CN104543679A

CN104543679A - Probiotic-containing health food and preparation method thereof

Info

Publication number: CN104543679A
Application number: CN201510033886.4A
Authority: CN
Inventors: 邵素英
Original assignee: SHAOXING SHANGYU HONGSHENG TECHNOLOGY TRANSFER SERVICES Co Ltd
Current assignee: SHAOXING SHANGYU HONGSHENG TECHNOLOGY TRANSFER SERVICES Co Ltd
Priority date: 2015-01-23
Filing date: 2015-01-23
Publication date: 2015-04-29

Abstract

The invention discloses a probiotic-containing health food and a preparation method thereof, wherein intestinal florae are adjusted in many aspects of increasing the types and quantity of probiotics, increasing the nutrient substances of the probiotics, inhibiting the growth of harmful fungi, enhancing human immunity and the like, so as to improve gastrointestinal functions. Probiotic powders of lactobacillus plantarum and the like are taken as main raw materials, and edible fungi extract, modified dietary fibres, a saussurea involucrata culture extract capable of remarkably enhancing the human immunity, resisting inflammation and easing pain and having anti-oxidization, anti-radiation, anti-fatigue functions and the like are scientifically compounded, and then the planting capacity and the planting time of endogenous and exogenous probiotics in human intestinal tracts are remarkably improved and prolonged, the growth and reproduction of intestinal harmful fungi, especially, Gram-negative bacterium, are effectively inhibited, and the composition of intestinal probiotic florae is adequately regulated while the types of the intestinal probiotic florae are increased; therefore, a health food with high probiotic property, capable of remarkably enhancing human immunity and containing probiotics is prepared.

Description

A kind of health food containing probio and preparation method thereof

Technical field

The present invention relates to health food, be specifically related to a kind of health food containing probio and preparation method thereof.

Background technology

Improve gastrointestinal function to comprise defaecation, regulating intestinal canal flora, promote digestion, have auxiliary protection four kinds of functions to gastric mucosa injure.The alternate balance of enteron aisle bacterium bacterium affects by the interaction etc. of the physiology of host, food, medicine and enteron aisle bacterium, and various factors all can destroy the alternate balance of bacterium, causes enteron aisle to be preponderated from beneficial bacterium and turns to harmful bacteria to preponderate, thus cause many intestines problems.Therefore, increasing the quantity of intestinal beneficial bacterium, reduce harmful bacteria quantity, is the key point ensureing the running of stomach health.

Flora imbalance (dysbacteriosis) refers to that the ratio in the normal flora of body a part between each bacterial classification occurs to change by a relatively large margin and exceeds the state of normal range (NR), consequent illness, is called that dysbacteriosis or flora replace disease (microbialselection and substitution).During flora imbalance, cause suprainfection or superinfection (superinfection) more, namely in the treatment of primary infection, there occurs the infection of another kind of new pathogenic bacteria.Being more common in of flora imbalance uses antibiotic and chronic wasting disease etc.After widely applying broad-spectrum antibiotic for a long time clinically, most of sensitive bacteria and normal flora suppressed or kill, but drug-fast bacteria then obtains survival advantage and amount reproduction causes a disease, as resistant Staphylococcus aureus causes diarrhoea, septicemia, thrush, vaginitis, enteron aisle and anal infection are caused to the insensitive candida albicans bacterium of antibiotic.

At present, the mode of adjustment gut flora is mainly with probiotic supplemented kind and quantity, probiotic supplemented desired nutritional material (the prebiotic factor), suppress or kill harmful intestinal tract bacteria (mainly Gram-negative bacteria), strengthen body immunity to improve macrophage to modes such as the phagocytosis of harmful bacteria to reach the object of gut flora balance, Chinese patent CN 10155908 A discloses the preparation method of the probiotics preparation of a kind of reducing blood lipid and regulating intestinal canal flora, Kefir lactobacillu plantarurn MA2 is carried out High Density Cultivation for (1) by its step, obtain zymotic fluid, its cell density is not less than 10 ⁸cfu/mL, (2) above-mentioned fermentation liquor collected by centrifugation obtains thalline, adds protective agent, is lyophilized into bacterium powder, (3) in the bacterium powder of freeze-drying, add oligosaccharide mixture, mix and obtain probiotics preparation finished product.Also there is while these product have hypolipemic function the physiological function of regulating intestinal canal flora, can treat and prevent hyperlipidemic conditions and regulating intestinal canal flora, and nontoxic, safety, long shelf-life, can be made into capsule, microcapsules, electuary, tablet etc.Chinese patent CN 102511707 B discloses health food of a kind of regulating intestinal canal and preparation method thereof, comprises the component of following weight portion: natural plant extracts 50-60%; FOS 8-12%; Probiotics probio 2-3%; Honey 20-30%; Wherein, described natural plant extracts comprises Fructus Fici extract, prune extract, tamarind extract, candied date extract, and its weight ratio is 10: 1: 1: 1.Natural plant extracts is pulverized; Take honey to be heated to boil in evaporating dish, treat that sweet surface rises bubble, takes off immediately; Load weighted natural plant extracts is mixed with FOS, probio, adds again when sweet temperature will be waited when mixing to be down to 30-40 DEG C, make doughy softwood; The dough softwood of having lived is placed, spice and honey component is fully mixed moist, make spheroidal pill.This product for primary raw material, regulates function of intestinal canal with pure natural plant extract, FOS, probiotics under the prerequisite not stimulating enteron aisle, reaches the effect of prophylactic treatment constipation.Chinese patent CN 103054938 A discloses a kind of oral formulations regulating Biology Barrier of Gastrointestinal Tract to lack of proper care, and it relates to a kind of oral formulations.The present invention will solve and widely apply now broad-spectrum antibiotic to regulate Biology Barrier of Gastrointestinal Tract imbalance clinically for a long time, produces the problem of drug resistance.Oral formulations of the present invention is made up of inulin, ginseng extract, Rhizoma Atractylodis Macrocephalae extract, tuckahoe extracts, licorice.The present invention is a kind of contributes to improving the oral formulations scientific formula of gastrointestinal function, raw material is easy to get, prepare conveniently, effect obviously, side effect is little, and is suitable for suitability for industrialized production.

Above-mentioned disclosed patent is adopted as traditional probio and the prebiotic factor and combination thereof, be not separated or turn out probiotic stronger probio, its probio biologically active is not also very desirable, the proliferation function of traditional prebiotic factor pair probio is delayed, and in suppression harmful intestinal tract bacteria, strengthen body immunity and then improve macrophage to the phagocytosis of harmful bacteria, to improve the less of beneficial bacteria of intestinal tract group aspect research.

Summary of the invention

Technical problem solved by the invention overcomes the existing defect improving gastrointestinal function health products, from increase probio kind and quantity, increase probio nutriment, suppress harmful bacteria growth and increase many-sided adjustment gut floras such as body immunity, improving gastrointestinal function.With probio pulvis such as Lactobacillus plantarums for primary raw material, science composite can repair cell tissue, there is antiulcer action, antioxidation, effect of scavenging radical and anti-aging effects, significantly can promote human body integral immunity and strengthen muscle power, improve the fungi extracts of gastrointestinal function; Modified dietary fiber containing high-load soluble cellulose; Body immunity can be significantly improved, anti-inflammatory and antalgic, the Saussurea involucrata culture extract etc. with functions such as anti-oxidant, radioresistance and antifatigues be raw material, and then while increase beneficial bacteria of intestinal tract flora kind, significantly enhance endogenous and that external source probio is in human body intestinal canal colonization ability and resident time, effectively inhibit the harmful intestinal tract bacteria especially growth of Gram-negative bacteria and breeding, fully have adjusted the composition of beneficial bacteria of intestinal tract flora, prepare a kind of probiotic strong, significantly improve body immunity, health food containing probio.

In order to achieve the above object, the present invention is by the following technical solutions:

Containing a health food for probio, prepared by the raw material of following parts by weight:

D-mannital 5-50 part, maltodextrin 5-30 part, microcrystalline cellulose 5-30 part, probio pulvis 5-25 part, modified dietary fiber 6-15 part, xylo-oligosaccharide 3-15 part, isomalt 3-15 part, WPC 3-15 part, fungi extracts 3-15 part, Saussurea involucrata culture extract 3-10 part, pectin decomposer 2-10 part, cyclohexaamylose 1-10 part, citric acid 0.1-3 part, dolomol 0.1-3 part, milk flavour 0.1-3 part, beta carotene 0.01-3 part;

Preferably, the described health food containing probio, prepared by the raw material of following parts by weight:

D-mannital 15-35 part, maltodextrin 10-20 part, microcrystalline cellulose 10-20 part, probio pulvis 10-20 part, modified dietary fiber 9-12 part, xylo-oligosaccharide 7-11 part, isomalt 7-11 part, WPC 7-11 part, fungi extracts 7-11 part, Saussurea involucrata culture extract 5-8 part, pectin decomposer 5-7 part, cyclohexaamylose 1.5-5 part, citric acid 1.5-2.5 part, dolomol 1.5-2.5 part, milk flavour 1.5-2.5 part, beta carotene 1-2 part;

More preferably, the described health food containing probio, prepared by the raw material of following parts by weight:

