CN106173613B - A method of fruits and vegetables or cereal or integration of drinking and medicinal herbs ferment health food are prepared using probiotics fermention - Google Patents

Abstract

The present invention relates to a kind of methods for preparing fruits and vegetables or cereal or integration of drinking and medicinal herbs ferment health food using probiotics fermention.Present inventor disclosed be suitable for fruits and vegetables or cereal or integration of drinking and medicinal herbs food materials etc. be ferment raw material produce ferment bacterial strain, optimize zymotechnique, the ferment fermenation raw liquid such as fruits and vegetables or cereal or integration of drinking and medicinal herbs food materials is obtained, obtains various ferment products through following process.Ferment obtained is a kind of ideal health food rich in ingredient exocellular polysaccharide, mannitol and SOD the effect of accumulation in fermentation process.

Description

It is a kind of to prepare fruits and vegetables or cereal or integration of drinking and medicinal herbs ferment health care food using probiotics fermention The method of product

Technical field

The present invention relates to food or field of health care products, more particularly it relates to which a kind of prepared using probiotics fermention The method of fruits and vegetables or cereal or integration of drinking and medicinal herbs ferment health food.

Background technique

Ferment class health food is to be made based on natural edible raw materials such as fresh fruit and vegetables using the fermentation of microorganism With a large amount of metabolites generated and accumulation is required are final to obtain the consumable products with special dietary and healthcare function.

Ferment is external very fashionable.In Japan and China Taiwan, ferment based food is as a kind of functional food It is very welcomed.According to incompletely statistics, only in Japan, existing brand just has several hundred kinds, has more than 20,000,000 people every year and eats, city Field reaction is very good.Taiwan ferment class health food has more than 40 years production histories, there is over one hundred kind of launch sale.Asia its His regional such as South Korea, is also in great demand to Indonesia at Malaysia very much.In the recent period, in Europe and the ground such as the U.S., Canada, Ferment class health food is also gradually fashionable.The ferment based food majority of domestic market sale is external import.

But traditional ferment product use aging process, fermentation period is long, usually 1 year or so even it is longer when Between.Aging process is not easy to the control of processing technology simultaneously, and product stability is relatively poor.Thus low efficiency, product price It is expensive.

Summary of the invention

The purpose of the present invention is to provide a kind of fruits and vegetables or cereal or integration of drinking and medicinal herbs ferment health food and preparation method thereof.

In the first aspect of the present invention, a kind of side for preparing fruits and vegetables or cereal or integration of drinking and medicinal herbs ferment fermenation raw liquid is provided Method, which comprises using fruits and vegetables or cereal or integration of drinking and medicinal herbs class food materials extract as fermentation substrate, with bulgarian milk bar Bacterium, streptococcus thermophilus, lactobacillus acidophilus and bifidobacterium adolescentis ferment as fermenting microbe, obtain fruits and vegetables or cereal or medicine Eat homologous ferment fermenation raw liquid.

In a preferred embodiment, the fruits and vegetables include but is not limited to: pawpaw, pineapple, apple, pears, orange, grape, west Melon, longan, dragon fruit, cherry, Kiwi berry, strawberry, lemon, shaddock, honey peach, orange, blueberry, banana, lichee, pomegranate, nothing Flowers and fruits, cumquat, ponkan, muskmelon, "Hami" melon, apricot, mulberry fruit, loquat, plum, red bayberry, mango, coconut milk, mangosteen, olive, Hu Luo Fore-telling, cucumber, wild cabbage, tomato, mushroom, kelp, agaric, wax gourd, Chinese yam, gumbo, asparagus etc.；Or

The cereal includes but is not limited to: oat, highland barley, wheat, barley, black rice, purple rice, mung bean, red bean, big Beans, Semen Coicis, corn etc.；Or

The integration of drinking and medicinal herbs food materials include but is not limited to: jujube, fructus lycii, longan, lily, lotus seeds, honey, hawthorn, Sea-buckthorn, Poria cocos, Gorgon fruit, honeysuckle, purple perilla, ginseng, rose etc..

In another preferred example, the fruits and vegetables or cereal or integration of drinking and medicinal herbs class food materials extract are fruits and vegetables or cereal or medicine It eats homologous class food materials (fruits and vegetables include that fruits and vegetables are dry or fresh fruit vegetable meat) and mixes homogenate with water, with cellulase and pectase enzyme Product after solution；With or without enzyme hydrolysis after homogenate；Or the fruits and vegetables extract is that the liquid that fruit press obtains produces The product of homogenate is mixed after object or fruits and vegetables are broken with water.

In another preferred example, the enzyme is decomposition of cellulose or the enzyme for decomposing pectic substance；Preferably cellulase And/or pectase；Or the enzyme is to promote amylolytic enzyme；Preferably α-amylase and/or carbohydrase.

In another preferred example, the ratio of the cellulase and pectase is 1:2~2:1 (activity ratio).

In another preferred example, when the cereal mixes homogenate with water, material-water ratio (mass ratio) is 1:5~15；Preferably Ground is 1:8~12.

It in another preferred example, further include the process of filtering after enzymatic hydrolysis；Preferably, be filtered with 200 mesh screens, Obtain filtrate.

In another preferred example, the fruits and vegetables or cereal or integration of drinking and medicinal herbs class food materials extract are used as after sterilization Fermentation substrate.

In another preferred example, the lactobacillus bulgaricus, streptococcus thermophilus, lactobacillus acidophilus and youth bifid bar Bacterium (according to inoculum concentration) is with (1~5): (1~5): (1~5): (1~5)；Preferably with (1~3): (1~3): (1~3): (1~ 3) ratio is mixed, for fermenting；Preferably, with (1 ± 0.1): (1 ± 0.1): (1 ± 0.1): (2 ± 0.2), (2 ± 0.2): (2 ± 0.2): (1 ± 0.1): (1 ± 0.1) or (1 ± 0.1): (1 ± 0.1): (1 ± 0.1): the ratio of (3 ± 0.3) into Row mixing, for fermenting；Preferably, with (1 ± 0.1): (1 ± 0.1): (1 ± 0.1): (2 ± 0.2) are mixed.

In another preferred example, the inoculum concentration of fermenting microbe is 4-12%；Preferably 8-10%.

In another preferred example, soluble solid content 8-15% in fermentation substrate；Preferably 8-11%.

In another preferred example, fermentation time 6~10 days；Preferably 7-9 days.

In another preferred example, after fermentation, (preferably low-temperature centrifugation) and film filtration sterilization are centrifuged.

In another aspect of this invention, a kind of fruits and vegetables or cereal or integration of drinking and medicinal herbs class food materials ferment fermenation raw liquid, institute are provided The fruits and vegetables or cereal or integration of drinking and medicinal herbs class food materials ferment fermenation raw liquid stated are prepared by the method.

In another aspect of this invention, the fruits and vegetables or cereal or integration of drinking and medicinal herbs class food materials ferment fermenation raw liquid are provided Purposes is used to prepare fruits and vegetables or cereal or integration of drinking and medicinal herbs ferment.

In another aspect of this invention, a kind of method preparing fruits and vegetables or cereal or integration of drinking and medicinal herbs ferment, the side are provided Method prepares fruits and vegetables or cereal or integration of drinking and medicinal herbs ferment including the use of the fruits and vegetables or cereal or integration of drinking and medicinal herbs ferment fermenation raw liquid.

In a preferred embodiment, the method also includes: also addition prebiotics, vitamin and mineral.

In another preferred example, the prebiotics include: oligoisomaltose and/or xylo-oligosaccharide etc..

In another preferred example, the vitamin includes: vitamin B₆, vitamin B₁₂, folic acid, inositol.

In another preferred example, the minerals include: L-AA potassium, calcium lactate.

In another aspect of this invention, a kind of fruits and vegetables or cereal or integration of drinking and medicinal herbs fermentoid or its process preparation are provided, In include the fruits and vegetables or cereal or integration of drinking and medicinal herbs ferment fermenation raw liquid；Preferably, the fruits and vegetables or cereal or medicine food are same Source ferment is prepared by the method；Preferably, in the ferment further include: prebiotics, vitamin and mineral Matter.

In a preferred embodiment, the prebiotics, vitamin and mineral include the following:

In another preferred example, the process preparation of the fruits and vegetables or cereal or integration of drinking and medicinal herbs ferment includes (but unlimited In): freeze dried powder, concentrate, granule, capsule, oral solution, tablet.

Other aspects of the invention are apparent to those skilled in the art due to this disclosure 's.

Detailed description of the invention

The preparation flow schematic diagram of Fig. 1, longan ferment fermenation raw liquid.

The schematic diagram of the process of Fig. 2, longan ferment finished product.

Fig. 3, glucose standard curve.

Fig. 4, DNS glucose standard curve.

Fig. 5, mannitol standard curve.

The influence of Fig. 6, inoculum concentration to residual sugar amount.

The influence of Fig. 7, inoculum concentration to exocellular polysaccharide.

The influence of Fig. 8, inoculum concentration to mannitol.

Fig. 9, inoculum concentration influence SOD.

The influence of Figure 10, concentration of substrate to residual sugar amount.

