CN117143795A - Method for improving oxidation resistance and promoting proliferation of probiotics and composition thereof - Google Patents
Method for improving oxidation resistance and promoting proliferation of probiotics and composition thereof Download PDFInfo
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- CN117143795A CN117143795A CN202311345212.9A CN202311345212A CN117143795A CN 117143795 A CN117143795 A CN 117143795A CN 202311345212 A CN202311345212 A CN 202311345212A CN 117143795 A CN117143795 A CN 117143795A
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- ZCIHMQAPACOQHT-ZGMPDRQDSA-N trans-isorenieratene Natural products CC(=C/C=C/C=C(C)/C=C/C=C(C)/C=C/c1c(C)ccc(C)c1C)C=CC=C(/C)C=Cc2c(C)ccc(C)c2C ZCIHMQAPACOQHT-ZGMPDRQDSA-N 0.000 description 1
- KBPHJBAIARWVSC-XQIHNALSSA-N trans-lutein Natural products CC(=C/C=C/C=C(C)/C=C/C=C(C)/C=C/C1=C(C)CC(O)CC1(C)C)C=CC=C(/C)C=CC2C(=CC(O)CC2(C)C)C KBPHJBAIARWVSC-XQIHNALSSA-N 0.000 description 1
- 235000004330 tyrosol Nutrition 0.000 description 1
- 235000021122 unsaturated fatty acids Nutrition 0.000 description 1
- 150000004670 unsaturated fatty acids Chemical class 0.000 description 1
- 235000015112 vegetable and seed oil Nutrition 0.000 description 1
- 235000019154 vitamin C Nutrition 0.000 description 1
- 239000011718 vitamin C Substances 0.000 description 1
- 235000019165 vitamin E Nutrition 0.000 description 1
- 229940046009 vitamin E Drugs 0.000 description 1
- 239000011709 vitamin E Substances 0.000 description 1
- FJHBOVDFOQMZRV-XQIHNALSSA-N xanthophyll Natural products CC(=C/C=C/C=C(C)/C=C/C=C(C)/C=C/C1=C(C)CC(O)CC1(C)C)C=CC=C(/C)C=CC2C=C(C)C(O)CC2(C)C FJHBOVDFOQMZRV-XQIHNALSSA-N 0.000 description 1
- 239000011701 zinc Substances 0.000 description 1
- 229910052725 zinc Inorganic materials 0.000 description 1
- OENHQHLEOONYIE-JLTXGRSLSA-N β-Carotene Chemical compound CC=1CCCC(C)(C)C=1\C=C\C(\C)=C\C=C\C(\C)=C\C=C\C=C(/C)\C=C\C=C(/C)\C=C\C1=C(C)CCCC1(C)C OENHQHLEOONYIE-JLTXGRSLSA-N 0.000 description 1
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
- C12N1/38—Chemical stimulation of growth or activity by addition of chemical compounds which are not essential growth factors; Stimulation of growth by removal of a chemical compound
-
- 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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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
- C12R2001/225—Lactobacillus
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
- C12R2001/225—Lactobacillus
- C12R2001/23—Lactobacillus acidophilus
Abstract
The application relates to a method for improving the antioxidation capability of probiotics and promoting proliferation and a composition thereof, belonging to the field of microbial culture. The method comprises the following steps: weighing a composition, wherein the composition is selected from one or two of apple peel powder, blueberry powder, blood orange powder, mulberry powder and red grape powder; preparing a special culture medium, wherein the raw materials comprise: yeast extract powder, peptone, naCl and KH 2 PO 4 、K 2 HPO 4 、CaCl 2 、MgSO 4 ·7H 2 O、NaHCO 3 Cysteine-acidsSalt, pig bile salt, tween 80; the preparation method comprises the following steps: weighing raw materials, dissolving the raw materials with distilled water, fixing the volume, and adding the composition into a culture medium after fixing the volume; inoculating probiotics, culturing and fermenting: inoculating the active probiotic seed solution into a special culture medium, and carrying out anaerobic fermentation on the inoculated culture medium. The scheme has excellent promotion effect on growth and proliferation of probiotics so as to resist oxidation.