D-mannital 25 parts, maltodextrin 15 parts, microcrystalline cellulose 15 parts, 15 parts, probio pulvis, modified dietary fiber 11 parts, xylo-oligosaccharide 9 parts, isomalt 9 parts, WPC 9 parts, fungi extracts 9 parts, Saussurea involucrata culture extract 7 parts, pectin decomposer 6 parts, cyclohexaamylose 3 parts, citric acid 2 parts, dolomol 2 parts, milk flavour 2 parts, beta carotene 1.5 parts;

Further, described probio pulvis include but not limited in following probio pulvis one or more: such as: Lactobacillus plantarum (Lactobacillus plantarum), bifidobacterium bifidum (Bifidobacterium bifidum), bifidobacterium infantis (B.infantis), long Bifidobacterium Bifidum (B.longum), short Bifidobacterium Bifidum (B.breve), bifidobacterium adolescentis (B.adolescentis), lactobacillus bulgaricus (Lactobacillus.Bulgaricus), lactobacillus acidophilus (L.acidophilus), Lactobacillus casei (Lactobacillus Casey), streptococcus thermophilus (Streptococcusthermophilus), also comprise for one or more in the fungi pulvis of health food, such as: saccharomyces cerevisiae (Saccharomyces cerevisiae), candida utili (Cadida atilis), Kluyveromyces lactis (Kluyveromyces lactis), saccharomyces carlsbergensis (Saccharomyces carlsbergensis), Paecilomyces hepiali chen (Paecilomyces hepiali Chen et Dai, sp.nov), Hirsutella hepiali Chen et Shen (Hirsutella hepiali Chenet Shen), glossy ganoderma (Ganoderma lucidum), purple sesame (Ganoderma sinensis), Ganoderma tsugae (Ganodermatsugae), monascus (Monacus anka), monascus purpureus (Monacus purpureus),

Described probio pulvis probiotics viable bacteria content is: 7x10 ¹²-9x10 ¹²cfu/g;

Preferably, described probio pulvis is formed by the pulvis Homogeneous phase mixing of following parts by weight: Lactobacillus plantarum 40-50 part, bifidobacterium longum 30-45 part, bifidobacterium bifidum 25-35 part, bifidobacterium adolescentis 15-20 part, lactobacillus bulgaricus 15-20 part, lactobacillus acidophilus 10-15 part, Lactobacillus casei 8-10 part, streptococcus thermophilus 8-10 part, saccharomyces cerevisiae 8-10 part, candida utili bacterium 4-6 part;

Preferably, described Lactobacillus plantarum pulvis is prepared by Lactobacillus plantarum (Lactobacillus plantarum) tlj-2014, this bacterial strain is preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center on July 2nd, 2014 and (is called for short CGMCC, address is: No. 3, Yard 1, BeiChen xi Road, Chaoyang District, Beijing City. Institute of Microorganism, Academia Sinica. and postcode: 100101), deposit number is CGMCC NO.9405, and Classification And Nomenclature is: Lactobacillus plantarum Lactobacillus plantarum.

Further, dietary fiber obtains through physics, chemistry or biological method process by described modified dietary fiber, Soluble Fiber cellulose content is high, biologically active strong, have cellulose that is important, positive role to beneficial bacteria of intestinal tract group, compared with full diet fiber, its biological action is more powerful: 1) Soluble Fiber cellulose content is high, more easily utilized by probio, probio can be improved in the growth of enteron aisle and fertility, increase kind and the quantity of probio flora, reduce large intestine pH value, improve intestine microenvironment; 2) powerful adsorption capacity, through modified, cellulosic specific area increases, and network is enriched, and absorption affinity strengthens, and the organic molecule ability of chelating, absorption cholesterol and bile acids is stronger, suppress human body to their absorption; 3) ion-exchange capacity strengthens, and to metallic element, particularly heavy metal element adsorption effect is stronger; 4) the stronger and impact of acid and alkali, alkali, salt of retentiveness, dilatancy, thickening property, 5) regulate and maintain the resident time of gut flora, strengthen the absorption and digestion ability of enteron aisle, improve body immunity; 6) promote gastrointestinal peristalsis, slow down and eliminate the bad reaction such as gasteremphraxis, abdominal distension; 7) bury function is strong, prevents external environment (oxygen, temperature, illumination, humidity etc.) factor from, on the impact of product quality, stabilizing the biologically active of product, extending the shelf-life of product;

Preferably, described modified fibre is obtained through biology enzyme enzymolysis by one or more in inulin, apple fiber, common oats fibre, Semen Tritici aestivi fiber;

More preferably, the preparation method of described modified dietary fiber comprises the following steps: by inulin, apple fiber, common oats fibre, Semen Tritici aestivi fiber 8-10:4-6:3-5:2-4 Homogeneous phase mixing in mass ratio, add its quality 3-7 water doubly, room temperature 200-300W, 35-40KHz condition ultrasonic extraction 10-15min, then at electric-field intensity 20-40kV/cm, burst length 400-500 μ s, carries out high-pressure pulse electric extraction under pulse frequency 200-300Hz condition; Be 4.5-6.5 by lactic acid adjust ph, add the biology enzyme of mixture quality 0.1-0.3%, in 45-55 DEG C of enzymolysis 20-48min; Enzymolysis liquid reduced pressure concentration, freeze drying, low-temperature grinding to particle diameter are that namely 0.1-0.3mm obtains modified dietary fiber;

Described biology enzyme is zytase, cellulase, laccase, pectase, tannase 5-9:2-4:2-4:2-4:1-3 Homogeneous phase mixing in mass ratio.

Further, described fungi extracts is with repair cell tissue, has antiulcer action, antioxidation, effect of scavenging radical and anti-aging effects, significantly can promote human body integral epidemic disease and exempt from power and strengthen edible mushroom that is physical, that improve gastrointestinal function to prepare through ultrasonic cleaning, high-pressure pulse electric process and biological enzymolysis;

Preferably, the preparation method of described fungi extracts comprises the steps: that Hericium erinaceus, ginkgo and black fungus being put into respectively the supersonic wave cleaning machine that 0.3-0.5% sodium bicarbonate solution is housed cleans 5-10min in 200W, 40KHz, drain, be cut into phase equal-specification, length and width is 5-10mm, the sheet of thickness 3-5mm or fourth, 8-10:5-7:4-6 Homogeneous phase mixing in mass ratio, then in-21--25 DEG C of freezing 10-30min, pulverize immediately, crushed material particle diameter is 0.2-1mm; Add crushed material quality 3-5 water doubly and obtain mixture, adjustment mixture pH is 4.5-6, at electric-field intensity 35-45kV/cm under room temperature, burst length 500-700 μ s, high-pressure pulse electric process is carried out under pulse frequency 200-400Hz condition, be warming up to 45 ~ 55 DEG C, add the mixed enzyme enzymolysis 4 ~ 6 hours of mixture weight 0.5-1.2%, obtain edible mushroom enzymolysis liquid, being evaporated to solid content is more than 30%, and freeze drying, low-temperature grinding to particle diameter are that namely 0.1-0.3mm obtains fungi extracts;

In described edible mushroom enzymolysis liquid, edible fungi polysaccharide content is up to 6.82%;

Described mixed enzyme parts by weight consist of: papain 20-30 part, bromelain 20-30 part, dextranase 10-15 part, zytase 10-15 part, pentosanase 10-15 part, ficin 5-10 part, middle temperature amylase 5-10 part, pectase 5-10 part, tannase 3-5 part.

Further, described Saussurea involucrata culture, purchased from Dalian Puruikang Biotechnology Co, is aubergine lumps particle, provenance system " Herba Saussureae Involueratae ";

Further, described Saussurea involucrata culture extract is that raw material obtains through the low temperature extractive technique such as freezing, pulverizing, the ultrasonic extraction of microwave radiation technology, low temperature enzymolysis, high-pressure pulse electric extraction with Saussurea involucrata culture, can retain Flavonoid substances and other anti-oxidant, volatile, thermal sensitivity active ingredients such as acacetin, dinatin, rutin, Quercetin to greatest extent;

Preferably, the preparation method of described Saussurea involucrata culture extract comprises the steps: to get Saussurea involucrata culture, it is in the container of the aseptic FOS aqueous solution of 0.9-1.1% that immersion fills mass percent concentration, takes out, then in-18--23 DEG C of freezing 5-10min; Be crushed to particle diameter 0.3-1mm immediately; To put in container and to add the water of 3-5 times of weight or the ethanol of 35-75%, obtaining mixed material, is 3.5-5.5 by lactic acid adjust ph, at electric-field intensity 25-35kV/cm under room temperature, burst length 300-500 μ s, carries out high-pressure pulse electric process under pulse frequency 200-300Hz condition; Then 30-40 DEG C is warming up to, insulation, Microwave Extraction is carried out under power 150-300W, frequency 2000Hz condition, wherein, each microwave irradiation total time 60-80s, carries out compartment irradiation: irradiation 10s, interval 10s, irradiation like this 10 times, carries out ultrasonic assistant extraction simultaneously under power 200-300W, frequency 30-40KHz condition; Continue to be warming up to 40-50 DEG C, insulation, then adds the compound enzyme enzymolysis 30-50min of mixed material gross weight 2.5-4%, filters, obtains filtrate; Filter residue 1-3 times of weight 76-80 DEG C of water rinse 3 times, rinsing liquid and filtrate merge, and Homogeneous phase mixing, at electric-field intensity 35-45kV/cm under room temperature, burst length 400-600 μ s, carries out high-pressure pulse electric process and namely obtains Saussurea involucrata culture extract under pulse frequency 200-300Hz condition;

Described compound enzyme is by pectase, cellulase, tannase, amylase and protease 3-7:3-5:2-4:1-3:1-2 Homogeneous phase mixing in mass ratio.

