CN108606329A - A kind of prebiotics, probiotics compound micro-ecological preparation and preparation method thereof - Google Patents

A kind of prebiotics, probiotics compound micro-ecological preparation and preparation method thereof Download PDF

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CN108606329A
CN108606329A CN201810412982.3A CN201810412982A CN108606329A CN 108606329 A CN108606329 A CN 108606329A CN 201810412982 A CN201810412982 A CN 201810412982A CN 108606329 A CN108606329 A CN 108606329A
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preparation
probiotics
prebiotics
bacillus subtilis
compound micro
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田木
王艳丹
赵琳琳
解慧
郝鑫
周晓梅
柴振宇
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BODA COLLEGE OF JILIN NORMAL UNIVERSITY
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    • AHUMAN NECESSITIES
    • A23FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23LFOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L33/00Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
    • A23L33/10Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
    • A23L33/135Bacteria or derivatives thereof, e.g. probiotics
    • AHUMAN NECESSITIES
    • A23FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23LFOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L33/00Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
    • A23L33/20Reducing nutritive value; Dietetic products with reduced nutritive value
    • A23L33/21Addition of substantially indigestible substances, e.g. dietary fibres
    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N1/00Microorganisms, 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
    • C12N1/14Fungi; Culture media therefor
    • C12N1/16Yeasts; Culture media therefor
    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N1/00Microorganisms, 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
    • C12N1/20Bacteria; Culture media therefor
    • AHUMAN NECESSITIES
    • A23FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23VINDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2002/00Food compositions, function of food ingredients or processes for food or foodstuffs

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  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Health & Medical Sciences (AREA)
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Abstract

The invention discloses a kind of prebiotics, probiotics compound micro-ecological preparation and preparation method thereof, and the application's mainly comprises the following steps:By choosing lactic acid bacteria, saccharomycete and the strain that bacillus subtilis is compound micro-ecological preparation, wherein lactic acid bacteria and saccharomycete come from Kefir granule, Kefir granule and bacillus subtilis are activated respectively, by Kefir granule and bacillus subtilis according to 3:1 ratio mixes; probiotics mixed bacteria is inoculated in culture medium by the 3~10% of fermentating liquid volume, fermented and cultured, adds prebiotics; skimmed milk powder, trehalose, sodium glutamate, glycerine are added simultaneously as protective agent, carries out low temperature spray drying after mixing and probiotics is made.It is characteristic of the invention that containing lactic acid bacteria in probiotics, saccharomycete and bacillus subtilis can adjust body microecological balance, while also contain prebiotic substance jointly, can preferably improve body health, improve immunity of organisms.