Description
Technical Field
The application relates to the field of microorganism culture, in particular to a method for improving the antioxidation capability of probiotics and promoting proliferation and a composition thereof.
Background
Biological oxidation is an important physiological process of a biological organism, when active oxygen and other free radicals in the organism exceed a certain number, the active oxygen and other free radicals attack cell membranes, react with serum anti-protease, even rob electrons with genes, influence cell activity, cause injury to the organism, cause tumors, arthritis, cardiovascular diseases and the like, and finally cause aging. Although the body normally has a free radical scavenging system, if the body is affected by certain factors, the free radical scavenging system fails to cause excessive production of free radicals in the body, proteins which act in the body can enhance the antioxidant capacity of the body by supplementing antioxidant substances in the diet.
Probiotics are living microorganisms that when ingested in sufficient quantities produce health benefits to the host. According to the list of strains for food and the list of strains for infant food issued by the national health management, strains which are likely to meet the definition of "probiotics" are mainly from the genus lactobacillus and bifidobacterium. The lactobacillus as resident flora of intestinal canal can directly play an antioxidant role in the key part of oxidative damage of organism, namely the intestinal canal, and the lactobacillus can be continuously proliferated after the intestinal canal is planted, so as to generate more lactobacillus and continuously remove free radical in the intestinal canal. Lactic acid bacteria are therefore becoming increasingly popular as a probiotic.
At present, probiotics selected for researching the antioxidant activity are mostly separated from fermented foods or natural environments. In-vitro and in-vivo experiments prove that probiotics have antioxidant activity, but researches and patents on how to improve the antioxidant activity of probiotics are rarely available, and most of documents screen strains through an in-vitro oxidation activity evaluation method or compound with other active ingredients with antioxidant capacity to improve the antioxidant performance of the composition.
In chinese patent No. 103582486B, a probiotic with antioxidant activity and its use, a composition for the treatment of oxidative stress is provided, comprising a mixture comprising at least one strain belonging to a species selected from the group comprising bifidobacterium lactis BS05 (ID 1666), lactobacillus acidophilus LA06 (ID 1683) and lactobacillus brevis LBR01 (ID 1685) and having antioxidant properties. The other components are selenium, zinc, magnesium, manganese, glutathione, superoxide dismutase (SOD), vitamin C, vitamin E, beta-carotene, lycopene, tomato seed oil, quercetin, tyrosol, resveratrol, hydroxytyrosol, oleuropein, lutein, spirulina, alpha-lipoic acid, omega-3 unsaturated fatty acids, berry extract, green tea extract, cactus extract, jerusalem artichoke extract, papaya extract, melon extract, apple extract, hops extract, camellia extract, red clover extract, elder extract, rosemary extract, cocoa extract, olive leaf extract, pine bark extract and oyster extract, and other plant extracts with polyphenol content of more than 1% by weight. The application does not improve the antioxidant capacity of probiotics, but only improves the antioxidant capacity of the composition by adding an antioxidant active ingredient additionally.
Based on the above circumstances, the present application provides a method for improving the antioxidant capacity of probiotics and promoting proliferation and a composition thereof.
Disclosure of Invention
In order to fill the gap in the prior art, the application provides a method for improving the antioxidation capability of probiotics and promoting proliferation and a composition thereof.
The first application provides a method for improving the antioxidation capability and promoting proliferation of probiotics, which adopts the following technical scheme:
a method for improving the antioxidant capacity and promoting proliferation of probiotics, comprising the steps of:
(1) Raw material preparation: weighing a composition, wherein the composition is selected from one or two of apple peel powder, blueberry fruit powder, blood orange powder, mulberry fruit powder and red grape fruit powder;
(2) Preparing a special culture medium:
the culture medium comprises the following raw materials in parts by weight: 150 to 250 parts of yeast extract powder, 150 to 250 parts of peptone, 0.5 to 1.5 parts of NaCl and 3 to 5 parts of KH 2 PO 4 3-5 parts of K 2 HPO 4 0.5-1.5 portions of CaCl 2 0.5 to 1.5 parts of MgSO 4 ·7H 2 O, 150-250 parts of NaHCO 3 40-60 parts of cysteine-acid salt, 40-60 parts of pig bile salt and 150-250 parts of Tween 80;
the preparation method of the culture medium comprises the following steps: weighing the raw materials, dissolving the raw materials with distilled water, fixing the volume, and adding 5-10 mg/mL of the composition into the culture medium after fixing the volume to obtain the composition;
(3) Inoculating probiotics, culturing and fermenting:
inoculating the active probiotic strain seed liquid into the special culture medium according to an inoculation proportion, and placing the inoculated culture medium into a fermentation device for anaerobic fermentation.