Described pectin decomposer has natural, soft agreeable to the taste tart flavour and significant antagonistic property, especially the strongest to Escherichia coli inhibitory action, also there is stronger non-oxidizability simultaneously, concentration can act on when reaching 0.01%, very strong eliminating effect is all shown to superoxide radical, hydroxy radical and DPPH free radical, when concentration reaches 1.0%, eradicating efficacy reaches 100%;

The science of described pectin decomposer is composite, effectively can improve the palatability of health food of the present invention, most importantly its significant antagonistic property and non-oxidizability avoid oxidation and the microorganism encroach of active ingredient, have stabilized and increased the biologically active of product, have extended the shelf-life of product;

Further, described pectin decomposer obtains pectin with waste residues such as the pericarps such as lemon, orange, shaddock, citrus, grape or hawthorn, beet, apples through ultrasonic cleaning, high-voltage pulse extraction, hot water extraction, alcohol precipitation, then obtained through pectinase enzymatic hydrolysis, preferably, the preparation method of described pectin decomposer, fresh hawthorn pomace, shaddock ped, pomace, megasse is comprised the steps: to put into the supersonic wave cleaning machine that 1.2% sodium bicarbonate solution is housed respectively and clean 5-10min in 200W, 40KHz, drain, be cut into phase equal-specification, length and width is 5-10mm, the sheet of thickness 3-5mm or fourth, 8-12:3-5:2-4:2-4 Homogeneous phase mixing in mass ratio, then in-21--25 DEG C of freezing 10-30min, pulverize immediately, crushed material particle diameter is 0.1-1mm, add crushed material quality 3-5 deionized water doubly, mix, be 2-3 by citric acid adjust ph, at electric-field intensity 35-45kV/cm under room temperature, burst length 500-700 μ s, high-pressure pulse electric process is carried out under pulse frequency 200-400Hz condition, then normal pressure boils 30-50min, Bag filter, filter residue its quality 1-3 filtrate normal pressure doubly boils 40-60min, Bag filter, merging filtrate, diatomite filtration, filtrate is clarified, at 75 DEG C, be evaporated to solid content is 6-10%, be cooled to room temperature rapidly, the pH value adding concentrate 1.5 times of volumes is 95% ethanol of 2-3, leave standstill 20-30min, pectin is precipitated out, the centrifugal 5-10min of 4000r/min, reclaim precipitation, with the absolute ethyl alcohol wash-out of 2 times of volumes, centrifugal, reclaim precipitation, operation like this 2 times, what obtain is deposited in 40-60 DEG C of vacuum drying 2-3h, obtain pectin, add pectin quality 4-5 40-50 DEG C of water doubly, stir, Homogeneous phase mixing, insulation, be 4.5-5.5 by lactic acid adjust ph, add the pectinase enzymatic hydrolysis 2-4h of 0.3-0.8U/mL, enzymolysis liquid filters, filtrate is gone out enzyme 10-15min in boiling water, be cooled to room temperature, ultrafiltration concentration, namely freeze drying obtains pectin decomposer.

The formulation of health food of the present invention is oral formulations, can be tablet, pulvis, capsule or granule, is preferably tablet.

Described tablets, transport, storage and easy to carry, proterties is stablized, and dosage is accurate, and cost is low.

The present invention can be prepared by the following method containing the health caring food tablet of probio, capsule or granule, comprises the steps:

1) according to formula rate, get D-mannital, maltodextrin, microcrystalline cellulose, xylo-oligosaccharide, isomalt, WPC, fungi extracts, Saussurea involucrata culture extract, milk flavour pulverizing respectively, cross 60-120 mesh sieve, then mix 3-15min, obtain mixed powder;

2) by pectin decomposer, citric acid, the beta carotene aqueous solution making 13-18% soluble in water, as adhesive;

3) in described mixed powder, add described adhesive and make softwood, granulate with 10-20 mesh sieve, obtain wet granular, put in power 3000W, frequency 2450MHz, temperature 55-65 DEG C dry 3-8min in microwave dryer, with the quick whole grain of 10-20 mesh sieve;

4) by modified dietary fiber, cyclohexaamylose and probiotic powder mixing 5-10min, probio pulvis must be embedded;

5) whole good particle and embedding probio pulvis, dolomol mixing 5-10min, after mixing, make tablet, capsule or granule.

Preferably, the preparation method of health caring food tablet of the present invention, comprises the steps:

5) whole good particle and embedding probio pulvis, dolomol mixing 5-10min, after mixing, compressing tablet, packs and obtains health caring food tablet.

The present invention, containing the preparation method of the health food pulvis of probio, comprises the steps:

1) by formula rate, by modified dietary fiber, cyclohexaamylose and probiotic powder mixing 5-10min, probio pulvis must be embedded;

2) according to formula rate, D-mannital, maltodextrin, microcrystalline cellulose, xylo-oligosaccharide, isomalt, WPC, fungi extracts, Saussurea involucrata culture extract, pectin decomposer, citric acid are pulverized, cross 60-120 mesh sieve, then namely obtain health food pulvis with embedding probio pulvis, beta carotene, milk flavour, dolomol mixing 3-15min.

Lactobacillus plantarum of the present invention (Lactobacillus plantarum) tlj-2014 is preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center on July 2nd, 2014 and (is called for short CGMCC, address is: No. 3, No. 1, North Star West Road, Chaoyang District, city of BeiJing, China institute. Institute of Microorganism, Academia Sinica. and postcode: 100101), deposit number is CGMCC NO.9405, and Classification And Nomenclature is: Lactobacillus plantarum Lactobacillus plantarum.

Described lactobacillus plantarum strain feature is as follows: examine under a microscope, and this bacterial strain is rod-short, and Gram's staining is positive, and atrichia does not produce gemma; On solid medium, this bacterium bacterium colony is white, and smooth surface is fine and close, and form is circular, and edge is more neat.

Physicochemical characteristics is: catalase (-), gelatin liquefaction (-), indoles experiment (+), motility (-), fermentation gas (-), nitrate reductase (-), fermentation gas (-), produce hydrogen sulfide gas (-), in pH4.0MRS culture medium, grow (+).

Described Lactobacillus plantarum adopts following flow process to carry out seed selection:

The original bacterial classification that sets out → test tube activation → dithyl sulfate (DES) mutagenesis → nitrosoguanidine (NTG) mutagenesis → plasma mutagenesis → dull and stereotyped primary dcreening operation → shaking flask sieves → mitotic stability test again.

Described starting strain is in MRS dextrose culture-medium, and the throughput rate of its lactic acid is 1.5g/L/d, almost stops growing when medium pH is 3.5, is 0.34mg/h/kg Chinese cabbage to the decomposition rate of natrium nitrosum.Starting strain is the greenfeed that Li Zheng is collected in Fattening Sheep field, Yanchi county Ningxia, acquisition time on September 15th, 2013.

In order to improve the decomposition rate of its production of lactic acid speed, acid-fast ability and nitrite, DES and NTG technology is adopted to carry out mutagenesis to this bacterial classification successively, after mutagenesis, bacterial strain adopts MRS calcium carbonate flat board to carry out primary dcreening operation, then 500mL shake flask fermentation is adopted, biosensor analysis instrument carries out multiple sieve to Producing Strain, the lactobacillus plantarum strain that seed selection is excellent, then does passage assays, evaluates its genetic stability.

Lactobacillus plantarum tlj-2014 genetic stability result shows: through continuous passage ten times, property indices is all more stable, and heredity is better, and proterties is not replied, therefore using the object bacterial strain that Lactobacillus plantarum tlj-2014 obtains as seed selection.

Empirical tests finds: the production of lactic acid speed of this mutagenic strain can reach 35g/L/d, and this bacterial strain lactic acid concn after 71 hours fermentation reaches 95g/L; Can survive under pH is the condition of 1.80.Degrading nitrite speed is fast, and capacity of decomposition reaches 9.8mg/h/kg (speed of spontaneous fermentation process nitrite accumulation is approximately 1.1mg/h/kg), can resistance to 1% cholate.

Therefore adopt this bacterial classification produce pickles, whole sweat nitrite concentration at below 5mg/kg, far below the content specified in standard GB/T 2714-2003 (20mg/kg).