The influence of Figure 11, concentration of substrate to exocellular polysaccharide.

The influence of Figure 12, concentration of substrate to mannitol.

The influence of Figure 13, fermentation time to residual sugar amount.

The influence of Figure 14, fermentation time to mannitol.

The influence of Figure 15, fermentation time to polysaccharide (LP).

Figure 16, fermentation time influence SOD.

Figure 17, longan ferment DPPH free radical scavenging ability.

Figure 18, longan ferment hydroxyl radical free radical Scavenging activity.

Figure 19, longan ferment ultra-oxygen anion free radical Scavenging activity.

Figure 20, jujube enzyme stoste preparation technology flow chart.

Figure 21, jujube ferment health-care food preparation technology flow chart.

Figure 22, Fructus Chaenomelis ferment stoste preparation technology flow chart.

The preparation technology flow chart of Figure 23, Fructus Chaenomelis ferment health food.

Figure 24, mixing pectase preparation technology flow chart.

Figure 25, mixing pectase powder health food preparation technology flow chart.

Figure 26, oat enzyme stoste preparation technology flow chart.

Figure 27, oat ferment preparation technology flow chart.

Figure 28, highland barley ferment preparation technology flow chart.

Figure 29, highland barley ferment health-care food preparation technology flow chart.

Figure 30, lactobacillus bulgaricus growth curve.

Figure 31, streptococcus thermophilus growth curve.

Figure 32, lactobacillus acidophilus growth curve.

Figure 33, bifidobacterium adolescentis growth curve.

Specific embodiment

The present inventor passes through in-depth study, suitable fermenting microbe is screened, with fruits and vegetables or cereal or integration of drinking and medicinal herbs class Food materials are fermentation raw material, optimize zymotechnique, fruits and vegetables or cereal or integration of drinking and medicinal herbs ferment fermenation raw liquid are obtained, through subsequent Processing obtains fruits and vegetables or cereal or integration of drinking and medicinal herbs ferment product.The fruits and vegetables or cereal or integration of drinking and medicinal herbs ferment are rich in fermentation process The effect of accumulation ingredient exocellular polysaccharide, mannitol and SOD, be a kind of ideal health food.

Term

As used herein, " fruits and vegetables or cereal or the integration of drinking and medicinal herbs ferment fermenation raw liquid " refer to fruits and vegetables or cereal or Integration of drinking and medicinal herbs class food materials extract is fermentation substrate, utilizes lactobacillus bulgaricus, streptococcus thermophilus, lactobacillus acidophilus and youth The hybrid bacterial strain of four kinds of bacterial strains of Bifidobacterium ferments, the tunning of acquisition.It is also called " tunning " for short.

As used herein, " fruits and vegetables or cereal or the integration of drinking and medicinal herbs ferment " refers to fruits and vegetables or cereal or integration of drinking and medicinal herbs The finished product obtained after the processing such as ferment fermenation raw liquid deployed, degerming, can be used as health care product or food.

As used herein, " the integration of drinking and medicinal herbs class food materials " refer to effect good for health and also often by as food A kind of food materials of product.

Production technology

The of the invention method for preparing fruits and vegetables or cereal or integration of drinking and medicinal herbs ferment fermenation raw liquid include: with fruits and vegetables or cereal or Integration of drinking and medicinal herbs class food materials extract is fermentation substrate, double with lactobacillus bulgaricus, streptococcus thermophilus, lactobacillus acidophilus and youth Discrimination bacillus is fermented as fermenting microbe, obtains fruits and vegetables or cereal or integration of drinking and medicinal herbs class food materials ferment fermenation raw liquid.Previous Technology in, production ferment is mainly based on spontaneous fermentation, and required time is longer and the quality of tunning is difficult to control.Cause This, the present inventor has investigated the beneficial bacterium of multiple types, finally screens lactobacillus bulgaricus, streptococcus thermophilus, acidophilus cream bar Bacterium and bifidobacterium adolescentis, using four kinds of bacterial strains as fermenting microbe to fruits and vegetables or cereal or integration of drinking and medicinal herbs class food materials extract It ferments, the tunning rich in beneficiating ingredients such as exocellular polysaccharide, mannitol and SOD can be obtained.Also, utilize this four kinds Bacterial strain ferments to fruits and vegetables or cereal or integration of drinking and medicinal herbs class food materials extract as fermentation strain, can be in the period of very short in (about 1 week) acquisition tunning significantly reduces the production time, so that fruit compared with using aging process production ferment The large-scale industrial production of vegetable or cereal or integration of drinking and medicinal herbs ferment is possibly realized.

As preferred embodiment of the invention, the lactobacillus bulgaricus, streptococcus thermophilus, lactobacillus acidophilus and youth Bifidobacterium is with (1~5): (1~5): (1~5): (1~5)；Preferably with (1~3): (1~3): (1~3): the ratio of (1~3) Example is mixed, for fermenting；Preferably, with (1 ± 0.1): (1 ± 0.1): (1 ± 0.1): (2 ± 0.2), (2 ± 0.2): (2 ± 0.2): (1 ± 0.1): (1 ± 0.1) or (1 ± 0.1): (1 ± 0.1): (1 ± 0.1): the ratio of (3 ± 0.3) is mixed, For fermenting；Preferably, with (1 ± 0.1): (1 ± 0.1): (1 ± 0.1): (2 ± 0.2) are mixed.It is mixed under said ratio It shares in fermentation, the wholesome nutriment such as more exocellular polysaccharide, mannitol and SOD can be obtained in tunning.

It about the initial inoculum of hybrid bacterial strain, is found after present invention research, using biggish inoculum concentration energy rapid synthesis Product, but if inoculum concentration is excessive, it is also possible to make that growth is too fast, nutriment largely consumes, and thallus early ageing occur, send out Zymotic fluid viscosity increases, and fermentation lacks of staying power, to influence the yield of lactic acid production and functional component.If inoculum concentration is very few, bacterium Body increasess slowly, and can extend fermentation period, is unfavorable for bacterial strain and adapts to enzymolysis liquid high yield.Therefore, as preferred embodiment of the invention, The inoculum concentration of the fermenting microbe is 4-12%；Preferably 8-10%.

According to the difference of raw material, processing appropriate need to be carried out to obtain fermentation substrate, such as abundant for water content And nutritional ingredient can be retained in the fruits and vegetables in the liquid after juicing, can obtain fermentation substrate using the method for squeezing；For Content of starch cereals abundant, then need to be liquefied or amylorrhexis.As preferred embodiment of the invention, in fermentation substrate Soluble solid content (i.e. fermentation substrate concentration) 8-11%.In the range, residual sugar amount and solid content ratio are lower, Illustrate that lactobacillus-fermented is fast, high to utilization of carbon source rate, exocellular polysaccharide and mannitol yield are relatively high.

As preferred embodiment of the invention, fermentation time 6~10 days.Preferably 7-9 days, within this time range, hair Exocellular polysaccharide, SOD, mannose generate more in zymotic fluid and residual sugar amount is lower.

The technique that the present invention optimizes can guarantee the processing stability of product, while substantially reduce fermentation period, significantly mention High effective component extraction rate.

After obtaining fruits and vegetables or cereal or integration of drinking and medicinal herbs ferment fermenation raw liquid, it can be deployed, degerming, to prepare Edible fruits and vegetables or cereal or integration of drinking and medicinal herbs ferment.

As preferred embodiment of the invention, after fermentation, by being centrifuged, preferably low-temperature centrifugation removal solid content after, Degerming is carried out by the method that film filters again.Degerming can be effectively realized using the technique, and more completely retain effectively at The activity divided.Film filtration sterilization technology is applied in ferment product processing technique by the present invention for the first time, substitutes conventionally pasturised The destruction generated by high temperature to nutritional components of the product is effectively reduced in method, improves product nutritive value and effective component benefit With rate.

As preferred embodiment of the invention, when preparing fruits and vegetables or cereal or integration of drinking and medicinal herbs ferment, in addition to fruits and vegetables or cereal Or other than integration of drinking and medicinal herbs ferment fermenation raw liquid, as needed, can also add wherein prebiotics (include: oligoisomaltose and/ Or xylo-oligosaccharide etc.) and vitamin, so that the fruits and vegetables or cereal or integration of drinking and medicinal herbs ferment have balanced effective nutrition Substance is beneficial to health.Depending on the type of the benefit materials such as prebiotics, the vitamin of addition can according to need, this In the range of skilled in the art realises that.

Fruits and vegetables or cereal or integration of drinking and medicinal herbs ferment product

The present invention also provides apply the method system for preparing fruits and vegetables or cereal or integration of drinking and medicinal herbs ferment fermenation raw liquid The standby fruits and vegetables obtained or cereal or integration of drinking and medicinal herbs ferment fermenation raw liquid, and fermented by the fruits and vegetables or cereal or integration of drinking and medicinal herbs ferment The fruits and vegetables or cereal or integration of drinking and medicinal herbs ferment of edible (or the taking) of the processed acquisition of stoste.The fruits and vegetables or cereal or medicine The process preparation for eating homologous ferment includes but is not limited to freeze dried powder, concentrate, granule, capsule, oral solution, tablet.