According to the scheme, the culture medium is prepared by specifically adopting the components such as cysteine-acid salt, pig bile salt and the like according to a specific dosage proportion, so that the probiotics powder is inoculated to ferment in the special culture medium, the nutrition components in the culture medium can promote the growth of the probiotics, the activity is poor or useless bacteria can be screened out in the fermentation process, so that the occupation of nutrient substances between the probiotics to fight against the growth space is improved, in addition, the added fruit powder composition in the culture medium is rich in plant polyphenols and polysaccharide substances, the polyphenols have strong oxidation resistance and physiological activity, free radicals in human bodies can be removed, the oxidation resistance of the polyphenols can protect the growth and propagation conditions of the probiotics, the survival rate of the probiotics is improved, the probiotics are regulated in the fermentation process, the mutual symbiosis and stable in structure are formed, the proliferation of the probiotics is promoted remarkably, and the oxidation resistance of the probiotics formed by fermentation in the special culture medium is also improved unexpectedly.
Preferably, the composition is selected from apple peel powder, blueberry powder and blood orange powder, and the mixing ratio is 1:1 (1-3).
Preferably, the fruit powder is obtained from powder obtained by crushing corresponding fruits after squeezing, low-temperature concentration and freeze-drying, and the mesh number is 80-100.
Preferably, the preparation method of the culture medium comprises the following steps: weighing the raw materials, dissolving the raw materials with distilled water, fixing the volume to 1L, sterilizing the raw materials for 20min at the temperature of 121 ℃, adding 3 to 5mL of hemin (5 mg/mL) and 0.5 to 1.5mL of vitamin K1 (10 mg/mL) under the aseptic condition, and deoxidizing the raw materials overnight in an anaerobic incubator.
Preferably, the culture medium comprises the following raw materials in parts by weight: 200 parts of yeast extract powder, 200 parts of peptone, 1 part of NaCl and 4 parts of KH 2 PO 4 4 parts of K 2 HPO 4 1 part CaCl 2 1 part of MgSO 4 ·7H 2 O, 200 parts of NaHCO 3 50 parts of cysteine-acid salt, 50 parts of pig bile salt and 200 parts of Tween 80; the preparation method of the culture medium comprises the following steps: weighing the raw materials, dissolving with distilled water, fixing volume to 1L, sterilizing at 121deg.C for 20min, adding 4mL of hemin (5 mg/mL) and 1mL of vitamin K1 (10 mg/mL) under aseptic condition, deoxidizing overnight in an anaerobic incubator, and adding 5mg/mL of the composition into the culture medium.
Preferably, the concentration of the active probiotic seed liquid is 15 hundred million strains per milliliter, and the inoculation proportion is that the active probiotic seed liquid: special medium = 1: (10-15).
Preferably, the anaerobic fermentation temperature is 35+/-0.5 ℃, and the rotating speed of the fermentation device is (120-180) r/min.
Preferably, after anaerobic fermentation has been completed for 24 hours, the temperature of the monitored medium is no longer increased.
Preferably, the fermentation process is suitable for culturing one or more of Lactobacillus acidophilus (La-14), bifidobacterium longum (BI-05), lactobacillus rhamnosus (Lr-G14), lactobacillus paracasei (Lpc-G110).
Preferably, the active probiotic seed liquid is obtained by activating and culturing probiotic powder, wherein the probiotic powder is one or more of lactobacillus acidophilus (La-14), bifidobacterium longum (BI-05), lactobacillus rhamnosus (Lr-G14) and lactobacillus paracasei (Lpc-G110).
