CN113621535B - Composite protective agent for improving survival rate of probiotics freeze-dried powder and preparation method thereof - Google Patents
Composite protective agent for improving survival rate of probiotics freeze-dried powder and preparation method thereof Download PDFInfo
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- CN113621535B CN113621535B CN202110792603.XA CN202110792603A CN113621535B CN 113621535 B CN113621535 B CN 113621535B CN 202110792603 A CN202110792603 A CN 202110792603A CN 113621535 B CN113621535 B CN 113621535B
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- 239000006041 probiotic Substances 0.000 title claims abstract description 61
- 235000018291 probiotics Nutrition 0.000 title claims abstract description 61
- 239000000843 powder Substances 0.000 title claims abstract description 41
- 239000003223 protective agent Substances 0.000 title claims abstract description 41
- 239000002131 composite material Substances 0.000 title claims abstract description 24
- 230000004083 survival effect Effects 0.000 title claims abstract description 23
- 238000002360 preparation method Methods 0.000 title claims abstract description 11
- 244000005700 microbiome Species 0.000 claims abstract description 61
- 150000004676 glycans Chemical class 0.000 claims abstract description 57
- 229920001282 polysaccharide Polymers 0.000 claims abstract description 57
- 239000005017 polysaccharide Substances 0.000 claims abstract description 57
- 239000000725 suspension Substances 0.000 claims abstract description 40
- 230000001580 bacterial effect Effects 0.000 claims abstract description 39
- 230000000529 probiotic effect Effects 0.000 claims abstract description 38
- 238000007710 freezing Methods 0.000 claims abstract description 28
- 239000001963 growth medium Substances 0.000 claims abstract description 24
- 230000008014 freezing Effects 0.000 claims abstract description 22
- 238000002156 mixing Methods 0.000 claims abstract description 16
- 239000010802 sludge Substances 0.000 claims abstract description 11
- HDTRYLNUVZCQOY-UHFFFAOYSA-N α-D-glucopyranosyl-α-D-glucopyranoside Natural products OC1C(O)C(O)C(CO)OC1OC1C(O)C(O)C(O)C(CO)O1 HDTRYLNUVZCQOY-UHFFFAOYSA-N 0.000 claims abstract description 10
- 108010010803 Gelatin Proteins 0.000 claims abstract description 10
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 claims abstract description 10
- 229930006000 Sucrose Natural products 0.000 claims abstract description 10
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 claims abstract description 10
- HDTRYLNUVZCQOY-WSWWMNSNSA-N Trehalose Natural products O[C@@H]1[C@@H](O)[C@@H](O)[C@@H](CO)O[C@@H]1O[C@@H]1[C@H](O)[C@@H](O)[C@@H](O)[C@@H](CO)O1 HDTRYLNUVZCQOY-WSWWMNSNSA-N 0.000 claims abstract description 10
- HDTRYLNUVZCQOY-LIZSDCNHSA-N alpha,alpha-trehalose Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@@H]1O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 HDTRYLNUVZCQOY-LIZSDCNHSA-N 0.000 claims abstract description 10
- 229920000159 gelatin Polymers 0.000 claims abstract description 10
- 239000008273 gelatin Substances 0.000 claims abstract description 10
- 235000019322 gelatine Nutrition 0.000 claims abstract description 10
- 235000011852 gelatine desserts Nutrition 0.000 claims abstract description 10
- 239000008103 glucose Substances 0.000 claims abstract description 10
- 235000020183 skimmed milk Nutrition 0.000 claims abstract description 10
- 239000005720 sucrose Substances 0.000 claims abstract description 10
- 239000006228 supernatant Substances 0.000 claims abstract description 9
- 230000004913 activation Effects 0.000 claims abstract description 5
- 238000000855 fermentation Methods 0.000 claims abstract description 3
- 230000004151 fermentation Effects 0.000 claims abstract description 3
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 31
- OAKJQQAXSVQMHS-UHFFFAOYSA-N Hydrazine Chemical compound NN OAKJQQAXSVQMHS-UHFFFAOYSA-N 0.000 claims description 16
- 241000756042 Polygonatum Species 0.000 claims description 16
- 239000000284 extract Substances 0.000 claims description 9
- 238000000605 extraction Methods 0.000 claims description 8
- 238000001914 filtration Methods 0.000 claims description 8
- 238000000643 oven drying Methods 0.000 claims description 8
- 238000010298 pulverizing process Methods 0.000 claims description 8