Fruits and vegetables or cereal or integration of drinking and medicinal herbs ferment of the invention can be prepared to food, health care product or drug.

Fruits and vegetables or cereal or integration of drinking and medicinal herbs ferment of the invention are when being used to take, general daily dose 50-200ml/ People is more suitable.Certainly, according to individual physique or according to the recommendation of health doctor, dose can also be carried out appropriate Adjustment.

Fruits and vegetables or cereal or integration of drinking and medicinal herbs ferment of the invention are that modern biotechnology is combined with traditional Chinese medical theory Product.Its not only maintain in fruits and vegetables or cereal or integration of drinking and medicinal herbs class food materials original plant Thick many candies, flavones and other effects at Point, also containing nutriments such as organic acid, prebiotics, vitamins, have the function of enhancing immunity of organisms, meanwhile, Ke Yigai Kind intestinal flora balance causes the environment for being unfavorable for pathogen existence, reduces pernicious bacteria in enteron aisle, stimulates the immune of body System improves its immunity.

Present invention will be further explained below with reference to specific examples.It should be understood that these embodiments are merely to illustrate the present invention Rather than it limits the scope of the invention.

One, test material

1, main raw material(s)

Longan (dried longan or fresh longan).

Oligoisomaltose (prebiotics).

2, experimental strain

Lactobacillus bulgaricus 6098, streptococcus thermophilus 6219, lactobacillus acidophilus 6085, bifidobacterium adolescentis 6176, It is provided by Chinese industrial Microbiological Culture Collection administrative center (CICC), cultivation temperature is 37 DEG C.

3, main medium

MRS culture medium (CICC guidance): peptone 10.0g, beef extract 5.0g, yeast powder 5.0g, glucose 5.0g, acetic acid Sodium 5g, dibasic ammonium citrate 2.0g, Tween 80 1.0g, K₂HPO₄2.0g, MgSO₄·7H₂O 0.2g, MnSO₄·H₂O 0.05g, CaCO₃20.0g, agar 15.0g, distilled water 1.0L, pH6.8.Medium sterilization condition: 121 DEG C, 15min.

4, main agents

Glucose, phenol, the concentrated sulfuric acid, concentrated hydrochloric acid, sodium metaperiodate, L- rhamnose, ammonium acetate, glacial acetic acid, acetylacetone,2,4-pentanedione, 3, 5- dinitrosalicylic acid, sodium hydroxide, sodium potassium tartrate tetrahydrate, re-distilled phenol；Sodium sulfite, trishydroxymethylaminomethane (Tris) are adjacent Benzenetriol, reagent are to analyze pure, traditional Chinese medicines chemical reagent Co., Ltd.

5, key instrument equipment

High speed freezing centrifuge Thermo；Microplate reader Thermo；- 80 DEG C of ultra low temperature freezer Thermo；HH-6 digital display constant temperature Water-bath Shanghai Bo Xun Industrial Co., Ltd.；PH meter plum Teller-support benefit instrument company；One two-sided purification work of SW-CJ-1F type Zuo Tai Purifying Equipment Co., Ltd., Suzhou；Vertical pressure steam sterilizer Shanghai Bo Xun Industrial Co., Ltd.；DHP-9162 type electricity Hot constant incubator；DF-101S heat collecting type constant temperature blender with magnetic force, electronic analytical balance, RE-52A Rotary Evaporators, Abbe folding Light instrument.

Two, analysis method

1, soluble solid content Brix is measured

Abbe's refractometer (hand-held).

2, fermentation liquid pH value measures

Accurate pH meter.

3, concentration of reduced sugar measures

The measurement of remaining sugar concentration uses DNS method in fermentation liquid:

(1) DNS reagent: 6.3g 3,5- dinitrosalicylic acid is dissolved in the sodium hydroxide solution of 262ml 2mol/L.By this Solution is mixed with the 500ml hot water for containing 182g sodium potassium tartrate tetrahydrate.5g re-distilled phenol and 5g sodium sulfite are added into the solution, It is sufficiently stirred and is allowed to dissolve, after solution is cooling, be diluted with water to 1000ml.Be stored in brown bottle (need to place in refrigerator It can use after a week).

(2) measurement of content of reducing sugar uses DNS method

It weighs the glucose greater than 1g and is placed in 98 DEG C of dryings of hot air drier to constant weight, accurately weigh 1.000g glucose, It is dissolved with distilled water and is settled to 1000ml.Configuration operation is carried out by table 3-1 respectively, heats and boils in boiling water bath after mixing well Boil 5min.Distilled water 4ml is added into each test tube respectively again after flowing water punching is cooling, mixes.It is blank control, 540nm wave with pipe 1 The long lower absorbance for surveying each pipe.Draw absorbance-glucose content curve.

4, polysaccharide (LP) assay

Using phend-sulphuric acid.

(1) standard curve making

Prepare 100 μ g/mL glucose standards.Respectively draw 0.0,0.1,0.2,0.3,0.4,0.5,0.6,0.7,0.8, 0.9,1.0mL solution is mended with distilled water to 2.0mL in test tube, and the phenol and the 5mL concentrated sulfuric acid of 1mL 6% is added in sequence, is added 6% phenol solution of 1.0mL adds the 5.0mL concentrated sulfuric acid after mixing and is reacted.After reaction mixture stands 10min 25min is placed in 25~30 DEG C of water-baths.After solution colour is stablized, it is control with reagent blank, measures extinction under 490nm Value.It is that coordinate, concentration of glucose are abscissa in length and breadth with light absorption value A, makes phend-sulphuric acid standard curve.

(2) measurement of exocellular polysaccharide

For longan fermented liquid after 200 mesh net filtrations, 4800rpm is centrifuged 20min, takes 5 milliliters of supernatant, 4 times of bodies are added Long-pending dehydrated alcohol is stayed overnight after sufficiently vibrating with precipitate polysaccharides.It is centrifuged 20min under 4800r/min revolving speed and obtains pale yellow precipitate. 20mL dehydrated alcohol is added in precipitating, is centrifuged 20min under 4800r/min after being sufficiently stirred.The precipitating of acquisition repeats the above behaviour Make 4-5 times, to remove the monosaccharide impurity in Thick many candies precipitating, until gained supernatant after centrifugation, is measured using sulfuric acid-phynol method It does not develop the color, to confirm the presence without glucide.

Polysaccharide precipitation carries out dissolution with water and is settled to 100mL, if concentration further takes out greatly 1mL and is settled to 50mL (i.e. dilution 50 × 100 times), and polyoses content is detected with sulfuric acid-phynol method.By the experimental procedure measurement light absorption value of standard curve making, then by Above-mentioned standard curvilinear equation calculates polyoses content.

5, the measurement of mannitol

Using colorimetric method.

The drafting of mannitol standard curve: taking 25mL colorimetric cylinder, the accurate mannitol standard that 50 μ gml-1 are added is molten respectively Liquid 0.0,0.2,0.4,0.6,0.8,1.0m1, then it is supplemented to 1ml respectively with distilled water, 1ml then is added in every colorimetric cylinder Sodium periodate solution mixes, puts 10min, rear that 2ml0.1%L- sandlwood sugar juice is added, and Nash reagent is added after mixing well 4ml, 53 DEG C of water-baths keep the temperature 15min, and taking-up is water-cooled, and with the cuvette of 10mm optical path, measures extinction under 412nm wavelength Degree maps to get standard curve to mannitol concentration with absorbance.

Sample measurement: accurate to draw after taking supernatant suitably to dilute after terminating the filtering fermentation liquor after reacting, centrifugation 1.0ml makes the content of wherein mannitol within the scope of 10~50 μ gml-1, then by the operation of measurement mannitol standard curve Measure absorbance.

6, superoxide dismutase (SOD) determination of activity

According to the method for GB/T 5009.171-2003.

7, total plate count detects: GB4789.2-2010.

8, enumeration of coliforms: GB4789.3-2010.

9, staphylococcus aureus detects: GB4789.10-2010.

10, growth of probiotics curve determination method

Configuration MRS fluid nutrient medium is sub-packed in 24 test tubes, and every test tube liquid amount is 10ml.It is living to be respectively connected to 10 μ l The four kinds of probiotics seed liquors changed, the stationary culture 48h in 37 DEG C of constant incubators.Since 0h, one bottle is taken out every 2h, Thallus OD value is measured under 600nm wavelength.Not connect the fluid nutrient medium of bacterium as control, while measuring when being just inoculated with strain Thallus OD value.Using the time as abscissa, thallus OD value is ordinate, draws the growth curve of strain.

11, probiotics seed fermentation liquid concentration mensuration

Probiotics seed fermentation liquid is made 10 respectively^-1, 10^-2, 10^-3, 10^-4, 10^-5, 10^-6, 10^{- 7,}10^-8, 10^-9, 10^-10It is dilute It releases.Take 10^-4, 10^-5, 10^-6, 10^{- 7,}10^-8The bacterium solution spread plate of extension rate, 37 DEG C of stationary cultures rs for 24 hours measure clump count, Calculate bacterial concentration.

Three, experimental method

1, the preservation of strain

It is cultivated using MRS culture medium, after passage three times, accesses MRS solid slope culture medium, periodically moved using low temperature Plant method is put into preservation in 4 DEG C of refrigerators.

2, the preparation of probiotics seed fermentation liquid

The well-grown thallus of a ring is scraped from culture presevation inclined-plane, access is equipped with 50ml liquid seed culture medium In 250ml triangular flask, 37 DEG C of stationary culture 15-24hrs.With 10% inoculum concentration, activated seed culture fluid is accessed It is spare after 37 DEG C of stationary culture 15-24hrs in 250ml triangular flask equipped with 100ml production medium.

Embodiment 1, the preparation of fermentation substrate and bacterial strain

The preparation flow of longan ferment fermenation raw liquid is as shown in Figure 1.

1, the preparation of longan water-soluble extractive

Longan dried fruit is weighed after shredding, and by material-water ratio 1:10 (mass ratio) plus water, after 90 DEG C of immersion 30min, uses high-speed set It knits bruisher to be beaten, cellulase and pectase be digested according to ratio (activity ratio) addition of 1:1, two kinds of enzymes Total additive amount 0.2% (mass ratio), 55 DEG C of hydrolysis temperature, enzymolysis time 120min, 90 DEG C of enzyme deactivation 10min after enzymatic hydrolysis, Enzymolysis liquid takes filtrate to get longan water-soluble extractive through 200 mesh net filtrations.

2, it sterilizes

Longan water-soluble extractive is through 95 DEG C of sterilization 15min, as fermentation substrate.

3, fermentation strain

Using fermentation substrate prepared above as substrate, pass through Large-scale Screening for a large amount of candidate strains, it is final to determine Four kinds of lactobacillus bulgaricus, streptococcus thermophilus, lactobacillus acidophilus, bifidobacterium adolescentis bacterial strain combinations are used to ferment.

Fermentation-scale are as follows: 2L shaking flask；Fermentation temperature is 37 DEG C, amphimicrobian culture, without ventilation.

Using exocellular polysaccharide, mannitol, superoxide dismutase, residual sugar amount etc. as evaluation index, best fermentation work is determined Skill condition.These beneficiating ingredients of exocellular polysaccharide, mannitol, superoxide dismutase are higher, then show the tunning function obtained Effective component content is higher.Strain is fermented using the glucide in fermentation substrate, and residual sugar amount is lower, usually illustrates that sugar is sharp More abundant, the degree for progress of fermenting is higher, and tunning is also more.

Embodiment 2, standard curve

1, exocellular polysaccharide measures standard curve

Determination of polysaccharide is carried out using phend-sulphuric acid.If front legal system makees standard curve, data such as table 1 is made.

Table 1, glucose standard curve make data

The glucose standard curve of acquisition such as Fig. 3.Light absorption value (A490nm) and concentration of glucose (× Csug, 100 μ g/mL) Relationship are as follows: y=0.0088x+0.097, coefficient R²=0.9958；Show glucose solution in 5-50 μ g/mL in good Linear relationship.

2, residual sugar amount standard curve determination

DNS standard curve is made, data such as table 2 is made.

Table 2, glucose standard are prepared

The absorbance value for measuring the glucose standards solution of various concentration, using glucose content X as abscissa, with light absorption value Y is ordinate, and the glucose solution standard curve made is as shown in Figure 4.

From fig. 4, it can be seen that the relationship at the wavelength 540nm between the absorbency Y and glucose content X of characteristic absorption is Y= 2.8473x+0.0266 (coefficient R²=0.9987), this glucose standard curve is used as the calculation basis of measurement result.

3, mannitol standard curve

The drafting of mannitol standard curve is carried out using colorimetric method.Make data such as table 3.

Table 3, mannitol standard curve making

As seen from Figure 5, the relationship at the wavelength 412nm between the absorbency Y and mannitol content X of characteristic absorption is Y= 0.0098x+0.13486 (coefficient R²=0.9999), show mannitol content in 10~50 μ gml^-1In good in range Linear relationship.

Embodiment 3, growth curve

1, lactobacillus bulgaricus growth curve

It can be seen that lactobacillus bulgaricus OD value from the growth curve of Figure 30 quickly to increase from 10h to 22hrs in logarithm Long, this stage is logarithmic growth phase.For 24 hours to 38h growth metabolic stability, thallus OD value is always held at 2.8 or so, this One stage was the stationary phase of growth.Thallus OD value starts to be gradually reduced after 40h, is the decline phase of thalli growth.

2, streptococcus thermophilus growth curve

It can be seen that streptococcus thermophilus thallus OD value from the growth curve of Figure 31 to increases slowly in 0h to 8h, it is raw for strain Long period of delay.Strain starts mass propagation in 10h to 22h, and thallus OD value is also in logarithm rapid growth, this stage is logarithm Growth period.22h is always held at 1.0 or so to 38h growth metabolic stability, thallus OD value, and stationary phase is relatively long.38h Thallus OD value starts fluctuation occurred afterwards.

3, lactobacillus acidophilus growth curve

It can be seen that lactobacillus acidophilus thallus OD value from the growth curve of Figure 32 to increases slowly in 0h to 8h, it is raw for strain Long period of delay.Strain starts mass propagation in 10h to 20h, and thallus OD value is also in logarithm rapid growth, this stage is logarithm Growth period.28h is always held at 2.0 or so to 38h growth metabolic stability, thallus OD value, this stage is growth Stationary phase.Thallus OD value starts to be gradually reduced after 40h, is the decline phase of thalli growth.

4, bifidobacterium adolescentis growth curve

As shown in figure 33, bifidobacterium adolescentis growth curve is different from the growth curve of other three kinds of lactic acid bacteria culturers, bacterium Four growth cycles of kind are than more significant.Thallus OD value increasess slowly in 0h to 5h, and strain constantly shakes down at this time, therefore This stage is the period of delay of growth.Strain rapidly increases in 5h to 15h mass propagation, thallus OD value, is logarithmic growth Phase.In 15h to 22h growth metabolic stability, this stage is the stationary phase of growth.After 22h thallus OD value start by Gradually decline, is the decline phase of thalli growth.

5, strain seed liquor concentration

The best incubation time of seed culture fluid is determined according to the growth curve of measurement.Generally select logarithmic growth phase Bacterial strain is as fermentation seed liquid.Work when four kinds of probiotics seed fermentation liquid logarithmic growth phases is measured using the method for plate culture count Bacterium number, four kinds of probiotics bacterial liquid viable counts are respectively as follows: lactobacillus bulgaricus 5.0 × 10 after measured⁸CFU/mL, streptococcus thermophilus 1.2×10⁸CFU/mL, lactobacillus acidophilus 4.2 × 10⁷CFU/mL, bifidobacterium adolescentis 1.7 × 10⁷CFU/mL.Meet leavening Bacterial concentration requirement.

Embodiment 4, longan ferment zymotechnique

1, the determination of strain inoculated amount and inoculative proportion

Setting strain ratio (streptococcus thermophilus, lactobacillus bulgaricus, lactobacillus acidophilus, bifidobacterium adolescentis=1:1: 1:1), fermentation substrate concentration soluble solid content (Brix)=10%, by probiotics according to inoculum concentration 2%, 4%, 6%, 8%, 10%, 12% (being v/v) is inoculated into fermentation substrate, is left to ferment under the conditions of 37 DEG C, and fermentation measured fermentation liquid after 7 days In exocellular polysaccharide, mannitol, SOD, determine optimum inoculation amount.

As a result, it has been found that using biggish inoculum concentration energy rapid synthesis product, but if inoculum concentration is excessive, it is also possible to make strain It grows too fast, nutriment largely to consume, and thallus early ageing occurs, fermentation liquid viscosity increases, and fermentation lacks of staying power, to influence The yield of lactic acid production and functional component.If inoculum concentration is very few, thallus increasess slowly, and can extend fermentation period, be unfavorable for bacterium Strain adapts to enzymolysis liquid high yield.

Lactic acid bacteria, which carries out fermentation using the carbon source in fermentation substrate, will be such that the content of reducing sugar in fermentation liquid reduces.By scheming Influence of 6 inoculum concentrations to residual sugar amount can be seen that inoculum concentration from 6% increase to 10% when, residual sugar amount with inoculum concentration increase and It reduces, when inoculum concentration is 10%, residual sugar amount is minimum, and when inoculum concentration increases to 12%, residual sugar amount increases instead.

Influence by Fig. 7, Fig. 8 inoculum concentration to exocellular polysaccharide and SOD the experimental results showed that, inoculum concentration increases to from 4% 10%, exocellular polysaccharide and SOD increase with the increase of inoculum concentration, and at 10%, exocellular polysaccharide and SOD reach most inoculum concentration Big value, when inoculum concentration continues to increase to 12%, exocellular polysaccharide and SOD are reduced instead.

Influence of Fig. 9 inoculum concentration to mannitol the experimental results showed that, when inoculum concentration is 8%, mannitol reaches maximum value 4.42mg/ml, inoculum concentration increase mannitol content and then reduce.The above reason, it may be possible to when inoculum concentration is excessive, so that thallus is preceding Phase growth is too fast, fermentation liquid viscosity is caused to increase, and thallus early ageing fermentation occurs and lacks of staying power, affects functional component yield.

Therefore, the preferable inoculum concentration of fermentation test is 4-12%；Also, determine that the inoculum concentration of 8-10% is sent out as probiotics The optimum inoculum concentration of the water-soluble substance of ferment longan.

2, fermentation liquid concentration of substrate

The fermentation substrate (4%, 6%, 8%, 10%, 12% (Brix)) for preparing various concentration, by hybrid bacterial strain (Bao Jiali Sub- lactobacillus, streptococcus thermophilus, lactobacillus acidophilus, bifidobacterium adolescentis=1:1:1:1) it is inoculated with according to 10% (v/v) inoculum concentration It into fermentation substrate, is left to ferment under the conditions of 37 DEG C, exocellular polysaccharide, the mannitol, SOD in fermentation liquid is measured after fermentation 7 days, really Fixed best concentration of substrate.

Such as Figure 10, influence of the concentration of substrate to residual sugar amount the result shows that, when concentration of substrate is between 8% to 10%, residual sugar Amount is lower with solid content ratio, illustrates that lactobacillus-fermented is fast, high to utilization of carbon source rate.

Concentration of substrate is close to the effect tendency of exocellular polysaccharide and mannitol content it can be seen from Figure 11 and Figure 12, bottom When object concentration is 8%, exocellular polysaccharide and mannitol yield highest.

Influence of the comprehensive concentration of substrate to residual sugar amount and exocellular polysaccharide and mannitol, determines soluble solid content 8- 10% as suitable concentration of substrate；Most preferably 8%.

3, strain ratio

Adjustment fermentation substrate concentration is 10% (Brix), inoculum concentration 10% (v/v), by lactobacillus bulgaricus, thermophilic chain Coccus, lactobacillus acidophilus, bifidobacterium adolescentis be inoculated into fermentation substrate according to different strain ratio.It is stood under the conditions of 37 DEG C Fermentation measures exocellular polysaccharide, mannitol, SOD in fermentation liquid, determines best strain ratio after fermentation 7 days.

As a result such as table 4.

Table 4, strain ratio are to probiotics fermention

By strain ratio to it can be seen from the water-soluble extract experimental result of probiotics fermention longan when strain ratio (protect plus Leah lactobacillus: streptococcus thermophilus: lactobacillus acidophilus: bifidobacterium adolescentis) at 1: 1: 1: 2,2: 2: 1: 1,1: 1: 1: 3, function It is relatively high to imitate component target exocellular polysaccharide, mannitol, superoxide dismutase.Wherein, ideal with 1: 1: 1: 2 effects.

4, fermentation time

Setting strain ratio (streptococcus thermophilus, lactobacillus bulgaricus, lactobacillus acidophilus, bifidobacterium adolescentis=1:1: 1:1), probiotics is inoculated into fermentation substrate according to inoculum concentration 10% (v/v) by fermentation substrate concentration Brix=10% (Brix) In, it is left to ferment under the conditions of 37 DEG C, investigates influence of the different fermentations time to exocellular polysaccharide, mannitol, SOD in fermentation liquid, Determine best fermentation time.

Influence of the fermentation time to probiotics fermention longan juice is as shown in Figure 13, Figure 14, Figure 15 and Figure 16, by Figure 15, figure 16 as can be seen that fermentation time increased to 8 days from 2 days, and exocellular polysaccharide, SOD increase, fermentation time as fermentation time increases When being 8 days, two indexes reach maximum value, and corresponding residual sugar amount reaches minimum (such as Figure 13).In Figure 14, fermentation time 6 days and 8 It when, there is peak in mannitol content.

Therefore, fermentation time was advisable at 6 days~10 days.Also, fermentation time is determined 8 days as probiotics fermention longan juice Optimum fermentation time.

5, the confirmatory experiment of the water-soluble extract optimal conditions of fermentation of longan

By orthogonal test to the water-soluble extract condition optimizing of lactobacillus-fermented longan after, it is determined that best zymotechnique item Part carries out confirmatory experiment with determining optimum combination experiment condition, investigates exocellular polysaccharide, mannitol and SOD, test combinations pair According to so that it is determined that best technological condition for fermentation.

On the basis of experiment of single factor, using the technique study probiotics fermention longan juice optimised process of orthogonal test. Inoculum concentration, 4 fermentation substrate, strain ratio and fermentation time factors are chosen, each factor takes 3 levels, using L9 (3⁴) just Test table is handed over, with exocellular polysaccharide, mannitol, superoxide dismutase (SOD) for evaluation index, is shown in Table 5.

Table 5, L₉(3⁴) orthogonal test table

Table 6, probiotics fermention longan orthogonal experiments

Found out by the factor pole size of the difference of table 6, using exocellular polysaccharide as evaluation index, influences longan ferment zymotechnique Factor primary and secondary sequence are as follows: B fermentation concentration of substrate > D fermentation time > A inoculum concentration > C strain ratio.Optimal conditions combines B₃D₂A₂C₁, inoculum concentration 9%, fermentation substrate concentration 9%, concentration strain ratio 1: 1: 1: 2, fermentation time 8 days.

Using mannitol as evaluation index, the factor primary and secondary sequence of longan ferment zymotechnique is influenced are as follows: B fermentation concentration of substrate > A inoculum concentration > C strain ratio > D fermentation time.Optimal conditions combination is B₂A₃C₂D₂, inoculum concentration 10%, fermentation substrate concentration 8%, concentration strain ratio 2: 2: 1: 1, fermentation time 8 days.

Using superoxide dismutase as evaluation index, the factor primary and secondary sequence of longan ferment zymotechnique is influenced are as follows: D fermentation Time > B fermentation concentration of substrate > A inoculum concentration > C strain ratio.Optimal conditions combination is D₃B₁A₂C₃, inoculum concentration 9%, fermentation substrate Concentration 7%, concentration strain ratio 1: 1: 1: 3, fermentation time 9 days.

It is different from the orthogonal experiments of superoxide dismutase as evaluation index using exocellular polysaccharide, mannitol.Three In index, exocellular polysaccharide is relatively stable with mannitol, and superoxide dismutase thermal stability is poor.The effect of this product ingredient It is determined as based on exocellular polysaccharide, optimum experimental condition is determined using exocellular polysaccharide as the orthogonal experiments of index, i.e. inoculum concentration 9%, fermentation substrate concentration 9%, concentration strain ratio 1: 1: 1: 2 fermentation time 8 days, prepares longan ferment hair with this condition Exocellular polysaccharide content 1.1086mg/mL in zymotic fluid.

6, filtering fermentation liquor, purifying

Fermentation ends, fermentation liquid are centrifuged 20min through 85 DEG C of sterilization 10min, 200 mesh screen initial filters, 4800rpm/min, then Through 30~50 Mm filters to get longan ferment fermenation raw liquid.

The preparation of embodiment 5, longan ferment product

The preparation flow of longan ferment product such as Fig. 2.

1, it deploys

The battalion such as prebiotics (oligoisomaltose or xylo-oligosaccharide etc.), the vitamin of proper proportion are added in enzyme stoste Support hardening agent.Additive amount is as shown in table 7.

Table 7

2, homogeneous

By the longan ferment product after allotment at 65 DEG C, homogeneous is carried out under the conditions of 25Mpa.

3, filling and sterilization

After 65 DEG C of sterilization 30min, hot filling is up to longan ferment finished product.

4, longan ferment product standard

Organoleptic indicator's such as table 8.

Table 8

Functional component:

Polyoses content >=800mg/L；

Prebiotics (oligoisomaltose) >=2.8g/100mL.

Physical and chemical index:

Total solids content: >=10%；

PH value: 3.6~4.2.

Microbiological indicator:

Total plate count cfu/mL < 100

Coliform, MPN/100mL < 3

Pathogenic bacteria are not detected.

Embodiment 6, the research of longan ferment antioxygenic property

1, DPPH free radical scavenging ability

40 μ L longan ferment bulk samples are added in 4mL 0.1mmol/L DPPH- methanol solution, add 450L 50mmol/L Tris-HCl buffer (7.4), water bath with thermostatic control 30min at 25 DEG C.Using deionized water as reference solution.In 517nm Lower measurement absorbance.

DPPH free radical scavenging ability (%)=[(A0- (A1-A2)/A0] × 100% formula (1)

In formula (1): A0 is the absorbance of blank control liquid；A1 is the absorbance that sample measures pipe；A2 is sample copy bottom tube Absorbance.

Contain the ingredients such as exocellular polysaccharide, mannitol, active enzyme and polyphenol, vitamin in longan ferment, therefore has anti- Oxidability.Such as Figure 17, longan ferment DPPH free radical scavenging ability enhances with the increase of longan ferment concentration, works as longan When ferment concentration increases to Brix16% from soluble solid content Brix 2%, DPPH free radical scavenging ability increases fast Speed, after longan ferment concentration reaches 16%, DPPH free radical scavenging ability increase tends towards stability.Reason may be when longan ferment To a certain extent, the ingredient that free radical is removed in longan ferment slows down with radical reaction speed for plain concentration increase.