The probiotics powder in the scheme is derived from a bacterial list which is issued by a health management department and can be used for foods and a bacterial list which is issued by an infant food, and the probiotics can obtain excellent antioxidant capacity and promotion effect of promoting proliferation in the special culture medium of the scheme.
In a second aspect, the application provides a composition with the functions of enhancing the antioxidation capability of probiotics and promoting proliferation, which adopts the following technical scheme:
a composition with effects of enhancing oxidation resistance and promoting proliferation of probiotics is prepared from one or two of apple peel powder, blueberry fruit powder, blood orange powder, mulberry fruit powder and red grape fruit powder, and can be used for culturing probiotics.
In summary, the application has the following beneficial technical effects:
the application adopts the self-made culture medium prepared by cysteine-acid salt, pig bile salt and other components according to the specific dosage proportion, has excellent promotion effect on the growth and proliferation of probiotics, and besides, the fruit powder composition innovatively added in the culture medium is rich in plant polyphenol substances, the antioxidation effect of the polyphenol substances not only protects the growth and proliferation conditions of the probiotics, but also further improves the survival rate of the probiotics, and the antioxidation activity of the probiotics formed by fermentation in the special culture medium is also unexpectedly promoted.
Detailed Description
The present application will be described in further detail with reference to examples.
Example 1
A method for improving the antioxidation capability and promoting proliferation of probiotics comprises the following steps:
(1) Raw material preparation: weighing a composition selected from apple peel powder, blueberry fruit powder and blood orange powder, and weighing 1mg/mL apple peel powder, 1mg/mL blueberry fruit powder and 3mg/mL blood orange powder according to the amount of a culture medium; the apple peel powder, the blueberry fruit powder and the blood orange powder are obtained by crushing corresponding fruits after squeezing, low-temperature concentration and freeze-drying, and the mesh number is 80-100, and the apple peel powder, the blueberry fruit powder and the blood orange powder are purchased from the market.
(2) Preparation of the culture medium:
the culture medium comprises the following raw materials in parts by weight: 200 parts of yeast extract powder, 200 parts of peptone, 1 part of NaCl and 4 parts of KH 2 PO 4 4 parts of K 2 HPO 4 1 part CaCl 2 1 part of MgSO 4 ·7H 2 O, 200 parts of NaHCO 3 50 parts of cysteine-acid salt, 50 parts of pig bile salt and 200 parts of Tween 8.
The preparation method of the culture medium comprises the following steps: sterilizing at 121deg.C for 20min, adding 4mL of hemin (5 mg/mL) and 1mL of VK1 (10 mg/mL) under aseptic condition, adding the above composition, and deoxidizing overnight in anaerobic incubator.
(3) Inoculating and culturing probiotics:
the probiotic powder purchased in the market is adopted, and is subjected to activation culture until the concentration is 15 hundred million strains per milliliter of active probiotic seed liquid, and the inoculation proportion is the active probiotic seed liquid: special medium = 1:15.
inoculating the active probiotic strain seed liquid into a special culture medium according to an inoculation proportion, placing the inoculated culture medium into a fermentation device for anaerobic fermentation, wherein the anaerobic fermentation temperature is 35+/-0.5 ℃, the rotating speed of the fermentation device is 120r/min, and after anaerobic fermentation is carried out for 24 hours, monitoring the internal temperature of the culture medium at regular time, and fermenting until the internal temperature of the culture medium is not increased any more.
Example 2
A method for improving the antioxidation capability and promoting proliferation of probiotics comprises the following steps:
(1) Raw material preparation: weighing the composition, wherein the composition is selected from apple peel powder, blueberry fruit powder and blood orange powder, and weighing 2mg/mL apple peel powder, 2mg/mL blueberry fruit powder and 1mg/mL blood orange powder according to the amount of the culture medium;
(2) Preparation of the culture medium:
the culture medium comprises the following raw materials in parts by weight: 200 parts of yeast extract powder, 200 parts of peptone, 1 part of NaCl and 4 parts of KH 2 PO 4 4 parts of K 2 HPO 4 1 part CaCl 2 1 part of MgSO 4 ·7H 2 O, 200 parts of NaHCO 3 50 parts ofCysteine-acid salt, 50 parts of pig bile salt and 200 parts of Tween 8.