Beneficial effect:

The present invention grows containing the health food of probio from increase probio kind and quantity, increase probio nutriment, suppression harmful bacteria and increases many-sided adjustment gut floras such as body immunity, improves gastrointestinal function.With probio pulvis such as Lactobacillus plantarums for primary raw material, science composite can repair cell tissue, there is antiulcer action, antioxidation, effect of scavenging radical and anti-aging effects, significantly can promote human body integral immunity and strengthen muscle power, improve the fungi extracts of gastrointestinal function; Modified dietary fiber containing high-load soluble cellulose; Body immunity can be significantly improved, anti-inflammatory and antalgic, the Saussurea involucrata culture extract etc. with functions such as anti-oxidant, radioresistance and antifatigues be raw material, and then while increase beneficial bacteria of intestinal tract flora kind, significantly enhance endogenous and that external source probio is in human body intestinal canal colonization ability and resident time, effectively inhibit the harmful intestinal tract bacteria especially growth of Gram-negative bacteria and breeding, fully have adjusted the composition of beneficial bacteria of intestinal tract flora, prepare a kind of probiotic strong, significantly improve body immunity, health food containing probio.Concrete know-why is:

1. the present invention is many containing the health food probio kind of probio, and Lactobacillus plantarum CGMCC NO.9405 production of lactic acid speed wherein can reach 35g/L/d, and this bacterial strain lactic acid concn after 71 hours fermentation reaches 95g/L; Can survive under pH is the condition of 1.80.Degrading nitrite speed is fast, and capacity of decomposition reaches 9.8mg/h/kg (speed of spontaneous fermentation process nitrite accumulation is approximately 1.1mg/h/kg), can resistance to 1% cholate, and its prebiotic performance is more superior.

2. ultrasonic extraction, high-pressure pulse electric extract and biological enzymolysis combination of sciences by the modified dietary fiber that prepared by the present invention, gained modified dietary fiber Soluble Fiber cellulose content is high, more easily utilized by probio, improve probio in the growth of enteron aisle and fertility, increase kind and the quantity of probio flora, reduce large intestine pH value, improve intestine microenvironment; High adsorption capacity, through modified, cellulosic specific area increases, and network is enriched, and absorption affinity strengthens, and the organic molecule ability of chelating, absorption cholesterol and bile acids is stronger, suppress human body to their absorption; Ion-exchange capacity strengthens, and to metallic element, particularly heavy metal element adsorption effect is stronger, effectively prevent body weight for humans metal poisoning; The stronger and impact of acid and alkali, alkali, salt of retentiveness, dilatancy, thickening property.Regulate and maintain the resident time of gut flora, strengthen the absorption and digestion ability of enteron aisle, improve body immunity; Effective promotion gastrointestinal peristalsis, slows down and eliminates the bad reaction such as gasteremphraxis, abdominal distension; Powerful bury function can prevent external environment (oxygen, temperature, illumination, humidity etc.) factor from, on the impact of product quality, stabilizing the biologically active of product, extending the shelf-life of product.

3. the fungi extracts that prepared by the present invention is with repair cell tissue, has antiulcer action, antioxidation, effect of scavenging radical and anti-aging effects, significantly can promote human body integral epidemic disease exempt from power and strengthen muscle power, improve that the edible mushroom of gastrointestinal function obtains through ultrasonic cleaning, high-pressure pulse electric process and biological enzymolysis, effective component extraction rate is high, in edible mushroom enzymolysis liquid, edible fungi polysaccharide content is up to 6.82%, biologically active is stronger, foodsafety is high, remains without agricultural chemicals and worm's ovum.

4. the Saussurea involucrata culture extract that prepared by the present invention has analgesia, anti-inflammatory, antirheumatic, Antiaggregating activity on platelet, reducing blood lipid, improve blood circulation, to immune system, there is regulating action, simultaneously anti-oxidant in addition, the Herba Saussureae Involueratae of many-sided pharmacodynamic action such as radioresistance and antifatigue is the Saussurea involucrata culture of provenance is that raw material is through freezing, pulverize, the ultrasonic extraction of microwave radiation technology, low temperature enzymolysis, low temperature extractive techniques such as high-pressure pulse electric extraction and obtaining, acacetin can be retained to greatest extent, dinatin, rutin, the Flavonoid substances such as Quercetin are anti-oxidant with other, volatile, thermal sensitivity active ingredient, active ingredient acquisition rate is high, technique is simple, time is short, improve raw material availability, reduce production cost, improve production efficiency.Test proves, Saussurea involucrata culture extract flavone yield prepared by the present invention can reach 97%, and solid yield can reach 65%.And saussurea involucrata cell culture is no longer containing colchicin human body being had to side effect; it is new resource food that the Ministry of Public Health ratifies Saussurea involucrata culture on May 20th, 2010, to the modernization of Chinese medicine of promotion China, protects wild plant resource, Development of Social Economy significant.

5. the pectin decomposer that prepared by the present invention has natural, soft agreeable to the taste tart flavour and significant antagonistic property, especially the strongest to Escherichia coli inhibitory action, also there is stronger non-oxidizability simultaneously, concentration can act on when reaching 0.01%, very strong eliminating effect is all shown to superoxide radical, hydroxy radical and DPPH free radical, when concentration reaches 1.0%, eradicating efficacy reaches 100%; The science of pectin decomposer is composite, effectively can improve the palatability of health food of the present invention, most importantly its significant antagonistic property and non-oxidizability avoid oxidation and the microorganism encroach of active ingredient, have stabilized and increased the biologically active of product, have extended the shelf-life of product.

6. the isomalt that science of the present invention is composite has low in calories, sugariness is natural pure, height endurability, non-cariogenic tooth, more rational negative heat of solution, high stability and non-hygroscopic, in the quality of improving product, extend the shelf life, promote that gastro-intestinal digestion and absorption aspect have excellent characteristic, simultaneously or a kind of excellent Bifidobacterium MF, although isomalt can not utilize by the enzyme system of human body and most microorganism, but can decompose by the Bifidobacterium in human body intestinal canal utilization, promote the growth and breeding of Bifidobacterium, maintain the microecological balance of enteron aisle, improve human body integral immunity, be conducive to the health of human body.

7. the present invention is the mutual synergistic result of each component containing the effect of the health food of probio, the not superposition of simple raw material function, the science of each raw material components is composite, and the effect of generation, considerably beyond the superposition of each single component function and effect, has advanced and practicality preferably.

8. the present invention is simple, easy to operate, low for equipment requirements containing preparation method's technique of the health food of probio; can industrialization and large-scale production; modified dietary fiber is finally mixed, has played the bury function that it is powerful, such that the proterties of product is more stable, the shelf-life is longer.

Detailed description of the invention

The present invention is described below by specific embodiment.Unless stated otherwise, technological means used in the present invention is method known in those skilled in the art.In addition, embodiment is interpreted as illustrative, but not limits the scope of the invention, and the spirit and scope of the invention only limited by claims.To those skilled in the art, under the prerequisite not deviating from essence of the present invention and scope, the various change carry out the material component in these embodiments and consumption or change also belong to protection scope of the present invention.

Prepared by embodiment 1 raw material

1. the preparation of Lactobacillus plantarum pulvis:

(1) first order seed is cultivated: access in 500 ml shake flasks by Lactobacillus plantarum CGMCC NO.9405 bacterial classification, seed culture medium loading amount 100 milliliters, cultivation temperature 37 DEG C, incubation time 24 hours;

(2) secondary seed cultivate: by first order seed according to 10% inoculum concentration access in 500 milliliters of secondary seed shaking flasks, condition of culture is identical with first order seed;

(3) three grades of seed culture: secondary seed is accessed in 5000 milliliters of three grades of seed flask with 10% inoculum concentration, seed culture medium loading amount 1000 milliliters, cultivation temperature 37 DEG C, incubation time 24 hours;

(4) first class seed pot is cultivated: the first class seed pot being 150L with 5% inoculum concentration access total measurement (volume) by three grades of seeds, fermentation medium loading amount 100L, cultivation temperature 37 DEG C, tank pressure 0.05MPa, incubation time 18 hours;

(5) fermentation tank culture: it is 3 tons of secondary seed tanks that first class seed pot bacterial classification is accessed total measurement (volume) with 5% inoculum concentration, fermentation medium loading amount 2 tons, condition of culture cultivation temperature 37 DEG C, tank pressure 0.05MPa, incubation time 22 hours.The complete zymotic fluid that ferments staticly settles, be centrifugally precipitated thing, and then add the carrier of sediment quality 0.8 times, mix, namely 50 DEG C of fluidized bed dryings obtain Lactobacillus plantarum pulvis, and vehicle weight consists of: sour milk powder 25 parts, 12 parts, dextrin.

Described seed culture medium quality group becomes: casein 1%, beef extract 1%, yeast extract 0.5%, glucose 0.5%, sodium acetate 0.5%, citric acid diamines 0.2%, Tween 800.1%, K ₂hPO ₄0.2%, MgSO ₄.7H ₂o 0.02%, MnSO ₄.H ₂o 0.005%, CaCO ₃2%, surplus is water, pH6.8.

Described fermentation medium quality group becomes: soybean protein 1%, beef extract 1%, yeast extract 0.5%, glucose 0.5%, sodium acetate 0.5%, citric acid diamines 0.2%, K ₂hPO ₄0.2%, MgSO ₄.7H ₂o 0.02%, MnSO ₄.H ₂o 0.005%, CaCO ₃2%, surplus is water, pH6.8.

In described Lactobacillus plantarum pulvis, viable bacteria content is 9x10 ¹²cfu/g.