- 238000007873 sieving Methods 0.000 claims description 8
- 238000005119 centrifugation Methods 0.000 claims description 4
- 239000002994 raw material Substances 0.000 claims description 3
- 238000001035 drying Methods 0.000 claims description 2
- 230000000813 microbial effect Effects 0.000 abstract description 9
- 239000000047 product Substances 0.000 abstract description 8
- 235000013305 food Nutrition 0.000 abstract description 3
- 230000000694 effects Effects 0.000 abstract description 2
- 239000003814 drug Substances 0.000 abstract 1
- 239000007788 liquid Substances 0.000 description 13
- 150000001875 compounds Chemical class 0.000 description 12
- 210000004027 cell Anatomy 0.000 description 7
- 230000003213 activating effect Effects 0.000 description 6
- 238000013329 compounding Methods 0.000 description 6
- 238000012258 culturing Methods 0.000 description 6
- 238000009630 liquid culture Methods 0.000 description 6
- 238000000034 method Methods 0.000 description 6
- 239000006286 aqueous extract Substances 0.000 description 2
- 210000002390 cell membrane structure Anatomy 0.000 description 2
- 238000007689 inspection Methods 0.000 description 2
- 239000008176 lyophilized powder Substances 0.000 description 2
- 230000003204 osmotic effect Effects 0.000 description 2
- 108090000623 proteins and genes Proteins 0.000 description 2
- 102000004169 proteins and genes Human genes 0.000 description 2
- 230000001256 tonic effect Effects 0.000 description 2
- 241000186015 Bifidobacterium longum subsp. infantis Species 0.000 description 1
- 241000186660 Lactobacillus Species 0.000 description 1
- 235000013960 Lactobacillus bulgaricus Nutrition 0.000 description 1
- 244000199885 Lactobacillus bulgaricus Species 0.000 description 1
- 241001468157 Lactobacillus johnsonii Species 0.000 description 1
- 240000006024 Lactobacillus plantarum Species 0.000 description 1
- 235000013965 Lactobacillus plantarum Nutrition 0.000 description 1
- 241000194035 Lactococcus lactis Species 0.000 description 1
- 235000014897 Streptococcus lactis Nutrition 0.000 description 1
- 241000194020 Streptococcus thermophilus Species 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 229940004120 bifidobacterium infantis Drugs 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 229940039696 lactobacillus Drugs 0.000 description 1
- 229940004208 lactobacillus bulgaricus Drugs 0.000 description 1
- 229940072205 lactobacillus plantarum Drugs 0.000 description 1
- 230000010534 mechanism of action Effects 0.000 description 1
- 235000013336 milk Nutrition 0.000 description 1
- 239000008267 milk Substances 0.000 description 1
- 210000004080 milk Anatomy 0.000 description 1
- 229940126680 traditional chinese medicines Drugs 0.000 description 1
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- 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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- 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
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Abstract
The invention discloses a composite protective agent for improving survival rate of probiotic freeze-dried powder and a preparation method thereof, wherein the composite protective agent comprises the following components: 1-10 parts of skimmed milk powder, 1-10 parts of trehalose, 1-10 parts of sucrose, 1-10 parts of glucose, 1-10 parts of gelatin and 1-10 parts of rhizoma polygonati polysaccharide; the preparation method of the probiotics freeze-dried powder comprises the following steps: s1: inoculating probiotic strains into a microorganism culture medium, placing the culture medium into a shaking table for fermentation culture and activation, and collecting activated microorganism suspension; s2: centrifuging the microorganism suspension in the step S1, discarding the supernatant, and collecting bacterial sludge; s3: adding a composite protective agent into the bacterial mud, and uniformly mixing the probiotic suspension; s4: and (3) pre-freezing the microbial suspension in the step (S3), and then performing vacuum freezing to obtain the probiotic freeze-dried powder. The invention improves the survival rate of the probiotic freeze-dried powder, and simultaneously increases the nourishing efficacy of the probiotic freeze-dried powder product by utilizing the nourishing effect of the rhizoma polygonati serving as a medicine and food resource.
Description
Technical Field
The invention relates to the technical field of traditional Chinese medicines, in particular to a composite protective agent for improving survival rate of probiotic freeze-dried powder and a preparation method thereof.
Background
For the existing probiotic products, the problem of low survival rate of probiotics exists, so that the effectiveness of the probiotic products is seriously inhibited, and the problem of how to improve the survival rate of the probiotics in the probiotic products is the technical problem to be solved by the invention.
Disclosure of Invention
Based on the technical problems in the background art, the invention provides the compound protective agent for improving the survival rate of the probiotic freeze-dried powder and the preparation method thereof, and the tonic efficacy of the probiotic freeze-dried powder product is improved by utilizing the tonic effect of the medical and edible dual-purpose resource polygonatum polysaccharide while improving the survival rate of the probiotic freeze-dried powder.
The invention provides a composite protective agent for improving survival rate of probiotics freeze-dried powder, which comprises the following raw materials in parts by weight: 1-10 parts of skimmed milk powder, 1-10 parts of trehalose, 1-10 parts of sucrose, 1-10 parts of glucose, 1-10 parts of gelatin and 1-10 parts of rhizoma polygonati polysaccharide.
Preferably, the polygonatum polysaccharide is an aqueous extract, and the preparation method comprises the following steps: oven drying rhizoma Polygonati, pulverizing, sieving, mixing with water, extracting at high temperature, and filtering to obtain rhizoma Polygonati polysaccharide water extract.
Preferably, the conditions of the drying are: the temperature is 50-70 ℃ and the time is 2-6h.
Preferably, the conditions of the high temperature extraction are: the mass volume ratio of rhizoma polygonati to water is 1:5-15, the temperature is 90-125 ℃ and the time is 0.5-2h.
The invention provides an application of a composite protective agent for improving the survival rate of probiotic freeze-dried powder in the probiotic freeze-dried powder.
Preferably, the preparation method of the probiotic freeze-dried powder comprises the following steps:
s1: inoculating probiotic strains into a microorganism culture medium, placing the culture medium into a shaking table for fermentation culture and activation, and collecting activated microorganism suspension;
s2: centrifuging the microorganism suspension in the step S1, discarding the supernatant, and collecting bacterial sludge;
s3: adding a composite protective agent into the bacterial mud, and uniformly mixing the probiotic suspension;
s4: and (3) pre-freezing the microbial suspension in the step (S3), and then performing vacuum freezing to obtain the probiotic freeze-dried powder.
Preferably, the conditions of activation in S1 are: the temperature is 30-38 ℃ and the time is 12-48h.
Preferably, the centrifugation time in S2 is 1-10min, and the rotation speed during centrifugation is 8000-12000rpm.
Preferably, the mass ratio of the bacterial sludge in the step S3 to the composite protective agent is 1:1-5.
Preferably, the pre-freezing condition in S4 is: the temperature is between minus 20 ℃ and minus 4 ℃ and the time is between 10 hours and 24 hours; the conditions of vacuum freezing are as follows: vacuum degree is 0.1-20Pa, cold hydrazine temperature is-180-45 ℃ and time is 12-24h.
Mechanism of action
The preparation method comprises the steps of mixing a protecting agent and a probiotic bacterial liquid, wherein the protecting agent is prepared from rhizoma polygonati polysaccharide serving as a raw material, and the rhizoma polygonati polysaccharide can be combined with probiotic cells to protect cell membrane structures, maintain stable cell osmotic pressure and protect protein structures in the probiotic cells, so that the survival rate of probiotics in probiotic products is improved.
Compared with the prior art, the invention has the beneficial technical effects that
The composite protective agent comprises the polygonatum polysaccharide which can be combined with the probiotic cells, so that the survival rate of the probiotics is improved by more than 10 percent compared with the survival rate of the probiotics without the polygonatum polysaccharide.
Drawings
FIG. 1 is a microscopic image of probiotics of example 1 (left) and its control group (right) as set forth in the present invention;
FIG. 2 is a microscopic image of probiotics of example 2 (left) and its control group (right) as proposed by the present invention;
FIG. 3 is a microscopic image of probiotics of example 3 (left) and its control group (right) as proposed by the present invention;
FIG. 4 is a microscopic image of probiotics of example 4 (left) and its control group (right) as set forth in the present invention;
FIG. 5 is a microscopic image of probiotics of example 5 (left) and its control group (right) as set forth in the present invention;
FIG. 6 is a microscopic image of probiotics of example 6 (left) and its control group (right) as set forth in the present invention;
Detailed Description
The skim milk powder of the present invention is purchased from Australian milk industry (China) Limited, trehalose is purchased from Japanese Kogyo Lin original Co., ltd, sucrose is purchased from Shanghai Yuan Ye Biotech Limited, glucose is purchased from national pharmaceutical group chemical reagent Limited, and gelatin is purchased from biological engineering (Shanghai) Co.
The products are all commercially available and are directly used without any treatment.
Example 1
The method for compounding the composite protective agent and the lactobacillus plantarum provided by the invention comprises the following steps:
(1) And (3) inoculating microorganisms into the microorganism liquid culture medium according to the volume ratio of the bacterial liquid to the microorganism culture medium of 1:100, placing the microorganism culture medium containing the bacterial liquid into a shaking table, fermenting, culturing and activating at 30 ℃, and collecting microorganism suspension after 12 hours.
(2) Centrifuging the microorganism suspension in a high-speed centrifuge at 12000rpm for 1min, discarding the supernatant, and collecting bacterial sludge.
(3) The microbial suspension containing the polygonatum polysaccharide protective agent is prepared by adding 1 part of skimmed milk powder, 1 part of trehalose, 1 part of sucrose, 1 part of glucose, 1 part of gelatin and 1 part of polygonatum polysaccharide into the bacterial mud according to the volume ratio of the compound protective agent to the bacterial mud of 1:1.
(4) Pre-freezing the microorganism suspension containing rhizoma Polygonati polysaccharide and protectant at-4deg.C for-12 hr, and freezing in vacuum freeze dryer for 24 hr to obtain microorganism lyophilized powder, wherein the vacuum freezing conditions are as follows: vacuum degree 20Pa, cold hydrazine temperature-180 deg.C, and time 24h.
The extraction method of the rhizoma polygonati polysaccharide comprises the following steps:
oven drying rhizoma Polygonati at 50deg.C for 2 hr, pulverizing, sieving, mixing with water, extracting at 90deg.C for 0.5 hr, and filtering to obtain rhizoma Polygonati polysaccharide water extract. Wherein the mass volume ratio of the rhizoma polygonati to the water is 1:5. Concentrating the water extractive solution to make the concentration of rhizoma Polygonati polysaccharide be 0.5g/L.
Example 2
The method for compounding the composite protective agent and the lactobacillus bulgaricus provided by the invention comprises the following steps:
(1) And (3) inoculating microorganisms into the microorganism liquid culture medium according to the volume ratio of the bacterial liquid to the microorganism culture medium of 1:100, placing the microorganism culture medium containing the bacterial liquid into a shaking table, fermenting, culturing and activating at 37 ℃, and collecting microorganism suspension after 24 hours.
(2) Centrifuging the microorganism suspension in a high-speed centrifugal machine at 8000rpm for 5min, discarding the supernatant, and collecting bacterial sludge.
(3) The microbial suspension containing the rhizoma polygonati polysaccharide protective agent is prepared by adding 3 parts of skimmed milk powder, 3 parts of trehalose, 1 part of sucrose, 5 parts of glucose, 3 parts of gelatin and 5 parts of rhizoma polygonati polysaccharide into bacterial mud according to the volume ratio of the compound protective agent to the bacterial mud of 1:3, and uniformly mixing.
(4) Pre-freezing the microorganism suspension containing the rhizoma polygonati polysaccharide protective agent at the temperature of-4 ℃ for 12 hours, and then placing the microorganism suspension in a vacuum freeze dryer for freezing for 24 hours to obtain rhizoma polygonati microorganism freeze-dried powder, wherein the vacuum freezing conditions are as follows: vacuum degree is 0.1Pa, cold hydrazine temperature is-45 ℃, and time is 12h.