The preparation method of the culture medium comprises the following steps: sterilizing at 121deg.C for 20min, adding 4mL of hemin (5 mg/mL) and 1mL of VK1 (10 mg/mL) under aseptic condition, adding the above composition, and deoxidizing overnight in anaerobic incubator.
(3) Inoculating and culturing probiotics:
inoculating the active probiotic strain seed liquid into a special culture medium according to an inoculation proportion, placing the inoculated culture medium into a fermentation device for anaerobic fermentation, wherein the anaerobic fermentation temperature is 35+/-0.5 ℃, the rotating speed of the fermentation device is 120r/min, and after anaerobic fermentation is carried out for 24 hours, monitoring the internal temperature of the culture medium at regular time, and fermenting until the internal temperature of the culture medium is not increased any more.
Example 3
A method for improving the antioxidation capability and promoting proliferation of probiotics comprises the following steps:
(1) Raw material preparation: weighing a composition selected from apple peel powder, and weighing 5mg/mL apple peel powder according to the amount of a culture medium;
(2) Preparation of the culture Medium
The culture medium comprises the following raw materials in parts by weight: 200 parts of yeast extract powder, 200 parts of peptone, 1 part of NaCl and 4 parts of KH 2 PO4, 4 parts K 2 HPO 4 1 part CaCl 2 1 part of MgSO 4 ·7H 2 O, 200 parts of NaHCO 3 50 parts of cysteine-acid salt, 50 parts of pig bile salt and 200 parts of Tween 80 are sterilized at 121 ℃ for 20min, 4mL of hemin (5 mg/mL) and 1mL of VK1 (10 mg/mL) are added under aseptic conditions, and the composition is added at the same time, and deoxygenated in an anaerobic incubator overnight for standby.
(3) Probiotic inoculation culture
Inoculating the active probiotic strain seed liquid into a special culture medium according to an inoculation proportion, placing the inoculated culture medium into a fermentation device for anaerobic fermentation, wherein the anaerobic fermentation temperature is 35+/-0.5 ℃, the rotating speed of the fermentation device is 120r/min, and after anaerobic fermentation is carried out for 24 hours, monitoring the internal temperature of the culture medium at regular time, and fermenting until the internal temperature of the culture medium is not increased any more.
Example 4
A method for improving the antioxidation capability and promoting proliferation of probiotics comprises the following steps:
(1) Raw material preparation: weighing a composition selected from blueberry fruit powder, and weighing 5mg/mL blueberry fruit powder according to the amount of a culture medium;
(2) Preparation of the culture Medium
The culture medium comprises the following raw materials in parts by weight: 200 parts of yeast extract powder, 200 parts of peptone, 1 part of NaCl and 4 parts of KH 2 PO4, 4 parts K 2 HPO 4 1 part CaCl 2 1 part of MgSO 4 ·7H 2 O, 200 parts of NaHCO 3 50 parts of cysteine-acid salt, 50 parts of pig bile salt and 200 parts of Tween 80 are sterilized at 121 ℃ for 20min, 4mL of hemin (5 mg/mL) and 1mL of VK1 (10 mg/mL) are added under aseptic conditions, and the composition is added at the same time, and deoxygenated in an anaerobic incubator overnight for standby.
(3) Probiotic inoculation culture
Inoculating the active probiotic strain seed liquid into a special culture medium according to an inoculation proportion, placing the inoculated culture medium into a fermentation device for anaerobic fermentation, wherein the anaerobic fermentation temperature is 35+/-0.5 ℃, the rotating speed of the fermentation device is 120r/min, and after anaerobic fermentation is carried out for 24 hours, monitoring the internal temperature of the culture medium at regular time, and fermenting until the internal temperature of the culture medium is not increased any more.
Example 5
A method for improving the antioxidation capability and promoting proliferation of probiotics comprises the following steps:
(1) Raw material preparation: weighing a composition selected from the group consisting of blood orange powder, weighing 5mg/mL of blood orange powder in an amount of culture medium;
(2) Preparation of the culture Medium
The culture medium comprises the following raw materials in parts by weight: 200 parts of yeast extract powder, 200 parts of peptone, 1 part of NaCl and 4 parts of KH 2 PO4, 4 parts K 2 HPO 4 1 part CaCl 2 1 part of MgSO 4 ·7H 2 O, 200 parts of NaHCO 3 50 parts of cysteine-acid salt, 50 parts of pig bile salt and 200 parts of Tween 80, sterilizing at 121 ℃ for 20min, and sterilizingUnder the condition of adding 4mL of hemin (5 mg/mL) and 1mL of VK1 (10 mg/mL), simultaneously adding the above composition, and deoxidizing overnight in an anaerobic incubator for later use.
(3) Probiotic inoculation culture
Inoculating the active probiotic strain seed liquid into a special culture medium according to an inoculation proportion, placing the inoculated culture medium into a fermentation device for anaerobic fermentation, wherein the anaerobic fermentation temperature is 35+/-0.5 ℃, the rotating speed of the fermentation device is 120r/min, and after anaerobic fermentation is carried out for 24 hours, monitoring the internal temperature of the culture medium at regular time, and fermenting until the internal temperature of the culture medium is not increased any more.
Comparative example
Comparative example 1
A probiotic fermentation process, comparative example 1 differs from example 1 only in that the fruit powder composition is replaced by 5mg/mL fructooligosaccharides.
Comparative example 2
A probiotic fermentation process, comparative example 2, differs from example 1 only in that the fruit powder composition is replaced with 5mg/mL of galactooligosaccharide.
Comparative example 3
A probiotic fermentation process, comparative example 3, differs from example 1 only in that the fruit powder composition was replaced with 5mg/mL of xylooligosaccharide.
Comparative example 4
Comparative example 4 differs from example 1 only in that no fruit powder composition was added.
Comparative example 5
A probiotic fermentation process, comparative example 5 differs from example 1 only in that no pig bile salts were added to the medium.
Comparative example 6
A probiotic fermentation process, comparative example 6, differs from example 1 only in that no porcine bile salt and no fruit powder composition were added to the medium.
Performance test
1. Detection of probiotic proliferation ability
The OD600 of each probiotic was measured by an ultraviolet spectrophotometer for the bacterial liquids before the culture of example 1, comparative example 2, comparative example 3, comparative example 4, comparative example 5, comparative example 6 and comparative example 6 after the culture was completed, and the detection results are shown in Table 1.
TABLE 1
2. Detection of hydroxyl radical HO-scavenging ability
The bacterial solutions obtained in each example and comparative example are freeze-dried to obtain probiotic powder, and oxidation resistance of each group of probiotic powder is tested:
1mL of salicylic acid-ethanol solution (5 mmol/L), 1mL of LFASO, was added to the tube 4 Solution (5 mmol/L), 1mLH 2 O 2 Solution (3 mmol/L) and 1mL of probiotic powder liquid (bacterial liquid concentration 1×10) 8 CFU/mL), and then constant volume to 10mL with double distilled water, water-bath at 37 ℃ for 15min, centrifuging at 4 ℃ at 6000rpm for 10min, and taking supernatant to measure absorbance at 510nm (with double distilled water as reference). HO-clearance (%) = [ (Ao-As)/Ao]The higher the HO-clearance rate, the stronger the clearance of active oxygen, i.e., the stronger the antioxidant capacity, the X100% (Ao is the OD value without a sample; as is the OD value with a sample). The detection results are shown in Table 2.
TABLE 2
From the analysis on the detection results of the table 1 and the table 2, the probiotics cultured by adopting the special culture medium of the scheme have obviously improved proliferation capacity and antioxidation capacity.
As is clear from the analysis of the detection results of example 1 and comparative examples 1-3, fructo-oligosaccharide, galacto-oligosaccharide or xylo-oligosaccharide is adopted to replace the fruit powder composition, the OD600 is improved compared with that before cultivation, but the rate of increase of the bacterial load of a special culture medium containing the fruit powder composition is not higher, and the hydroxy free radical scavenging capacity is not stronger than that of probiotics of the special culture medium, so that the probiotics in the special culture medium are rapidly increased, and the probiotics have excellent growth and propagation conditions due to the fact that the probiotics are rich in polysaccharides, polyphenols and the like instead of the fructo-oligosaccharide, galacto-oligosaccharide or xylo-oligosaccharide in the fruit powder.