Description

A kind of prebiotics, probiotics compound micro-ecological preparation and preparation method thereof
Technical field
The invention belongs to technical field of microbial fermentation, be related to a kind of prebiotics, probiotics compound micro-ecological preparation and its Preparation method.
Background technology
Nineteen forties add antibiotic to promote fast growth of animal in feed, but with antibiosis Plain long-term extensive use, will produce drug resistance, medicament residue, the drawbacks such as animal body immunity degradation.Probiotics with The advantages that its green, safety, drug residue free, has gradually replaced antibiotic and has been widely used.With traditional feed addictive phase Than probiotics micro-ecological formulation can effectively inhibit the growth and breeding of the pathogenic bacteria and pathogenic microorganism in alimentary canal, and make benefit Raw bacterium is colonized in enteron aisle, and adjustment animal intestinal tract balance has the effect of the prevention of Animal diseases certain.Currently, the world is each State gradually increases and widelys popularize to the attention degree of probiotics.Prebiotics, probiotics compound micro-ecological preparation are with micro- Ecology is used as theoretical foundation, is made by special processing process, micro- in animal body to improve containing viable bacteria and its metabolite The ecological balance and play prebiotic effect, improve animal immunizing power and the general level of the health active bacteria formulation.
A kind of replenishers of diet of prebiotics pass through the growth and work of the one or more of probiotics of the stimulation of selectivity Property, so as to improve substance of the host health without being digested absorption.Common prebiotics have oligofructose, inulin, hedgehog hydnum Mushroom polysaccharide, Agaricus Blazei Murrill polysaccharide and lentinan etc..Prebiotics cannot be utilized directly by body and be played a role, but pass through probiotics It plays a role jointly.Do not have to decompose the substance of prebiotics in body alimentary canal, therefore its most of prebiotic substance can pass through machine Body small intestine and stomach are utilized without being decomposed, and are directly entered big enteral and are utilized by probiotics such as Bifidobacterium, lactic acid bacterias.
Kefir fermenting agent be by lactic acid bacteria, saccharomycete, a variety of probiotic groups such as acetic acid bacteria at bacterial strain, lactic acid therein Bacterium and its metabolite there is strong inhibiting effect, saccharomycete therein can generate unique aroma pathogenic microbes Taste, thus Kefir grains are also known as champagne in acidified milk.It is a kind of leavening containing more bacterial strains.
The main probiotics preparation of domestic and international market is bacillus subtilis formulation and lactobacillus preparation at present, and majority is more Kind single culture is prepared, and strain constitutes relatively simple.The present invention is using Kefir fermenting agent as lactic acid bacteria and saccharomycete Source is mixed with bacillus subtilis, and cultivation and fermentation, addition hericium erinaceum polysaccharide, Agaricus Blazei Murrill polysaccharide and mushroom are mixed through high density One or more of polysaccharide passes through low temperature spray drying technology, and prebiotics, probiotics compound micro-ecological preparation is made.Micro- life State additive has total viable count high, can adjust the gastrointestinal tract balance of body, promote body growth, enhancing immunity of organisms etc. Effect.
Invention content
The purpose of the present invention is to provide a kind of prebiotics, probiotics compound micro-ecological preparation and preparation method thereof, prepare Prebiotics, probiotics compound micro-ecological preparation not only contain the prebiotic substance for improving host health, while there are many also containing Probiotics and total viable count height, the effect of playing and adjust body gastrointestinal tract balance, enhance immunity of organisms.
The present invention is realized by following technological means:A kind of prebiotics, probiotics compound micro-ecological preparation, it is compound Probiotics are formed by prebiotics and probiotic combinations, and the probiotics is by lactic acid bacteria, saccharomycete and bacillus subtilis Mixed fermentation forms, and lactic acid bacteria and saccharomycete are provided by Kefir granule compound strain, and total viable count is more than 5,000,000,000 in preparation Cfu/g, the prebiotics are one or more of hericium erinaceum polysaccharide, Agaricus Blazei Murrill polysaccharide and lentinan composition, additive amount It is 0.3%~1.5%.
The preparation method of prebiotics, probiotics compound micro-ecological preparation:Compound micro-ecological preparation is mixed using a variety of strains It ferments, after High Density Cultivation, adds prebiotic substance, be made by low temperature spray drying.By probiotics mixed bacteria by fermentation The 3~10% of liquid product are inoculated in culture medium, and the wherein inoculative proportion of Kefir granule and bacillus subtilis is 3:1,28~36 Under the conditions of DEG C, ferment 16~24 hours.
