freeze drier Freeze and produce Paula enlightening saccharomycete bacterium powder.
The Paula enlightening saccharomycete freeze drying protectant of the present embodiment, freeze drying protectant is added during bacterium powder is prepared, can The freezing tolerance of thalline is improved, so as to improve the lyophilized survival rate of bacterium powder, prepared Paula enlightening saccharomycete bacterium powder freezes Survival rate reaches 80.23%, and bacterium powder viable count is 1.84 × 1010CFU/g。
Embodiment 2：
The Paula compound freeze drying protectant of enlightening saccharomycete of the present embodiment includes：2g xylo-oligosaccharide, 5g synanthrin, 10g FOS, 0.3g sodium acid carbonate, 5g sodium glutamate, 20g lactose, 0.8g ascorbic acid sodium and 100mL distilled water.
Its preparation method is as follows：
1) 0.8g ascorbic acid sodium first, is taken to be dissolved in 20mL, 0.9% sterile saline, then by 0.22 μm of filter Membrane filtration is degerming to obtain solution A；
2) 2g xylo-oligosaccharide, 5g synanthrin, 10g FOS, 0.3g sodium acid carbonate, 5g paddy ammonia secondly, are taken Sour sodium, 20g lactose are dissolved in 80mL distilled water, and being then cooled to room temperature after 113 DEG C of sterilizings obtains solution B；
3) solution A and solution B finally, are well mixed the compound freeze drying protectant of get Bao Ladi saccharomycete.
The application process of the compound freeze drying protectant of Paula enlightening saccharomycete of the present invention is：By Paula enlightening saccharomycete and Paula enlightening The compound freeze drying protectant of saccharomycete is with 1：60min is balanced after 2 mass ratio is well mixed, is then placed in vacuum freeze drier Freeze and produce Paula enlightening saccharomycete bacterium powder.
The Paula enlightening saccharomycete freeze drying protectant of the present embodiment, freeze drying protectant is added during bacterium powder is prepared, can The freezing tolerance of thalline is improved, so as to improve the lyophilized survival rate of bacterium powder, prepared Paula enlightening saccharomycete bacterium powder freezes Survival rate reaches 82.63%, and bacterium powder viable count is 1.89 × 1010CFU/g。
Embodiment 3：
The Paula compound freeze drying protectant of enlightening saccharomycete of the present embodiment includes：3g xylo-oligosaccharide, 8g synanthrin, 10g FOS, 0.6g sodium acid carbonate, 5g sodium glutamate, 22g lactose, 0.8g ascorbic acid sodium and 100mL distilled water.
Its preparation method is as follows：
1) 0.8g ascorbic acid sodium first, is taken to be dissolved in 20mL, 0.9% sterile saline, then by 0.22 μm of filter Membrane filtration is degerming to obtain solution A；
2) 3g xylo-oligosaccharide, 8g synanthrin, 10g FOS, 0.6g sodium acid carbonate, 5g paddy ammonia secondly, are taken Sour sodium, 22g lactose are dissolved in 80mL distilled water, and being then cooled to room temperature after 118 DEG C of sterilizings obtains solution B；
3) solution A and solution B finally, are well mixed the compound freeze drying protectant of get Bao Ladi saccharomycete.
The application process of the compound freeze drying protectant of Paula enlightening saccharomycete of the present invention is：By Paula enlightening saccharomycete and Paula enlightening The compound freeze drying protectant of saccharomycete is with 1：40min is balanced after 2 mass ratio is well mixed, is then placed in vacuum freeze drier Freeze and produce Paula enlightening saccharomycete bacterium powder.
The Paula enlightening saccharomycete freeze drying protectant of the present embodiment, freeze drying protectant is added during bacterium powder is prepared, can The freezing tolerance of thalline is improved, so as to improve the lyophilized survival rate of bacterium powder, prepared Paula enlightening saccharomycete bacterium powder freezes Survival rate reaches 84.36%, and bacterium powder viable count is 1.97 × 1010CFU/g。
Embodiment 4：
The Paula compound freeze drying protectant of enlightening saccharomycete of the present embodiment includes：4g xylo-oligosaccharide, 7g synanthrin, 12g FOS, 0.5g sodium acid carbonate, 6g sodium glutamate, 23g lactose, 1.0g ascorbic acid sodium and 100mL distilled water.
Its preparation method is as follows：
1) 1.0g ascorbic acid sodium first, is taken to be dissolved in 20mL, 0.9% sterile saline, then by 0.22 μm of filter Membrane filtration is degerming to obtain solution A；
2) 4g xylo-oligosaccharide, 7g synanthrin, 12g FOS, 0.5g sodium acid carbonate, 6g paddy ammonia secondly, are taken Sour sodium, 23g lactose are dissolved in 80mL distilled water, and being then cooled to room temperature after 115 DEG C of sterilizings obtains solution B；
3) solution A and solution B finally, are well mixed the compound freeze drying protectant of get Bao Ladi saccharomycete.
The application process of the compound freeze drying protectant of Paula enlightening saccharomycete of the present invention is：By Paula enlightening saccharomycete and Paula enlightening The compound freeze drying protectant of saccharomycete is with 1：45min is balanced after 2 mass ratio is well mixed, is then placed in vacuum freeze drier Freeze and produce Paula enlightening saccharomycete bacterium powder.
The Paula enlightening saccharomycete freeze drying protectant of the present embodiment, freeze drying protectant is added during bacterium powder is prepared, can The freezing tolerance of thalline is improved, so as to improve the lyophilized survival rate of bacterium powder, prepared Paula enlightening saccharomycete bacterium powder freezes Survival rate reaches 86.26%, and bacterium powder viable count is 2.13 × 1010CFU/g。