In addition, from the analysis of the test results of comparative examples 4 to 6, it is known that the special culture medium has excellent proliferation ability and antioxidation ability for probiotics, and is an effect that is contributed by the pig bile salt and fruit powder composition added in the special culture medium. According to the scheme, the pig bile salt and other components are added specifically to prepare the culture medium according to a specific dosage proportion, so that useless bacteria or weak bacteria in the bacterial liquid are screened out in the fermentation process, plant polyphenols and other components which are rich in the fruit powder composition are provided for the probiotics by replacing prebiotics in the simulated intestinal environment of the special culture medium, the effect of improving the survival rate of the screened probiotics is achieved, so that a mutually symbiotic and structurally stable flora is formed in the fermentation process, and the bacterial amount among the probiotics such as lactobacillus acidophilus, bifidobacterium longum, lactobacillus rhamnosus and lactobacillus paracasei is increased uniformly, thereby improving the proliferation efficiency of the probiotics as a whole and improving the antioxidation capability of the probiotics.
In addition, in further intensive studies, it was found that the improvement of the proliferation efficiency of bifidobacterium longum in the case of adding a single fruit powder was not very remarkable, probably due to the growth characteristics of the bifidobacterium longum in a lump, resulting in a decrease in the nutrition absorption efficiency of bifidobacterium longum, a high concentration of metabolites of the flora, which is unfavorable for the proliferation and growth thereof, and particularly when no pig bile salt is added, the aseptic bacteria become dominant bacteria in a local area, resulting in a certain degree of inhibition of the growth of bifidobacterium longum. The test in examples 1-5 shows that the special added fruit powder composition is selected to be apple peel powder, blueberry fruit powder and blood orange powder which are mixed according to the ratio of 1:1:3, so that the antioxidation capability and proliferation efficiency of each probiotic can be enhanced, the stacking aggregation condition of bifidobacterium longum can be controlled by the special fruit powder ratio, and the bacterial load growth of bifidobacterium longum can be improved more obviously.
The present embodiment is only for explanation of the present application and is not to be construed as limiting the present application, and modifications to the present embodiment, which may not creatively contribute to the present application as required by those skilled in the art after reading the present specification, are all protected by patent laws within the scope of claims of the present application.
Claims (10)
1. A method for improving the antioxidant capacity and promoting proliferation of probiotics, comprising the steps of:
(1) Raw material preparation: weighing a composition, wherein the composition is selected from one or two of apple peel powder, blueberry fruit powder, blood orange powder, mulberry fruit powder and red grape fruit powder;
(2) Preparing a special culture medium:
the culture medium comprises the following raw materials in parts by weight: 150 to 250 parts of yeast extract powder, 150 to 250 parts of peptone, 0.5 to 1.5 parts of NaCl and 3 to 5 parts of KH 2 PO 4 3-5 parts of K 2 HPO 4 0.5-1.5 portions of CaCl 2 0.5 to 1.5 parts of MgSO 4 ·7H 2 O, 150-250 parts of NaHCO 3 40-60 parts of cysteine-acid salt, 40-60 parts of pig bile salt and 150-250 parts of Tween 80;
the preparation method of the culture medium comprises the following steps: weighing the raw materials, dissolving the raw materials with distilled water, fixing the volume, and adding 5-10 mg/mL of the composition into the culture medium after fixing the volume to obtain the composition;
(3) Inoculating probiotics, culturing and fermenting:
inoculating the active probiotic strain seed liquid into the special culture medium according to an inoculation proportion, and placing the inoculated culture medium into a fermentation device for anaerobic fermentation.
2. The method for improving the antioxidant capacity and promoting proliferation of probiotics according to claim 1, wherein the composition is prepared by mixing apple peel powder, blueberry fruit powder and blood orange powder in a mixing ratio of 1:1 (1-3).