In above-mentioned preparation, the culture medium contains following components by weight percentage:Concentrated whey protein powder 2~ 10%, glucose 1~10%, yeast powder 0.5~1%, calcium carbonate 0.4~1%, magnesium sulfate 0.01~0.05%, manganese sulfate 0.001~0.005%, whey powder 1~10%.Mentioned component is dissolved in distilled water, and it is 6.5~7.5 to adjust medium pH, Sterilization conditions keep the temperature 10~30min under the conditions of being 85~115 DEG C, are cooled to 20~30 DEG C.
In above-mentioned preparation, the mixed fermentation condition is:Probiotics mixed bacteria is pressed the 3~10% of fermentating liquid volume It is inoculated in culture medium, the wherein inoculative proportion of Kefir fermenting agent and bacillus subtilis is 3:Under the conditions of 1,28~36 DEG C, hair Ferment 16~24 hours, the prebiotics and protective agent for adding 0.3%-1.5% are made by spray drying.
The prebiotics, the specific preparation method of probiotics compound micro-ecological preparation include the following steps:
1) bacillus subtilis is activated, and prepares Kefir fermenting agent.
2) probiotics mixed bacteria is inoculated in by the 3~10% of fermentating liquid volume in high-density culture medium, wherein opening phenanthrene That leavening and the inoculative proportion of bacillus subtilis are:Kefir fermenting agent: bacillus subtilis=3:1, it is inoculated in culture In base, under the conditions of 28~36 DEG C, ferment 16~24 hours.
3) after fermentation, stirring centrifugation centrifuges 15~30min under the conditions of 3000~6000r/min, collects concentration Thalline.
4) prebiotics of 0.3%-1.5% are added into concentration thalline, while skimmed milk powder 10~20%, trehalose is added 1~10%, sodium glutamate 1~10%, glycerine 1~5% are used as protective agent, carry out low temperature spray drying after mixing, control Inlet air temperature is 80~120 DEG C, 50~70 DEG C of leaving air temp, to ensure to contain a certain amount of total viable count in the preparation prepared.
The beneficial effects of the present invention are:
1) Kefir fermenting agent and bacillus subtilis are carried out mixed culture fermentation, the probiotics of preparation by the present invention Strain is various, and total viable count is up to 5,000,000,000 cfu/g or more;2) the composite bacteria of the invention addition pair after mixed culture is fermented The beneficial prebiotic substance of body adjusts body gastrointestinal tract balance to preferably play, promotes body digestion, increase body body Weight, enhancing immunity of organisms and other effects.
Specific implementation mode
1) preparation of probiotics leaven:
The preparation of Kefir fermenting agent:The degreasing milk solution of configuration 10%, by 85~95 DEG C, after 10~30min sterilizations, 20~30 DEG C are cooled to, Kefir grains throw type leaven is added according to the 0.1~0.5% of skimmed milk volume, at 23~28 DEG C Under the conditions of 14~18h of fermented and cultured, it is spare under the conditions of being stored in 4 DEG C.
The preparation of bacillus subtilis:The bacillus subtilis frozen is subjected to scribing line culture, colonies typical is isolated and connects Kind shake culture 18h under the conditions of 37 DEG C in fluid nutrient medium, the standard type of bacillus subtilis is met through microscopy, grows shape State is good, and being inoculated in fluid nutrient medium by inoculum concentration 3~6% is activated, 37 DEG C, and shake culture 18~for 24 hours, it is stored in 4 DEG C Under the conditions of it is spare.
2) mixed fermentation:
The configuration of culture medium:10~20g of whey protein concentrate, 6~8g of glucose, yeast are added into 500mL distilled water 3~5g of powder, 3~6g of calcium carbonate, 0.1~0.3g of magnesium sulfate, 0.01~0.03g of manganese sulfate, 10~15g of whey powder fully dissolve It is 6.5~7.5 to adjust medium pH afterwards, and sterilization conditions keep the temperature 10~30min under the conditions of being 85~115 DEG C, are cooled to 20~30 It is DEG C spare.To culture medium inoculated after cooling, by 3~6% inoculum concentrations, wherein Kefir fermenting agent:Bacillus subtilis=3: 1,28~36 DEG C, fermented and cultured 18~for 24 hours.
3) preparation of compound micro-ecological preparation:
Fermentation broth agitation obtained is uniform, under the conditions of 4~10 DEG C, 3000~6000 revs/min of rotating speed, the time 15~ 30min centrifugal treatings collect concentration thalline;Based on thalline mass percent, the prebiotic substance of 0.3%-1.5% is added, is taken off Fat milk powder 10~15%, trehalose 1~5%, sodium glutamate 1~5%, glycerine 1~3% are uniformly mixed, and it is dry to carry out cold nebulization Dry, control inlet air temperature is 80~120 DEG C, and 50~70 DEG C of leaving air temp obtains prebiotics, probiotics compound micro-ecological preparation.
The following examples are further illustrations of the invention, rather than limiting the invention.
Embodiment 1:
1) preparation of probiotics leaven
The preparation of Kefir fermenting agent:The degreasing milk solution of configuration 10% after 30min sterilizations, is cooled to 20 by 85 DEG C DEG C, Kefir grains throw type leaven is added according to the 0.1% of skimmed milk volume, fermented and cultured 14h under conditions of 28 DEG C is preserved It is spare under the conditions of 4 DEG C.
The preparation of bacillus subtilis:The bacillus subtilis frozen is subjected to scribing line culture, colonies typical is isolated and connects Kind shake culture 18h under the conditions of 37 DEG C in fluid nutrient medium, the standard type of bacillus subtilis is met through microscopy, grows shape State is good, and being inoculated in fluid nutrient medium by inoculum concentration 6% is activated, 37 DEG C, shake culture 18h, standby under the conditions of being stored in 4 DEG C With.