The extraction method of the rhizoma polygonati polysaccharide comprises the following steps:
oven drying rhizoma Polygonati at 60deg.C for 4 hr, pulverizing, sieving, mixing with water, extracting at 108deg.C for 1 hr, and filtering to obtain rhizoma Polygonati polysaccharide water extract. Wherein the mass volume ratio of the rhizoma polygonati to the water is 1:10. Concentrating the water extractive solution to make the concentration of rhizoma Polygonati polysaccharide be 0.5g/L.
Example 3
The method for compounding the compound protective agent and lactobacillus johnsonii provided by the invention comprises the following steps:
(1) And (3) inoculating microorganisms into the microorganism liquid culture medium according to the volume ratio of the bacterial liquid to the microorganism culture medium of 1:100, placing the microorganism culture medium containing the bacterial liquid into a shaking table, fermenting, culturing and activating at 37 ℃, and collecting microorganism suspension after 48 hours.
(2) The microorganism suspension was centrifuged in a high-speed centrifuge at 120000rpm for 1min, the supernatant was discarded, and the bacterial sludge was collected.
(3) The microbial suspension containing the polygonatum polysaccharide protective agent is prepared by adding 5 parts of skimmed milk powder, 10 parts of trehalose, 10 parts of sucrose, 2 parts of glucose, 3 parts of gelatin and 10 parts of polygonatum polysaccharide into the bacterial mud according to the volume ratio of the compound protective agent to the bacterial mud of 1:1, wherein the formula and the weight parts of the compound protective agent are uniformly mixed.
(4) Pre-freezing the microorganism suspension containing rhizoma Polygonati polysaccharide and protectant at-4deg.C for-12 hr, and freezing in vacuum freeze dryer for 24 hr to obtain rhizoma Polygonati microorganism lyophilized powder, wherein the vacuum freezing conditions are as follows: 10Pa, and the temperature of the cold hydrazine is minus 90 ℃ for 18 hours.
The extraction method of the rhizoma polygonati polysaccharide comprises the following steps:
oven drying rhizoma Polygonati at 70deg.C for 6 hr, pulverizing, sieving, mixing with water, extracting at 125deg.C for 2 hr, and filtering to obtain rhizoma Polygonati polysaccharide water extract. Wherein the mass volume ratio of the rhizoma polygonati to the water is 1:15. Concentrating the water extractive solution to make the concentration of rhizoma Polygonati polysaccharide be 0.5g/L.
Example 4
The method for compounding the compound protective agent and the lactococcus lactis provided by the invention comprises the following steps:
(1) And (3) inoculating microorganisms into the microorganism liquid culture medium according to the volume ratio of the bacterial liquid to the microorganism culture medium of 1:100, placing the microorganism culture medium containing the bacterial liquid into a shaking table, fermenting, culturing and activating at 37 ℃, and collecting microorganism suspension after 24 hours.
(2) Centrifuging the microorganism suspension in a high-speed centrifugal machine at 8000rpm for 5min, discarding the supernatant, and collecting bacterial sludge.
(3) The microbial suspension containing the rhizoma polygonati polysaccharide protective agent is prepared by adding 3 parts of skimmed milk powder, 3 parts of trehalose, 1 part of sucrose, 5 parts of glucose, 3 parts of gelatin and 5 parts of rhizoma polygonati polysaccharide into bacterial mud according to the volume ratio of the compound protective agent to the bacterial mud of 1:3, and uniformly mixing.
(4) Pre-freezing the microorganism suspension containing the rhizoma polygonati polysaccharide protective agent at the temperature of-4 ℃ for 12 hours, and then placing the microorganism suspension in a vacuum freeze dryer for freezing for 24 hours to obtain rhizoma polygonati microorganism freeze-dried powder, wherein the vacuum freezing conditions are as follows: vacuum degree 5Pa, cold hydrazine temperature-60 deg.C, and time 18h.