3. The method for improving the oxidation resistance and promoting proliferation of probiotics according to claim 1, wherein the fruit powder is obtained from powder obtained by crushing corresponding fruits after squeezing, low-temperature concentration and freeze-drying, and the mesh number is 80-100.
4. The method for improving the antioxidant capacity and promoting proliferation of probiotics according to claim 1, wherein the preparation method of the special culture medium is as follows: weighing the raw materials, dissolving the raw materials with distilled water, fixing the volume to 1L, sterilizing the raw materials for 20min at the temperature of 121 ℃, adding 3-5 mL of hemin (5 mg/mL) and 0.5-1.5 mL of vitamin K1 (10 mg/mL) under the aseptic condition, and deoxidizing overnight in an anaerobic incubator for later use.
5. The method for improving the antioxidant capacity and promoting proliferation of probiotics according to claim 1, wherein the raw materials of the special culture medium comprise, in parts by weight: 200 parts of yeast extract powder, 200 parts of peptone, 1 part of NaCl and 4 parts of KH 2 PO 4 4 parts of K 2 HPO 4 1 part CaCl 2 1 part of MgSO 4 ·7H 2 O, 200 parts of NaHCO 3 50 parts of cysteine-acid salt, 50 parts of pig bile salt and 200 parts of Tween 80; the preparation method of the culture medium comprises the following steps: weighing the raw materials, dissolving with distilled water, fixing volume to 1L, sterilizing at 121deg.C for 20min, adding 4mL of hemin (5 mg/mL) and 1mL of vitamin K1 (10 mg/mL) under aseptic condition, deoxidizing overnight in an anaerobic incubator, and adding 5mg/mL of the composition into the culture medium.
6. The method for improving the antioxidant capacity and promoting proliferation of probiotics according to claim 1, wherein the concentration of the active probiotic seed liquid is 15 hundred million strains per milliliter, and the inoculation proportion is that the active probiotic seed liquid: special medium = 1: (10-15).
7. The method for improving the oxidation resistance and promoting proliferation of probiotics according to claim 1, wherein the anaerobic fermentation temperature is 35+/-0.5 ℃, and the rotating speed of the fermentation device is (120-180) r/min.
8. The method for improving the antioxidant capacity and promoting proliferation of probiotics according to claim 1, wherein the fermentation is completed when the temperature of the monitored culture medium is not increased after the anaerobic fermentation is completed for 24 hours.
9. The method for improving the antioxidant capacity and promoting proliferation of probiotics according to claim 1, wherein the fermentation method is suitable for culturing one or more of lactobacillus acidophilus (La-14), bifidobacterium longum (BI-05), lactobacillus rhamnosus (Lr-G14) and lactobacillus paracasei (Lpc-G110).
10. A composition having an enhanced antioxidant capacity of probiotics and a proliferation promoting effect, characterized in that it is a mixture of fruit powders selected from one or two of apple peel powder, blueberry fruit powder, blood orange powder, mulberry fruit powder and red grape fruit powder, which is used in the method for enhancing antioxidant capacity and proliferation promoting effect of probiotics according to any one of claims 1 to 9.
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Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR20190002036A (en) * | 2017-06-29 | 2019-01-08 | 수원대학교산학협력단 | Prebiotics comprising blueberry extract for proliferating intestinal probiotics the use thereof |
| CN111493315A (en) * | 2020-05-26 | 2020-08-07 | 广东时代食品与生命健康研究有限公司 | Memory-improving antioxidant composition and application thereof |
| CN115197870A (en) * | 2022-05-19 | 2022-10-18 | 微康益生菌(苏州)股份有限公司 | Preparation method and application of lactobacillus with high antioxidant stress performance |
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| KR20190002036A (en) * | 2017-06-29 | 2019-01-08 | 수원대학교산학협력단 | Prebiotics comprising blueberry extract for proliferating intestinal probiotics the use thereof |
| CN111493315A (en) * | 2020-05-26 | 2020-08-07 | 广东时代食品与生命健康研究有限公司 | Memory-improving antioxidant composition and application thereof |
| CN115197870A (en) * | 2022-05-19 | 2022-10-18 | 微康益生菌(苏州)股份有限公司 | Preparation method and application of lactobacillus with high antioxidant stress performance |
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