2, hydroxyl radical free radical Scavenging activity

135 μ L longan ferment bulk samples are added to 1.4mL 6mmol/L H₂O₂, 0.6mL20mmol/L water is then added Poplar acid sodium and 2mL 1.5mmol/L ferrous sulfate, water bath with thermostatic control 1h at 37 DEG C.Using deionized water as reference solution.At 562nm Measure absorbance.Hydroxyl radical free radical Scavenging activity is calculated according to formula (1).

Longan ferment hydroxyl radical free radical Scavenging activity is as shown in figure 18, when longan ferment Brix amount increases to 14% from 2% When, hydroxyl radical free radical Scavenging activity increases rapidly, and after concentration reaches 14%, Hydroxyl radical-scavenging ability increases slowly, tends to Stablize.

3, ultra-oxygen anion free radical Scavenging activity measures

The TrisHCI buffer 4.5ml for taking 0.05mol/L pH8.2, is placed in 25 DEG C of water-baths and preheats 20min, respectively The longan ferment stoste of 1ml various concentration and the pyrogallol solution of 0.4ml 25mmol/L is added, in 25 DEG C of water-baths after mixing Middle reaction 5min is added 8mol/L HCl1.0ml and terminates reaction, make reference with Tris-HCI buffer, measures and inhale at 299nm Luminosity calculates clearance rate.Blank control group replaces each processing of sample to do three repetitions with 1ml sample solvent.Clearance rate Calculation formula: ultra-oxygen anion free radical clearance rate (%)=100% (A1-A2)/A1, wherein A1 is the average absorbance of blank Degree, A2 are the mean light absorbency of sample.

Assay NBT photoreduction surveys longan ferment ultra-oxygen anion free radical Scavenging activity, the experimental results showed that, work as osmanthus Circle ferment concentration is in 10% to 20% range, and when Brix is 16%, ultra-oxygen anion free radical Scavenging activity is most strong (Figure 19).

The experimental results showed that longan ferment stoste has DPPH free radical, Hydroxyl radical-scavenging ability, superoxide anion is certainly By base Scavenging activity, therefore there is high antioxygenic property.

The preparation of embodiment 7, jujube ferment and corresponding health food

1, jujube ferment preparation process

Experimental raw: jujube, pure water.

Enzyme preparation: cellulase, pectase, Novi believe that biological (China) Investment Co., Ltd provides.

Experimental strain: lactobacillus bulgaricus 6098, streptococcus thermophilus 6219, lactobacillus acidophilus 6085, youth bifid bar Bacterium 6176 is provided by Chinese industrial Microbiological Culture Collection administrative center (CICC), and cultivation temperature is 37 DEG C.

Main medium: with used in longan ferment.

Main agents: with used in longan ferment.

Key instrument equipment: when film filters, using 0.4 μm of filter membrane, the dynamic pump (beauty of Masterflex L/S Easy-Load spiral shell Millipore company, state)；It is used in other same longan ferments.

Identification and analysis method: with used in longan ferment.

Jujube enzyme stoste preparation technology flow chart such as Figure 20.Main flow is as follows:

(1) preparation of jujube Aqueous extracts

It weighs after jujube stoning, adds water by material-water ratio 1:10, after 90 DEG C of immersion 30min, carried out with high-speed tissue mashing machine Mashing digests cellulase and pectase according to the ratio addition of 1:1, the total additive amount 800U/g of enzyme, hydrolysis temperature 55 DEG C, enzymolysis time 120min, 90 DEG C of enzyme deactivation 10min after enzymatic hydrolysis, enzymolysis liquid is through 100 mesh net filtrations to get jujube water Extract, adjustment soluble solid content to 10%.

(2) it sterilizes

Jujube Aqueous extracts are through 85 DEG C of sterilization 15min, as fermentation substrate.

(3) probiotics fermention jujube Aqueous extracts

Jujube Aqueous extracts are carried out using lactobacillus bulgaricus, streptococcus thermophilus, lactobacillus acidophilus, bifidobacterium adolescentis Inoculation fermentation, fermentation substrate concentration 10%, lactobacillus bulgaricus: streptococcus thermophilus: lactobacillus acidophilus: bifidobacterium adolescentis is 1: 1: 1: 2 (v/v), inoculum concentration 9% (v/v), fermentation time 8 days.

Analyze jujube ferment exocellular polysaccharide, mannitol, superoxide dismutase SOD and other functional ingredient content.

(4) filtering fermentation liquor, purifying

Fermentation ends, fermentation liquid are centrifuged 20min through 4800rpm/min, take supernatant under 0.20Mpa pressure through 0.4 μm Film aseptic filtration is to get jujube enzyme stoste.

2, jujube enzyme stoste evaluation result

(1) functional component is analyzed

Functional component analyzes result such as table 9.

Table 9

(2) sensory evaluation

Jujube enzyme stoste is in peony, no suspended substance；Fragrance with jujube；Mouthfeel is strong, sour-sweet tasty and refreshing, has red The distinctive flavour of jujube fermentation liquid.

3, health food is prepared using jujube enzyme stoste

The process flow for preparing health food using jujube ferment is as shown in figure 21.Include the following:

(1) it deploys

After jujube enzyme stoste is suitably concentrated, the prebiotics (oligoisomaltose or xylo-oligosaccharide) of proper proportion are added And the nutrition fortifiers such as vitamin, minerals prepare jujube ferment health food, formula is as shown in table 10.

Table 10, jujube ferment Ricipe for health care food

(2) homogeneous

By the jujube ferment product after allotment at 55 DEG C, homogeneous is carried out under the conditions of 25Mpa.

(3) degerming, filling

Through 0.4 μm of film aseptic filtration under 0.20Mpa pressure after homogeneous, sterile filling is up to jujube ferment health food.

The jujube ferment health food of acquisition: being in dark red liquid, no suspended substance；With the peculiar fragrance of jujube, free from extraneous odour； Sour-sweet tasty and refreshing, mouthfeel is strong, has the distinctive flavour of jujube ferment.Its polyoses content >=100mg/100mL；Prebiotics are (oligomeric Isomaltose) >=2.8g/100mL；Superoxide dismutase >=500U/100mL；Total dissoluble solid content: >=10%；pH Value: 3.6~4.2；Microbe colony sum cfu/mL < 100；Coliform MPN/100mL < 3；No pathogenic bacteria detection.

The preparation of embodiment 8, Fructus Chaenomelis ferment and corresponding health food

1, Fructus Chaenomelis ferment preparation process

Experimental raw: pawpaw, pure water.

Experimental strain: with used in longan ferment.

Main medium: with used in longan ferment.

Main agents: with used in longan ferment.

Key instrument equipment: film filtering uses 0.4 μm of filter membrane, the dynamic pump (U.S. of Masterflex L/S Easy-Load spiral shell Millipore company)；It is used in other same longan ferments.

Identification and analysis method: with used in longan ferment.

Fructus Chaenomelis ferment stoste preparation process flow such as Figure 22.Main flow is as follows:

(1) preparation of Chinese flowering quince juice

Pawpaw is cleaned, peeled and pulped rear stripping and slicing, weighing, 100 mesh net filtrations are after juicing up to Chinese flowering quince juice.

(2) it sterilizes

Chinese flowering quince juice is through 85 DEG C of sterilization 15min, as fermentation substrate.

(3) probiotics fermention Chinese flowering quince juice

Chinese flowering quince juice is inoculated with using lactobacillus bulgaricus, streptococcus thermophilus, lactobacillus acidophilus, bifidobacterium adolescentis Fermentation, fermentation substrate concentration 10% (Brix), lactobacillus bulgaricus: streptococcus thermophilus: lactobacillus acidophilus: bifidobacterium adolescentis For 1: 1: 1: 2 (v/v), inoculum concentration 9% (v/v), fermentation time 8 days.Analyze Fructus Chaenomelis ferment exocellular polysaccharide, mannitol, super oxygen Object mutase SOD and other functional ingredient content.

(4) filtering fermentation liquor, purifying

Fermentation ends, fermentation liquid are centrifuged 20min through 4800rpm/min, take supernatant under 0.20Mpa pressure through 0.4 μm Film aseptic filtration is to get Fructus Chaenomelis ferment stoste.

2, Fructus Chaenomelis ferment stoste functional component

(1) functional component is analyzed

Functional component analyzes result such as table 11.

Table 11

(2) sensory evaluation

Fructus Chaenomelis ferment stoste is in Chinese red, has the distinctive fragrance of pawpaw, and sweet mouthfeel is tasty and refreshing, has Fructus Chaenomelis ferment fermentation The distinctive flavour of liquid.

3, health food is prepared using Fructus Chaenomelis ferment

The process flow for preparing health food using Fructus Chaenomelis ferment is as shown in figure 23.Include the following:

(1) it deploys

After Fructus Chaenomelis ferment stoste is suitably concentrated, the prebiotics (oligoisomaltose or xylo-oligosaccharide) of proper proportion are added And the nutrition fortifiers such as vitamin, minerals prepare Fructus Chaenomelis ferment health food, formula is as shown in table 12.

Table 12, Fructus Chaenomelis ferment Ricipe for health care food

(2) homogeneous

By the Fructus Chaenomelis ferment product after allotment at 55 DEG C, homogeneous is carried out under the conditions of 25Mpa.

(3) degerming, filling

Through 0.4 μm of film aseptic filtration under 0.20Mpa pressure, sterile filling is up to Fructus Chaenomelis ferment health food.

The Fructus Chaenomelis ferment health food of acquisition: being in Chinese red liquid, no suspended substance；With the peculiar fragrance of pawpaw, free from extraneous odour； Sour-sweet tasty and refreshing, mouthfeel is strong, has the distinctive flavour of Fructus Chaenomelis ferment.Its polyoses content >=100mg/100mL；Prebiotics are (oligomeric Isomaltose) >=2.8g/100mL；Superoxide dismutase >=2000U/100mL；Total solids content: >=10%；PH value: 3.6~4.2.Microbe colony sum cfu/mL < 100, coliform MPN/100mL < 3；No pathogenic bacteria detection.

Embodiment 9, mixing pectase

1, pectase preparation process is mixed

Experimental raw: apple, pears, sweet orange, grape, shaddock, watermelon, fresh longan, lichee, dragon fruit, cherry, Kiwi berry, Strawberry, pawpaw, lemon, carrot, cucumber, tomato.

Experimental strain: with used in longan ferment.

Main medium: with used in longan ferment.

Main agents: with used in longan ferment.

Key instrument equipment: film filtering uses 0.4 μm of filter membrane, the dynamic pump (U.S. of Masterflex L/S Easy-Load spiral shell Millipore company), 10ND FREEZE DRYER, 2XZ-2 type suspension formula vacuum pump, S22-2 constant temperature blender with magnetic force；It is other With used in longan ferment.

Identification and analysis method: with used in longan ferment.

Mix pectase stoste preparation process flow such as Figure 24.Main flow is as follows:

(1) preparation of mixed fruit and vegetable juice

By apple, pears, sweet orange, grape, shaddock, watermelon, fresh longan, lichee, dragon fruit, cherry, Kiwi berry, strawberry, wood The fruit and vegetable materials such as melon, lemon, carrot, cucumber, tomato, clean, stoning or peeling, stripping and slicing, etc. weight weigh, 100 after juicing Mixed fruit and vegetable juice is obtained after mesh net filtration.

(2) it sterilizes

Mixed fruit and vegetable juice is through 85 DEG C of sterilization 15min, as fermentation substrate.

(3) probiotics inoculation fermentation

Mixed fruit and vegetable juice is carried out using lactobacillus bulgaricus, streptococcus thermophilus, lactobacillus acidophilus, bifidobacterium adolescentis Inoculation fermentation, fermentation substrate concentration 9% (Brix), lactobacillus bulgaricus: streptococcus thermophilus: lactobacillus acidophilus: youth bifid Bacillus is 1: 1: 1: 2 (v/v), and inoculum concentration 9% (v/v), fermentation 8 days up to mixing pectase stoste.

2, pectase stoste functional component is mixed

(1) functional component is analyzed

Functional component analyzes result such as table 13.

Table 13

(2) sensory evaluation

Mixing pectase stoste is in pale red, has the mixing distinctive fragrance of fruits and vegetables, and sweet mouthfeel is tasty and refreshing, has fruits and vegetables The distinctive flavour of ferment fermentation liquid.

3, pectase application example is mixed

The process flow for preparing health food using mixing pectase is as shown in figure 25.Include the following:

(1) it deploys

By the nutrition fortifiers such as prebiotics (oligoisomaltose or xylo-oligosaccharide), vitamin, minerals and maltodextrin, After the auxiliary materials such as cycloheptaamylose, sucrose ester are irradiated sterilization, aseptically it is added in mixing pectase stoste, formula As shown in table 14.It is sealed after all raw material mixing, it is uniform to feed liquid through magnetic stirring apparatus stirring 30min.

Table 14, mixing pectase Ricipe for health care food

(2) it is freeze-dried

Mixing pectase after allotment is freeze-dried 48hrs under the conditions of -40 DEG C, pectase powder must be mixed.

(3) it packs

It is under 20-26 DEG C of temperature, the gnotobasis of humidity < 70%, mixing pectase powder progress is aseptic subpackaged, i.e., Obtain pectase powder health food.

The pectase powder health food (containing probiotics) of acquisition: being in pale red powder；With mixing the peculiar fragrance of fruits and vegetables, Free from extraneous odour；Warm water Instant Drinks are sour-sweet tasty and refreshing, have the distinctive flavour of pectase.Its polyoses content >=50mg/100mg；Prebiotics (oligoisomaltose) >=2.8g/100mg；Superoxide dismutase >=500U/100mg；Total amount of probiotics cfu/g > 10⁶；Greatly Intestinal flora MPN/100mg < 3；No pathogenic bacteria detection.

Embodiment 10, oat ferment

1, oat enzyme stoste preparation process

Experimental raw: oat, pure water.

Experimental strain: with used in longan ferment.

Main medium: with used in longan ferment.

Main agents: with used in longan ferment.

Key instrument equipment: with used in longan ferment.

Identification and analysis method: with used in longan ferment.

Oat enzyme stoste preparation process flow such as Figure 26.Main flow is as follows:

(1) preparation of oat slurry

Oat 500g is taken, 5L dissolved in purified water is added and obtains oat slurry.

(2) boiling

Oat slurry is cooling after 121 DEG C of boiling 15min, as fermentation substrate.

(3) probiotics fermention

Oat slurry is inoculated with using lactobacillus bulgaricus, streptococcus thermophilus, lactobacillus acidophilus, bifidobacterium adolescentis Fermentation, fermentation substrate concentration 10%, lactobacillus bulgaricus: streptococcus thermophilus: lactobacillus acidophilus: bifidobacterium adolescentis ratio is 1: 1: 1: 2 (v/v), inoculum concentration 9% (v/v), fermentation time 8 days.Analyze oat ferment exocellular polysaccharide, mannitol, superoxides Mutase SOD and other functional ingredient content.

(4) filtering fermentation liquor, purifying

Fermentation ends, fermentation liquid is after 85 DEG C of sterilization 10min, 4800rpm/min centrifugation 20min through 200 mesh net filtrations Up to oat ferment fermenation raw liquid.

2, oat enzyme stoste functional component is analyzed

(1) functional component is analyzed

Functional component analyzes result such as table 15.

Table 15

(2) sensory evaluation

Oat enzyme stoste is creamy white, and has the distinctive fragrance of cereal, and sweet mouthfeel is tasty and refreshing, has cereal fermentation liquor special Some flavours.

3, oat ferment application example

The process flow for preparing health food using oat ferment is as shown in figure 27.

(1) it deploys

After oat enzyme stoste is suitably concentrated, the prebiotics (oligoisomaltose or xylo-oligosaccharide) of proper proportion are added And the nutrition fortifiers such as vitamin, minerals prepare oat ferment health food, formula is as shown in table 16.

Table 16, oat ferment Ricipe for health care food

2, homogeneous

By the oat ferment product after allotment at 65 DEG C, homogeneous is carried out under the conditions of 25Mpa.

3, filling and sterilization

After 65 DEG C of sterilization 30min, hot filling to obtain the final product

Oat ferment health food.

The oat ferment health food of acquisition: be creamy white liquid；With the peculiar fragrance of fermented grain, free from extraneous odour；It is sour-sweet Tasty and refreshing, mouthfeel is strong, has the peculiar flavour of cereal ferment.Its polyoses content >=30mg/100mL；Prebiotics (oligomeric different malt Sugar) >=2.8g/100mL；Total solids content >=10%；PH value: 3.6~4.2；Microbe colony sum cfu/mL < 100；Greatly Intestinal flora MPN/100mL < 3；Pathogenic bacteria are without detection.

Embodiment 11, highland barley ferment

1, highland barley enzyme stoste preparation process

Experimental raw: highland barley, honey, pure water.

Experimental strain: with used in longan ferment.

Main medium: with used in longan ferment.

Main agents: with used in longan ferment.

Key instrument equipment: with used in longan ferment.

Identification and analysis method: with used in longan ferment.

Highland barley enzyme stoste preparation process flow such as Figure 28.Main flow is as follows:

(1) preparation of highland barley fermentation substrate

1kg highland barley raw material is taken, 2L pure water is added and impregnates, makes starch water swelling, is easy to be gelatinized.After impregnating 12hrs, Remaining water out.Add 1L pure water, stir evenly after be put into boiling in autoclave (temperature be 121 DEG C, time 40min), boiling After be supplemented 2L moisture.α-amylase Clearflow AA is added according to the additive amount of 0.32kg/tonnes, in 95 DEG C of thermostatted water 2hrs is placed in bath.Liquefaction terminates, and adjusts pH value 4.4-4.8, terminates enzymatic hydrolysis.After liquefaction, when temperature drops to 60 DEG C or less Carbohydrase OPTIMAX4060DHP is added according to 0.80kg/tonnes additive amount, places rs for 24 hours in 60 DEG C of waters bath with thermostatic control；Sugar After changing rs for 24 hours, enzyme deactivation 10min is in 90 DEG C of water-baths to get highland barley enzymolysis liquid.

(2) it sterilizes

The honey that 5% (v/v) is added in highland barley enzymolysis liquid, through 85 DEG C of sterilization 15min, as fermentation substrate.

(3) it is inoculated with probiotics fermention

Using lactobacillus bulgaricus, streptococcus thermophilus, lactobacillus acidophilus, bifidobacterium adolescentis to highland barley fermentation substrate into Row inoculation fermentation, fermentation substrate concentration 6%, lactobacillus bulgaricus: streptococcus thermophilus: lactobacillus acidophilus: bifidobacterium adolescentis Ratio is 1: 1: 1: 2 (v/v), inoculum concentration 9% (v/v), fermentation time 8 days.It analyzes highland barley ferment exocellular polysaccharide, mannitol, surpass Superoxide dismutase SOD and other functional ingredient content.

(4) filtering fermentation liquor, purifying

Fermentation ends, fermentation liquid is after 85 DEG C of sterilization 10min, 4800rpm/min centrifugation 20min through 200 mesh net filtrations Up to highland barley enzyme stoste.

2, highland barley enzyme stoste functional component is analyzed

(1) functional component is analyzed

Functional component analyzes result such as table 17.

Table 17

(2) sensory evaluation

Highland barley enzyme stoste has the distinctive fragrance of cereal in faint yellow, and sweet mouthfeel has cereal fermentation liquor distinctive Flavour.

3, health food is prepared using highland barley ferment

The process flow for preparing health food using highland barley enzyme stoste is as shown in figure 29.

(1) it deploys

After highland barley enzyme stoste is suitably concentrated, the prebiotics (oligoisomaltose or xylo-oligosaccharide) of proper proportion are added And the nutrition fortifiers such as vitamin, minerals prepare highland barley ferment health food, formula is as shown in table 18.

Table 18, highland barley ferment Ricipe for health care food

2, homogeneous

By the highland barley ferment product after allotment at 65 DEG C, homogeneous is carried out under the conditions of 25Mpa.

3, filling and sterilization

After 65 DEG C of sterilization 30min, hot filling is up to highland barley ferment health food.

The highland barley ferment health food of acquisition: being in weak yellow liquid；With the peculiar fragrance of fermented grain, free from extraneous odour；It is sour-sweet Tasty and refreshing, mouthfeel is strong, has the peculiar flavour of cereal ferment.Its polyoses content >=100mg/100mL；Prebiotics (oligomeric different malt Sugar) >=3.0g/100mL；Total solids content >=10%；PH value: 3.6~4.2；Microbe colony sum cfu/mL < 100；Greatly Intestinal flora MPN/100mL < 3；Pathogenic bacteria are not detected.

Embodiment 12, ferment shelf-life experiment (by taking longan ferment as an example)

Health food stability test refers to that health food passes through the test of certain procedures and method, investigates the sense of product Official, chemistry, physics and biology situation of change.According to product characteristic difference, stability test can be divided into accelerated test, long-term Test and short term tests.It is three months that accelerated test, which generally investigates the time, i.e., to place 0 month, January, 2 months, sample in March examine It examines.Data can be replaced with same batch products hygiene test result within 0 month.Accelerated test should be placed in 37 ± 2 DEG C of temperature, relatively wet Degree RH75 ± 5% avoids storing three months under conditions of light direct projection, is approximately equivalent to the product room temperature lower shelf-life 2 years.For Longan ferment finished product is placed in 37 DEG C of incubators and carries out shelf-life experiment three months by this, and every month is to microbiological indicator, function Effect ingredient is analyzed, while carrying out sensory evaluation to product.Experimental result is as shown in table 19.

Table 19, longan ferment health drink shelf-life experimental result

The longan ferment shelf-life the experimental results showed that, within the accelerated test phase, product sanitary microorganism index meets food Hygienic requirements, for functional component almost without loss, product sensory evaluation result is good, according to accelerated test as a result, the shelf-life is 2 Year.

Embodiment 13, ferment safety toxicology test experience result

Carry out according to " health food Toxicological evaluation program and method of inspection specification " (Ministry of Public Health's version in 2003) The first stage of ferment product and the safety toxicology detection of second stage, the safety toxicology of longan ferment detect knot Fruit.As shown in table 20, acute toxicity testing result is nontoxic；Micronucleus test, mouse inbred strain and 30 days are fed Feeding test is feminine gender.Toxicology testing result provides foundation for ferment product edible safety.

Table 20, longan ferment safety toxicology testing result

It should be understood that after reading the above teachings of the present invention, those skilled in the art can make the present invention Various changes or modification, these equivalent forms also fall within the scope of the appended claims of the present application.

Claims (19)

1. a kind of method for preparing cereal or integration of drinking and medicinal herbs ferment fermenation raw liquid, which is characterized in that the described method includes: with cereal Or integration of drinking and medicinal herbs class food materials extract is fermentation substrate, with lactobacillus bulgaricus, streptococcus thermophilus, lactobacillus acidophilus and youth Bifidobacterium is fermented as fermenting microbe, obtains cereal or integration of drinking and medicinal herbs ferment fermenation raw liquid；Wherein, the Bao Jiali Sub- lactobacillus, streptococcus thermophilus, lactobacillus acidophilus and bifidobacterium adolescentis are with (1 ± 0.1): (1 ± 0.1): (1 ± 0.1): (2 ± 0.2) it is mixed.

2. the method as described in claim 1, which is characterized in that the cereal includes: oat, highland barley, wheat, barley, black Rice, purple rice, mung bean, red bean, soybean, Semen Coicis, corn, rice, millet etc.；Or

The integration of drinking and medicinal herbs class food materials include: jujube, fructus lycii, longan, lily, lotus seeds, honey, hawthorn, sea-buckthorn, Poria cocos, Gorgon euryale Reality, honeysuckle, purple perilla, ginseng, rose, olive etc..

3. the method as described in claim 1, which is characterized in that the cereal or integration of drinking and medicinal herbs class food materials extract is cereal Or integration of drinking and medicinal herbs class food materials mix the product of homogenate with water；With or without enzyme hydrolysis after homogenate.

4. method as claimed in claim 3, which is characterized in that the enzyme is decomposition of cellulose or the enzyme for decomposing pectic substance； Or

The enzyme is to promote amylolytic enzyme.

5. method as claimed in claim 4, which is characterized in that the decomposition of cellulose decomposes the enzyme of pectic substance as fiber Plain enzyme and/or pectase；Or

The amylolytic enzyme of the promotion is α-amylase and/or carbohydrase.

6. method as claimed in claim 5, which is characterized in that the ratio of the cellulase and pectase is 1:2~2: 1。

7. the method as described in claim 1, which is characterized in that the inoculum concentration of fermenting microbe is 4-12%；Preferably 8- 10%.

8. the method as described in claim 1, which is characterized in that soluble solid content 8-15% in fermentation substrate.

9. the method as described in claim 1, which is characterized in that fermentation time 6~10 days.

10. the method as described in claim 1, which is characterized in that after fermentation, carry out centrifugation and film filtration sterilization；It is described from The heart is low-temperature centrifugation.

11. a kind of cereal or integration of drinking and medicinal herbs class food materials ferment fermenation raw liquid, which is characterized in that the cereal or integration of drinking and medicinal herbs class Food materials ferment fermenation raw liquid is prepared by any method of claim 1-10.

12. the purposes of cereal described in claim 11 or integration of drinking and medicinal herbs class food materials ferment fermenation raw liquid, be used to prepare cereal or Integration of drinking and medicinal herbs class food materials ferment.

13. a kind of method for preparing cereal or integration of drinking and medicinal herbs class food materials ferment, which is characterized in that the method includes utilizing right It is required that cereal described in 11 or integration of drinking and medicinal herbs class food materials ferment fermenation raw liquid prepare cereal or integration of drinking and medicinal herbs class food materials ferment.

14. method as claimed in claim 13, which is characterized in that the method also includes: also addition prebiotics, vitamin and Minerals.

15. a kind of cereal or integration of drinking and medicinal herbs class food materials ferment or its process preparation, which is characterized in that the cereal or medicine food are same Include cereal or integration of drinking and medicinal herbs class food materials ferment fermenation raw liquid described in claim 11 in the class food materials ferment of source.

16. cereal as claimed in claim 15 or integration of drinking and medicinal herbs class food materials ferment or its process preparation, which is characterized in that described Cereal or integration of drinking and medicinal herbs class food materials ferment be to be prepared by the method for claim 13 or 14.

17. cereal as claimed in claim 16 or integration of drinking and medicinal herbs class food materials ferment or its process preparation, which is characterized in that described Ferment in further include: prebiotics, vitamin and mineral.

18. cereal as claimed in claim 17 or integration of drinking and medicinal herbs class food materials ferment, which is characterized in that the prebiotics, dimension Raw element and minerals include the following:

19. cereal as claimed in claim 16 or integration of drinking and medicinal herbs class food materials ferment, which is characterized in that the process preparation packet It includes: freeze dried powder, concentrate, granule, capsule, oral solution, tablet.