2. the preparation of modified dietary fiber

The preparation method of described modified dietary fiber comprises the following steps: by inulin, apple fiber, common oats fibre, Semen Tritici aestivi fiber 9:5:4:3 Homogeneous phase mixing in mass ratio, add the water of its quality 5 times, room temperature 300W, 40KHz condition ultrasonic extraction 12min, then at electric-field intensity 30kV/cm, burst length 500 μ s, carries out high-pressure pulse electric extraction under pulse frequency 300Hz condition; Be 5.0 by lactic acid adjust ph, add the biology enzyme of mixture quality 0.2%, in 50 DEG C of enzymolysis 35min; Enzymolysis liquid reduced pressure concentration, freeze drying, low-temperature grinding to particle diameter are that namely 0.2mm obtains modified dietary fiber;

Described biology enzyme is zytase, cellulase, laccase, pectase, tannase 7:3:3:3:2 Homogeneous phase mixing in mass ratio.

3. the preparation of fungi extracts

The preparation method of described fungi extracts comprises the steps: that Hericium erinaceus, ginkgo and black fungus being put into respectively the supersonic wave cleaning machine that 0.4% sodium bicarbonate solution is housed cleans 8min in 200W, 40KHz, drain, be cut into phase equal-specification, length and width is 5-10mm, the sheet of thickness 3-5mm or fourth, in mass ratio 9:6:5 Homogeneous phase mixing, then in-23 DEG C of freezing 20min, pulverize immediately, crushed material particle diameter is 0.5mm; The water adding crushed material quality 4 times obtains mixture, adjustment mixture pH is 5.2, at electric-field intensity 40kV/cm under room temperature, burst length 600 μ s, carries out high-pressure pulse electric process under pulse frequency 300Hz condition, is warming up to 50 DEG C, add the mixed enzyme enzymolysis 5 hours of mixture weight 0.8%, obtain edible mushroom enzymolysis liquid, being evaporated to solid content is more than 30%, and freeze drying, low-temperature grinding to particle diameter are that namely 0.2mm obtains fungi extracts;

Described mixed enzyme parts by weight consist of: papain 25 parts, bromelain 25 parts, dextranase 12 parts, zytase 12 parts, pentosanase 12 parts, ficin 8 parts, middle temperature amylase 8 part, pectase 7 parts, tannase 4 parts.

4. the preparation of Saussurea involucrata culture extract

The preparation method of described Saussurea involucrata culture extract comprises the steps: to get Saussurea involucrata culture, and it is in the container of the aseptic FOS aqueous solution of 1.0% that immersion fills mass percent concentration, takes out, then in-20 DEG C of freezing 8min; Be crushed to particle diameter 0.5mm immediately; Putting in container and to add the ethanol of 40% of 4 times of weight, obtaining mixed material, is 4.5 by lactic acid adjust ph, and at electric-field intensity 30kV/cm under room temperature, burst length 400 μ s, carries out high-pressure pulse electric process under pulse frequency 250Hz condition; Then 35 DEG C are warming up to, insulation, Microwave Extraction is carried out under power 200W, frequency 2000Hz condition, wherein, each microwave irradiation total time 70s, carries out compartment irradiation: irradiation 10s, interval 10s, irradiation like this 10 times, carries out ultrasonic assistant extraction simultaneously under power 250W, frequency 35KHz condition; Continue to be warming up to 45 DEG C, insulation, then adds the compound enzyme enzymolysis 40min of mixed material gross weight 3.3%, filters, obtains filtrate; Filter residue 2 times of weight, 78 DEG C of water rinses 3 times, rinsing liquid and filtrate merge, Homogeneous phase mixing, and at electric-field intensity 40kV/cm under room temperature, burst length 500 μ s, carries out high-pressure pulse electric process and namely obtain Saussurea involucrata culture extract under pulse frequency 250Hz condition;

Described compound enzyme is by pectase, cellulase, tannase, amylase and protease 5:4:3:2:1.5 Homogeneous phase mixing in mass ratio.

5. the preparation of pectin decomposer

The preparation method of described pectin decomposer, fresh hawthorn pomace, shaddock ped, pomace, megasse is comprised the steps: to put into the supersonic wave cleaning machine that 1.2% sodium bicarbonate solution is housed respectively and clean 8min in 200W, 40KHz, drain, be cut into phase equal-specification, length and width is 5-10mm, the sheet of thickness 3-5mm or fourth, in mass ratio 10:4:3:3 Homogeneous phase mixing, then in-23 DEG C of freezing 20min, pulverize immediately, crushed material particle diameter is 0.5mm, add the deionized water of crushed material quality 4 times, mix, be 2.5 by citric acid adjust ph, at electric-field intensity 40kV/cm under room temperature, burst length 600 μ s, high-pressure pulse electric process is carried out under pulse frequency 300Hz condition, then normal pressure boils 40min, Bag filter, the filter residue filtrate normal pressure of its quality 2 times boils 50min, Bag filter, merging filtrate, diatomite filtration, filtrate is clarified, at 75 DEG C, be evaporated to solid content is 8%, be cooled to room temperature rapidly, the pH value adding concentrate 1.5 times of volumes is 95% ethanol of 2.5, leave standstill 25min, pectin is precipitated out, the centrifugal 8min of 4000r/min, reclaim precipitation, with the absolute ethyl alcohol wash-out of 2 times of volumes, centrifugal, reclaim precipitation, operation like this 2 times, what obtain is deposited in 50 DEG C of vacuum drying 2.5h, obtain pectin, add 45 DEG C of water of pectin quality 4.5 times, stir, Homogeneous phase mixing, insulation, be 5 by lactic acid adjust ph, add the pectinase enzymatic hydrolysis 3h of 0.5U/mL, enzymolysis liquid filters, filtrate is gone out enzyme 12min in boiling water, be cooled to room temperature, ultrafiltration concentration, namely freeze drying obtains pectin decomposer.

The Lactobacillus plantarum pulvis that following examples use, modified dietary fiber, fungi extracts, Saussurea involucrata culture extract, pectin decomposer are embodiment 1 to be prepared, and other raw material is commercial commodity.

Embodiment 2

Containing a health caring food tablet for probio, prepared by the raw material of following parts by weight:

Described probio pulvis is formed by the pulvis Homogeneous phase mixing of following parts by weight: Lactobacillus plantarum 45 parts, bifidobacterium longum 38 parts, bifidobacterium bifidum 30 parts, bifidobacterium adolescentis 18 parts, lactobacillus bulgaricus 18 parts, lactobacillus acidophilus 12 parts, Lactobacillus casei 9 parts, streptococcus thermophilus 9 parts, saccharomyces cerevisiae 9 parts, candida utili bacterium 5 parts;

Described probio pulvis probiotics viable bacteria content is: 9x10 ¹²cfu/g;

Preparation method, comprises the steps:

1) according to formula rate, get D-mannital, maltodextrin, microcrystalline cellulose, xylo-oligosaccharide, isomalt, WPC, fungi extracts, Saussurea involucrata culture extract, milk flavour pulverizing respectively, cross 80 mesh sieves, then mix 10min, obtain mixed powder;

2) by soluble in water to pectin decomposer, citric acid, beta carotene make 15% the aqueous solution, as adhesive;

3) in described mixed powder, add described adhesive and make softwood, granulate with 16 mesh sieves, obtain wet granular, put in power 3000W, frequency 2450MHz, the dry 5min of temperature 60 C in microwave dryer, with the quick whole grain of 16 mesh sieve;

4) by modified dietary fiber, cyclohexaamylose and probiotic powder mixing 8min, probio pulvis must be embedded;

5) whole good particle and embedding probio pulvis, dolomol mixing 8min, after mixing, compressing tablet, packs and obtains health caring food tablet.

Embodiment 3

D-mannital 15 parts, maltodextrin 10 parts, microcrystalline cellulose 10 parts, 10 parts, probio pulvis, modified dietary fiber 9 parts, xylo-oligosaccharide 7 parts, isomalt 7 parts, WPC 7 parts, fungi extracts 7 parts, Saussurea involucrata culture extract 5 parts, pectin decomposer 5 parts, cyclohexaamylose 1.5 parts, citric acid 1.5 parts, dolomol 1.5 parts, milk flavour 1.5 parts, beta carotene 1 part;

Described probio pulvis is formed by the pulvis Homogeneous phase mixing of following parts by weight: Lactobacillus plantarum 40 parts, bifidobacterium longum 30 parts, bifidobacterium bifidum 25 parts, bifidobacterium adolescentis 15 parts, lactobacillus bulgaricus 15 parts, lactobacillus acidophilus 10 parts, Lactobacillus casei 8 parts, streptococcus thermophilus 8 parts, saccharomyces cerevisiae 8 parts, candida utili bacterium 4 parts;

Described probio pulvis probiotics viable bacteria content is: 7x10 ¹²cfu/g;

Preparation method, comprises the steps:

1) according to formula rate, get D-mannital, maltodextrin, microcrystalline cellulose, xylo-oligosaccharide, isomalt, WPC, fungi extracts, Saussurea involucrata culture extract, milk flavour pulverizing respectively, cross 60 mesh sieves, then mix 3min, obtain mixed powder;

2) by soluble in water to pectin decomposer, citric acid, beta carotene make 13% the aqueous solution, as adhesive;

3) in described mixed powder, add described adhesive and make softwood, granulate with 10 mesh sieves, obtain wet granular, put in power 3000W, frequency 2450MHz, temperature 55 DEG C of dry 3min in microwave dryer, with the quick whole grain of 10 mesh sieve;

4) by modified dietary fiber, cyclohexaamylose and probiotic powder mixing 5min, probio pulvis must be embedded;

5) whole good particle and embedding probio pulvis, dolomol mixing 5min, after mixing, compressing tablet, packs and obtains health caring food tablet.