2) mixed fermentation
The configuration of culture medium:Whey protein concentrate 10g, glucose 8g, yeast powder 5g, carbon are added into 500mL distilled water Sour calcium 3g, magnesium sulfate 0.3g, manganese sulfate 0.01g, whey powder 15g, it is 6.5 that medium pH is fully adjusted after dissolving, sterilization conditions Keep the temperature 30min under the conditions of being 85 DEG C, be cooled to 20 DEG C it is spare.To culture medium inoculated after cooling, by 3% inoculum concentration, wherein open Fei Er leavenings:Bacillus subtilis=3:1,28 DEG C, fermented and cultured is for 24 hours.
3) preparation of compound micro-ecological preparation
Fermentation broth agitation obtained is uniform, and under the conditions of 4 DEG C, 3000 revs/min of rotating speed, time 30min centrifugal treating is received Collection concentration thalline;Based on thalline mass percent, 0.3% prebiotic substance, skimmed milk powder 15%, trehalose 1%, paddy are added Propylhomoserin sodium 5%, glycerine 1% are uniformly mixed, and carry out low temperature spray drying, and control inlet air temperature is 80 DEG C, 50 DEG C of leaving air temp, Obtain prebiotics, probiotics compound micro-ecological preparation.
Embodiment 2
1) preparation of probiotics leaven
The preparation of Kefir fermenting agent:The degreasing milk solution of configuration 10% after 20min sterilizations, is cooled to 25 by 90 DEG C DEG C, Kefir grains throw type leaven is added according to the 0.3% of skimmed milk volume, fermented and cultured 18h under conditions of 23 DEG C is preserved It is spare under the conditions of 4 DEG C.
The preparation of bacillus subtilis:The bacillus subtilis frozen is subjected to scribing line culture, colonies typical is isolated and connects Kind shake culture 18h under the conditions of 37 DEG C in fluid nutrient medium, the standard type of bacillus subtilis is met through microscopy, grows shape State is good, and being inoculated in fluid nutrient medium by inoculum concentration 3% is activated, 37 DEG C, and shake culture is for 24 hours, standby under the conditions of being stored in 4 DEG C With.
2) mixed fermentation
The configuration of culture medium:Whey protein concentrate 20g, glucose 6g, yeast powder 3g, carbon are added into 500mL distilled water Sour calcium 6g, magnesium sulfate 0.2g, manganese sulfate 0.02g, whey powder 10g, it is 7.5 that medium pH is fully adjusted after dissolving, sterilization conditions Keep the temperature 10min under the conditions of being 115 DEG C, be cooled to 30 DEG C it is spare.To culture medium inoculated after cooling, by 6% inoculum concentration, wherein Kefir fermenting agent:Bacillus subtilis=3:1,36 DEG C, fermented and cultured 18h.
3) preparation of compound micro-ecological preparation
Fermentation broth agitation obtained is uniform, and under the conditions of 8 DEG C, 4500 revs/min of rotating speed, time 25min centrifugal treating is received Collection concentration thalline;Based on thalline mass percent, 0.9% prebiotic substance, skimmed milk powder 12%, trehalose 3%, paddy are added Propylhomoserin sodium 1%, glycerine 2% are uniformly mixed, and carry out low temperature spray drying, and control inlet air temperature is 120 DEG C, 70 DEG C of leaving air temp, Obtain prebiotics, probiotics compound micro-ecological preparation.
Embodiment 3
1) preparation of probiotics leaven
The preparation of Kefir fermenting agent:The degreasing milk solution of configuration 10% after 10min sterilizations, is cooled to 30 by 95 DEG C DEG C, Kefir grains throw type leaven is added according to the 0.5% of skimmed milk volume, fermented and cultured 16h under conditions of 25 DEG C is preserved It is spare under the conditions of 4 DEG C.
The preparation of bacillus subtilis:The bacillus subtilis frozen is subjected to scribing line culture, colonies typical is isolated and connects Kind shake culture 18h under the conditions of 37 DEG C in fluid nutrient medium, the standard type of bacillus subtilis is met through microscopy, grows shape State is good, and being inoculated in fluid nutrient medium by inoculum concentration 5% is activated, 37 DEG C, shake culture 20h, standby under the conditions of being stored in 4 DEG C With.
2) mixed fermentation
The configuration of culture medium:Whey protein concentrate 15g, glucose 7g, yeast powder 4g, carbon are added into 500mL distilled water Sour calcium 4g, magnesium sulfate 0.1g, manganese sulfate 0.03g, whey powder 12g, it is 7.0 that medium pH is fully adjusted after dissolving, sterilization conditions Keep the temperature 20min under the conditions of being 100 DEG C, be cooled to 25 DEG C it is spare.To culture medium inoculated after cooling, by 4% inoculum concentration, wherein Kefir fermenting agent:Bacillus subtilis=3:1,32 DEG C, fermented and cultured 20h.
3) preparation of compound micro-ecological preparation
Fermentation broth agitation obtained is uniform, under the conditions of 10 DEG C, 6000 revs/min of rotating speed, time 15min centrifugal treating, Collect concentration thalline;Based on thalline mass percent, add 1.5% prebiotic substance, skimmed milk powder 10%, trehalose 5%, Sodium glutamate 3%, glycerine 3% are uniformly mixed, and carry out low temperature spray drying, and control inlet air temperature is 100 DEG C, leaving air temp 60 DEG C, obtain prebiotics, probiotics compound micro-ecological preparation.