The extraction method of the rhizoma polygonati polysaccharide comprises the following steps:
oven drying rhizoma Polygonati at 50deg.C for 4 hr, pulverizing, sieving, mixing with water, extracting at 98deg.C for 1.5 hr, and filtering to obtain rhizoma Polygonati polysaccharide water extract. Wherein the mass volume ratio of the rhizoma polygonati to the water is 1:8. Concentrating the water extractive solution to make the concentration of rhizoma Polygonati polysaccharide be 0.5g/L.
Example 5
The method for compounding the compound protective agent and the bifidobacterium infantis provided by the invention comprises the following steps:
(1) And (3) inoculating microorganisms into the microorganism liquid culture medium according to the volume ratio of the bacterial liquid to the microorganism culture medium of 1:100, placing the microorganism culture medium containing the bacterial liquid into a shaking table, fermenting, culturing and activating at 37 ℃, and collecting microorganism suspension after 24 hours.
(2) Centrifuging the microorganism suspension in a high-speed centrifugal machine at 8000rpm for 5min, discarding the supernatant, and collecting bacterial sludge.
(3) The microbial suspension containing the rhizoma polygonati polysaccharide protective agent is prepared by adding 3 parts of skimmed milk powder, 3 parts of trehalose, 1 part of sucrose, 5 parts of glucose, 3 parts of gelatin and 5 parts of rhizoma polygonati polysaccharide into bacterial mud according to the volume ratio of the compound protective agent to the bacterial mud of 1:3, and uniformly mixing.
(4) Pre-freezing the microorganism suspension containing the rhizoma polygonati polysaccharide protective agent at the temperature of-4 ℃ for 12 hours, and then placing the microorganism suspension in a vacuum freeze dryer for freezing for 24 hours to obtain rhizoma polygonati microorganism freeze-dried powder, wherein the vacuum freezing conditions are as follows: vacuum degree is 0.1Pa, cold hydrazine temperature is-45 ℃, and time is 12h.
The extraction method of the rhizoma polygonati polysaccharide comprises the following steps:
oven drying rhizoma Polygonati at 62deg.C for 5 hr, pulverizing, sieving, mixing with water, extracting at 115deg.C for 1.5 hr, and filtering to obtain rhizoma Polygonati polysaccharide water extract. Wherein the mass volume ratio of the rhizoma polygonati to the water is 1:12. Concentrating the water extractive solution to make the concentration of rhizoma Polygonati polysaccharide be 0.5g/L.
Example 6
The method for compounding the compound protective agent and streptococcus thermophilus provided by the invention comprises the following steps:
(1) And (3) inoculating microorganisms into the microorganism liquid culture medium according to the volume ratio of the bacterial liquid to the microorganism culture medium of 1:100, placing the microorganism culture medium containing the bacterial liquid into a shaking table, fermenting, culturing and activating at 37 ℃, and collecting microorganism suspension after 24 hours.
(2) Centrifuging the microorganism suspension in a high-speed centrifugal machine at 8000rpm for 5min, discarding the supernatant, and collecting bacterial sludge.
(3) The microbial suspension containing the rhizoma polygonati polysaccharide protective agent is prepared by adding 3 parts of skimmed milk powder, 3 parts of trehalose, 1 part of sucrose, 5 parts of glucose, 3 parts of gelatin and 5 parts of rhizoma polygonati polysaccharide into bacterial mud according to the volume ratio of the compound protective agent to the bacterial mud of 1:3, and uniformly mixing.
(4) Pre-freezing the microorganism suspension containing the rhizoma polygonati polysaccharide protective agent at the temperature of-4 ℃ for 12 hours, and then placing the microorganism suspension in a vacuum freeze dryer for freezing for 24 hours to obtain rhizoma polygonati microorganism freeze-dried powder, wherein the vacuum freezing conditions are as follows: vacuum degree is 0.1Pa, cold hydrazine temperature is-45 ℃, and time is 12h.
The extraction method of the rhizoma polygonati polysaccharide comprises the following steps:
oven drying rhizoma Polygonati at 60deg.C for 5 hr, pulverizing, sieving, mixing with water, extracting at 120deg.C for 1.5 hr, and filtering to obtain rhizoma Polygonati polysaccharide water extract. Wherein the mass volume ratio of the rhizoma polygonati to the water is 1:10. Concentrating the water extractive solution to make the concentration of rhizoma Polygonati polysaccharide be 0.5g/L.