Embodiment 4

Containing a health-care edible capsule agent for probio, prepared by the raw material of following parts by weight:

D-mannital 35 parts, maltodextrin 20 parts, microcrystalline cellulose 20 parts, 20 parts, probio pulvis, modified dietary fiber 12 parts, xylo-oligosaccharide 11 parts, isomalt 11 parts, WPC 11 parts, fungi extracts 11 parts, Saussurea involucrata culture extract 8 parts, pectin decomposer 7 parts, cyclohexaamylose 5 parts, citric acid 2.5 parts, dolomol 2.5 parts, milk flavour 2.5 parts, beta carotene 2 parts;

Described probio pulvis is formed by the pulvis Homogeneous phase mixing of following parts by weight: Lactobacillus plantarum 50 parts, bifidobacterium longum 45 parts, bifidobacterium bifidum 35 parts, bifidobacterium adolescentis 20 parts, lactobacillus bulgaricus 20 parts, lactobacillus acidophilus 15 parts, Lactobacillus casei 10 parts, streptococcus thermophilus 10 parts, saccharomyces cerevisiae 10 parts, candida utili bacterium 6 parts;

Described probio pulvis probiotics viable bacteria content is: 8x10 ¹²cfu/g;

Preparation method, comprises the steps:

5) whole good particle and embedding probio pulvis, dolomol mixing 8min, after mixing, dress grain, selects capsule, polishing, makes health-care edible capsule agent.

Embodiment 5

Containing a health food granule for probio, prepared by the raw material of following parts by weight:

D-mannital 5 parts, maltodextrin 5 parts, microcrystalline cellulose 5 parts, 5 parts, probio pulvis, modified dietary fiber 6 parts, xylo-oligosaccharide 3 parts, isomalt 3 parts, WPC 3 parts, fungi extracts 3 parts, Saussurea involucrata culture extract 3 parts, pectin decomposer 2 parts, cyclohexaamylose 1 part, citric acid 0.1 part, dolomol 0.1 part, milk flavour 0.1 part, beta carotene 0.01 part;

Described probio pulvis probiotics viable bacteria content is: 9x10 ¹²cfu/g;

Preparation method, comprises the steps:

5) whole good particle and embedding probio pulvis, dolomol mixing 8min, after mixing, pack, seal and obtain health food granule.

Embodiment 6

Containing a health food pulvis for probio, prepared by the raw material of following parts by weight:

D-mannital 50 parts, maltodextrin 30 parts, microcrystalline cellulose 30 parts, 25 parts, probio pulvis, modified dietary fiber 15 parts, xylo-oligosaccharide 15 parts, isomalt 15 parts, WPC 15 parts, fungi extracts 15 parts, Saussurea involucrata culture extract 10 parts, pectin decomposer 10 parts, cyclohexaamylose 10 parts, citric acid 3 parts, dolomol 3 parts, milk flavour 3 parts, beta carotene 3 parts;

Described probio pulvis probiotics viable bacteria content is: 9x10 ¹²cfu/g;

Preparation method, comprises the steps:

1) by formula rate, by modified dietary fiber, cyclohexaamylose and probiotic powder mixing 8min, probio pulvis must be embedded;

2) according to formula rate, D-mannital, maltodextrin, microcrystalline cellulose, xylo-oligosaccharide, isomalt, WPC, fungi extracts, Saussurea involucrata culture extract, pectin decomposer, citric acid are pulverized, cross 100 mesh sieves, then namely obtain health food pulvis with embedding probio pulvis, beta carotene, milk flavour, dolomol mixing 8min.

Embodiment 7 the present invention is containing the sensory evaluation test of the health food of probio

The health food of inviting 24 personnel to prepare embodiment of the present invention 2-6 and commercially available two kinds of health foods containing probio are judged, sense organ is given a mark, wherein specialty and each 12 of layman, professional is young, middle aged, each 4 of old age, men and women half and half, layman is juvenile, young, middle aged, each 3 of old age, and men and women half and half; Marking comprises outward appearance (20 points), quality (25 points), local flavor (30 points), mouthfeel (25 points) four aspects, and marking personnel independently carry out, and are independent of each other, accurate to ensure judging result.Add up judging result, equal score value gets approximation, retains integer, specifically in table 1:

Table 1 sensory evaluation statistics

Note: show significant difference (P < 0.05) with marking different lowercase alphabet in a line, mark different capitalization and represent that difference extremely significantly (P < 0.01), indicate same letter and represent difference not significantly (P > 0.05).

Above result shows, any one all obviously will be better than the commercially available health food containing probio to health food prepared by the present invention from outward appearance, quality, local flavor and mouthfeel, particularly local flavor is fabulous, and also the consumer of applicable Different age group, different hierarchy of consumption eats simultaneously.

Embodiment 8 the present invention is containing the probiotic test of the health food of probio

By health caring food tablet prepared by the embodiment of the present invention 2, being prepared into viable count with sterilized water is 2 × 10 ¹⁰the probiotic solution of CFU/mL, saves backup in 4 DEG C;

1, the 10mL probiotic solution kept of going bail for is injected in test tube 1, adopts ten times of stepwise dilutions to 10 ^-8, get 1mL dilution on flat board, the MRS agar medium being cooled to 45 DEG C poured in dull and stereotyped (sterilizing), shake up rapidly after sterilizing.Again the test tube 2 that 10mL probiotic solution is housed is placed in 80-90 DEG C of water-bath and heats 15-25min, get the probiotic solution after heating and carry out ten times of stepwise dilutions to 10 ^-8, get 1mL dilution on flat board, the MRS agar medium being cooled to 45 DEG C poured in dull and stereotyped (sterilizing) upper and shake up rapidly after sterilizing.Finally the flat board before heating and after heating is all cultivated 24h under 35 DEG C of conditions, calculate the quantity before and after heating.

Result shows, Viable detection reaches 88%.

2, the resistance test of SGF and intestinal juice: the hydrochloric acid 16.4mL adding distil water dilution of getting 100g/L, pH value is made to be respectively 1.5,2.5 and 3.5, get 100mL dilute hydrochloric acid solution, add 1g pepsin respectively, it is made fully to dissolve, obtain SGF, miillpore filter degerming (0.22 μm) is for subsequent use.Get potassium dihydrogen phosphate 6.8g, the 500mL that adds water makes dissolving, by 0.1moL/L sodium hydroxide solution adjust ph to 6.8; Separately get trypsase 10g, the 100mL that adds water makes dissolving, and after two liquid mixing, be diluted with water to 1000ml, obtain simulated intestinal fluid, miillpore filter degerming (0.22 μm) is for subsequent use.Get the probiotic solution that 1mL keeps and join (i.e. ten times of stepwise dilutions) in the SGF of 9mL, and fully mix on the oscillator rapidly, be then placed in 30-45 DEG C of quiescent culture 2-4h.Take out nutrient solution when 1h, 2h, 3h, 4h respectively and count remaining viable count immediately, compare with former viable count, result shows, Viable detection is 97%.Then each 1mL of the nutrient solution digesting different time is taken in simulated gastric fluid, being inoculated in 9mL pH value is respectively in the simulated intestinal fluid of 6.8, be placed in 30-45 DEG C of quiescent culture 2-4h, and respectively 0,3,6,24h sampling, measure its viable count, compare with former viable count, result shows that Viable detection is 99%.

3, simulating the resistance test of cholate: the solution making 1g/L with pancreatin, and the pig cholate adding 0.8% in the solution, is 8.0 with the NaOH adjustment pH of 10%, then uses 0.45 μm of micro-filtrate membrane filtration and degerming.The probiotic solution kept by 0.5mL is inoculated into 4.5mL and simulates in cholate, obtains nutrient solution, count remaining viable count after cultivating 24h.By nutrient solution ten times of stepwise dilutions to 10 in sterile saline ^-8, and pour into MRS, be then placed in 35 DEG C of quiescent culture 24h.Result shows that Viable detection is 99%.

Above result of the test shows, the probio in health food of the present invention probiotic (heat resistance, resistance to pH, bile tolerance) is comparatively strong, is very applicable to human intestines and stomach's environment, and in simulated gastrointestinal environments, survival rate is large, effectively can improve gastrointestinal function.

It should be noted that: health food prepared by embodiment of the present invention 3-6 has above-mentioned experiment effect equally, between each embodiment and little with above-mentioned experiment effect otherness.

Embodiment 9 health food mouse intestinal performance test

Choose common Kunming small white mouse 60, male and female half and half, 18-20g, conventional word is supported.Therefrom random choose 40,9:00 gavage Lincomycin Hydrochloride 0.2mL every morning (20mg)/only, other as a control group, every day same time gavage equivalent sterile saline, continuous one week, prepares the mouse model of intestinal bacilli illness.Model group mouse diet declines, and do not occur dead and phenomenon of significantly suffering from diarrhoea, arrange soft excrement, profile normal aqueous divides more, and bedding and padding are moist.By 40 intestinal bacilli illness mouse, be divided into 2 groups at random, one group 20 only as treatment group, the probiotic solution 0.5ml (2 × 10 that every day, gavage was kept ¹⁰cfu/ml)/only, another 20 as natural recovering group, every day same time gavage equivalent sterile saline, continuous two weeks.21 days whole experimental periods, observe growth and the defecation situation of small white mouse every day, weigh in the 8th, 21 day mouse to probiotic tablet treatment group and natural recovering group, calculate each group of weight average growth rate, result is as table 1; Within every 5 days, survey each group of stool in mice Escherichia coli quantity, calculate average, result is as table 2.Get stool in mice and be about 0.1g, in aseptic operating platform, add 3 beades (adding 0.5mL dilution with 0.1g excrement sample), dilute and inoculate maconkey agar culture medium, calculate every gram of coliform count wet just.