Claims (3)

1. a kind of prebiotics, probiotics compound micro-ecological preparation, it is characterised in that:Compound micro-ecological preparation is by prebiotics and prebiotic Bacterium is composed, and the probiotics is formed by lactic acid bacteria, saccharomycete and bacillus subtilis mixed fermentation, lactic acid bacteria and yeast Bacterium is provided by Kefir granule compound strain, and total viable count is more than 5,000,000,000 cfu/g in preparation, and the prebiotics are that Hericium erinaceus is more One or more of sugar, Agaricus Blazei Murrill polysaccharide and lentinan composition, additive amount are 0.3%~1.5%.
2. the preparation method of prebiotics according to claim 1, probiotics compound micro-ecological preparation, which is characterized in that system Standby compound micro-ecological preparation uses a variety of strain mixed fermentations, after High Density Cultivation, prebiotic substance is added, by low-temp. spraying Mist drying is made.
3. the preparation method of prebiotics according to claim 2, probiotics compound micro-ecological preparation, it is characterised in that:It will Probiotics mixed bacteria is inoculated in culture medium by the 3~10% of fermentating liquid volume, wherein Kefir granule and bacillus subtilis Inoculative proportion is 3:Under the conditions of 1,28~36 DEG C, ferment 16~24 hours.
CN201810412982.3A 2018-05-03 2018-05-03 A kind of prebiotics, probiotics compound micro-ecological preparation and preparation method thereof Pending CN108606329A (en)

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Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN112457990A (en) * 2020-12-18 2021-03-09 中国科学院合肥物质科学研究院 Freeze-drying protective agent and application thereof
CN113181220A (en) * 2021-05-18 2021-07-30 郭云 Bacillus subtilis spray and preparation method thereof
CN113789276A (en) * 2021-09-15 2021-12-14 千宠堂(福建)生物科技有限公司 Lactobacillus plantarum for producing beta-nicotinamide mononucleotide and application thereof
CN113841901A (en) * 2021-09-15 2021-12-28 广西大学 Preparation method of high-activity synbiotics preparation freeze-dried powder

Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN112457990A (en) * 2020-12-18 2021-03-09 中国科学院合肥物质科学研究院 Freeze-drying protective agent and application thereof
CN113181220A (en) * 2021-05-18 2021-07-30 郭云 Bacillus subtilis spray and preparation method thereof
CN113789276A (en) * 2021-09-15 2021-12-14 千宠堂(福建)生物科技有限公司 Lactobacillus plantarum for producing beta-nicotinamide mononucleotide and application thereof
CN113841901A (en) * 2021-09-15 2021-12-28 广西大学 Preparation method of high-activity synbiotics preparation freeze-dried powder

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