Claims (5)

  1. A kind of 1. compound freeze drying protectant of Paula enlightening saccharomycete, it is characterised in that：The chrysanthemum of xylo-oligosaccharide, 5-8g including 2-4g Sugar, 8-12g FOS, 0.3-0.6g sodium acid carbonate, 4-6g sodium glutamate, 20-23g lactose, 0.7g-1g it is anti- The distilled water of bad sour sodium and 100mL.
  2. A kind of 2. preparation method of the compound freeze drying protectant of Paula enlightening saccharomycete, it is characterised in that：
    1) first, take 0.7g-1g ascorbic acid sodium to be dissolved in and filtered in 20mL sterile salines degerming obtain solution A；
    2) 2-4g xylo-oligosaccharide, 5-8g synanthrin, 8-12g FOS, 0.3-0.6g sodium acid carbonate, 4- secondly, are taken 6g sodium glutamate and 20-23g lactose are dissolved in 80mL distilled water, are then cooled to room temperature after 100-118 DEG C of sterilizing Obtain solution B；
    3) solution A and solution B finally, are well mixed the compound freeze drying protectant of get Bao Ladi saccharomycete.
  3. 3. the preparation method of the compound freeze drying protectant of Paula enlightening saccharomycete according to claim 2, it is characterised in that：It is described The sterile saline of step 1) use 0.9% sterile saline.
  4. 4. the preparation method of the compound freeze drying protectant of Paula enlightening saccharomycete according to claim 2, it is characterised in that：It is described Step 1) filtration sterilization using 0.22 μm of membrane filtration it is degerming.
  5. A kind of 5. application process of the compound freeze drying protectant of Paula enlightening saccharomycete as claimed in claim 1, it is characterised in that：Will Paula enlightening saccharomycete is with the Paula compound freeze drying protectant of enlightening saccharomycete with 1：40-60min is balanced after 2 mass ratio is well mixed, It is then placed in vacuum freeze drier to freeze and produces Paula enlightening saccharomycete bacterium powder.
CN201710964587.1A 2017-10-17 2017-10-17 A kind of compound freeze drying protectant of Paula enlightening saccharomycete and its preparation and application method Pending CN107746813A (en)

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Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN109504614A (en) * 2018-12-29 2019-03-22 山东景芝酒业股份有限公司 A kind of preparation method of native country saccharomyces cerevisiae dry powder
CN112522119A (en) * 2020-11-20 2021-03-19 江苏大学 Preparation method and application of combined microbial inoculum based on Klebsiella citrullina and Candida intermedia
CN113621519A (en) * 2021-08-11 2021-11-09 河南农业大学 Composite yeast freeze-drying protective agent

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CN102358884A (en) * 2011-10-13 2012-02-22 陕西农产品加工技术研究院 Bifidobacterium bifidum lyophilization composite protection agent and application method thereof

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Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN109504614A (en) * 2018-12-29 2019-03-22 山东景芝酒业股份有限公司 A kind of preparation method of native country saccharomyces cerevisiae dry powder
CN112522119A (en) * 2020-11-20 2021-03-19 江苏大学 Preparation method and application of combined microbial inoculum based on Klebsiella citrullina and Candida intermedia
CN113621519A (en) * 2021-08-11 2021-11-09 河南农业大学 Composite yeast freeze-drying protective agent
CN113621519B (en) * 2021-08-11 2023-07-18 河南农业大学 Composite yeast freeze-drying protective agent

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