Table 2 mouse Gain weight

Grouping	Average starting weight (g/ only)	Average end heavy (g/ only)	Average growth rate (%)
				Natural recovering group	20.69±1.33	27.34±1.59	32.14 ^a
Treatment group	20.41±1.45	32.58±1.62	59.63 ^b

The situation of coliform count in table 3 stool in mice

Health food treatment group Mouse Weight average growth rate (59.63%) is significantly higher than natural recovering group (32.14%); Solution rear intestinal Escherichia coli quantity of feeding significantly declines, reduce by 82.62%, significantly lower than natural recovering group (24.78%), show the probio rapid field planting in small white mouse enteron aisle in health caring food tablet of the present invention, form dominant microflora, and effectively suppress the growth and breeding of the pathogens such as Escherichia coli, and resident time is long, continues, effectively improve enteron aisle performance.

Embodiment 10 health food of the present invention is on the impact of immunity of organisms

1 experiment purpose

By exercise tolerance test (mouse forced swimming), verify raising immunity, the antifatigue effect of health food of the present invention.

2 experiment materials and reagent

2.1 for reagent thing:

The commercially available health food (G1) containing probio; The commercially available health food (G2) containing probio; Health food (G3-G7) prepared by embodiment of the present invention 2-6.

2.2 reagent:

Liver/muscle glycogen testing cassete, builds up institute of biological products purchased from Nanjing; The concentrated sulfuric acid (AR), Nanjing Chemistry Reagent Co., Ltd.; Physiological saline, Shangdong Changfu Jiejing Pharmaceutical Industry Co., Ltd..

3. animal used as test

ICR mouse, ♂, cleaning grade, body weight 18-22g, is provided by Yangzhou University's comparative medicine center, the quality certification number: SCXK (Soviet Union) 2007 – 0001, the free diet of experimental session mouse.

4. key instrument

Aluminum swimming trunk (50cm × 50cm × 40cm), galvanized wire, low-temperature and high-speed centrifuge: 5804R type, Eppendrof company; Water-bath: DK-S26 type, the grand experimental facilities Co., Ltd of upper Nereid; Electronic scale: BS224S type, Sartorius company; Stopwatch, thermometer

5. experiment grouping

5.1 dosage groupings and given the test agent give the time and at random mouse are divided into 8 groups, often organize 10,1st group to the 7th group respectively to the medicine of G1 ~ G7,8th group is blank group, give isopyknic distilled water, the often every average daily gavage of group 1 time, gavage volume is 0.2ml/10g, gives given the test agent continuously 30 days.

5.2 sample preparations the 1st group to the 7th group: take 2.25g drug sample, be assigned to 150ml with distilled water; Blank group: distilled water 150ml.

6. experimental technique

After 6.1 swimming with a load attached to the body experiment last administration 30min, put mouse in swimming trunk, the depth of water is no less than 30cm, water temperature 25 ± 1 DEG C, the sheet lead of mouse root of the tail portion load 5% body weight, and the swimming of record mouse starts to the dead time, as mouse swimming time.

After 6.2 mice serum urea measure last administration 30min, be not swimming with a load attached to the body 90min in the water of 30 DEG C in temperature, eyeball blood sampling 0.5mL (not adding anti-coagulants) is plucked after rest 60min, put 4 DEG C of refrigerator 3h, the centrifugal 15min of 2000r/min after blood clotting, gets serum and send clinical laboratory of Lianyungang First People's Hospital to detect.

After the mensuration last administration 30min of 6.3 hepatic glycogen, be not swimming with a load attached to the body 90min in the water of 25 ± 1 DEG C in temperature, cervical dislocation puts to death mouse, clean with physiological saline, and with after filter paper suck dry moisture, accurately taking liver 100mg, hepatic glycogen detection kit detects Mouse Liver glycogen content.

Take a blood sample after the mensuration last administration 30min of 6.4 blood lactase acid, then do not bear a heavy burden stops after temperature is the water went swimming 10min of 30 DEG C.Lactic acid instrument assay method: after rest 20min, each blood sampling 20 μ L add in 40 μ L rupture of membranes liquid before swimming, after swimming, after swimming respectively, the smudge cells lactic acid instrument that fully vibrates immediately measures.(blood lactase acid TG-AUC=5 × (the blood lactase acid value of the rear 20min of blood lactase acid value+2 × swimming of the rear 0min of front blood lactase acid value+3 × swimming that swims)

7. observation index walking weight load, blood lactase acid, urea, glycogen initial value

8. statistical method experimental data is used represent, employing t inspection is compared between organizing

9. experimental result

9.1 health foods of the present invention are on the impact of Mouse Weight

Each group of mouse is after giving G1 ~ G7 medicine, before, in, shown in post-weight sees the following form respectively, the original body mass of each group mouse and increase weight body weight and the more equal no difference of science of statistics of control group (P > 0.05), show G1 ~ G7 medicine all without obvious toxicity.Experimental result refers to table 4.

The original body mass of table 4 swimming with a load attached to the body experiment mice, mid-term body weight and terminate body weight ( )

9.2 health foods of the present invention are on the impact of mice burden swimming time

After per os gives mouse G1 ~ G7 medicine, G1 ~ G2 medicine compares with blank group, can obviously extend the mice burden swimming time, there is significant difference (P < 0.05), health food G3 ~ G7 medicine of the present invention compares with blank group, can the significant prolongation mice burden swimming time, there is pole significant difference (P < 0.01), and be obviously better than G1 ~ G2 medicine.The results detailed in Table 5.

Table 5 health food of the present invention on the impact of mice burden swimming time ( )

" * " p<0.05vs blank;

" * * " p<0.01vs blank;

9.3 health foods of the present invention are on the impact of blood lactase acid before and after mouse movement

After per os gives mouse health food of the present invention, health food G3 ~ G7 medicine of the present invention to compare with control group blood lactase acid TG-AUC after mouse movement significant difference (P < 0.05), decrease though G1 ~ G2 medicine group Mouse Blood lactic acid TG-AUC compares with control group, and no difference of science of statistics (P > 0.05).The results are shown in Table 6.

Table 6 health food of the present invention on the impact of blood lactase acid level before and after mouse movement ( )

" * " p<0.05vs blank;

9.4 health foods of the present invention are on the impact of Mouse Liver glycogen

After per os gives mouse G1 ~ G7 medicine, G1 ~ G2 medicine compares with blank group, Mouse Liver glycogen content all has obvious rising, there is significant difference (P < 0.05), health food G3 ~ G7 medicine of the present invention compares with blank group, Mouse Liver glycogen content all has obvious rising, has pole significant difference (P < 0.01), and is obviously better than G1 ~ G2 medicine.The results detailed in Table 7.

Table 7 health food of the present invention on the impact of Mouse Liver glycogen content ( )

" * " p<0.05vs blank;

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9.5 health foods of the present invention are on the impact of mice serum urea

After per os gives mouse G1 ~ G7 medicine, G1 ~ G2 medicine group compares with blank group, after mouse movement, serum urea content all has obvious reduction, there is significant difference (P < 0.05), health food G3 ~ G7 medicine of the present invention compares with blank group, after mouse movement, serum urea content all has obvious reduction all to have obvious reduction, there is pole significant difference (P < 0.01), and be obviously better than G1 ~ G2 medicine.The results detailed in Table 8.

Table 8 health food of the present invention on the impact of mice serum urea content ( )

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10. experiment conclusion

This experiment is mainly through mice burden swimming experiment, and the deposit simultaneously detecting Mouse Liver glycogen observes raising immunity, the antifatigue effect of health food of the present invention.Preliminary Results shows below:

1, G3 ~ G7 health food of the present invention all can extend the mice burden swimming time (P < 0.01), and successful is better than the analysis of functional food sold of other G1 ~ G2.

2, biochemistry detection aspect display, the lactic acid content that after each dosage group of G3 ~ G7 health food of the present invention all can reduce motion, in mice serum, glucose anerobic glycolysis produces, compare with control group and have significant difference (P < 0.05), and although the analysis of functional food sold of other G1 ~ G2 also can reduce the lactic acid content that in the rear mice serum of motion, glucose anerobic glycolysis produces, but compare with control group, no difference of science of statistics (P > 0.05);

3, each dosage group of G3 ~ G7 health food of the present invention all can significantly improve the deposit (P < 0.01) of glycogen in mouse liver, and successful is better than the analysis of functional food sold of other G1 ~ G2;

4, high lithemia model finds, G3 ~ G7 health food of the present invention significantly can reduce the content (P < 0.01) of the rear urea in serum of mouse swimming, and successful is better than other G1 ~ G2 analysis of functional food sold;

11. conclusions

Above-mentioned experiment proves that health food of the present invention can significantly improve immunity of organisms, improve muscle power and the endurance of mouse, the content of urea in serum and lactic acid after reduction mouse movement, and the deposit of glycogen in mouse liver can be significantly improved, contribute to the fatigue that alleviation exercise load causes; The time that mice burden swimming to power exhausts can be extended.

