CN105455083A - Compounded bee pollen and preparation method thereof - Google Patents

Abstract

The invention discloses compounded bee pollen and a preparation method thereof. Bee pollen is adopted as a main raw material, natural plant source enzymes with high enzyme activity and rich enzyme systems and pupa intestinal tract extract including queen bee larva and drone pupa saliva with the most original bee pollen forming process and the most complete enzyme systems and intestinal tract enzyme systems are scientifically compounded, the microwave high-temperature instant puffing technology and the probiotic fermentation technology are adopted, fast, effective and deep wall breaking is carried out on the bee pollen through special biological enzymolysis, microwave high-temperature instant low-temperature puffing and probiotic fermentation, wall-broken bee pollen is prepared and then is scientifically compounded with one or more functional auxiliary materials, the healthcare bee pollen with the high wall breaking rate, high efficiency, low cost and high functionality is prepared, and the natural flavor and nutritional ingredients of the bee pollen are kept to the maximum extent.

Description

A kind of composite melissa powder and preparation method thereof

Technical field

The present invention relates to Bee Pollen, be specifically related to a kind of composite melissa powder and preparation method thereof.

Background technology

The pollen load taken back when Bee Pollen refers to honeybee producting honey, at the pollen formed after storage and fermentation in honeycomb.Honeybee is when gathering honey, and corbiculate leg can collect pollen, and form pollen load, after entering honeycomb, pollen load can be in store.Its main dietotherapy composition is: protein, amino acid, vitamin, Bee Pollen element, trace element, organized enzyme, flavone compound, lipid, nucleic acid, brassin, phytic acid etc.Wherein amino acid content and ratio are the amino acid patterns recommended closest to FAO (Food and Agriculture Organization of the United Nation) (FAO), and this is extremely rare in wholefood.Bee Pollen is the concentrate that the nutritious material be worth with drug effect forms, and it is containing protein, carbohydrate, mineral matter, vitamin and other active material.Bee Pollen is fabulous natural nutrient food, is also a kind of desirable invigorant simultaneously, and has certain medical function.

Bee Pollen derives from the Nature, the pollen grain that to be honeybee gather in phanerogam (nectariferous plant and plant of pollen) pistil, and add special glandular secretion thing (nectar and saliva) and mixed a kind of irregular discoid shape thing.Bee Pollen has unique natural health effect and medical treatment and its cosmetic values, 's be familiar with by increasing people, it is a kind of high protein and low fat nutritional health food, being described as " all-round nutraceutical " " concentrated natural Drug Storage " " cosmetics for oral administration " etc., is the rarity in mankind's wholefood.

Compared with other natural products, the structure comparison of pollen is special.Pollen grain be by pollen wall, germinal aperature and plasm three part form.Pollen wall, also known as sporoderm, can be divided into inner and outer wall, and inwall is the innermost one deck of sporoderm, smooth, thin and flexible.Outer wall is thicker, hard and lack flexibility, and be one deck that in sporoderm, structure is more complicated, main component is sporopollenin, and this material has the resistance effect of physics, chemistry and enzyme, acidproof, alkaline-resisting, heatproof, withstand voltage, to hydrochloric acid in gastric juice and other digestive system enzyme also highly stable.Research proves: the nutriment in non-broken pollen is not easily digested, and broken wall can make the active material of pollen fully discharge, thus effect of its pollen product and health care are all increased substantially.Pollen wall both have impact on absorbing of pollens nutrition composition, also prevented the release of nutriment during extraction, greatly limit deep development and the utilization of pollen, and therefore, the broken wall of Bee Pollen is very important.

The method of pollen broken wall is mainly divided into mechanical breaking-wall method, physical wall breaking and biological wall breaking both at home and abroad at present.

Mechanical breaking-wall method mainly relies on the effect such as extruding, shearing of machinery, and pollen wall and interior membrane vesicle are broken, and the content release of pollen, common have whirlwind comminuting method and ultramicro grinding method.Hao Xiaoliang etc. adopt whirlwind comminuting method to have studied the broken wall of pollen pini, and Cao Longkui etc. adopt the ultrafine powder technical research broken wall of zasiokaurin, make pollen particles particle diameter reach less than 8 μm, and epigranular are reasonable.Chinese patent CN103181511 discloses a kind of preparation method of ultramicro safflower bee pollen, it is characterized in that safflower bee pollen in the freezer of temperature-20 ~-30 DEG C after freezing 5 ~ 8 days, moving on to temperature is place 20 ~ 30 minutes in the insulating box of 65 ~ 75 DEG C or greenhouse, moving on to temperature is again 45 ~ 50 DEG C of drying box dryings 3 ~ 5 hours, move on to after dry cooling in water-cooled grinding micronizer and pulverize, coolant outlet water temperature controls below 25 DEG C, smashing fineness controls at 1500 ~ 2000 orders, after the ultramicro safflower bee pollen sterilization ground, pack by foil sealing, it is stand-by in-5 ~ 7 DEG C of freezers for being housed in temperature.Use the super-micro wall-broken safflower bee pollen prepared of the inventive method, the absorptivity of safflower bee pollen effective ingredient can be improved, give full play to safflower bee pollen to cardiovascular and cerebrovascular disease, improve effect that blood circulation has prevention and therapy.Chinese patent CN101449754B discloses a kind of production method of broker wall bee pollen granules, belongs to the processing technology of Bee Pollen.The sporoderm-broken rate that current existing broken wall of melissa pollen technology has is low, and some equipment is complicated, and some broken wall speed is slow etc.The invention provides a kind of broken wall efficiency high, sterilization effect is good, the simple broker wall bee pollen granules production method of use equipment, comprise sterilizing, broken wall, granulation and inspection process, it is characterized in that: described wall-breaking machine is vibro-grinding wall-breaking machine, and the vibration frequency of this vibro-grinding wall-breaking machine is between 1200-1600cpm, and Oscillation Amplitude is between 3.5-5mm, it is 100kg that described Bee Pollen often criticizes inventory, and broken time is 2 hours.But the extruding that mechanical breaking-wall method sporoderm-broken rate energy consumption is high and strong, shearing may cause the loss of nutritional labeling.

Physical wall breaking by physical actions such as radiation, swelling, infiltration, ultrasonic wave, microwave, difference variation, pollen wall is broken, and mainly contains temperature differential method, radiation method, hydration broken wall method, supercritical ultrasonics technology, osmotic pressure broken wall method, ultralow temperature adds microwave frequency measurment method and liquid nitrogen quenching broken wall method.Chinese patent CN102114058B is used for the treatment of the production method of the bee pollen tablet of fatty liver, relate to product tablet production technology, the Bee Pollen of removal of impurities is immersed in normal pressure and temperature water after air-flow crushing broken wall treatment, after ultrasonic wave process, through the refrigerated centrifuge centrifugal treating that rotating speed is 8000 ~ 10000 turns, get supernatant, then by supernatant under the environment temperature of-5 ~ 5 DEG C, cross the molecular sieve of below 2800D, isolate the Bee Pollen hydrotrope that molecular weight is less than 2800D; Bee Pollen hydrotrope molecular weight being less than 2800D after vacuum freezedrying, obtain water content lower than 4% melissa freeze-dried powder; By described melissa freeze-dried powder and water soluble starch mixing film-making.Chinese patent CN101416763B discloses a kind of bee pollen beverage and preparation method thereof, comprising the broken wall treatment method of pretreatment being carried out to Bee Pollen, difference Poluo wall combines with mechanical breaking-wall method, refrigerate standing, centrifugal, filter, melissa powder that constant volume, the multiple working procedure such as filling obtain clarification.But single physical method sporoderm-broken rate is lower and high to some equipment requirement.

Biological wall breaking is because sporoderm-broken rate is high, shell-broken effect is remarkable; Action condition is gentle, little to heat-sensitive nutrition destructiveness; Also Bee Pollen absorption and digestion can be promoted while broken wall, produce the plurality of advantages such as functional metabolic product and be widely used in technology for broken wall of melissa pollen and relevant Bee Pollen product, the fermentation that biological wall breaking is large mainly comprises breaking wall by fermentation and enzymolysis broken wall.

Enzymolysis broken wall utilizes enzyme to make some ingredient breakdown on pollen wall destroy pollen wall, and germinal aperature is opened, and pollen content flows out.Enzyme conventional in current enzymolysis broken wall has cellulase, hemicellulase, pectase, protease, amylase, complex enzyme etc.Chinese patent CN102746413B discloses a kind of bee pollen polysaccharide enzymolysis broken wall in conjunction with the ultrasonic leach extraction method of hot water, comprises the following steps: by Bee Pollen drying, removal of impurities, pulverize, sieve; Enzymolysis broken wall; Heat ultrasonic lixiviate; Dehydration; Perchloric acid removing protein; DEAE column chromatography, wash-out; CTAB is separated containing acidic polysaccharose precipitation with containing neutral polysaccharide precipitation; The pure bee pollen polysaccharide of high-purity is obtained through dehydration, vacuum refrigeration, drying after dissolving polysaccharide is deep.Chinese patent CN101401821B discloses one and Bee Pollen is become polypeptide through protease hydrolyzed, tablet made by polypeptide and preparation method thereof.Be will pulverize through screening, impurity elimination, dried Bee Pollen, reach requirement aft-loaded airfoil skill water, stir, be positioned over less than-18 DEG C freezing 24-48 hour.The fresh water (FW) adding 100 DEG C after taking-up immediately stirs and dissolves, and with homogenizer homogeneous 20-60 minute, puts into enzymatic vessel and adds cellulase and carry out enzymolysis 0.5-2 hour, add protease and carry out enzymolysis 3-6 hour after rear alkali lye adjusted to ph.Immediately bee pollen peptide liquid is heated after enzymolysis and carry out going out enzyme.Take starch as excipient, carry out granulation, compressing tablet and get final product.Feature is 1) selectivity: protein, pectin, cellulose, the amylum body macromolecular substances at protease, pectase, cellulase, the amylase single-minded pollen enzymolysis wall of energy and germinal aperature place; 2) high efficiency: enzyme digestion reaction shell-broken effect is good; 3) reaction condition is gentle, is conducive to the preservation of nutrient substance; But due to the single-minded characteristic of Bee Pollen sporoderm complicated component and enzyme, existing enzyme preparation and composite, not can solve enzymolysis broken wall problem, meanwhile, due to yielding poorly of enzyme, price is more expensive, and enzymolysis cost is high.

Breaking wall by fermentation method mainly utilizes the various enzymes produced in the beneficial bacterium sweat such as yeast, aspergillus, lactic acid bacteria, and the various enzymes that pollen inside is contained, utilize the effect of these enzymes to get through the passage portion (germinal aperature, ditch) of pollen inside and outside wall, thus make the nutritional labeling stripping in inwall.Bacterial classification conventional in current breaking wall by fermentation has aspergillus, yeast, lactic acid bacteria, flat mushroom, mushroom etc.Chinese patent CN101756091B discloses a kind of pollen natto tablet, comprises the component of following weight portion: pollen 4 ~ 5 parts, natto 3 ~ 4 parts, 1 ~ 2 part, red colouring agent for food, also used as a Chinese medicine, compound sugar 1 ~ 2 part.Described pollen take Bee Pollen as raw material, obtains: first use 50 ~ 100PPMClO through following preparation method ₂the aqueous solution sprays the airtight sterilization of raw material Bee Pollen 30 minutes, and then adjusting humidity is 25%, and inoculating starter ferments, temperature 30 ~ 37 DEG C, ferments 12 hours, inspection acidity, and as pH=4.5, low temperature drying, obtains pollen; Described leavening is streptococcus lactis and Bacillus acidi lactici.The fermentation process of Chinese patent CN104286623A Bee Pollen and Bee Pollen, adopt probio mixed solid fermentation Bee Pollen, production cost is low, and the fund of investment is less, convenient in downstream, pollutes little.The fermentation Bee Pollen produced, can regulate human body function of intestinal canal.The protein contained in Bee Pollen is broken down into little peptide and free amino acid, is conducive to digesting and assimilating of stomach, removes anaphylactogen.Described probio adopts bifidobacterium lactis or Lactobacillus plantarum or lactobacillus acidophilus.The viable count of described probio is 1 × 10 ⁹-5 × 10 ⁹cFU/g.The manufacture method of a Chinese patent CN104059842A Bee Pollen wine, the step of the manufacture method of described Bee Pollen wine is as follows: get the Bee Pollen 0.8-1 weight portion that sterilization treatment crosses and be cooled to 30-35 DEG C, the black clothing song adding 1.5-2 weight portions adds the softening cold boiling water of 0.8--1 weight portion again, put into batching basin to stir all, to put in wine vat that sterilization treatment crosses sealing and fermenting 40-48 hours; Add the softening cold boiling water of 10-15 weight portions again, add the honey of 10-15 weight portion 40-41 concentration, add longan benevolence 1.5-2.5 weight portion processed, add respectively in feed liquid, stir, blanking is complete to be sealed, temperature between regulation and control yeast phase, remain between temperature 20-25 DEG C, the wine vat that falls is 30-35 days to fermentation dwell time; Extract the wine liquid after filtering, water proof heat sterilization temperature 85-90 DEG C, 20-30 minutes, seals precipitation up for safekeeping 15-20 days after cooling, filling finished product can be waited to drink.Chinese patent CN102389069A discloses a kind of broken wall of melissa pollen method, it is characterized in that comprising the steps: to add culture propagation in Bee Pollen, makes broken wall of melissa pollen; The yeast added in Bee Pollen counts 0.5 ~ 3% of Bee Pollen weight with dry weight; After adding yeast in Bee Pollen, then add the water of Bee Pollen volume 1.5 ~ 2.5 times, then in 28 ~ 34 DEG C of fermentations 2 ~ 4 days, finally in 36 ~ 38 DEG C of heating 4 ~ 8h, obtain the Bee Pollen solution after broken wall.Chinese patent CN103598654B discloses a kind of pollen active probiotic drink and preparation method thereof.This pollen active probiotic drink is raw material with Bee Pollen, is equipped with a certain proportion of maltose and/or galactolipin, through lactobacillus BL1 and lactobacillus BL2 two kinds of probiotic mixed fermentations, then forms through allotment.The present invention is directed to Bee Pollen raw material acidity comparatively strong, not easily by the feature that microorganism utilizes, preferably two acid resistances are strong and have the bacterial strain of multiple merit, make number of live bacteria of probiotics in product 30 days remain on 10 by controlling zymotechnique ¹¹more than cfu/L, and a large amount of flavone aglycone can be obtained by biological conversion Bee Pollen flavonoid glycoside, improve the bioavailability of Bee Pollen flavones.Be rich in other Bee Pollen and probiotics fermention product, nutritious, aromatic flavour, sour and sweet palatability, has the functions such as beautifying face and moistering lotion, develop immunitypty, reduction cholesterol, regulating intestinal canal flora and Constipation simultaneously simultaneously.Aspergillus fermentation method weak flavor in above-mentioned breaking wall by fermentation method; When oyster mushroom fermentation method and the inoculation of mushroom ferment method, easy miscellaneous bacteria infects, mycelial growth is slow, antibacterial ability is poor, sporoderm-broken rate is low; Fermented by lactic acid bacteria and yeast fermentation method sporoderm-broken rate is high, nutrient component damages is little, amino acid content is high but the time is long, speed is slow after broken wall.

To sum up, seek one and keep the natural flavour mountaineous and nutritional labeling of Bee Pollen to greatest extent, the broken wall of melissa pollen method that sporoderm-broken rate is high, efficiency is high, cost is low, necessary with the field of deep expanding Bee Pollen.

Summary of the invention

Technical problem solved by the invention is the defect overcoming existing broken wall of melissa pollen technology, take Bee Pollen as primary raw material, enzyme activity is high, the natural plant enzyme that enzyme system is abundant and the most original containing Bee Pollen forming process, the pupa worm enteron aisle extract science of the queen bee nit that enzyme system is the most complete and drone pupae saliva and enteron aisle enzyme system is composite, and adopt the instantaneous puffing technique of microwave high-temperature and probiotics fermention technology, by Specialty bio enzymolysis, the instantaneous low temperature expanding of microwave high-temperature and probiotics fermention carry out fast to Bee Pollen, effectively and degree of depth broken wall, obtained broker wall bee pollen, then composite with one or more functional auxiliary material science, obtained one keeps the natural flavour mountaineous and nutritional labeling of Bee Pollen to greatest extent, sporoderm-broken rate is high, efficiency is high, cost is low, functional health care composite melissa powder by force.

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

A kind of composite melissa powder, mainly contains the raw material preparation of following parts by weight:

Bee Pollen 100-130 part, plant extracts 10-30 part, pupa worm enteron aisle extract 10-20 part, modified dietary fiber 10-20 part, compound sugar 7-15 part, Lactobacillus plantarum powder 6-10 part, hydroxyl isomaltulose 2-6 part, mango powder 2-6 part;

Preferably, described composite melissa powder, mainly contains the raw material preparation of following parts by weight:

Bee Pollen 110-120 part, plant extracts 15-25 part, pupa worm enteron aisle extract 13-17 part, modified dietary fiber 13-17 part, compound sugar 9-13 part, Lactobacillus plantarum powder 7-9 part, hydroxyl isomaltulose 3-5 part, mango powder 3-5 part;

More preferably, described composite melissa powder, mainly contains the raw material preparation of following parts by weight:

Bee Pollen 115 parts, plant extracts 20 parts, pupa worm enteron aisle extract 15 parts, modified dietary fiber 15 parts, compound sugar 11 parts, 8 parts, Lactobacillus plantarum powder, hydroxyl isomaltulose 4 parts, mango powder 4 parts;

Further, described composite melissa powder also comprises one or more auxiliary materials of following parts by weight;

Algae 0.6-1.2 part, corn oligopeptide powder 0.5-1 part, pueraria root powder 0.5-1 part, fish oil 0.2-0.8 part, phytosterol 0.2-0.8 part, phosphatidyl serine 0.1-0.7 part, chitosan oligosaccharide 0.1-0.7 part, Marine fishbone collagen oligomeric Gly-His-Lys 0.1-0.7 part, Chinese herbal medicine 0.1-0.6 part, Cordyceps militaris 0.1-0.5 part; Soybean protein isolate 0.1-0.5 part;

Preferably, described composite melissa powder also comprises one or more auxiliary materials of following parts by weight;

Algae 0.8-1.0 part, corn oligopeptide powder 0.7-0.9 part, pueraria root powder 0.7-0.9 part, fish oil 0.4-0.6 part, phytosterol 0.4-0.6 part, phosphatidyl serine 0.3-0.5 part, chitosan oligosaccharide 0.3-0.5 part, Marine fishbone collagen oligomeric Gly-His-Lys 0.3-0.5 part, Chinese herbal medicine 0.2-0.4 part, Cordyceps militaris 0.2-0.4 part; Soybean protein isolate 0.2-0.4 part;

More preferably, described composite melissa powder also comprises one or more auxiliary materials of following parts by weight;

0.9 part, algae, corn oligopeptide powder 0.8 part, pueraria root powder 0.8 part, 0.5 part, fish oil, phytosterol 0.5 part, phosphatidyl serine 0.4 part, chitosan oligosaccharide 0.4 part, the oligomeric Gly-His-Lys of Marine fishbone collagen 0.4 part, 0.3 part, Chinese herbal medicine, Cordyceps militaris 0.3 part; Soybean protein isolate 0.3 part;

Further, described Bee Pollen is any one in bee pollen of maize, sesame Bee Pollen, bee pollen form cole, bee's pollen from buckwheat;

Further, not only containing the abundant various plants enzyme such as plant rennet, amylase, hemicellulase, esterase, oxidoreducing enzyme but also containing the nutriment such as plant polyose and monose, plant amylum, vegetable protein in described plant extracts, not only can be the phytoenzyme that composite melissa powder provides comprehensive, natural, also can be the nutriment that probio provides comprehensively, enriches, composite with pupa worm enteron aisle extract science, sporoderm-broken rate is higher, better effects if;

Preferably, described plant extracts adopts the low temperature extractive techniques such as ultrasonic cleaning, the ultrasonic extraction of microwave radiation technology and high-pressure pulse electric extraction, reduced pressure concentration, effectively improves raw material availability, phytoenzyme activity and productive rate; Effectively ensure that the foodsafety of plant extracts;

More preferably, the preparation method of described plant extracts is: by fructus hordei germinatus and wheat malt 8-10:1-3 Homogeneous phase mixing in mass ratio, be crushed to granularity 0.5-1mm, obtain pulverizing Fructus Hordei Germinatus, then by pawpaw, pineapple, fig respectively in supersonic wave cleaning machine at power 200W, ultrasonic cleaning 5-10min under frequency 30KHz condition, drain, granularity 0.5-1mm is crushed under room temperature, and 7-9:1-3:1-2 Homogeneous phase mixing in mass ratio, add mixture quality 3-5 pulverizing Fructus Hordei Germinatus doubly and obtain raw mixture, add raw mixture quality 1-3 water doubly, be 3-4 by citric acid adjust ph, at power 150-300W, Microwave Extraction is carried out under frequency 2000Hz condition, wherein, each microwave irradiation total time 60-80s, carry out compartment irradiation: irradiation 10s, interval 10s, control temperature 20-35 DEG C, irradiation like this 10 times, simultaneously at power 200-300W, ultrasonic assistant extraction is carried out under frequency 30-40KHz condition, insulation 1-3h, then, Microwave Extraction is carried out under power 200-400W, frequency 2000Hz condition, wherein, each microwave irradiation total time 90-105s, carry out compartment irradiation: irradiation 15s, interval 10s, control temperature 40-60 DEG C, irradiation like this 10 times, simultaneously at power 300-500W, carry out ultrasonic assistant extraction under frequency 40-50KHz condition, last Temperature fall to room temperature, in electric-field intensity 25-35kV/cm, burst length 400-600 μ s, carries out high-pressure pulse electric and extracts 15-20min under pulse frequency 200-300Hz condition, extracting liquid filtering obtains the first filtrate, adds the water rinse of filter residue 4 times, filters to obtain the second filtrate, and by the first filtrate and the second filtrate 1:3-5 Homogeneous phase mixing in mass ratio, being evaporated to solid content is more than 20%, obtains plant extracts,

Preferably, in described supersonic wave cleaning machine, cleaning fluid is the sodium bicarbonate solution of 0.3-0.5%.

Further, described pupa worm enteron aisle extract be with the drone pupae enteron aisle of the queen bee nit enteron aisle of 2-3 age in days and 11-12 age in days for raw material, obtain through freeze proof protection, freezing crushing, biological demulsifying and centrifugation;

Preferably, the preparation method of described pupa worm enteron aisle extract, comprise the steps: the drone pupae enteron aisle 3-5:1-3 mixing in mass ratio by the queen bee nit enteron aisle of 2-3 age in days and 11-12 age in days, add the sericin peptide taken solution that mixture quality 1-2 mass percent is doubly 8-12%, stir, put-18--22 DEG C of freezing 5-10min, broken, grind to form homogenate, add the biological demulsifying agent breakdown of emulsion 20-40min of its quality 0.03-0.05%, at 18000-25000g, centrifugal 20-30min under 4 DEG C of conditions, middle level liquid after collection first time is centrifugal, then at 18000-25000g, centrifugal 20-30min under 4 DEG C of conditions, middle level liquid after collection second time is centrifugal, obtain pupa worm enteron aisle extract,

Preferably, the pH value of described sericin peptide taken solution is 6-7;

Preferably, described biological demulsifying agent is that glycolipid class, lipopeptid class, cell wall are in conjunction with the one or more combination in class biological demulsifying agent;

Preferably, the quality group of described biological demulsifying agent becomes: glycolipid class: lipopeptid class: cell wall is in conjunction with class=5-7:4-6:2-4;

Preferably, described glycolipid class biological demulsifying agent is one or both combinations in rhamnolipid, APG;

More preferably, the quality group of described glycolipid class biological demulsifying agent becomes: rhamnolipid: APG=3-5:1-2.

Further, described modified dietary fiber be dietary fiber is obtained through physics, chemistry or biological method process there is strong retentiveness, Soluble Fiber cellulose content that dilatancy, thickening property, adsorptivity and network are enriched is high, biologically active strong, have cellulose that is important, positive role to human body beneficial flora, compared with full diet fiber, its biological action is more powerful, also greatly can extend the shelf-life of composite melissa powder;

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, comprise the following steps: by inulin, Semen Tritici aestivi fiber, common oats fibre 5-7:2-4:1-3 Homogeneous phase mixing in mass ratio, add its quality 3-7 water doubly, room temperature 100-300W, 35-40KHz condition ultrasonic extraction 10-15min, then at electric-field intensity 20-40kV/cm, burst length 300-500 μ s, carries out high-pressure pulse electric process 10-15min under pulse frequency 200-400Hz 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 filters, and filtrate reduced in volume, freeze drying to moisture are that namely 5-8% obtains modified dietary fiber;

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

Further, described compound sugar is at least one in FOS, stachyose, gossypose, xylo-oligosaccharide, galactooligosaccharide, soyabean oligosaccharides, oligoisomaltose, oligomeric maltose;

Preferably, described compound sugar parts by weight consist of FOS 40-50 part, oligomeric maltose 30-40 part, gossypose 20-40 part, soyabean oligosaccharides 16-18 part, galactooligosaccharide 10-15 part, xylo-oligosaccharide 10-15 part, oligoisomaltose 10-15 part, stachyose 8-12 part.

Further, described Lactobacillus plantarum powder is with Lactobacillus plantarum CGMCCNO.11763 for starting strain is prepared according to a conventional method, and its viable bacteria content is: 7 × 10 ¹²-9 × 10 ¹²cfu/g; Wherein freeze drying process adopt cryoprotector with the animal or plant containing antifreeze protein for raw material, obtained through high-pressure pulse electric extraction, ultrasonic assistant Microwave Extraction and compound enzyme enzymolysis, effectively can improve Lactobacillus plantarum powder at freezing dry process Viable detection;

Preferably, described protectant preparation method, comprise the steps: winter rye, Ammopiptanthus mongolicus, sharkskin collagen cleans respectively, drain, 8-10:3-5:2-4 Homogeneous phase mixing in mass ratio, adding mixed material quality 0.1-1 pH value is doubly that the lactic acid of 3.8-4.5 soaks 3-8h, pulverize immediately after-18--22 DEG C of freezing 1-2h, freezing thickness of feed layer 3-5cm, crushed material particle diameter 0.5-3mm, then crushed material quality 10-20 water is doubly added, be 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, high-pressure pulse electric process 20-30min is carried out under pulse frequency 200-300Hz condition, then under power 150-300W condition, carry out microwave irradiation in room temperature and extract 15-20min, under power 200-300W, frequency 30-40KHz condition, carry out ultrasonic assistant extraction simultaneously, add the compound enzyme of extract quality 1-2%, in 45-55 DEG C of enzymolysis 30-50min, enzymolysis liquid filters, filtrate is concentrated, low-temperature grinding to particle diameter is that namely 0.1-0.3mm obtains protective agent,

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

Further, described algae is any one in haematococcus pluvialis, salt algae, blunt top spirulina.

Further, described Chinese herbal medicine is prepared through ultramicro grinding with one or more in chrysanthemum, sophora flower, ginseng, lily, the root of kudzu vine, matrimony vine, dandelion, Chinese yam, Poria cocos, sea-buckthorn, rose, honeysuckle, peppermint, Momordica grosvenori, donkey-hide gelatin, red date, mulberry leaf, balsam pear, Agricus blazei.

Another object of the present invention is to provide the preparation method of above-mentioned composite melissa powder, comprises the steps:

1) each supplementary material is taken according to formula;

2) Bee Pollen is put in the supersonic wave cleaning machine that 0.3-0.5% sodium bicarbonate solution is housed, in 200W under room temperature, 40KHz cleans 3-5min, rinsing, drain, be soak 5-7min in the sericin peptide taken solution of 13-18% in mass percent, take out, put-18--22 DEG C of freezing 5-10min, broken, immediately at power 3-5Kw, frequency 2450MHz, temperature 120-140 DEG C of heating using microwave 5-7s, add former Bee Pollen quality 3-5 water doubly, be 4-6 by lactic acid adjust ph, in electric-field intensity 20-40kV/cm, burst length 400-600 μ s, high-pressure pulse electric process 10-15min is carried out under pulse frequency 200-400Hz room temperature condition, then room temperature 400W, 40KHz condition ultrasonic process 10-15min obtains Bee Pollen pretreatment fluid,

3) Bee Pollen pretreatment fluid is warming up to 40-50 DEG C, adds plant extracts and pupa worm enteron aisle extract wherein, stir, insulation, enzymolysis 10-15min, obtains Bee Pollen enzymolysis liquid;

4) in Bee Pollen enzymolysis liquid, add the modified dietary fiber of compound sugar, 20-30%, abundant dissolving 5-10min, Temperature fall is to 40-45 DEG C, add the 50-60% ferment at constant temperature 3-5h of Lactobacillus plantarum opaque amount, then with the ramp of 0.8-1.0 DEG C/min to 50-55 DEG C, add the 40-50% ferment at constant temperature 1.5-3.5h of Lactobacillus plantarum opaque amount, finally be cooled to 40-45 DEG C with the speed of 0.6-0.8 DEG C/min, continue fermentation 0.5-1h, fermentation liquor 100-300 eye mesh screen filters, and namely filtrate obtain broker wall bee pollen through ultrafiltration, reduced pressure concentration, freeze drying;

5) broker wall bee pollen and hydroxyl isomaltulose, mango powder and auxiliary material are added V-type blending tank Homogeneous phase mixing 25-35min, speed of agitator 20-40r/min, then residue modified dietary fiber Homogeneous phase mixing 12-18min is added, speed of agitator 40-60r/min, namely sterile filling, sealing, packaging obtain composite melissa powder;

The broker wall bee pollen sporoderm-broken rate adopting said method to prepare can reach more than 98%, and in composite melissa powder, Lactobacillus plantarum viable count can reach 6 × 10 ¹¹-9 × 10 ¹¹cfu/g.

Lactobacillus plantarum of the present invention (Lactobacillusplantarum) XH is preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center (being called for short CGMCC) on November 30th, 2015, preserving number is CGMCCNO.11763, preservation address is: No. 3, No. 1, North Star West Road, Chaoyang District, city of BeiJing, China institute, Institute of Microorganism, Academia Sinica, postcode: 100101.

Lactobacillus plantarum probiotic properties is as follows:

Lactobacillus plantarum CGMCCNO.11763 provided by the present invention finds to survive under pH is the condition of 1.50 through experiment, after 1% cholate cultivates 4 hours, be still in existing state; Lactobacillus plantarum CGMCCNO.11763 degrading nitrite speed is fast, and capacity of decomposition reaches 10.9mg/h/kg, and this bacterial classification is when producing pickles, and whole sweat nitrite concentration is at below 4.8mg/kg; CGMCCNO.11763, after fermentation 60h hour, can reach 64.76% to degrading rate of cholesterol.CGMCCNO.11763 Adhering capacity measure from aggegation rate be 95.71%.

Lactobacillus plantarum CGMCCNO.11763 is to cholesterol degradation capability study and mensuration:

Get MRS cholesterol fluid nutrient medium (the cholesterol level 0.1mg/ml that 1mlCGMCCNO.11763 mother liquor is inoculated in 10mL, pH6.2) in, the constant temperature of 37 DEG C leaves standstill cultivates 20h respectively, 40h, 60h is for subsequent use, to access the MRS cholesterol culture medium of 1mL sterilized water for contrast, get bacterium liquid sample and each 1ml of contrast liquid of above cultivation different time, 9000r/min, centrifugal 10min at 4 DEG C, obtain fermented supernatant fluid, o-phthalaldehyde method measures cholesterol level in supernatant and (is specially: get each supernatant 0.1ml in corresponding test tube, add glacial acetic acid 0.3ml, the OPA 0.15ml of 1mg/ml, slowly add concentrated sulfuric acid 1.0ml, mix.Room temperature leaves standstill 10min, under 550nm, survey light absorption value).Each processes 3 repetitions, in kind makes cholesterol calibration curve, calculates cholesterol level and degradation rate in supernatant, the results are shown in Table 1.Known, CGMCCNO.11763 has good degradation to cholesterol, and after fermentation 60h hour, degradation rate can reach 64.76%.

The degraded situation of table 1 pair cholesterol.

Degradation time (h)	0	20h	40h	60h
					Cholesterol level (mg/ml)	0.2273±0.0058	0.1356±0.0018	0.1011±0.0094	0.801±0.0231
Degrading rate of cholesterol %		40.34％	55.52％	64.76％

The bile tolerance test of Lactobacillus plantarum CGMCCNO.11763 bacterial strain:

Get CGMCCNO.11763 bacterium liquid 1mL and inoculate bacterial classification in the 10mLMRS fluid nutrient medium (PH=6.4) containing different cholate (concentration gradients is 0.0%, 0.2%, 0.4%, 0.6%, 0.8%, 1%), cultivate 0 at being placed in 37 DEG C respectively, 2,4h, each process 3 repetition.Respectively get 1ml sample bacterium liquid to mix in 9ml physiological saline, prepare dilution factor solution, get 0.1ml dilution to be coated with in MRS, (each dilution factor do 3 parallel) record of being inverted cultivation 48 hours in 37 DEG C of biochemical cultivation cases calculates the several number of bacterium on flat board.The results are shown in Table 2.The increment of this bacterium known bacterium after gallbladder salinity is 1% process 4h still

Reach 0.59 ± 0.92 × 10 ⁷(cfu/ml), good bile tolerance ability is had.

Table 2 bile tolerance ability detects [(± s) × 10 ⁷cfu/ml]

The acid resistance test of Lactobacillus plantarum CGMCCNO.11763 bacterial strain

Get HLX37 mother liquor and inoculate bacterial classification in the 10mLMRS fluid nutrient medium of different pH value (pH gradient is 1.5,2.0,2.5,3.0,3.5,4.0) by 1ml, at being placed in 37 DEG C, cultivate 0 respectively, 2,4h, each processes 3 repetitions.Respectively get 1ml sample bacterium liquid to mix in 9ml physiological saline, prepare dilute solution, get 0.1ml dilution and be coated with in MRS, the bacterium colony number on (each dilution factor do 3 parallel) record flat board of being inverted cultivation 48 hours in 37 DEG C of biochemical cultivation cases.The results are shown in Table 3.Illustrate that this bacterium has very strong acid-fast ability.

Table 3 acid-fast ability detects [(± s) × 10 ⁷cfu/ml]

The Adhering capacity of Lactobacillus plantarum CGMCCNO.11763 measures

Cultivation CGMCCNO.11763 (MRS fluid nutrient medium), bacillus coli DH 5 alpha (LB fluid nutrient medium) 24h obtain zymotic fluid, be placed in 3000r/min respectively, centrifugal 10min at 4 DEG C, collect bacterium mud, use the sterile phosphate buffer of pH=7.0 (PBS) to wash bacterium mud respectively and (namely in bacterium colony, add PBS 2 times, after concussion mixes, be placed in 3000r/min, centrifugal 10min at 4 DEG C, collect thalline).From aggegation rate (%): the suspension bacteria liquid and the bacteria suspension that with the light absorption value that bacterium mud CGMCCNO.11763 to be formed in wavelength 600nm place by aseptic PBS are 0.4 ± 0.1 (A0), measuring light absorption value A24 after leaving standstill 24h, is (A0-A24)/A0 from aggegation rate (%) formula.; His aggegation rate (%): being adjusted to the outstanding bacterium liquid of CGMCCNO.11763 and bacillus coli DH 5 alpha at the light absorption value at wavelength 600nm place is the mix suspending bacterium liquid of 0.6 ± 0.1 (A0).Measure light absorption value A24 after leaving standstill 24H, his aggegation rate (%) formula is (A0-A24)/A0.Measurement result is in table 5, and known CGMCCNO.11763's is 95.71% from aggegation rate, has very strong Adhering capacity.

Table 4 Adhering capacity table

The bacterial strain physiological property of Lactobacillus plantarum CGMCCNO.11763

Described Lactobacillus plantarum (Lactobacillusplantarum) XH is preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center (being called for short CGMCC) on November 30th, 2015, preserving number is CGMCCNO.11763, preservation address is: No. 3, No. 1, North Star West Road, Chaoyang District, city of BeiJing, China institute, Institute of Microorganism, Academia Sinica, postcode: 100101.

This bacterial 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 (+).Be accredited as Lactobacillus plantarum (Lactobacillusplantarum) through Physiology and biochemistry, called after Lactobacillus plantarum (Lactobacillusplantarum) XH.

Bacterial strain can at 57 DEG C well-grown, glucose tolerance is 275g/L.

Lactobacillus plantarum of the present invention, by gathering people Li Jianshu, is separated in Yoghourt and obtains, acquisition time on June 2nd, 2015 from Xinjiang Uygur fellow-villager family.

Lactobacillus plantarum CGMCCNO.11763 of the present invention finds to survive under pH is the condition of 1.50 through experiment, after 1% cholate cultivates 4 hours, be still in existing state; Lactobacillus plantarum CGMCCNO.11763 degrading nitrite speed is fast, and capacity of decomposition reaches 10.9mg/h/kg, and this bacterial classification is when producing pickles, and whole sweat nitrite concentration is at below 4.8mg/kg; CGMCCNO.11763, after fermentation 60h hour, can reach 64.76% to degrading rate of cholesterol.CGMCCNO.11763 Adhering capacity measure from aggegation rate be 95.71%, bacterial strain can at 57 DEG C well-grown, glucose tolerance is 275g/L.

Beneficial effect:

The present invention take Bee Pollen as primary raw material, enzyme activity is high, the natural plant enzyme that enzyme system is abundant and the most original containing Bee Pollen forming process, the pupa worm enteron aisle extract science of the queen bee nit that enzyme system is the most complete and drone pupae saliva and enteron aisle enzyme system is composite, and adopt the instantaneous puffing technique of microwave high-temperature and probiotics fermention technology, by Specialty bio enzymolysis, the instantaneous low temperature expanding of microwave high-temperature and probiotics fermention carry out fast to Bee Pollen, effectively and degree of depth broken wall, obtained broker wall bee pollen, then composite with one or more functional auxiliary material science, obtained one keeps the natural flavour mountaineous and nutritional labeling of Bee Pollen to greatest extent, sporoderm-broken rate is high, efficiency is high, cost is low, functional health care composite melissa powder by force.Concrete test effect is shown in embodiment seven to ten two, and concrete component know-why is as follows:

1. the broker wall bee pollen that prepared by the present invention take Bee Pollen as raw material, first adopts ultrasonic cleaning to carry out desinsection to Bee Pollen and to go out ovum, killing microorganisms, removal residues of pesticides and heavy metal ion etc., substantially increase the foodsafety of composite melissa powder; Soak in containing the aqueous solution of sericin peptide taken, can reduce to greatest extent through the freezing and loss of bee pollen activity material that is that cause, improve the recovery rate of Bee Pollen active ingredient, also established solid foundation for follow-up alternating temperature probiotics fermention simultaneously; Adopt microwave high-temperature instantaneous expanded make through freezing Bee Pollen appropriateness expanded, enhance between the inside and outside wall of Bee Pollen and inclusion with being separated of the own structure of matter, fluffy, facilitate the formation of netted result, enhance the permeability of inside and outside mass exchange, follow-up enzyme molecule, lactic acid bacteria infiltrate into inside and play a role, to reach the maximization accumulation of Bee Pollen active ingredient; The low temperature extractive techniques such as high-pressure pulse electric, ultrasonic, biological enzymolysis are organically combined, functional materials and the active ingredient of Bee Pollen can be retained to greatest extent; The animal ferment source science that the phytoenzyme enrich, enzyme system high containing multiple enzyme activity and Bee Pollen are formed is composite, can effectively to degrade various macromolecular substances in Bee Pollen, its abundant enzyme system is that commercially available complex enzyme is incomparable, more meet the original natural rule of " take from nature, be used for nature " of the formation of Bee Pollen material and degraded; With functional plants lactobacillus powder for leavening, adopt temperature-variable fermentation and gradation vaccination ways can obtain the maximum accumulation of proliferation of probiotics and functional metabolic product to greatest extent while carrying out further degree of depth broken wall to Bee Pollen.The broker wall bee pollen sporoderm-broken rate adopting said method to prepare can reach more than 98%.

2. the queen bee nit enteron aisle of the composite 2-3 age in days of pupa worm enteron aisle extract science prepared of the present invention and the drone pupae enteron aisle of 11-12 age in days are raw material, soak in containing the aqueous solution of sericin peptide taken, can reduce to greatest extent through loss that is freezing and the pupa worm active material caused, improve the recovery rate of pupa worm active ingredient; Use biological demulsifying agent breakdown of emulsion, can promote the miscible materials such as the fat in pupa worm enteron aisle with containing being separated of hydrolyzate of enriching enteron aisle enzyme, the maximization that can obtain enteron aisle enzyme is extracted, relative to existing direct centrifugation or add the separation method such as buffer solution, biochemicals, enteron aisle enzyme recovery rate is high, technique is simple, foodsafety is high, environmentally friendly.Meanwhile, pupa worm enteron aisle extract, through follow-up enzymolysis, also contains queen bee nit polypeptide and drone pupae polypeptide, for composite melissa powder adds more functional bioactive material.

3. Lactobacillus plantarum CGMCCNO.11763 of the present invention finds to survive under pH is the condition of 1.50 through experiment, after 1% cholate cultivates 4 hours, be still in existing state; Lactobacillus plantarum CGMCCNO.11763 degrading nitrite speed is fast, and capacity of decomposition reaches 10.9mg/h/kg, and this bacterial classification is when producing pickles, and whole sweat nitrite concentration is at below 4.8mg/kg; CGMCCNO.11763, after fermentation 60h hour, can reach 64.76% to degrading rate of cholesterol.CGMCCNO.11763 Adhering capacity measure from aggegation rate be 95.71%.

4. the cryoprotector that adopts when preparing Lactobacillus plantarum pulvis of the present invention is by composite containing the higher winter rye of antifreeze protein, Ammopiptanthus mongolicus and sharkskin collagen science; obtain through high-pressure pulse electric extraction, ultrasonic assistant Microwave Extraction and biological enzymolysis; omnidistance low temperature extracts; antifreeze protein recovery rate is high; loss is few; the protective agent antifreeze peptide content obtained is high, kind is complete, functional strong, cryoprotective effects is good, improves the viable bacteria content in Lactobacillus plantarum pulvis.

5. not only contain the abundant various plants enzyme such as plant rennet, amylase, hemicellulase, esterase, oxidoreducing enzyme but also contain the nutriment such as plant polyose and monose, plant amylum, vegetable protein, Soluble Fiber in the plant extracts that prepared by the present invention, not only can be the phytoenzyme that composite melissa powder provides comprehensive, natural, also can be the nutriment that Lactobacillus plantarum provides comprehensively, enriches; After the low temperature such as ultrasonic cleaning, the ultrasonic extraction of microwave radiation technology and high-pressure pulse electric extraction, reduced pressure concentration extract, its effect is more remarkable, and effectively improves raw material availability, phytoenzyme activity and productive rate; Effectively ensure that the foodsafety of plant extracts.

6. ultrasonic extraction, high-pressure pulse electric extract and biological enzymolysis combination of sciences by the modified dietary fiber that prepared by the present invention, the stronger and impact of acid and alkali, alkali, salt of gained modified dietary fiber retentiveness, dilatancy, thickening property, Soluble Fiber cellulose content is high, more easily utilized by lactic acid bacteria, improve lactic acid bacteria in the growth of human body intestinal canal and fertility, increase kind and the quantity of probio flora, reduce human body intestinal canal pH value, improve human body intestinal canal micro-ecological environment; 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; Regulate and maintain the resident time of gut flora, strengthen the absorption and digestion ability of enteron aisle, improve immunity of organisms; Effective promotion gastrointestinal peristalsis, slows down and eliminates the bad reaction such as gasteremphraxis, abdominal distension; Powerful bury function can prevent environment (oxygen, temperature, illumination, water activity etc.) factor from, on the impact of product quality, stabilizing the biologically active of composite melissa powder, extending the shelf-life of composite melissa powder.

7. the present invention by compound sugar and modified dietary fiber science composite, the nutriment providing comprehensively to probio, enrich, not only achieve the maximization accumulation of the maximization proliferation and function metabolite of probio, and effectively improve mouthfeel and the stability of finished product composite melissa powder.

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

It should be noted that the technique effect of composite melissa powder of the present invention is each component technical characteristic and method characteristic collaborative, interactional result mutually, the not superposition of simple technical characteristic (component function), the combination of each component technical characteristic and the collaborative effect produced, considerably beyond the superposition of each monotechnics feature functionality and effect, have advanced and practicality preferably.

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.

Embodiment one:

Prepared by raw material

1, the preparation of plant extracts

Its preparation method is:

By fructus hordei germinatus and wheat malt 9:2 Homogeneous phase mixing in mass ratio, be crushed to granularity 0.8mm, obtain pulverizing Fructus Hordei Germinatus, then by pawpaw, pineapple, fig respectively in supersonic wave cleaning machine at power 200W, ultrasonic cleaning 8min under frequency 30KHz condition, drain, granularity 0.8mm is crushed under room temperature, and 8:2:1.5 Homogeneous phase mixing in mass ratio, the pulverizing Fructus Hordei Germinatus adding mixture quality 4 times obtains raw mixture, add the water of raw mixture quality 2 times, be 3.5 by citric acid adjust ph, at power 225W, Microwave Extraction is carried out under frequency 2000Hz condition, wherein, each microwave irradiation total time 70s, carry out compartment irradiation: irradiation 10s, interval 10s, control temperature 30 DEG C, irradiation like this 10 times, simultaneously at power 250W, ultrasonic assistant extraction is carried out under frequency 35KHz condition, insulation 2h, then, carries out Microwave Extraction under power 300W, frequency 2000Hz condition, wherein, each microwave irradiation total time 100s, carries out compartment irradiation: irradiation 15s, interval 10s, control temperature 50 DEG C, irradiation like this 10 times, simultaneously at power 400W, carry out ultrasonic assistant extraction under frequency 45KHz condition, last Temperature fall to room temperature, in electric-field intensity 30kV/cm, burst length 500 μ s, carries out high-pressure pulse electric and extracts 18min under pulse frequency 250Hz condition, extracting liquid filtering obtains the first filtrate, adds the water rinse of filter residue 4 times, filters to obtain the second filtrate, and by the first filtrate and the second filtrate 1:4 Homogeneous phase mixing in mass ratio, being evaporated to solid content is 30%, obtains plant extracts,

In above-mentioned supersonic wave cleaning machine, cleaning fluid is the sodium bicarbonate solution of 0.4%.

2, the preparation of pupa worm enteron aisle extract

Its preparation method comprises the steps:

By the 4:2 mixing in mass ratio of the drone pupae enteron aisle of the queen bee nit enteron aisle of 3 ages in days and 12 ages in days, the mass percent adding mixture quality 1.5 times is the sericin peptide taken solution of 10%, stir, put-20 DEG C of freezing 8min, broken, grind to form homogenate, add the biological demulsifying agent breakdown of emulsion 30min of its quality 0.04%, at 22000g, centrifugal 25min under 4 DEG C of conditions, the middle level liquid after collection first time is centrifugal, then at 22000g, centrifugal 25min under 4 DEG C of conditions, the middle level liquid after collection second time is centrifugal, obtains pupa worm enteron aisle extract;

The pH value of above-mentioned sericin peptide taken solution is 6.5;

The quality group of above-mentioned biological demulsifying agent becomes: glycolipid class: lipopeptid class: cell wall is in conjunction with class=6:5:3;

Wherein the quality group of glycolipid class biological demulsifying agent becomes: rhamnolipid: APG=4:1.5.

3, the preparation of modified dietary fiber

The preparation method of modified dietary fiber, comprises the following steps:

By inulin, Semen Tritici aestivi fiber, common oats fibre 6:3:2 Homogeneous phase mixing in mass ratio, add the water of its quality 5 times, room temperature 200W, 38KHz condition ultrasonic extraction 13min, then at electric-field intensity 30kV/cm, burst length 400 μ _s, under pulse frequency 300Hz condition, carry out high-pressure pulse electric process 13min; Be 5.5 by lactic acid adjust ph, add the biology enzyme of mixture quality 0.2%, in 50 DEG C of enzymolysis 34min; Enzymolysis liquid filters, and filtrate reduced in volume, freeze drying to moisture are 6.5% namely obtain modified dietary fiber;

Above-mentioned biology enzyme is cellulase, zytase, laccase, pectase 2:2:1.5:1.5 Homogeneous phase mixing in mass ratio.

4, the preparation of Lactobacillus plantarum powder

Lactobacillus plantarum powder is with Lactobacillus plantarum CGMCCNO.11763 for starting strain is prepared according to a conventional method, and its viable bacteria content is: 9 × 10 ¹²cfu/g; Wherein freeze drying process adopt cryoprotector with the animal or plant containing antifreeze protein for raw material, obtained through high-pressure pulse electric extraction, ultrasonic assistant Microwave Extraction and compound enzyme enzymolysis, effectively can improve Lactobacillus plantarum powder at freezing dry process Viable detection;

Protectant preparation method, comprises the steps:

Winter rye, Ammopiptanthus mongolicus, sharkskin collagen are cleaned respectively, drain, 9:4:3 Homogeneous phase mixing in mass ratio, the pH value adding mixed material quality 0.5 times be 4.2 lactic acid soak 5h, pulverize immediately after-20 DEG C of freezing 1.5h, freezing thickness of feed layer 4cm, crushed material particle diameter 2mm, then the water of crushed material quality 15 times is added, be 4.5 by lactic acid adjust ph, at electric-field intensity 30kV/cm under room temperature, burst length 400 μ s, carries out high-pressure pulse electric process 25min under pulse frequency 250Hz condition; Then under power 225W condition, carry out microwave irradiation in room temperature and extract 18min, under power 250W, frequency 35KHz condition, carry out ultrasonic assistant extraction simultaneously; Add the compound enzyme of extract quality 1.5%, in 50 DEG C of enzymolysis 40min; Enzymolysis liquid filters, filtrate is concentrated, low-temperature grinding to particle diameter is that namely 0.2mm obtains protective agent;

Above-mentioned compound enzyme is cellulase, protease, amylase, pectase, tannase 3:2:2:1.5:1.5 Homogeneous phase mixing in mass ratio.

Following examples two to six plant extracts used, pupa worm enteron aisle extract, modified dietary fiber, Lactobacillus plantarum powder are prepared by embodiment one.

Embodiment two:

A kind of haematococcus pluvialis pollen, mainly contains the raw material preparation of following parts by weight:

Bee Pollen 115 parts, plant extracts 20 parts, pupa worm enteron aisle extract 15 parts, modified dietary fiber 15 parts, compound sugar 11 parts, 8 parts, Lactobacillus plantarum powder, hydroxyl isomaltulose 4 parts, mango powder 4 parts, haematococcus pluvialis 0.9 part;

Wherein Bee Pollen is sesame Bee Pollen;

A preparation method for haematococcus pluvialis pollen, comprises the steps:

1) each supplementary material is taken according to formula;

2) Bee Pollen is put in the supersonic wave cleaning machine that 0.4% sodium bicarbonate solution is housed, in 200W under room temperature, 40KHz cleans 4min, rinsing, drain, be soak 6min in the sericin peptide taken solution of 16% in mass percent, take out, put-20 DEG C of freezing 8min, broken, immediately at power 4Kw, frequency 2450MHz, temperature 130 DEG C of heating using microwave 6s, add the water of former Bee Pollen quality 4 times, be 5 by lactic acid adjust ph, in electric-field intensity 30kV/cm, burst length 500 μ s, high-pressure pulse electric process 13min is carried out under pulse frequency 300Hz room temperature condition, then room temperature 400W, 40KHz condition ultrasonic process 13min obtains Bee Pollen pretreatment fluid,

3) Bee Pollen pretreatment fluid is warming up to 45 DEG C, adds plant extracts and pupa worm enteron aisle extract wherein, stir, insulation, enzymolysis 13min, obtains Bee Pollen enzymolysis liquid;

4) add in Bee Pollen enzymolysis liquid compound sugar, 25% modified dietary fiber, abundant dissolving 8min, Temperature fall to 43 DEG C, add 55% ferment at constant temperature 4h of Lactobacillus plantarum opaque amount, then with the ramp to 53 DEG C of 0.9 DEG C/min, add 45% ferment at constant temperature 2.5h of Lactobacillus plantarum opaque amount, finally be cooled to 43 DEG C with the speed of 0.7 DEG C/min, continue fermentation 0.8h, fermentation liquor 200 eye mesh screen filters, and namely filtrate obtain broker wall bee pollen through ultrafiltration, reduced pressure concentration, freeze drying;

5) broker wall bee pollen and hydroxyl isomaltulose, mango powder and haematococcus pluvialis are added V-type blending tank Homogeneous phase mixing 30min, speed of agitator 30r/min, then residue modified dietary fiber Homogeneous phase mixing 15min is added, speed of agitator 50r/min, namely sterile filling, sealing, packaging obtain haematococcus pluvialis pollen;

The broker wall bee pollen sporoderm-broken rate adopting said method to prepare can reach 99%, and in haematococcus pluvialis pollen, Lactobacillus plantarum viable count can reach 7.5 × 10 ¹¹cfu/g.

Embodiment three:

A kind of fish oil pollen, mainly contains the raw material preparation of following parts by weight:

Bee Pollen 110 parts, plant extracts 15 parts, pupa worm enteron aisle extract 13 parts, modified dietary fiber 13 parts, compound sugar 9 parts, 7 parts, Lactobacillus plantarum powder, hydroxyl isomaltulose 3 parts, mango powder 3 parts, 0.5 part, fish oil;

Wherein Bee Pollen is bee pollen of maize;

A preparation method for fish oil pollen, comprises the steps:

1) each supplementary material is taken according to formula;

2) Bee Pollen is put in the supersonic wave cleaning machine that 0.3% sodium bicarbonate solution is housed, in 200W under room temperature, 40KHz cleans 3min, rinsing, drain, be soak 5min in the sericin peptide taken solution of 13% in mass percent, take out, put-18 DEG C of freezing 5min, broken, immediately at power 3Kw, frequency 2450MHz, temperature 120 DEG C of heating using microwave 5s, add the water of former Bee Pollen quality 3 times, be 4 by lactic acid adjust ph, in electric-field intensity 20kV/cm, burst length 400 μ s, high-pressure pulse electric process 10min is carried out under pulse frequency 200Hz room temperature condition, then room temperature 400W, 40KHz condition ultrasonic process 10min obtains Bee Pollen pretreatment fluid,

3) Bee Pollen pretreatment fluid is warming up to 40 DEG C, adds plant extracts and pupa worm enteron aisle extract wherein, stir, insulation, enzymolysis 10min, obtains Bee Pollen enzymolysis liquid;

4) add in Bee Pollen enzymolysis liquid compound sugar, 20% modified dietary fiber, abundant dissolving 5min, Temperature fall to 40 DEG C, add 50% ferment at constant temperature 3h of Lactobacillus plantarum opaque amount, then with the ramp to 50 DEG C of 0.8 DEG C/min, add 40% ferment at constant temperature 1.5h of Lactobacillus plantarum opaque amount, finally be cooled to 40 DEG C with the speed of 0.6 DEG C/min, continue fermentation 0.5h, fermentation liquor 100 eye mesh screen filters, and namely filtrate obtain broker wall bee pollen through ultrafiltration, reduced pressure concentration, freeze drying;

5) broker wall bee pollen and hydroxyl isomaltulose, mango powder and fish oil are added V-type blending tank Homogeneous phase mixing 25min, speed of agitator 20r/min, then add residue modified dietary fiber Homogeneous phase mixing 12min, speed of agitator 40r/min, namely sterile filling, sealing, packaging obtain fish oil pollen;

The broker wall bee pollen sporoderm-broken rate adopting said method to prepare can reach 98.5%, and in fish oil pollen, Lactobacillus plantarum viable count can reach 8.5 × 10 ¹¹cfu/g.

Embodiment four:

A kind of agate coffee pollen, mainly contains the raw material preparation of following parts by weight:

Bee Pollen 120 parts, plant extracts 25 parts, pupa worm enteron aisle extract 17 parts, modified dietary fiber 17 parts, compound sugar 13 parts, 9 parts, Lactobacillus plantarum powder, hydroxyl isomaltulose 5 parts, mango powder 5 parts, pueraria root powder 0.8 part;

Wherein Bee Pollen is bee pollen form cole;

A preparation method for agate coffee pollen, comprises the steps:

1) each supplementary material is taken according to formula;

2) Bee Pollen is put in the supersonic wave cleaning machine that 0.5% sodium bicarbonate solution is housed, in 200W under room temperature, 40KHz cleans 5min, rinsing, drain, be soak 7min in the sericin peptide taken solution of 18% in mass percent, take out, put-22 DEG C of freezing 10min, broken, immediately at power 5Kw, frequency 2450MHz, temperature 140 DEG C of heating using microwave 7s, add the water of former Bee Pollen quality 5 times, be 6 by lactic acid adjust ph, in electric-field intensity 40kV/cm, burst length 600 μ s, high-pressure pulse electric process 15min is carried out under pulse frequency 400Hz room temperature condition, then room temperature 400W, 40KHz condition ultrasonic process 15min obtains Bee Pollen pretreatment fluid,

3) Bee Pollen pretreatment fluid is warming up to 50 DEG C, adds plant extracts and pupa worm enteron aisle extract wherein, stir, insulation, enzymolysis 15min, obtains Bee Pollen enzymolysis liquid;

4) add in Bee Pollen enzymolysis liquid compound sugar, 30% modified dietary fiber, abundant dissolving 10min, Temperature fall to 45 DEG C, add 60% ferment at constant temperature 5h of Lactobacillus plantarum opaque amount, then with the ramp to 55 DEG C of 1.0 DEG C/min, add 50% ferment at constant temperature 3.5h of Lactobacillus plantarum opaque amount, finally be cooled to 45 DEG C with the speed of 0.8 DEG C/min, continue fermentation 1h, fermentation liquor 300 eye mesh screen filters, and namely filtrate obtain broker wall bee pollen through ultrafiltration, reduced pressure concentration, freeze drying;

5) broker wall bee pollen and hydroxyl isomaltulose, mango powder and pueraria root powder are added V-type blending tank Homogeneous phase mixing 35min, speed of agitator 40r/min, then add residue modified dietary fiber Homogeneous phase mixing 18min, speed of agitator 60r/min, namely sterile filling, sealing, packaging obtain agate coffee pollen;

The broker wall bee pollen sporoderm-broken rate adopting said method to prepare can reach 99.5%, and in agate coffee pollen, Lactobacillus plantarum viable count can reach 6 × 10 ¹¹cfu/g.

Embodiment five:

One plant sterols pollen, mainly contains the raw material preparation of following parts by weight:

Bee Pollen 100 parts, plant extracts 10 parts, pupa worm enteron aisle extract 10 parts, modified dietary fiber 10 parts, compound sugar 7 parts, 6 parts, Lactobacillus plantarum powder, hydroxyl isomaltulose 2 parts, mango powder 2 parts, phytosterol 0.5 part;

Wherein Bee Pollen is bee pollen of maize;

The preparation method of one plant sterols pollen, comprises the steps:

1) each supplementary material is taken according to formula;

2) Bee Pollen is put in the supersonic wave cleaning machine that 0.3% sodium bicarbonate solution is housed, in 200W under room temperature, 40KHz cleans 5min, rinsing, drain, be soak 7min in the sericin peptide taken solution of 13% in mass percent, take out, put-18 DEG C of freezing 10min, broken, immediately at power 3Kw, frequency 2450MHz, temperature 140 DEG C of heating using microwave 5s, add the water of former Bee Pollen quality 5 times, be 4 by lactic acid adjust ph, in electric-field intensity 40kV/cm, burst length 400 μ s, high-pressure pulse electric process 10min is carried out under pulse frequency 400Hz room temperature condition, then room temperature 400W, 40KHz condition ultrasonic process 15min obtains Bee Pollen pretreatment fluid,

3) Bee Pollen pretreatment fluid is warming up to 40 DEG C, adds plant extracts and pupa worm enteron aisle extract wherein, stir, insulation, enzymolysis 15min, obtains Bee Pollen enzymolysis liquid;

4) add in Bee Pollen enzymolysis liquid compound sugar, 20% modified dietary fiber, abundant dissolving 10min, Temperature fall to 40 DEG C, add 60% ferment at constant temperature 3h of Lactobacillus plantarum opaque amount, then with the ramp to 50 DEG C of 1.0 DEG C/min, add 50% ferment at constant temperature 1.5h of Lactobacillus plantarum opaque amount, finally be cooled to 40 DEG C with the speed of 0.8 DEG C/min, continue fermentation 1h, fermentation liquor 100 eye mesh screen filters, and namely filtrate obtain broker wall bee pollen through ultrafiltration, reduced pressure concentration, freeze drying;

5) broker wall bee pollen and hydroxyl isomaltulose, mango powder and phytosterol are added V-type blending tank Homogeneous phase mixing 35min, speed of agitator 20r/min, then residue modified dietary fiber Homogeneous phase mixing 18min is added, speed of agitator 40r/min, namely sterile filling, sealing, packaging obtain phytosterol pollen;

The broker wall bee pollen sporoderm-broken rate adopting said method to prepare can reach 99%, and in phytosterol pollen, Lactobacillus plantarum viable count can reach 9 × 10 ¹¹cfu/g.

Embodiment six:

A kind of Cordyceps militaris pollen, mainly contains the raw material preparation of following parts by weight:

Bee Pollen 130 parts, plant extracts 30 parts, pupa worm enteron aisle extract 20 parts, modified dietary fiber 20 parts, compound sugar 15 parts, 10 parts, Lactobacillus plantarum powder, hydroxyl isomaltulose 6 parts, mango powder 6 parts, Cordyceps militaris 0.3 part;

Wherein Bee Pollen is bee pollen form cole;

A preparation method for Cordyceps militaris pollen, comprises the steps:

1) each supplementary material is taken according to formula;

2) Bee Pollen is put in the supersonic wave cleaning machine that 0.5% sodium bicarbonate solution is housed, in 200W under room temperature, 40KHz cleans 3min, rinsing, drain, be soak 5min in the sericin peptide taken solution of 18% in mass percent, take out, put-22 DEG C of freezing 5min, broken, immediately at power 5Kw, frequency 2450MHz, temperature 120 DEG C of heating using microwave 7s, add the water of former Bee Pollen quality 3 times, be 6 by lactic acid adjust ph, in electric-field intensity 20kV/cm, burst length 600 μ s, high-pressure pulse electric process 15min is carried out under pulse frequency 200Hz room temperature condition, then room temperature 400W, 40KHz condition ultrasonic process 10min obtains Bee Pollen pretreatment fluid,

3) Bee Pollen pretreatment fluid is warming up to 50 DEG C, adds plant extracts and pupa worm enteron aisle extract wherein, stir, insulation, enzymolysis 10min, obtains Bee Pollen enzymolysis liquid;

4) add in Bee Pollen enzymolysis liquid compound sugar, 30% modified dietary fiber, abundant dissolving 5min, Temperature fall to 45 DEG C, add 50% ferment at constant temperature 5h of Lactobacillus plantarum opaque amount, then with the ramp to 55 DEG C of 0.8 DEG C/min, add 40% ferment at constant temperature 3.5h of Lactobacillus plantarum opaque amount, finally be cooled to 45 DEG C with the speed of 0.6 DEG C/min, continue fermentation 0.5h, fermentation liquor 300 eye mesh screen filters, and namely filtrate obtain broker wall bee pollen through ultrafiltration, reduced pressure concentration, freeze drying;

5) broker wall bee pollen and hydroxyl isomaltulose, mango powder and Cordyceps militaris are added V-type blending tank Homogeneous phase mixing 25min, speed of agitator 40r/min, then add residue modified dietary fiber Homogeneous phase mixing 12min, speed of agitator 60r/min, namely sterile filling, sealing, packaging obtain Cordyceps militaris pollen;

The broker wall bee pollen sporoderm-broken rate adopting said method to prepare can reach 99.5%, and in Cordyceps militaris pollen, Lactobacillus plantarum viable count can reach 8 × 10 ¹¹cfu/g.

Experimental example seven eats the change of T-CHOL after haematococcus pluvialis pollen

Select the adult 48 of T-CHOL 180mg/dl-250mg/dl, men and women half and half, is divided into three groups at random; 150 milliliters of mineral waters are drunk in first group of dinner every day; Second group of common commercially available haematococcus pluvialis pollen 150g of dinner every day Instant Drinks, the haematococcus pluvialis pollen 150g of the 3rd group of dinner every day Instant Drinks embodiment of the present invention 2, every day period eats same food, and food comprises meat, egg, vegetables and fruit.Respectively at the previous day of experiment beginning and the blood of the 20th, 40,60 day acquisition test person, measure the total cholesterol level in blood, result is as table 1:

Table 1: total cholesterol level testing result in blood

After the haematococcus pluvialis pollen of the Instant Drinks embodiment of the present invention 2 as seen from the above table, the content generation significant change of the T-CHOL in adult's blood.Compared with common commercially available haematococcus pluvialis pollen, the content of haematococcus pluvialis pollen of the present invention to the T-CHOL in adult's blood significantly reduces, and the content of the T-CHOL that mineral water forms in year human blood significantly increases, although commercially available haematococcus pluvialis pollen decreases, but compared with product of the present invention, effect is remarkable, it can thus be appreciated that the haematococcus pluvialis pollen that the present invention adopts the specific bacterial strain with reduction cholesterol characteristic to prepare has the health-care effect well reducing cholesterol.

It should be noted that: the composite melissa powder prepared by embodiment of the present invention 3-6 has above-mentioned technique effect equally, between each embodiment, otherness is not remarkable.

Embodiment eight haematococcus pluvialis pollen of the present invention shelf-life implants living preparation of lactobacillus stable content is tested

Get haematococcus pluvialis pollen prepared by the embodiment of the present invention 2 in room temperature 22-25 DEG C, store 3 months, 6 months, 12 months, 24 months, 36 months respectively under ventilation condition and measure Lactobacillus plantarum viable bacteria content, result is as table 2:

Table 2: shelf-life implants living preparation of lactobacillus content detection result

Above result shows: the storage stability of haematococcus pluvialis pollen of the present invention is better, environment resistant (temperature, appropriateness, oxygen, illumination, moisture etc.) influence factor ability is large, shelf-life Lactobacillus plantarum viable bacteria content loss late maximum (36 months) is 15%, higher than existing like product viable bacteria content, loss late is low, long shelf-life, reflects that other bioactive ingredients of haematococcus pluvialis pollen has same storage stability simultaneously.

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

The sensory evaluation test of embodiment nine haematococcus pluvialis pollen of the present invention

24 personnel are invited to judge with the haematococcus pluvialis pollen of commercially available two kinds of similar identical dates of manufacture haematococcus pluvialis pollen prepared by the embodiment of the present invention 2, 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 3:

Table 3: 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 commercially available haematococcus pluvialis pollen to haematococcus pluvialis pollen prepared by the present invention from outward appearance, quality, local flavor and mouthfeel, particularly outward appearance, local flavor and mouthfeel are fabulous, and also the consumer of applicable Different age group, different hierarchy of consumption eats simultaneously.

It should be noted that: composite melissa powder 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.

The probiotic test of embodiment ten haematococcus pluvialis of the present invention pollen

The haematococcus pluvialis pollen embodiment of the present invention 2 prepared, being prepared into viable count with sterilized water is 2 × 10 ¹⁰the Lactobacillus plantarum solution of CFU/mL, saves backup in 4 DEG C;

1, the 10mL Lactobacillus plantarum 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 Lactobacillus plantarum solution is housed is placed in 80-90 DEG C of water-bath and heats 15-25min, get the Lactobacillus plantarum 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 more than 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 Lactobacillus plantarum 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 98%.Then each 1mL of the nutrient solution digesting different time is taken in simulated gastric fluid, being inoculated in 9mLpH 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 Lactobacillus plantarum solution inoculum kept by 0.5mL is simulated in cholate to 4.5mL, 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, Lactobacillus plantarum in haematococcus pluvialis pollen of the present invention probiotic (heat resistance, resistance to pH, bile tolerance) is stronger, very be applicable to human intestines and stomach's environment, in simulated gastrointestinal environments, survival rate is large, effectively can improve gastrointestinal function.

It should be noted that: composite melissa powder 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 11 haematococcus pluvialis pollen of the present invention mouse intestinal performance test

The haematococcus pluvialis pollen embodiment of the present invention 2 prepared, being prepared into viable count with sterilized water is 2 × 10 ¹⁰the Lactobacillus plantarum solution of CFU/mL, saves backup in 4 DEG C;

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 Lactobacillus plantarum 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 haematococcus pluvialis pollen treatment group and natural recovering group, calculate each group of weight average growth rate, result is as table 4; Within every 5 days, survey each group of stool in mice Escherichia coli quantity, calculate average, result is as table 5.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 4: 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	34.28±1.62	67.96 b

Table 5: the situation of coliform count in stool in mice

Haematococcus pluvialis pollen treatment group Mouse Weight average growth rate (67.96%) is significantly higher than natural recovering group (32.14%); Solution rear intestinal Escherichia coli quantity of feeding significantly declines, reduce by 97.07%, significantly lower than natural recovering group (24.78%), show the Lactobacillus plantarum rapid field planting in small white mouse enteron aisle in haematococcus pluvialis pollen 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.

It should be noted that: composite melissa powder 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 12 composite melissa powder 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 composite melissa powder of the present invention.

2 experiment materials and reagent

2.1 for reagent thing:

Commercially available composite melissa powder (G1); Commercially available composite melissa powder (G2); Composite melissa powder (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 Ningxia Medical University's comparative medicine center, 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 Affiliated Hospital of Ningxia Medical University 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 composite melissa powders of the present invention are on the impact of Mouse Weight

Each group of mouse is after giving G1 ~ G9 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 ~ G9 medicine all without obvious toxicity.Experimental result refers to table 6.

The original body mass of table 6 swimming with a load attached to the body experiment mice, mid-term body weight and terminate body weight

9.2 composite melissa powders 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), composite melissa powder 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 7.

Table 7 composite melissa powder is on the impact of mice burden swimming time

" * " p<0.05vs blank;

" * * " p<0.01vs blank;

9.3 composite melissa powders of the present invention are on the impact of blood lactase acid before and after mouse movement

After per os gives mouse composite melissa powder of the present invention, composite melissa powder 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 8.

Table 8 composite melissa powder of the present invention is on the impact of blood lactase acid level before and after mouse movement

" * " p<0.05vs blank;

9.4 composite melissa powders 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), composite melissa powder 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 9.

Table 9 composite melissa powder of the present invention is on the impact of Mouse Liver glycogen content

" * " p<0.05vs blank;

" * * " p<0.01vs blank;

9.5 composite melissa powders 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), composite melissa powder G3 ~ G7 medicine of the present invention compares with blank group, after mouse movement, serum urea content all has obvious reduction, has pole significant difference (P < 0.01), and is obviously better than G1 ~ G2 medicine.The results detailed in Table 10.

Table 10 composite melissa powder of the present invention is on the impact of mice serum urea content

" * " p<0.05vs blank;

" * * " p<0.01vs blank;

10. experiment conclusion

This experiment is mainly through mice burden swimming experiment, and the deposit simultaneously detecting Mouse Liver glycogen observes the effect that composite melissa powder of the present invention improves immunity, antifatigue.Preliminary Results shows below:

1, G3 ~ G7 composite melissa powder of the present invention all can extend the mice burden swimming time (P < 0.01), and successful is better than the composite melissa powder of other G1 ~ G2.

2, biochemistry detection aspect display, the lactic acid content that after each dosage group of G3 ~ G7 composite melissa powder 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 composite melissa powder 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 composite melissa powder of the present invention all can significantly improve the deposit (P < 0.01) of glycogen in mouse liver, and successful is better than the composite melissa powder of other G1 ~ G2;

4, high lithemia model finds, G3 ~ G7 composite melissa powder 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 composite melissa powder;

11. conclusions

Above-mentioned experiment proves that composite melissa powder 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 (10)

1. a composite melissa powder, mainly contain the raw material preparation of following parts by weight: Bee Pollen 100-130 part, plant extracts 10-30 part, pupa worm enteron aisle extract 10-20 part, modified dietary fiber 10-20 part, compound sugar 7-15 part, Lactobacillus plantarum powder 6-10 part, hydroxyl isomaltulose 2-6 part, mango powder 2-6 part;

Described composite melissa powder also comprises one or more auxiliary materials of following parts by weight; Algae 0.6-1.2 part, corn oligopeptide powder 0.5-1 part, pueraria root powder 0.5-1 part, fish oil 0.2-0.8 part, phytosterol 0.2-0.8 part, phosphatidyl serine 0.1-0.7 part, chitosan oligosaccharide 0.1-0.7 part, Marine fishbone collagen oligomeric Gly-His-Lys 0.1-0.7 part, Chinese herbal medicine 0.1-0.6 part, Cordyceps militaris 0.1-0.5 part; Soybean protein isolate 0.1-0.5 part;

Described Lactobacillus plantarum powder is for starting strain is prepared according to a conventional method with Lactobacillus plantarum CGMCCNO.11763.

2. composite melissa powder as claimed in claim 1, it is characterized in that, the preparation method of described pupa worm enteron aisle extract, comprise the steps: the drone pupae enteron aisle 3-5:1-3 mixing in mass ratio by the queen bee nit enteron aisle of 2-3 age in days and 11-12 age in days, add the sericin peptide taken solution that mixture quality 1-2 mass percent is doubly 8-12%, stir, put-18--22 DEG C of freezing 5-10min, broken, grind to form homogenate, add the biological demulsifying agent breakdown of emulsion 20-40min of its quality 0.03-0.05%, at 18000-25000g, centrifugal 20-30min under 4 DEG C of conditions, middle level liquid after collection first time is centrifugal, then at 18000-25000g, centrifugal 20-30min under 4 DEG C of conditions, middle level liquid after collection second time is centrifugal, obtain pupa worm enteron aisle extract.

3. composite melissa powder as claimed in claim 2, it is characterized in that, the pH value of described sericin peptide taken solution is 6-7.

4. composite melissa powder as claimed in claim 2, it is characterized in that, the quality group of described biological demulsifying agent becomes: glycolipid class: lipopeptid class: cell wall is in conjunction with class=5-7:4-6:2-4.

5. composite melissa powder as claimed in claim 4, it is characterized in that, the quality group of described glycolipid class biological demulsifying agent becomes: rhamnolipid: APG=3-5:1-2.

6. composite melissa powder as claimed in claim 1, it is characterized in that, the preparation method of described plant extracts is: by fructus hordei germinatus and wheat malt 8-10:1-3 Homogeneous phase mixing in mass ratio, be crushed to granularity 0.5-1mm, obtain pulverizing Fructus Hordei Germinatus, then by pawpaw, pineapple, fig respectively in supersonic wave cleaning machine at power 200W, ultrasonic cleaning 5-10min under frequency 30KHz condition, drain, granularity 0.5-1mm is crushed under room temperature, and 7-9:1-3:1-2 Homogeneous phase mixing in mass ratio, add mixture quality 3-5 pulverizing Fructus Hordei Germinatus doubly and obtain raw mixture, add raw mixture quality 1-3 water doubly, be 3-4 by citric acid adjust ph, at power 150-300W, Microwave Extraction is carried out under frequency 2000Hz condition, wherein, each microwave irradiation total time 60-80s, carry out compartment irradiation: irradiation 10s, interval 10s, control temperature 20-35 DEG C, irradiation like this 10 times, simultaneously at power 200-300W, ultrasonic assistant extraction is carried out under frequency 30-40KHz condition, insulation 1-3h, then, Microwave Extraction is carried out under power 200-400W, frequency 2000Hz condition, wherein, each microwave irradiation total time 90-105s, carry out compartment irradiation: irradiation 15s, interval 10s, control temperature 40-60 DEG C, irradiation like this 10 times, simultaneously at power 300-500W, carry out ultrasonic assistant extraction under frequency 40-50KHz condition, last Temperature fall to room temperature, in electric-field intensity 25-35kV/cm, burst length 400-600 μ s, carries out high-pressure pulse electric and extracts 15-20min under pulse frequency 200-300Hz condition, extracting liquid filtering obtains the first filtrate, adds the water rinse of filter residue 4 times, filters to obtain the second filtrate, and by the first filtrate and the second filtrate 1:3-5 Homogeneous phase mixing in mass ratio, being evaporated to solid content is more than 20%, obtains plant extracts.

7. composite melissa powder as claimed in claim 1, it is characterized in that, the preparation method of described modified dietary fiber, comprise the following steps: by inulin, Semen Tritici aestivi fiber, common oats fibre 5-7:2-4:1-3 Homogeneous phase mixing in mass ratio, add its quality 3-7 water doubly, room temperature 100-300W, 35-40KHz condition ultrasonic extraction 10-15min, then at electric-field intensity 20-40kV/cm, burst length 300-500 μ s, carries out high-pressure pulse electric process 10-15min under pulse frequency 200-400Hz 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 filters, and filtrate reduced in volume, freeze drying to moisture are that namely 5-8% obtains modified dietary fiber;

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

8. composite melissa powder as claimed in claim 1, it is characterized in that, described Chinese herbal medicine is prepared through ultramicro grinding with one or more in chrysanthemum, sophora flower, ginseng, lily, the root of kudzu vine, matrimony vine, dandelion, Chinese yam, Poria cocos, sea-buckthorn, rose, honeysuckle, peppermint, Momordica grosvenori, donkey-hide gelatin, red date, mulberry leaf, balsam pear, Agricus blazei.

9. the preparation method of composite melissa powder as claimed in claim 1, is characterized in that, comprise the steps:

1) each supplementary material is taken according to formula;

2) Bee Pollen is put in the supersonic wave cleaning machine that 0.3-0.5% sodium bicarbonate solution is housed, in 200W under room temperature, 40KHz cleans 3-5min, rinsing, drain, be soak 5-7min in the sericin peptide taken solution of 13-18% in mass percent, take out, put-18--22 DEG C of freezing 5-10min, broken, immediately at power 3-5Kw, frequency 2450MHz, temperature 120-140 DEG C of heating using microwave 5-7s, add former Bee Pollen quality 3-5 water doubly, be 4-6 by lactic acid adjust ph, in electric-field intensity 20-40kV/cm, burst length 400-600 μ s, high-pressure pulse electric process 10-15min is carried out under pulse frequency 200-400Hz room temperature condition, then room temperature 400W, 40KHz condition ultrasonic process 10-15min obtains Bee Pollen pretreatment fluid,

3) Bee Pollen pretreatment fluid is warming up to 40-50 DEG C, adds plant extracts and pupa worm enteron aisle extract wherein, stir, insulation, enzymolysis 10-15min, obtains Bee Pollen enzymolysis liquid;

4) in Bee Pollen enzymolysis liquid, add the modified dietary fiber of compound sugar, 20-30%, abundant dissolving 5-10min, Temperature fall is to 40-45 DEG C, add the 50-60% ferment at constant temperature 3-5h of Lactobacillus plantarum opaque amount, then with the ramp of 0.8-1.0 DEG C/min to 50-55 DEG C, add the 40-50% ferment at constant temperature 1.5-3.5h of Lactobacillus plantarum opaque amount, finally be cooled to 40-45 DEG C with the speed of 0.6-0.8 DEG C/min, continue fermentation 0.5-1h, fermentation liquor 100-300 eye mesh screen filters, and namely filtrate obtain broker wall bee pollen through ultrafiltration, reduced pressure concentration, freeze drying;

5) broker wall bee pollen and hydroxyl isomaltulose, mango powder and auxiliary material are added V-type blending tank Homogeneous phase mixing 25-35min, speed of agitator 20-40r/min, then residue modified dietary fiber Homogeneous phase mixing 12-18min is added, speed of agitator 40-60r/min, namely sterile filling, sealing, packaging obtain composite melissa powder.

10. the preparation method of composite melissa powder as claimed in claim 9, it is characterized in that, the preparation method of cryoprotector in described Lactobacillus plantarum powder preparation process, comprise the steps: winter rye, Ammopiptanthus mongolicus, sharkskin collagen cleans respectively, drain, 8-10:3-5:2-4 Homogeneous phase mixing in mass ratio, adding mixed material quality 0.1-1 pH value is doubly that the lactic acid of 3.8-4.5 soaks 3-8h, pulverize immediately after-18--22 DEG C of freezing 1-2h, freezing thickness of feed layer 3-5cm, crushed material particle diameter 0.5-3mm, then crushed material quality 10-20 water is doubly added, be 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, high-pressure pulse electric process 20-30min is carried out under pulse frequency 200-300Hz condition, then under power 150-300W condition, carry out microwave irradiation in room temperature and extract 15-20min, under power 200-300W, frequency 30-40KHz condition, carry out ultrasonic assistant extraction simultaneously, add the compound enzyme of extract quality 1-2%, in 45-55 DEG C of enzymolysis 30-50min, enzymolysis liquid filters, filtrate is concentrated, low-temperature grinding to particle diameter is that namely 0.1-0.3mm obtains protective agent,

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