The number of probiotics in the rhizoma polygonati microbial freeze-dried powder prepared in examples 1-6 is detected and microscopic observed by referring to food safety national standard food microbiology inspection lactobacillus inspection (GB 4789.35-2010), and meanwhile, rhizoma polygonati polysaccharide is not added as a control group of each experimental group, wherein the control group is different from the corresponding experimental group in that rhizoma polygonati polysaccharide is not added, and the rest conditions are the same as the experimental group, and the results are shown in table 1.
TABLE 1 results of probiotic viable count detection
As can be seen from table 1, the survival number of probiotics added with the composite protectant of the polygonatum polysaccharide is improved by more than 10% compared with that of probiotics without the composite protectant of the polygonatum polysaccharide, because the polygonatum polysaccharide can be combined with probiotic cells to protect cell membrane structures, maintain stable cell osmotic pressure and protect the protein structure in the probiotic cells, thereby improving the survival rate of the probiotics in the probiotic products. In addition, for protecting probiotics by the polygonatum polysaccharide, not only the polygonatum polysaccharide aqueous extract of the application can be used, but also other forms of polygonatum polysaccharide can be adopted, and the scheme is also in the protection scope of the invention.
Claims (7)
1. The composite protective agent for improving the survival rate of the probiotics freeze-dried powder is characterized by comprising the following raw materials in parts by weight: 1-10 parts of skimmed milk powder, 1-10 parts of trehalose, 1-10 parts of sucrose, 1-10 parts of glucose, 1-10 parts of gelatin and 1-10 parts of rhizoma polygonati polysaccharide;
the Polygonatum polysaccharide is Polygonatum polysaccharide water extract, and the preparation method comprises the following steps: oven drying rhizoma Polygonati, pulverizing, sieving, mixing with water, extracting at high temperature, and filtering to obtain rhizoma Polygonati polysaccharide water extract;
the drying conditions are as follows: the temperature is 50-70 ℃ and the time is 2-6h;
the conditions of the high-temperature extraction are as follows: the mass volume ratio of rhizoma polygonati to water is 1:5-15, the temperature is 90-125 ℃ and the time is 0.5-2h.
2. The use of a composite protectant for improving survival rate of probiotic freeze-dried powder according to claim 1 in probiotic freeze-dried powder.
3. The use of the composite protectant for improving survival rate of the probiotics freeze-dried powder according to claim 2, characterized in that the preparation method of the probiotics freeze-dried powder comprises the following steps:
s1: inoculating probiotic strains into a microorganism culture medium, placing the culture medium into a shaking table for fermentation culture and activation, and collecting activated microorganism suspension;
s2: centrifuging the microorganism suspension in the step S1, discarding the supernatant, and collecting bacterial sludge;
s3: adding a composite protective agent into the bacterial mud, and uniformly mixing to obtain a probiotic suspension;
s4: pre-freezing the probiotic suspension in the step S3, and then performing vacuum freezing on the probiotic freeze-dried powder.
4. The use of a composite protectant for improving survival rate of probiotic freeze-dried powder according to claim 3, wherein the activation condition in S1 is: the temperature is 30-38 ℃ and the time is 12-48h.
5. The use of a composite protectant for improving survival rate of probiotic freeze-dried powder according to claim 3, wherein the centrifugation time in S2 is 1-10min, and the rotation speed during centrifugation is 8000-12000rpm.
6. The application of the composite protective agent for improving the survival rate of the freeze-dried probiotic powder according to claim 3, wherein the mass ratio of the bacterial sludge in S3 to the composite protective agent is 1:1-5.
7. The use of a composite protectant for improving survival rate of probiotic freeze-dried powder according to claim 3, wherein the pre-freezing condition in S4 is: the temperature is-20 to-4 ℃ and the time is 10-24 hours; the conditions of vacuum freezing are as follows: vacuum degree is 0.1-20Pa, cold hydrazine temperature is-180 to-45 ℃ and time is 12-24h.
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