Claims

1. the health food containing probio, prepared by the raw material of following parts by weight: D-mannital 5-50 part, maltodextrin 5-30 part, microcrystalline cellulose 5-30 part, probio pulvis 5-25 part, modified dietary fiber 6-15 part, xylo-oligosaccharide 3-15 part, isomalt 3-15 part, WPC 3-15 part, fungi extracts 3-15 part, Saussurea involucrata culture extract 3-10 part, pectin decomposer 2-10 part, cyclohexaamylose 1-10 part, citric acid 0.1-3 part, dolomol 0.1-3 part, milk flavour 0.1-3 part, beta carotene 0.01-3 part;

Described probio pulvis is formed by the pulvis Homogeneous phase mixing of following parts by weight: Lactobacillus plantarum 40-50 part, bifidobacterium longum 30-45 part, bifidobacterium bifidum 25-35 part, bifidobacterium adolescentis 15-20 part, lactobacillus bulgaricus 15-20 part, lactobacillus acidophilus 10-15 part, Lactobacillus casei 8-10 part, streptococcus thermophilus 8-10 part, saccharomyces cerevisiae 8-10 part, candida utili bacterium 4-6 part;

Described Lactobacillus plantarum pulvis is prepared by Lactobacillus plantarum tlj-2014, this bacterial strain is preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center on July 2nd, 2014, deposit number is CGMCC NO.9405, and Classification And Nomenclature is: Lactobacillus plantarum Lactobacillus plantarum.

2. the health food containing probio as claimed in claim 1, it is characterized in that, prepared by the raw material of following parts by weight: D-mannital 15-35 part, maltodextrin 10-20 part, microcrystalline cellulose 10-20 part, probio pulvis 10-20 part, modified dietary fiber 9-12 part, xylo-oligosaccharide 7-11 part, isomalt 7-11 part, WPC 7-11 part, fungi extracts 7-11 part, Saussurea involucrata culture extract 5-8 part, pectin decomposer 5-7 part, cyclohexaamylose 1.5-5 part, citric acid 1.5-2.5 part, dolomol 1.5-2.5 part, milk flavour 1.5-2.5 part, beta carotene 1-2 part.

3. the health food containing probio as claimed in claim 1, it is characterized in that, prepared by the raw material of following parts by weight: D-mannital 25 parts, maltodextrin 15 parts, microcrystalline cellulose 15 parts, 15 parts, probio pulvis, modified dietary fiber 11 parts, xylo-oligosaccharide 9 parts, isomalt 9 parts, WPC 9 parts, fungi extracts 9 parts, Saussurea involucrata culture extract 7 parts, pectin decomposer 6 parts, cyclohexaamylose 3 parts, citric acid 2 parts, dolomol 2 parts, milk flavour 2 parts, beta carotene 1.5 parts.

4. the health food containing probio as claimed in claim 1, it is characterized in that, the preparation method of described modified dietary fiber comprises the following steps: by inulin, apple fiber, common oats fibre, Semen Tritici aestivi fiber 8-10:4-6:3-5:2-4 Homogeneous phase mixing in mass ratio, add its quality 3-7 water doubly, room temperature 200-300W, 35-40KHz condition ultrasonic extraction 10-15min, then at electric-field intensity 20-40kV/cm, burst length 400-500 μ s, carries out high-pressure pulse electric extraction under pulse frequency 200-300Hz condition; Be 4.5-6.5 by lactic acid adjust ph, add the biology enzyme of mixture quality 0.1-0.3%, in 45-55 DEG C of enzymolysis 20-48min; Enzymolysis liquid reduced pressure concentration, freeze drying, low-temperature grinding to particle diameter are that namely 0.1-0.3mm obtains modified dietary fiber;

5. the health food containing probio as claimed in claim 1, it is characterized in that, the preparation method of described fungi extracts comprises the steps: that Hericium erinaceus, ginkgo and black fungus being put into respectively the supersonic wave cleaning machine that 0.3-0.5% sodium bicarbonate solution is housed cleans 5-10min in 200W, 40KHz, drain, be cut into phase equal-specification, length and width is 5-10mm, the sheet of thickness 3-5mm or fourth, 8-10:5-7:4-6 Homogeneous phase mixing in mass ratio, then in-21--25 DEG C of freezing 10-30min, pulverize immediately, crushed material particle diameter is 0.2-1mm; Add crushed material quality 3-5 water doubly and obtain mixture, adjustment mixture pH is 4.5-6, at electric-field intensity 35-45kV/cm under room temperature, burst length 500-700 μ s, high-pressure pulse electric process is carried out under pulse frequency 200-400Hz condition, be warming up to 45 ~ 55 DEG C, add the mixed enzyme enzymolysis 4 ~ 6 hours of mixture weight 0.5-1.2%, obtain edible mushroom enzymolysis liquid, being evaporated to solid content is more than 30%, and freeze drying, low-temperature grinding to particle diameter are that namely 0.1-0.3mm obtains fungi extracts;

6. the health food containing probio as claimed in claim 1, it is characterized in that, the preparation method of described Saussurea involucrata culture extract comprises the steps: to get Saussurea involucrata culture, it is in the container of the aseptic FOS aqueous solution of 0.9-1.1% that immersion fills mass percent concentration, take out, then in-18--23 DEG C of freezing 5-10min; Be crushed to particle diameter 0.3-1mm immediately; To put in container and to add the water of 3-5 times of weight or the ethanol of 35-75%, obtaining mixed material, is 3.5-5.5 by lactic acid adjust ph, at electric-field intensity 25-35kV/cm under room temperature, burst length 300-500 μ s, carries out high-pressure pulse electric process under pulse frequency 200-300Hz condition; Then 30-40 DEG C is warming up to, insulation, Microwave Extraction is carried out under power 150-300W, frequency 2000Hz condition, wherein, each microwave irradiation total time 60-80s, carries out compartment irradiation: irradiation 10s, interval 10s, irradiation like this 10 times, carries out ultrasonic assistant extraction simultaneously under power 200-300W, frequency 30-40KHz condition; Continue to be warming up to 40-50 DEG C, insulation, then adds the compound enzyme enzymolysis 30-50min of mixed material gross weight 2.5-4%, filters, obtains filtrate; Filter residue 1-3 times of weight 76-80 DEG C of water rinse 3 times, rinsing liquid and filtrate merge, and Homogeneous phase mixing, at electric-field intensity 35-45kV/cm under room temperature, burst length 400-600 μ s, carries out high-pressure pulse electric process and namely obtains Saussurea involucrata culture extract under pulse frequency 200-300Hz condition;

7. the health food containing probio as claimed in claim 1, it is characterized in that, the preparation method of described pectin decomposer comprises the steps: fresh hawthorn pomace, shaddock ped, pomace, megasse to put into the supersonic wave cleaning machine that 1.2% sodium bicarbonate solution is housed respectively and clean 5-10min in 200W, 40KHz, drain, be cut into phase equal-specification, length and width is 5-10mm, the sheet of thickness 3-5mm or fourth, 8-12:3-5:2-4:2-4 Homogeneous phase mixing in mass ratio, then in-21--25 DEG C of freezing 10-30min, pulverize immediately, crushed material particle diameter is 0.1-1mm, add crushed material quality 3-5 deionized water doubly, mix, be 2-3 by citric acid adjust ph, at electric-field intensity 35-45kV/cm under room temperature, burst length 500-700 μ s, high-pressure pulse electric process is carried out under pulse frequency 200-400Hz condition, then normal pressure boils 30-50min, Bag filter, filter residue its quality 1-3 filtrate normal pressure doubly boils 40-60min, Bag filter, merging filtrate, diatomite filtration, filtrate is clarified, at 75 DEG C, be evaporated to solid content is 6-10%, be cooled to room temperature rapidly, the pH value adding concentrate 1.5 times of volumes is 95% ethanol of 2-3, leave standstill 20-30min, pectin is precipitated out, the centrifugal 5-10min of 4000r/min, reclaim precipitation, with the absolute ethyl alcohol wash-out of 2 times of volumes, centrifugal, reclaim precipitation, operation like this 2 times, what obtain is deposited in 40-60 DEG C of vacuum drying 2-3h, obtain pectin, add pectin quality 4-5 40-50 DEG C of water doubly, stir, Homogeneous phase mixing, insulation, be 4.5-5.5 by lactic acid adjust ph, add the pectinase enzymatic hydrolysis 2-4h of 0.3-0.8U/mL, enzymolysis liquid filters, filtrate is gone out enzyme 10-15min in boiling water, be cooled to room temperature, ultrafiltration concentration, namely freeze drying obtains pectin decomposer.

8., containing the preparation method of the health food of probio as described in as arbitrary in claim 1-7, it is characterized in that, comprise the steps:

9., as claimed in claim 8 containing the preparation method of the health food of probio, it is characterized in that, comprise the steps:

10., containing the preparation method of the health food of probio as described in as arbitrary in claim 1-7, it is characterized in that, comprise the steps: