CN105831665A

CN105831665A - Bee pollen being capable of reducing cholesterol and preparation method thereof

Info

Publication number: CN105831665A
Application number: CN201610178523.4A
Authority: CN
Inventors: 邵素英
Original assignee: Individual
Current assignee: Individual
Priority date: 2016-03-25
Filing date: 2016-03-25
Publication date: 2016-08-10

Abstract

The invention discloses bee pollen being capable of reducing cholesterol and a preparation method thereof. With bee pollen as a main raw material, a natural plant-sourced enzyme being high in enzyme activity and abundant in enzyme system is compounded with a pupa intestine extract, which contains the most original and the most comprehensive enzyme system during formation of the bee pollen and is composed of saliva and intestines of queen bee larvas and drone pupae, wherein the bee pollen is quickly, effectively and deeply subjected to cell wall disruption with microwave high-temperature instant puffing technology and probiotic fermentation technology for the special biological enzymolysis, microwave high-temperature instant puffing and probiotic fermentation, thereby preparing the cell-disruption bee pollen. The bee pollen is then compounded with one or more functional auxiliary materials scientifically to prepare the bee pollen being capable of reducing cholesterol, which maintains the natural flavor and nutrients of the bee pollen as most as possible, has high cell disruption rate, is high in efficiency and is strong in functions.

Description

A kind of bee pollen reducing cholesterol and preparation method thereof

Technical field

The present invention relates to bee pollen, be specifically related to a kind of bee pollen reducing cholesterol and preparation method thereof.

Background technology

Bee pollen refers to the pollen load taken back during honeybee producting honey, at the pollen formed after storage and fermentation in Nidus Vespae.Apis is when gathering honey, and corbiculate leg can collect pollen, forms pollen load, and after entering Nidus Vespae, pollen load can be stored.Its main dietetic therapy composition is: protein, aminoacid, vitamin, bee pollen element, trace element, organized enzyme, flavone compound, lipid, nucleic acid, brassin, phytic acid etc..Wherein amino acid content and ratio are closest to the amino acid pattern that FAO (Food and Agriculture Organization of the United Nation) (FAO) is recommended, and this is the most rare in wholefood.Bee pollen is the concentrate that the nutritious material being worth with drug effect is formed, and it is containing protein, carbohydrate, mineral, vitamin and other active substance.Bee pollen is fabulous natural nutrient food, is also a kind of preferably tonic simultaneously, and has certain medical function.

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

Compared with other natural product, the structure comparison of pollen is special.Pollen grain is to be made up of pollen wall, germinal aperature and protoplasm three part.Pollen wall, also known as sporoderm, can be divided into inner and outer wall, and inwall is innermost one layer of sporoderm, smooth, thin and flexible.Outer wall is thicker, hard and lack flexibility, be that in sporoderm, structure is more complicated one layer, and main component is sporopollenin, and this material has a resistance effect of physics, chemistry and enzyme, acidproof, alkaline-resisting, heatproof, pressure, the most highly stable to gastric acid and other digestive system enzyme.Research proves: the nutrient substance in non-pollen (processed by breaking wall) is the most easily digested, and the active substance that breaking cellular wall can make pollen fully discharges, so that effect of its pollen product and health care all increase substantially.Pollen wall both have impact on absorbing of pollens nutrition composition, the release of nutrient substance when also preventing extraction, greatly limit deep development and the utilization of pollen, and therefore, the breaking cellular wall of bee pollen is critically important.

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

Mechanical breaking-wall method mainly relies on the effects such as the extruding of machinery, shearing, makes pollen wall and interior membrane vesicle rupture, and the content release of pollen, common have whirlwind comminuting method and micronizing method.Hao Xiaoliang etc. use whirlwind comminuting method to have studied the breaking cellular wall of Pollen Pini, and Cao Longkui etc. uses the ultrafine powder technical research breaking cellular wall of Pollen Maydis, makes pollen particles particle diameter reach below 8 μm, and epigranular is reasonable.Chinese patent CN 103181511 discloses the preparation method of a kind of ultramicro safflower bee pollen, it is characterized in that safflower bee pollen is in the freezer of temperature-20～-30 DEG C after freezing 5～8 days, move on to calorstat that temperature is 65～75 DEG C or greenhouse are placed 20～30 minutes, moving on to temperature again is that 45～50 DEG C of drying baker are dried 3～5 hours, move on to water-cooled after being dried cooling and grind pulverizing in super micron mill, coolant outlet water temperature controls below 25 DEG C, smashing fineness controls at 1500～2000 mesh, after the ultramicro safflower bee pollen sterilization ground, pack by foil sealing, it is housed in temperature for stand-by in-5～7 DEG C of freezers.Use the super-micro wall-broken safflower bee pollen prepared of the inventive method, the absorbance of safflower bee pollen active ingredient can be improved, give full play to safflower bee pollen to cardiovascular and cerebrovascular disease, improve blood circulation and have prevention and effect for the treatment of.Chinese patent CN 101449754 B discloses the production method of a kind of broker wall bee pollen granules, belongs to the processing technique 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 breaking cellular wall speed is slow etc..The present invention provides a kind of breaking cellular wall efficiency high, sterilization effect is good, use equipment simple broker wall bee pollen granules production method, including sterilizing, breaking cellular wall, pelletize and inspection process, it is characterized in that: described wall-breaking machine is breaking cellular wall by vibrating and grinding machine, and the frequency of vibration of this breaking cellular wall by vibrating and grinding machine is between 1200-1600cpm, and Oscillation Amplitude is between 3.5-5mm, every batch of inventory of described bee pollen is 100kg, and broken time is 2 hours.But, extruding that mechanical breaking-wall method sporoderm-broken rate energy consumption is high and strong, shear the loss being likely to result in nutritional labeling.

Physical wall breaking is made by the physical actions such as radiation, swelling, infiltration, ultrasound wave, microwave, difference variation makes pollen wall rupture, and mainly has temperature differential method, radiation method, hydration breaking cellular wall method, supercritical ultrasonics technology, osmotic pressure breaking cellular wall method, ultralow temperature to add microwave frequency measurment method and liquid nitrogen quenching breaking cellular wall method.Chinese patent CN 102114058 B is for treating the production method of the bee pollen tablet of fatty liver, relate to product tablet production technology, the bee pollen of remove impurity is immersed in normal pressure and temperature water after comminution by gas stream broken wall treatment, after ultrasonic Treatment, through the refrigerated centrifuger centrifugal treating that rotating speed is 8000～10000 turns, take supernatant, then by supernatant under the ambient temperature of-5～5 DEG C, cross the molecular sieve of below 2800D, isolate the molecular weight bee pollen hydrotrope less than 2800D；After the molecular weight chilled vacuum drying of the bee pollen hydrotrope less than 2800D, obtain the water content Herba leucadis ciliatae lyophilized powder less than 4%；By described Herba leucadis ciliatae lyophilized powder and water soluble starch mixing film-making.Chinese patent CN 101416763 B discloses a kind of bee pollen beverage and preparation method thereof, and including bee pollen is carried out pretreatment, broken wall treatment method that breaking cellular wall by temperature difference combines with mechanical breaking-wall method, cold preservation stand, are centrifuged, filtration, constant volume, the multiple working procedure such as fill prepare the Herba leucadis ciliatae powder of clarification.But, single physical method sporoderm-broken rate is relatively low and high to some equipment requirements.

Biological wall breaking is because sporoderm-broken rate is high, shell-broken effect is notable；Action condition is gentle, little to heat-sensitive nutrition destructiveness；While breaking cellular wall, may additionally facilitate bee pollen digestion and absorb, producing 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, from the point of view of the fermentation that biological wall breaking is big, mainly include breaking wall by fermentation and enzymolysis breaking cellular wall.

Enzymolysis breaking cellular wall is to utilize enzyme to make some ingredient breakdown on pollen wall destroy pollen wall, makes germinal aperature open, and pollen content flows out.In enzymolysis breaking cellular wall, conventional enzyme has cellulase, hemicellulase, pectase, protease, amylase, compound enzyme etc. at present.Chinese patent CN 102746413 B discloses a kind of bee pollen polysaccharide enzymolysis breaking cellular wall and combines the ultrasonic leach extraction method of hot water, comprises the following steps: is dried by bee pollen, remove impurity, pulverizes, sieve；Enzymolysis breaking cellular wall；Heat ultrasonic extraction；Dehydration；Perchloric acid removing protein；DEAE column chromatography, eluting；CTAB separates containing acidic polysaccharose precipitation and precipitates containing neutral polysaccharide；Through dehydration, vacuum freezing after dissolving polysaccharide is deep, it is dried to obtain the pure bee pollen polysaccharide of high-purity.Chinese patent CN 101401821 B discloses one and through protease hydrolyzed, bee pollen is become polypeptide, tablet made by polypeptide and preparation method thereof.It is will to pulverize through screening, roguing, dried bee pollen, reaches requirement post-treatment skill water, stir, be positioned over less than-18 DEG C freezing 24-48 hour.The fresh water (FW) stirring adding 100 DEG C after taking-up immediately is dissolved, and with homogenizer homogenizing 20-60 minute, puts into and adds cellulase in enzymatic vessel and carry out enzymolysis 0.5-2 hour, after adjust pH value with alkali liquor after add protease and carry out enzymolysis 3-6 hour.Immediately bee pollen peptide liquid is heated after enzymolysis and carry out enzyme denaturing.With starch as excipient, carry out pelletize, tabletting and get final product.Feature is 1) specificity: the protein at protease, pectase, cellulase, the amylase single-minded pollen enzymolysis wall of energy and germinal aperature, pectin, cellulose, starch grain macromolecular substances；2) high efficiency: enzyme digestion reaction shell-broken effect is good；3) reaction condition is gentle, the beneficially preservation of nutrient substance；But due to bee pollen sporoderm complicated component and the single-minded characteristic of enzyme, existing enzyme preparation and compounding, it is impossible to well solving enzymolysis breaking cellular wall problem, simultaneously as yielding poorly of enzyme, expensive, enzymolysis cost is high.

Breaking wall by fermentation method is mainly by the various enzymes produced in the probiotics sweats such as yeast, aspergillosis, lactic acid bacteria, and the various enzymes contained inside pollen, the passage portion (germinal aperature, ditch) of pollen inside and outside wall is got through in the effect utilizing these enzymes, so that the nutritional labeling dissolution in inwall.In breaking wall by fermentation, conventional strain has aspergillosis, yeast, lactic acid bacteria, Pleurotus ostreatus, Lentinus Edodes etc. at present.Chinese patent CN 101756091 B discloses a kind of pollen natto tablet, including the component of following weight portion: pollen 4～5 parts, natto 3～4 parts, Monas cuspurpureus Went 1～2 parts, oligosaccharide 1～2 parts.Described pollen is with bee pollen as raw material, obtains through following preparation method: first with 50～100PPM ClO₂Aqueous 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, checks acidity, as pH=4.5, cold drying, obtains pollen；Described leaven is streptococcus acidi lactici and lactobacillus.Chinese patent CN 104286623 A bee pollen and the fermentation process of bee pollen, use probiotic bacteria mixed solid fermentation bee pollen, and production cost is low, and the fund of investment is less, and it is convenient to process in downstream, pollutes little.The fermentation bee pollen produced, can regulate human body intestinal canal function.The protein contained in bee pollen is broken down into digesting and assimilating of little peptide and free amino acid, beneficially the intestines and stomach, removes anaphylactogen.Described probiotic bacteria uses Bifidobacterium lactis or Lactobacillus plantarum or bacillus acidophilus.The viable count of described probiotic bacteria is 1 × 10⁹-5×10⁹CFU/g.The manufacture method of a Chinese patent CN 104059842 A bee pollen wine, the step of the manufacture method of described bee pollen wine is as follows: takes the bee pollen 0.8-1 weight portion that sterilization treatment crosses and is cooled to 30-35 DEG C, add the cold boiled water of softening that the black clothing song of 1.5 2 weight portions adds 0.8--1 weight portion, put into the stirring of dispensing basin all, put into sealing and fermenting 40 48 hours in the wine vat that sterilization treatment is crossed；Add the cold boiled water of softening of 10 15 weight portions, add the Mel of 10 15 weight portion 40 41 concentration, Arillus Longan core 1.5 2.5 weight portion that addition processed, it is separately added in feed liquid, being stirred uniformly, blanking is complete to be sealed, temperature during regulation and control fermentation, being maintained between temperature 20 25 DEG C, the wine vat that falls is 30 35 days to fermentation dwell time；Extracting the wine liquid after filtering, water proof heat sterilization temperature 85 90 DEG C, 20 30 minutes, seal precipitation up for safekeeping 15 20 days after cooling, fill finished product can be waited to drink.Chinese patent CN 102389069 A discloses a kind of broken wall of melissa pollen method, it is characterised in that comprises the steps: to add culture propagation in bee pollen, makes broken wall of melissa pollen；The yeast added in bee pollen is calculated as the 0.5～3% of bee pollen weight with dry weight；After bee pollen adds yeast, add bee pollen volume 1.5～the water of 2.5 times, then ferment 2～4 days in 28～34 DEG C, finally heat 4～8h in 36～38 DEG C, obtain the bee pollen solution after breaking cellular wall.Chinese patent CN 103598654 B discloses a kind of pollen active probiotic drink and preparation method thereof.This pollen active probiotic drink is with bee pollen as raw material, is equipped with a certain proportion of maltose and/or galactose, through lactobacillus BL1 and two kinds of probiotic mixed fermentations of lactobacillus BL2, more formulated forms.The present invention is directed to bee pollen raw material acidity relatively strong, be difficult to the feature utilized by microorganism, preferably two acid resistances are strong and have the bacterial strain of multiple merit, are maintained at 10 by number of live bacteria of probiotics in controlling fermentation technology and making product 30 days¹¹More than cfu/L, and a large amount of flavone aglycone can be obtained by bioconversion bee pollen flavonoid glycoside, improve the bioavailability of bee pollen flavone.Simultaneously rich in other bee pollen and probiotics fermention product, nutritious, aromatic flavor, sour and sweet palatability, there is the functions such as looks improving and the skin nourishing, enhancing immunity, reduction cholesterol, regulating intestinal canal flora and Constipation simultaneously.Aspergillosis 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 are high, nutrient component damages is little, amino acid content is high but time length, speed are slow after breaking cellular wall.

To sum up, seeking one and keep bee pollen natural flavour mountaineous to greatest extent and nutritional labeling, sporoderm-broken rate is high, efficiency is high, the broken wall of melissa pollen method of low cost, necessary to expand the field of deep of bee pollen.

Summary of the invention

Solved by the invention technical problem is that the defect overcoming existing broken wall of melissa pollen technology, with bee pollen as primary raw material, enzyme activity is high, natural plant enzyme that enzyme system is abundant and the most original containing bee pollen forming process, the pupa worm intestinal extract science of queen bee nit that enzyme system is the most complete and drone pupa saliva and intestinal enzyme system compounds, and use the instantaneous puffing technique of microwave high-temperature and probiotics fermention technology, by Specialty bio enzymolysis, bee pollen is carried out quickly by the instantaneous low temperature expanding of microwave high-temperature and probiotics fermention, effectively and degree of depth breaking cellular wall, prepare broker wall bee pollen, then compound with one or more functional auxiliary material science, prepared one keeps bee pollen natural flavour mountaineous and nutritional labeling to greatest extent, sporoderm-broken rate is high, efficiency is high, low cost, the bee pollen of functional strong reduced cholesterol.

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

A kind of bee pollen reducing cholesterol, mainly has the raw material of following parts by weight to prepare:

Bee pollen 150-180 part, plant extract 10-30 part, pupa worm intestinal extract 10-20 part, modified dietary fiber 10-20 part, oligosaccharide 20-30 part, Lactobacillus plantarum powder 6-10 part, hydroxyl isomaltulose 2-6 part, mango powder 2-6 part；

Preferably, the described bee pollen reducing cholesterol, mainly have the raw material of following parts by weight to prepare:

Bee pollen 160-170 part, plant extract 15-25 part, pupa worm intestinal extract 13-17 part, modified dietary fiber 13-17 part, oligosaccharide 23-27 part, Lactobacillus plantarum powder 7-9 part, hydroxyl isomaltulose 3-5 part, mango powder 3-5 part；

It is highly preferred that the described bee pollen reducing cholesterol, the raw material of following parts by weight is mainly had to prepare:

Bee pollen 165 parts, plant extract 20 parts, pupa worm intestinal extract 15 parts, modified dietary fiber 15 parts, oligosaccharide 25 parts, 8 parts of Lactobacillus plantarum powder, hydroxyl isomaltulose 4 parts, mango powder 4 parts；

Further, the described bee pollen reducing cholesterol also includes one or more adjuvants 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, plant sterol 0.2-0.8 part, phosphatidyl serine 0.1-0.7 part, oligochitosan 0.1-0.7 part, marine fishbone collagen oligopeptide powder 0.1-0.7 part, Chinese herbal medicine 0.1-0.6 part, Cordyceps militaris (L.) Link. 0.1-0.5 part；Soybean protein isolate 0.1-0.5 part；

Preferably, the described bee pollen reducing cholesterol also includes one or more adjuvants 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, plant sterol 0.4-0.6 part, phosphatidyl serine 0.3-0.5 part, oligochitosan 0.3-0.5 part, marine fishbone collagen oligopeptide powder 0.3-0.5 part, Chinese herbal medicine 0.2-0.4 part, Cordyceps militaris (L.) Link. 0.2-0.4 part；Soybean protein isolate 0.2-0.4 part；

It is highly preferred that the described bee pollen reducing cholesterol also includes one or more adjuvants of following parts by weight；

0.9 part of algae, corn oligopeptide powder 0.8 part, pueraria root powder 0.8 part, 0.5 part of fish oil, plant sterol 0.5 part, phosphatidyl serine 0.4 part, oligochitosan 0.4 part, marine fishbone collagen oligopeptide powder 0.4 part, Chinese herbal medicine 0.3 part, Cordyceps militaris (L.) Link. 0.3 part；Soybean protein isolate 0.3 part；

Further, any one during described bee pollen is bee pollen of maize, Semen Sesami bee pollen, Brassica campestris L pollen, pollen of Semen Fagopyri Esculenti；

Further, described plant extract does not contain only the various plants enzymes such as abundant plant rennet, amylase, hemicellulase, esterase, oxidoreductase and containing the nutrient substance such as vegetable polysaccharides and monosaccharide, plant amylum, vegetable protein, can be not only that the bee pollen that can reduce cholesterol provides comprehensive, natural phytoenzyme, can be also that probiotic bacteria provides nutrient substance comprehensive, abundant, compound with pupa worm intestinal extract science, sporoderm-broken rate is higher, and effect is more preferable；

Preferably, described plant extract uses the extract at low temperature technology such as ultrasonic cleaning, microwave-assisted supersound extraction and high-pressure pulse electric extraction, concentrating under reduced pressure, is effectively increased raw material availability, phytoenzyme activity and productivity；The foodsafety of plant extract has been effectively ensured；

It is highly preferred that the preparation method of described plant extract is: Fructus Hordei Germinatus and Fructus Tritici aestivi 8-10:1-3 in mass ratio are uniformly mixed, is crushed to granularity 0.5-1mm, obtains pulverizing Fructus Hordei Germinatus；nullThen by Fructus Chaenomelis、Fructus Ananadis comosi、Fructus Fici in ultrasonic washing unit at power 200W、Ultrasonic cleaning 5-10min under the conditions of frequency 30KHz，Drain，It is crushed to granularity 0.5-1mm under room temperature，And 7-9:1-3:1-2 uniformly mixes in mass ratio，The pulverizing Fructus Hordei Germinatus adding mixture quality 3-5 times obtains raw mixture，Add the water of raw mixture quality 1-3 times，It is 3-4 with Fructus Citri Limoniae acid for adjusting pH value，At power 150-300W、Microwave extraction is carried out under the conditions of frequency 2000Hz，Wherein，Microwave exposure total time 60-80s every time,Carry out compartment irradiation: irradiation 10s，Interval 10s，Control temperature 20-35 DEG C，So irradiation 10 times，Simultaneously at power 200-300W,Ultrasonic assistant extraction is carried out under the conditions of frequency 30-40KHz；Insulation 1-3h, then, microwave extraction is carried out under the conditions of power 200-400W, frequency 2000Hz, wherein, microwave exposure total time 90-105s every time, carry out compartment irradiation: irradiation 15s, interval 10s, controls temperature 40-60 DEG C, such irradiation 10 times, simultaneously at power 300-500W, carry out ultrasonic assistant extraction under the conditions of frequency 40-50KHz, be finally naturally cooling 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 the conditions of pulse frequency 200-300Hz；Extracting solution filters to obtain the first filtrate, adds the water rinsing of filtering residue 4 times, filters to obtain the second filtrate, the first filtrate and the second filtrate 1:3-5 is in mass ratio uniformly mixed, and being evaporated to solid content is more than 20%, obtains plant extract；

Preferably, in described ultrasonic washing unit, cleanout fluid is the sodium bicarbonate solution of 0.3-0.5%.

Further, described pupa worm intestinal extract is with the queen bee nit intestinal of 2-3 age in days and the drone pupa intestinal of 11-12 age in days as raw material, obtains through freeze proof protection, freezing crushing, biological demulsifying and centrifugation；

nullPreferably，The preparation method of described pupa worm intestinal extract，Comprise the steps: the drone pupa intestinal 3-5:1-3 in mass ratio mixing of the queen bee nit intestinal by 2-3 age in days and 11-12 age in days，Add the sericin peptide taken solution that mass percent is 8-12% of mixture quality 1-2 times，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 the conditions of 4 DEG C，Collect the middle level liquid after being centrifuged for the first time，Then at 18000-25000g，Centrifugal 20-30min under the conditions of 4 DEG C，Collect the middle level liquid after second time is centrifuged，Obtain pupa worm intestinal 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 combine 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 combines class=5-7:4-6:2-4；

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

It is highly preferred that the quality group of described glycolipid class biological demulsifying agent becomes: rhamnolipid: alkyl polyglucoside=3-5:1-2.

Further, described modified dietary fiber is that through physics, chemical or biological method having of processing and obtain, the abundant soluble fiber cellulose content of strong retentiveness, dilatancy, thickening property, adsorptivity and network is high, biological activity is strong, have important, the cellulose of positive role to human body beneficial flora by dietary fiber, compared with full diet fiber, its biological action is more powerful, also can be greatly prolonged the shelf-life of the bee pollen that can reduce cholesterol；

Preferably, described modified fibre is to be obtained through enzyme enzymolysis by one or more in inulin, apple fiber, Herba avenae fatuae fiber, Semen Tritici aestivi fiber；

More preferably, the preparation method of described modified dietary fiber, comprise the following steps: inulin, Semen Tritici aestivi fiber, Herba avenae fatuae fiber 5-7:2-4:1-3 in mass ratio is uniformly mixed, add the water of its quality 3-7 times, room temperature 100-300W, 35-40KHz condition supersound extraction 10-15min, then in electric field intensity 20-40kV/cm, burst length 300-500 μ s, carry out high voltage pulse electric field processing 10-15min under the conditions of pulse frequency 200-400Hz；It is 4.5-6.5 with breast acid for adjusting pH value, adds the enzyme of mixture quality 0.1-0.3%, in 45-55 DEG C of enzymolysis 20-48min；Enzymolysis solution filters, and filtrate reduced in volume, lyophilization to moisture are that 5-8% i.e. obtains modified dietary fiber；

Described enzyme is that cellulase, xylanase, laccase, pectase 1-3:1-3:1-2:1-2 in mass ratio uniformly mixes.

Further, during described oligosaccharide is oligofructose, stachyose, Raffinose, oligomeric xylose, oligomeric galactose, soybean oligo saccharide, oligomeric isomaltose, Oligomeric maltose at least one；

Preferably, described oligosaccharide parts by weight consist of oligofructose 40-50 part, Oligomeric maltose 30-40 part, Raffinose 20-40 part, soybean oligo saccharide 16-18 part, oligomeric galactose 10-15 part, oligomeric xylose 10-15 part, oligomeric isomaltose 10-15 part, stachyose 8-12 part.

Further, described Lactobacillus plantarum powder is prepared with Lactobacillus plantarum CGMCC NO.11763 according to a conventional method for starting strain, and its viable bacteria content is: 7 × 10¹²-9×10¹²cfu/g；The cryoprotective agent that wherein freeze drying process uses is with the animal or plant containing antifreeze protein as raw material; prepared through high-pressure pulse electric extraction, ultrasonic assistant microwave extraction and compound enzyme enzymolysis, Lactobacillus plantarum powder can be effectively improved at freezing dry process Viable detection；

Preferably, described protectant preparation method, comprise the steps: winter rye, Caulis et Folium Ammopiptanthi Mongolici, sharkskin collagen protein is respectively washed, drain, 8-10:3-5:2-4 in mass ratio uniformly mixes, add the lactic acid moistening 3-8h that pH value is 3.8-4.5 of mixed material quality 0.1-1 times, pulverize immediately after-18--22 DEG C of freezing 1-2h, freezing thickness of feed layer 3-5cm, ground product particle diameter 0.5-3mm, it is subsequently added into the water of ground product quality 10-20 times, it is 3.5-5.5 with breast acid for adjusting pH value, at room temperature in electric field intensity 25-35kV/cm, burst length 300-500 μ s, high voltage pulse electric field processing 20-30min is carried out under the conditions of pulse frequency 200-300Hz；Then under the conditions of power 150-300W, carry out microwave irradiation and extraction 15-20min in room temperature, simultaneously at power 200-300W, under the conditions of frequency 30-40KHz, carry out ultrasonic assistant extraction；Add the compound enzyme of extracting solution quality 1-2%, in 45-55 DEG C of enzymolysis 30-50min；Enzymolysis solution filters, filtrate concentrates, low-temperature grinding to particle diameter is that 0.1-0.3mm i.e. obtains protective agent；

Described compound enzyme is that cellulase, protease, amylase, pectase, tannase 2-4:1-3:1-3:1-2:1-2 in mass ratio uniformly mixes.

Further, any one during described algae is Haematocoocus Pluvialls, salt alga, spirulina plalensis.

Further, described Chinese herbal medicine is to prepare through micronizing with one or more in Flos Chrysanthemi, Flos Sophorae, Radix Ginseng, Bulbus Lilii, Radix Puerariae, Fructus Lycii, Herba Taraxaci, Rhizoma Dioscoreae, Poria, Fructus Hippophae, Flos Rosae Rugosae, Flos Lonicerae, Herba Menthae, Fructus Momordicae, Colla Corii Asini, Fructus Jujubae, Folium Mori, Fructus Momordicae charantiae, Agaricus blazei Murrill.

Another object of the present invention is to provide the preparation method of the above-mentioned bee pollen reducing cholesterol, comprises the steps:

1) each supplementary material is weighed according to formula；

null2) bee pollen is put in the ultrasonic washing unit equipped with 0.3-0.5% sodium bicarbonate solution，In 200W under room temperature、40KHz cleans 3-5min，Rinsing，Drain，5-7min is soaked in the sericin peptide taken solution that mass percent is 13-18%，Take out，Put-18-22 DEG C of freezing 5-10min，Broken，Immediately at power 3-5Kw,Frequency 2450MHz,Temperature 120-140 DEG C microwave heating 5-7s,Add the water of former bee pollen quality 3-5 times，It is 4-6 with breast acid for adjusting pH value，In electric field intensity 20-40kV/cm，Burst length 400-600 μ s,High voltage pulse electric field processing 10-15min is carried out under pulse frequency 200-400Hz room temperature condition，Then room temperature 400W、40KHz condition supersound process 10-15min obtains bee pollen pretreatment fluid；

3) bee pollen pretreatment fluid is warming up to 40-50 DEG C, is added thereto to plant extract and pupa worm intestinal extract, stirs, insulation, enzymolysis 10-15min, obtain bee pollen enzymolysis solution；

4) in bee pollen enzymolysis solution, oligosaccharide, the modified dietary fiber of 20-30% are added, fully dissolve 5-10min, it is naturally cooling 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 it is 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 filtrate i.e. obtains broker wall bee pollen through ultrafiltration, concentrating under reduced pressure, lyophilization；

5) broker wall bee pollen is added V-type blending tank with hydroxyl isomaltulose, mango powder and adjuvant and uniformly mix 25-35min, speed of agitator 20-40r/min, it is subsequently adding residue modified dietary fiber and uniformly mixes 12-18min, speed of agitator 40-60r/min, sterile filling, seals, packs and i.e. obtain the bee pollen that can reduce cholesterol；

Use broker wall bee pollen sporoderm-broken rate prepared by said method up to more than 98%, it is possible to decrease in the bee pollen of cholesterol, Lactobacillus plantarum viable count is up to 6 × 10¹¹-9×10¹¹cfu/g。

Lactobacillus plantarum of the present invention (Lactobacillus plantarum) 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 CGMCC NO.11763, preservation address is: North Star West Road 1, Chaoyang District, city of BeiJing, China institute 3, Institute of Microorganism, Academia Sinica, postcode: 100101.

Lactobacillus plantarum probiotic properties is as follows:

Lactobacillus plantarum CGMCC NO.11763 provided by the present invention is found under conditions of pH is 1.50 survive, still in existing state after 1% cholate is cultivated 4 hours through experiment；Lactobacillus plantarum CGMCC NO.11763 degrading nitrite speed is fast, and capacity of decomposition reaches 10.9mg/h/kg, and this strain is when producing Pickles, and whole sweat nitrite concentration is at below 4.8mg/kg；CGMCC NO.11763, after fermentation 60h hour, can reach 64.76% to degrading rate of cholesterol.CGMCC NO.11763 Adhering capacity measure from coagulation rate be 95.71%.

Lactobacillus plantarum CGMCC NO.11763 is to cholesterol degradation capability study and mensuration:

Take 1ml CGMCC NO.11763 mother solution and be inoculated in MRS cholesterol fluid medium (the cholesterol level 0.1mg/ml of 10mL, pH 6.2) in, the constant temperature of 37 DEG C stands and cultivates 20h respectively, 40h, 60h is standby, with access 1mL sterilized water MRS cholesterol culture medium for comparison, take bacteria liquid sample and the comparison each 1ml of liquid of above cultivation different time, 9000r/min, centrifugal 10min at 4 DEG C, obtain fermented supernatant fluid, in o-phthalaldehyde method mensuration supernatant, cholesterol level is (particularly as follows: take each supernatant 0.1ml in corresponding test tube, add glacial acetic acid 0.3ml, o-phthalaldehyde(OPA) 0.15ml of 1mg/ml, it is slowly added into concentrated sulphuric acid 1.0ml, mix homogeneously.Room temperature stands 10min, surveys light absorption value under 550nm).Each process 3 repetition, in kind makes cholesterol standard curve, calculates cholesterol level and degradation rate in supernatant, the results are shown in Table 1.Understanding, CGMCCNO.11763 has good Degradation to cholesterol, and after fermentation 60h hour, degradation rate can reach 64.76%.

The table 1 degraded situation to 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 CGMCC NO.11763 bacterial strain:

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

Reach 0.59 ± 0.92 × 10⁷(cfu/ml), there is good bile tolerance ability.

Table 2 bile tolerance ability detection [(± s) × 10⁷cfu/ml]

The acid resistance test of Lactobacillus plantarum CGMCC NO.11763 bacterial strain

Take HLX37 mother solution by 1ml inoculation strain in the 10mLMRS fluid medium of different pH value (pH gradient is 1.5,2.0,2.5,3.0,3.5,4.0), be placed at 37 DEG C and cultivate 0 respectively, 2,4h, each process 3 repetition.Respectively take 1ml sample bacterium solution to mix in 9ml normal saline, prepare dilute solution, take 0.1ml diluent and be coated with in MRS, in 37 DEG C of biochemical cultivation cases, be inverted the bacterium colony number cultivated on 48 hours (each dilution factor do 3 parallel) record flat board.The results are shown in Table 3.Illustrate that this bacterium has the strongest acid-fast ability.

Table 3 acid-fast ability detection [(± s) × 10⁷cfu/ml]

The Adhering capacity of Lactobacillus plantarum CGMCC NO.11763 measures

Cultivate CGMCC NO.11763 (MRS fluid medium), bacillus coli DH 5 alpha (LB fluid medium) 24h obtains fermentation liquid, it is respectively placed in 3000r/min, centrifugal 10min at 4 DEG C, collect bacterium mud, (in bacterium colony, PBS is i.e. added 2 times respectively with sterile phosphate buffer (PBS) the washing bacterium mud of pH=7.0, after concussion mix homogeneously, it is placed in 3000r/min, centrifugal 10min at 4 DEG C, collects thalline).From coagulation rate (%): with aseptic PBS, bacterium mud CGMCC NO.11763 is formed in the suspension bacteria liquid and bacteria suspension that the light absorption value at wavelength 600nm is 0.4 ± 0.1 (A0), measure light absorption value A24 after standing 24h, be (A0 A24)/A0 from coagulation rate (%) formula.；His coagulation rate (%): the outstanding bacterium solution of CGMCC NO.11763 and bacillus coli DH 5 alpha is adjusted to the mix suspending bacterium solution that the light absorption value at wavelength 600nm is 0.6 ± 0.1 (A0).Measuring light absorption value A24 after standing 24H, his coagulation rate (%) formula is (A0 A24)/A0.Measurement result is shown in Table 4, it is known that CGMCC NO.11763 is 95.71% from coagulation rate, has the strongest Adhering capacity.

Table 4 Adhering capacity table

The bacterial strain physiological property of Lactobacillus plantarum CGMCC NO.11763

Described Lactobacillus plantarum (Lactobacillus plantarum) 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 CGMCC NO.11763, preservation address is: North Star West Road 1, Chaoyang District, city of BeiJing, China institute 3, 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, atrichia, does not produce spore；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 (-), indole experiment (+), mobility (-), fermentation gas (-), nitrate reductase (-), fermentation gas (-), product hydrogen sulfide gas (-), pH4.0MRS culture medium grows (+).It is accredited as Lactobacillus plantarum (Lactobacillus plantarum), named Lactobacillus plantarum (Lactobacillus plantarum) XH through Physiology and biochemistry.

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, isolated in Yoghourt from Xinjiang Uygur fellow-villager family, acquisition time on June 2nd, 2015.

Lactobacillus plantarum CGMCC NO.11763 of the present invention is found under conditions of pH is 1.50 survive, still in existing state after 1% cholate is cultivated 4 hours through experiment；Lactobacillus plantarum CGMCC NO.11763 degrading nitrite speed is fast, and capacity of decomposition reaches 10.9mg/h/kg, and this strain 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.CGMCC NO.11763 Adhering capacity measure from coagulation rate be 95.71%, bacterial strain can at 57 DEG C well-grown, glucose tolerance is 275g/L.

Beneficial effect:

The present invention is with bee pollen as primary raw material, enzyme activity is high, natural plant enzyme that enzyme system is abundant and the most original containing bee pollen forming process, the pupa worm intestinal extract science of queen bee nit that enzyme system is the most complete and drone pupa saliva and intestinal enzyme system compounds, and use the instantaneous puffing technique of microwave high-temperature and probiotics fermention technology, by Specialty bio enzymolysis, bee pollen is carried out quickly by the instantaneous low temperature expanding of microwave high-temperature and probiotics fermention, effectively and degree of depth breaking cellular wall, prepare broker wall bee pollen, then compound with one or more functional auxiliary material science, prepared one keeps bee pollen natural flavour mountaineous and nutritional labeling to greatest extent, sporoderm-broken rate is high, efficiency is high, low cost, the bee pollen of functional strong reduced cholesterol.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, with bee pollen as raw material, carries out parasite killing initially with ultrasonic cleaning and goes out ovum, killing microorganisms, removal pesticide residues and heavy metal ion etc., substantially increase the foodsafety of the bee pollen that can reduce cholesterol bee pollen；Aqueous solution containing sericin peptide taken soaks, the loss of bee pollen activity material that is chilled and that cause can be reduced to greatest extent, improve the extraction ratio of bee pollen effective ingredient, also establish solid foundation for follow-up alternating temperature probiotics fermention simultaneously；Use that microwave high-temperature is instantaneous expanded makes chilled bee pollen appropriateness expanded, enhance between the inside and outside wall of bee pollen and inclusions and the separation of the structure of matter own, fluffy, promote 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 effective ingredient；The extract at low temperature technology such as high-pressure pulse electric, ultrasonic, biological enzymolysis are organically combined, functional materials and the effective ingredient of bee pollen can be retained to greatest extent；The animal ferment source science that the phytoenzyme high containing multiple enzyme activity, enzyme system is abundant and bee pollen are formed is compounded, can effectively degrade various macromolecular substances in bee pollen, its abundant enzyme system is that commercially available compound enzyme is incomparable, more meet the original natural rule of " the taking from nature, be used for nature " of the formation of bee pollen material and degraded；With functional plants lactobacillus powder as leaven, use temperature-variable fermentation and vaccination ways can obtain proliferation of probiotics while bee pollen is carried out further degree of depth breaking cellular wall to greatest extent and the maximum of functional metabolic product accumulates by several times.Use broker wall bee pollen sporoderm-broken rate prepared by said method up to more than 98%.

2. the drone pupa intestinal of queen bee nit intestinal and 11-12 age in days that the pupa worm intestinal extract science that prepared by the present invention compounds 2-3 age in days is raw material, aqueous solution containing sericin peptide taken soaks, the loss of pupa worm active substance that is chilled and that cause can be reduced to greatest extent, improve the extraction ratio of pupa worm effective ingredient；Use biological demulsifying agent breakdown of emulsion, can promote that the miscible materials such as the fat in pupa worm intestinal separate with the hydrolyzed solution containing abundant intestinal enzyme, the maximization that can obtain intestinal enzyme is extracted, relative to existing direct centrifugation or the addition separation method such as buffer, biochemicals, intestinal enzyme extraction rate is high, technique is simple, foodsafety is high, environmentally friendly.Meanwhile, pupa worm intestinal extract, through follow-up enzymolysis, possibly together with queen bee nit polypeptide and drone pupa polypeptide, adds more functional bioactive material for reducing the bee pollen of cholesterol.

3. the Lactobacillus plantarum CGMCC NO.11763 of the present invention is found under conditions of pH is 1.50 survive, still in existing state after 1% cholate is cultivated 4 hours through experiment；Lactobacillus plantarum CGMCC NO.11763 degrading nitrite speed is fast, and capacity of decomposition reaches 10.9mg/h/kg, and this strain is when producing Pickles, and whole sweat nitrite concentration is at below 4.8mg/kg；CGMCC NO.11763, after fermentation 60h hour, can reach 64.76% to degrading rate of cholesterol.CGMCC NO.11763 Adhering capacity measure from coagulation rate be 95.71%.

4. the winter rye higher containing antifreeze protein, Caulis et Folium Ammopiptanthi Mongolici and sharkskin collagen protein science are compounded by the cryoprotective agent that the present invention uses when preparing Lactobacillus plantarum powder; prepare through high-pressure pulse electric extraction, ultrasonic assistant microwave extraction and biological enzymolysis; omnidistance extract at low temperature; antifreeze protein extraction ratio is high; loss is few; the protective agent antifreeze peptide content obtained is high, kind is complete, functional by force, cryoprotective effects is good, improves the viable bacteria content in Lactobacillus plantarum powder.

5. the plant extract that prepared by the present invention does not contain only the various plants enzymes such as abundant plant rennet, amylase, hemicellulase, esterase, oxidoreductase and containing the nutrient substance such as vegetable polysaccharides and monosaccharide, plant amylum, vegetable protein, soluble fiber, can be not only that the bee pollen that can reduce cholesterol provides comprehensive, natural phytoenzyme, can be also Lactobacillus plantarum provide comprehensively, abundant nutrient substance；After the extract at low temperature such as ultrasonic cleaning, microwave-assisted supersound extraction and high-pressure pulse electric extraction, concentrating under reduced pressure, its effect is more significantly, and is effectively increased raw material availability, phytoenzyme activity and productivity；The foodsafety of plant extract has been effectively ensured.

6. supersound extraction, high-pressure pulse electric are extracted and biological enzymolysis combination of sciences by the modified dietary fiber that prepared by the present invention, gained modified dietary fiber retentiveness, dilatancy, thickening property be higher and acid and alkali, alkali, the impact of salt, soluble fiber cellulose content is high, it is easier to be 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 probiotic bacteria flora, reduce human body intestinal canal pH value, improve human body intestinal canal microecological environment；High adsorption capacity, after modified, the specific surface area of cellulose increases, and network is enriched, and absorption affinity strengthens, and the organic molecule ability chelating, adsorbing cholesterol and bile acids is higher, the suppression human body absorption to them；Ion-exchange capacity strengthens, and to metallic element, particularly heavy metal element adsorption effect is higher, effectively prevent body weight for humans metal poisoning；Regulation and the resident time of maintenance intestinal microbial population, strengthen digestion and the absorbability of intestinal, improve immunity of organisms；Effectively facilitate gastrointestinal peristalsis, slow down and eliminate the untoward reaction such as flatulence, abdominal distention；Powerful embedding effect can prevent environment (oxygen, temperature, illumination, the water activity etc.) factor impact on product quality, stabilizes the biological activity of the bee pollen that can reduce cholesterol, extends the shelf-life of the bee pollen that can reduce cholesterol.

7. oligosaccharide and modified dietary fiber science are compounded by the present invention, nutrient substance comprehensive, abundant is provided to probiotic bacteria, not only achieve the maximization accumulation maximizing propagation and functional metabolic product of probiotic bacteria, and effectively improve finished product and can reduce the mouthfeel of bee pollen and the stability of cholesterol.

8. the preparation method preparation method technique of the present invention is simple, easy to operate, low for equipment requirements, can industrialization and large-scale production, modified dietary fiber is finally mixed, has played its powerful embedding effect so that the character of product is more stable, the shelf-life is longer.

It should be noted that the present invention can reduce the result having the technical effect that each component technical characteristic and method feature are mutually collaborative, interacting of the bee pollen of cholesterol, the superposition of not simple technical characteristic (component function), the combination of each component technical characteristic and the collaborative effect produced, considerably beyond each monotechnics feature functionality and the superposition of effect, have the most advanced and practicality.

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.It addition, embodiment is interpreted as illustrative, and unrestricted the scope of the present invention, the spirit and scope of the invention are limited only by the claims that follow.To those skilled in the art, on the premise of without departing substantially from spirit and scope of the present invention, the various changes or the change that carry out the material component in these embodiments and consumption fall within protection scope of the present invention.

Embodiment one:

Prepared by raw material

1, the preparation of plant extract

Its preparation method is:

Fructus Hordei Germinatus and Fructus Tritici aestivi 9:2 in mass ratio are uniformly mixed, is crushed to granularity 0.8mm, obtains pulverizing Fructus Hordei Germinatus；Then by Fructus Chaenomelis, Fructus Ananadis comosi, Fructus Fici in ultrasonic washing unit at power 200W, ultrasonic cleaning 8min under the conditions of frequency 30KHz, drain, it is crushed to granularity 0.8mm under room temperature, and 8:2:1.5 uniformly mixes 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, it is 3.5 with Fructus Citri Limoniae acid for adjusting pH value, at power 225W, microwave extraction is carried out under the conditions of frequency 2000Hz, wherein, microwave exposure total time 70s every time, carry out compartment irradiation: irradiation 10s, interval 10s, control temperature 30 DEG C, so irradiation 10 times, simultaneously at power 250W, ultrasonic assistant extraction is carried out under the conditions of frequency 35KHz；Insulation 2h, then, carries out microwave extraction under the conditions of power 300W, frequency 2000Hz, wherein, each microwave exposure total time 100s, compartment irradiation is carried out: irradiation 15s, interval 10s, controls temperature 50 C, such irradiation 10 times, simultaneously at power 400W, carry out ultrasonic assistant extraction under the conditions of frequency 45KHz, be finally naturally cooling to room temperature, in electric field intensity 30kV/cm, burst length 500 μ s, carries out high-pressure pulse electric and extracts 18min under the conditions of pulse frequency 250Hz；Extracting solution filters to obtain the first filtrate, adds the water rinsing of filtering residue 4 times, filters to obtain the second filtrate, the first filtrate and the second filtrate 1:4 is in mass ratio uniformly mixed, and being evaporated to solid content is 30%, obtains plant extract；

In above-mentioned ultrasonic washing unit, cleanout fluid is the sodium bicarbonate solution of 0.4%.

2, the preparation of pupa worm intestinal extract

Its preparation method comprises the steps:

By queen bee nit intestinal and the drone pupa intestinal 4:2 in mass ratio mixing of 12 ages in days of 3 ages in days, add the sericin peptide taken solution that mass percent is 10% of mixture quality 1.5 times, 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, under the conditions of 4 DEG C, centrifugal 25min, collects the middle level liquid after being centrifuged for the first time, then at 22000g, centrifugal 25min under the conditions of 4 DEG C, collects the middle level liquid after second time is centrifuged, obtains pupa worm intestinal 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 combines class=6:5:3；

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

3, the preparation of modified dietary fiber

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

Inulin, Semen Tritici aestivi fiber, Herba avenae fatuae fiber 6:3:2 in mass ratio is uniformly mixed, add the water of its quality 5 times, room temperature 200W, 38KHz condition supersound extraction 13min, then in electric field intensity 30kV/cm, burst length 400 μ s, carries out high voltage pulse electric field processing 13min under the conditions of pulse frequency 300Hz；It is 5.5 with breast acid for adjusting pH value, adds the enzyme of mixture quality 0.2%, in 50 DEG C of enzymolysis 34min；Enzymolysis solution filters, and filtrate reduced in volume, lyophilization to moisture are 6.5% i.e. to obtain modified dietary fiber；

Above-mentioned enzyme is that cellulase, xylanase, laccase, pectase 2:2:1.5:1.5 in mass ratio uniformly mixes.

4, the preparation of Lactobacillus plantarum powder

Lactobacillus plantarum powder is prepared with Lactobacillus plantarum CGMCC NO.11763 according to a conventional method for starting strain, and its viable bacteria content is: 9 × 10¹²cfu/g；The cryoprotective agent that wherein freeze drying process uses is with the animal or plant containing antifreeze protein as raw material; prepared through high-pressure pulse electric extraction, ultrasonic assistant microwave extraction and compound enzyme enzymolysis, Lactobacillus plantarum powder can be effectively improved at freezing dry process Viable detection；

Protectant preparation method, comprises the steps:

Winter rye, Caulis et Folium Ammopiptanthi Mongolici, sharkskin collagen protein are respectively washed, drain, 9:4:3 in mass ratio uniformly mixes, add the lactic acid moistening 5h that pH value is 4.2 of mixed material quality 0.5 times, pulverize immediately after-20 DEG C of freezing 1.5h, freezing thickness of feed layer 4cm, ground product particle diameter 2mm, it is subsequently added into the water of ground product quality 15 times, it is 4.5 with breast acid for adjusting pH value, at room temperature in electric field intensity 30kV/cm, burst length 400 μ s, carries out high voltage pulse electric field processing 25min under the conditions of pulse frequency 250Hz；Then under the conditions of power 225W, carry out microwave irradiation and extraction 18min in room temperature, simultaneously at power 250W, under the conditions of frequency 35KHz, carry out ultrasonic assistant extraction；Add the compound enzyme of extracting solution quality 1.5%, in 50 DEG C of enzymolysis 40min；Enzymolysis solution filters, filtrate concentrates, low-temperature grinding to particle diameter is that 0.2mm i.e. obtains protective agent；

Above-mentioned compound enzyme is that cellulase, protease, amylase, pectase, tannase 3:2:2:1.5:1.5 in mass ratio uniformly mixes.

Used by following example two to six, plant extract, pupa worm intestinal extract, modified dietary fiber, Lactobacillus plantarum powder are prepared by embodiment one.

Embodiment two:

A kind of Haematocoocus Pluvialls bee pollen reducing cholesterol, mainly has the raw material of following parts by weight to prepare:

Bee pollen 165 parts, plant extract 20 parts, pupa worm intestinal extract 15 parts, modified dietary fiber 15 parts, oligosaccharide 25 parts, 8 parts of Lactobacillus plantarum powder, hydroxyl isomaltulose 4 parts, mango powder 4 parts, Haematocoocus Pluvialls 0.9 part；

Wherein bee pollen is Semen Sesami bee pollen；

The preparation method of a kind of Haematocoocus Pluvialls bee pollen reducing cholesterol, comprises the steps:

1) each supplementary material is weighed according to formula；

2) bee pollen is put in the ultrasonic washing unit equipped with 0.4% sodium bicarbonate solution, in 200W under room temperature, 40KHz cleans 4min, rinsing, drain, 6min is soaked in the sericin peptide taken solution that mass percent is 16%, take out, put-20 DEG C of freezing 8min, broken, immediately at power 4Kw, frequency 2450MHz, 130 DEG C of microwave heating 6s of temperature, add the water of former bee pollen quality 4 times, it is 5 with breast acid for adjusting pH value, in electric field intensity 30kV/cm, burst length 500 μ s, high voltage pulse electric field processing 13min is carried out under pulse frequency 300Hz room temperature condition, then room temperature 400W, 40KHz condition supersound process 13min obtains bee pollen pretreatment fluid；

3) bee pollen pretreatment fluid is warming up to 45 DEG C, is added thereto to plant extract and pupa worm intestinal extract, stirs, insulation, enzymolysis 13min, obtain bee pollen enzymolysis solution；

4) in bee pollen enzymolysis solution, oligosaccharide, the modified dietary fiber of 25% are added, fully dissolve 8min, it is naturally cooling to 43 DEG C, add 55% ferment at constant temperature 4h of Lactobacillus plantarum opaque amount, then with the ramp of 0.9 DEG C/min to 53 DEG C, add 45% ferment at constant temperature 2.5h of Lactobacillus plantarum opaque amount, finally it is 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 filtrate i.e. obtains broker wall bee pollen through ultrafiltration, concentrating under reduced pressure, lyophilization；

5) broker wall bee pollen is added V-type blending tank with hydroxyl isomaltulose, mango powder and Haematocoocus Pluvialls and uniformly mix 30min, speed of agitator 30r/min, it is subsequently adding residue modified dietary fiber and uniformly mixes 15min, speed of agitator 50r/min, sterile filling, seals, packs and i.e. obtain the Haematocoocus Pluvialls bee pollen that can reduce cholesterol；

Using broker wall bee pollen sporoderm-broken rate prepared by said method up to 99%, in Haematocoocus Pluvialls bee pollen, Lactobacillus plantarum viable count is up to 7.5 × 10¹¹cfu/g。

Embodiment three:

A kind of fish oil bee pollen reducing cholesterol, mainly has the raw material of following parts by weight to prepare:

Bee pollen 160 parts, plant extract 15 parts, pupa worm intestinal extract 13 parts, modified dietary fiber 13 parts, oligosaccharide 23 parts, 7 parts of Lactobacillus plantarum powder, hydroxyl isomaltulose 3 parts, mango powder 3 parts, 0.5 part of fish oil；

Wherein bee pollen is bee pollen of maize；

The preparation method of a kind of fish oil bee pollen reducing cholesterol, comprises the steps:

1) each supplementary material is weighed according to formula；

2) bee pollen is put in the ultrasonic washing unit equipped with 0.3% sodium bicarbonate solution, in 200W under room temperature, 40KHz cleans 3min, rinsing, drain, 5min is soaked in the sericin peptide taken solution that mass percent is 13%, take out, put-18 DEG C of freezing 5min, broken, immediately at power 3Kw, frequency 2450MHz, 120 DEG C of microwave heating 5s of temperature, add the water of former bee pollen quality 3 times, it is 4 with breast acid for adjusting pH value, in electric field intensity 20kV/cm, burst length 400 μ s, high voltage pulse electric field processing 10min is carried out under pulse frequency 200Hz room temperature condition, then room temperature 400W, 40KHz condition supersound process 10min obtains bee pollen pretreatment fluid；

3) bee pollen pretreatment fluid is warming up to 40 DEG C, is added thereto to plant extract and pupa worm intestinal extract, stirs, insulation, enzymolysis 10min, obtain bee pollen enzymolysis solution；

4) in bee pollen enzymolysis solution, oligosaccharide, the modified dietary fiber of 20% are added, fully dissolve 5min, it is naturally cooling to 40 DEG C, add 50% ferment at constant temperature 3h of Lactobacillus plantarum opaque amount, then with the ramp of 0.8 DEG C/min to 50 DEG C, add 40% ferment at constant temperature 1.5h of Lactobacillus plantarum opaque amount, finally it is 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 filtrate i.e. obtains broker wall bee pollen through ultrafiltration, concentrating under reduced pressure, lyophilization；

5) broker wall bee pollen is added V-type blending tank with hydroxyl isomaltulose, mango powder and fish oil and uniformly mix 25min, speed of agitator 20r/min, it is subsequently adding residue modified dietary fiber and uniformly mixes 12min, speed of agitator 40r/min, sterile filling, seals, packs and i.e. obtain the fish oil bee pollen that can reduce cholesterol；

Using broker wall bee pollen sporoderm-broken rate prepared by said method up to 98.5%, in fish oil bee pollen, Lactobacillus plantarum viable count is up to 8.5 × 10¹¹cfu/g。

Embodiment four:

A kind of Lepidinm meyenii Walp bee pollen reducing cholesterol, mainly has the raw material of following parts by weight to prepare:

Bee pollen 170 parts, plant extract 25 parts, pupa worm intestinal extract 17 parts, modified dietary fiber 17 parts, oligosaccharide 27 parts, 9 parts of Lactobacillus plantarum powder, hydroxyl isomaltulose 5 parts, mango powder 5 parts, pueraria root powder 0.8 part；

Wherein bee pollen is Brassica campestris L pollen；

The preparation method of a kind of Lepidinm meyenii Walp bee pollen reducing cholesterol, comprises the steps:

1) each supplementary material is weighed according to formula；

2) bee pollen is put in the ultrasonic washing unit equipped with 0.5% sodium bicarbonate solution, in 200W under room temperature, 40KHz cleans 5min, rinsing, drain, 7min is soaked in the sericin peptide taken solution that mass percent is 18%, take out, put-22 DEG C of freezing 10min, broken, immediately at power 5Kw, frequency 2450MHz, 140 DEG C of microwave heating 7s of temperature, add the water of former bee pollen quality 5 times, it is 6 with breast acid for adjusting pH value, in electric field intensity 40kV/cm, burst length 600 μ s, high voltage pulse electric field processing 15min is carried out under pulse frequency 400Hz room temperature condition, then room temperature 400W, 40KHz condition supersound process 15min obtains bee pollen pretreatment fluid；

3) bee pollen pretreatment fluid is warming up to 50 DEG C, is added thereto to plant extract and pupa worm intestinal extract, stirs, insulation, enzymolysis 15min, obtain bee pollen enzymolysis solution；

4) in bee pollen enzymolysis solution, oligosaccharide, the modified dietary fiber of 30% are added, fully dissolve 10min, it is naturally cooling to 45 DEG C, add 60% ferment at constant temperature 5h of Lactobacillus plantarum opaque amount, then with the ramp of 1.0 DEG C/min to 55 DEG C, add 50% ferment at constant temperature 3.5h of Lactobacillus plantarum opaque amount, finally it is cooled to 45 DEG C with the speed of 0.8 DEG C/min, continuing fermentation 1h, fermentation liquor 300 eye mesh screen filters, and filtrate i.e. obtains broker wall bee pollen through ultrafiltration, concentrating under reduced pressure, lyophilization；

5) broker wall bee pollen is added V-type blending tank with hydroxyl isomaltulose, mango powder and pueraria root powder and uniformly mix 35min, speed of agitator 40r/min, it is subsequently adding residue modified dietary fiber and uniformly mixes 18min, speed of agitator 60r/min, sterile filling, seals, packs and i.e. obtain the Lepidinm meyenii Walp bee pollen that can reduce cholesterol；

Using broker wall bee pollen sporoderm-broken rate prepared by said method up to 99.5%, in Lepidinm meyenii Walp bee pollen, Lactobacillus plantarum viable count is up to 6 × 10¹¹cfu/g。

Embodiment five:

A kind of plant sterol bee pollen reducing cholesterol, mainly has the raw material of following parts by weight to prepare:

Bee pollen 150 parts, plant extract 10 parts, pupa worm intestinal extract 10 parts, modified dietary fiber 10 parts, oligosaccharide 20 parts, 6 parts of Lactobacillus plantarum powder, hydroxyl isomaltulose 2 parts, mango powder 2 parts, plant sterol 0.5 part；

Wherein bee pollen is bee pollen of maize；

The preparation method of a kind of plant sterol bee pollen reducing cholesterol, comprises the steps:

1) each supplementary material is weighed according to formula；

2) bee pollen is put in the ultrasonic washing unit equipped with 0.3% sodium bicarbonate solution, in 200W under room temperature, 40KHz cleans 5min, rinsing, drain, 7min is soaked in the sericin peptide taken solution that mass percent is 13%, take out, put-18 DEG C of freezing 10min, broken, immediately at power 3Kw, frequency 2450MHz, 140 DEG C of microwave heating 5s of temperature, add the water of former bee pollen quality 5 times, it is 4 with breast acid for adjusting pH value, in electric field intensity 40kV/cm, burst length 400 μ s, high voltage pulse electric field processing 10min is carried out under pulse frequency 400Hz room temperature condition, then room temperature 400W, 40KHz condition supersound process 15min obtains bee pollen pretreatment fluid；

3) bee pollen pretreatment fluid is warming up to 40 DEG C, is added thereto to plant extract and pupa worm intestinal extract, stirs, insulation, enzymolysis 15min, obtain bee pollen enzymolysis solution；

4) in bee pollen enzymolysis solution, oligosaccharide, the modified dietary fiber of 20% are added, fully dissolve 10min, it is naturally cooling to 40 DEG C, add 60% ferment at constant temperature 3h of Lactobacillus plantarum opaque amount, then with the ramp of 1.0 DEG C/min to 50 DEG C, add 50% ferment at constant temperature 1.5h of Lactobacillus plantarum opaque amount, finally it is cooled to 40 DEG C with the speed of 0.8 DEG C/min, continuing fermentation 1h, fermentation liquor 100 eye mesh screen filters, and filtrate i.e. obtains broker wall bee pollen through ultrafiltration, concentrating under reduced pressure, lyophilization；

5) broker wall bee pollen is added V-type blending tank with hydroxyl isomaltulose, mango powder and plant sterol and uniformly mix 35min, speed of agitator 20r/min, it is subsequently adding residue modified dietary fiber and uniformly mixes 18min, speed of agitator 40r/min, sterile filling, seals, packs and i.e. obtain the plant sterol bee pollen that can reduce cholesterol；

Using broker wall bee pollen sporoderm-broken rate prepared by said method up to 99%, in plant sterol bee pollen, Lactobacillus plantarum viable count is up to 9 × 10¹¹cfu/g。

Embodiment six:

A kind of Cordyceps militaris (L.) Link. bee pollen reducing cholesterol, mainly has the raw material of following parts by weight to prepare:

Bee pollen 180 parts, plant extract 30 parts, pupa worm intestinal extract 20 parts, modified dietary fiber 20 parts, oligosaccharide 30 parts, 10 parts of Lactobacillus plantarum powder, hydroxyl isomaltulose 6 parts, mango powder 6 parts, Cordyceps militaris (L.) Link. 0.3 part；

Wherein bee pollen is Brassica campestris L pollen；

The preparation method of a kind of Cordyceps militaris (L.) Link. bee pollen reducing cholesterol, comprises the steps:

1) each supplementary material is weighed according to formula；

2) bee pollen is put in the ultrasonic washing unit equipped with 0.5% sodium bicarbonate solution, in 200W under room temperature, 40KHz cleans 3min, rinsing, drain, 5min is soaked in the sericin peptide taken solution that mass percent is 18%, take out, put-22 DEG C of freezing 5min, broken, immediately at power 5Kw, frequency 2450MHz, 120 DEG C of microwave heating 7s of temperature, add the water of former bee pollen quality 3 times, it is 6 with breast acid for adjusting pH value, in electric field intensity 20kV/cm, burst length 600 μ s, high voltage pulse electric field processing 15min is carried out under pulse frequency 200Hz room temperature condition, then room temperature 400W, 40KHz condition supersound process 10min obtains bee pollen pretreatment fluid；

3) bee pollen pretreatment fluid is warming up to 50 DEG C, is added thereto to plant extract and pupa worm intestinal extract, stirs, insulation, enzymolysis 10min, obtain bee pollen enzymolysis solution；

4) in bee pollen enzymolysis solution, oligosaccharide, the modified dietary fiber of 30% are added, fully dissolve 5min, it is naturally cooling to 45 DEG C, add 50% ferment at constant temperature 5h of Lactobacillus plantarum opaque amount, then with the ramp of 0.8 DEG C/min to 55 DEG C, add 40% ferment at constant temperature 3.5h of Lactobacillus plantarum opaque amount, finally it is 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 filtrate i.e. obtains broker wall bee pollen through ultrafiltration, concentrating under reduced pressure, lyophilization；

5) broker wall bee pollen is added V-type blending tank with hydroxyl isomaltulose, mango powder and Cordyceps militaris (L.) Link. and uniformly mix 25min, speed of agitator 40r/min, it is subsequently adding residue modified dietary fiber and uniformly mixes 12min, speed of agitator 60r/min, sterile filling, seals, packs and i.e. obtain the Cordyceps militaris (L.) Link. bee pollen that can reduce cholesterol；

Using broker wall bee pollen sporoderm-broken rate prepared by said method up to 99.5%, in Cordyceps militaris (L.) Link. bee pollen, Lactobacillus plantarum viable count is up to 8 × 10¹¹cfu/g。

Experimental example seven eats the change of T-CHOL after Haematocoocus Pluvialls bee pollen

Selecting the adult 48 of T-CHOL 180mg/dl-250mg/dl, men and women half and half, is randomly divided into three groups；150 milliliters of mineral waters are drunk in first group of dinner every day；Second group of dinner every day Instant Drinks common commercially available Haematocoocus Pluvialls bee pollen 150g, Haematocoocus Pluvialls bee pollen 150g of the 3rd group of dinner every day Instant Drinks embodiment of the present invention 2, eat same food every day period, and food includes meat, egg, vegetable and fruit.Gather the blood of experimenter respectively in the previous day and the 20th, 40,60 days that experiment starts, measure the total cholesterol level in blood, result such as table 5:

Table 5: total cholesterol level testing result in blood

Time	0 day	20 days	40 days	60 days
					First group (mg/dl)	202.3	203.8	206.2	209.3
Second group (mg/dl)	207.6	206.5	203.2	202.1
					3rd group (mg/dl)	211.8	202.9	191.6	179.3

As seen from the above table after the Haematocoocus Pluvialls bee pollen of the Instant Drinks embodiment of the present invention 2, the content generation significant change of the T-CHOL in adult's blood.Compared with common commercially available Haematocoocus Pluvialls bee pollen, the content of the T-CHOL in adult's blood is significantly reduced by Haematocoocus Pluvialls bee pollen of the present invention, and the content of the T-CHOL in mineral water composition year human blood significantly increases, although commercially available Haematocoocus Pluvialls bee pollen decreases, but compared with product of the present invention, effect is the most notable, it follows that the Haematocoocus Pluvialls bee pollen that the present invention uses the specific bacterial strain with reduction cholesterol characteristic to prepare has the health-care effect well reducing cholesterol.

It should be understood that the bee pollen of the reduced cholesterol prepared by embodiment of the present invention 3-6 has above-mentioned technique effect equally, between each embodiment, diversity is the most notable.

Embodiment eight Haematocoocus Pluvialls of the present invention bee pollen shelf-life implants living preparation of lactobacillus stable content is tested

The Haematocoocus Pluvialls bee pollen taking the embodiment of the present invention 2 preparation stores 3 months, 6 months, 12 months, 24 months, 36 months and measure Lactobacillus plantarum viable bacteria content under room temperature 22-25 DEG C, ventilation condition respectively, result such as table 6:

Table 6: shelf-life implants living preparation of lactobacillus content detection result

Result above shows: the storage stability of Haematocoocus Pluvialls bee pollen of the present invention is preferable, environment resistant (temperature, appropriateness, oxygen, illumination, moisture etc.) influence factor's ability is big, shelf-life Lactobacillus plantarum viable bacteria content loss rate maximum (36 months) is 15%, higher than existing like product viable bacteria content, loss rate is low, long shelf-life, reflects that other bioactive ingredients of Haematocoocus Pluvialls bee pollen has same storage stability simultaneously.

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

The sensory evaluation test of embodiment nine Haematocoocus Pluvialls of the present invention bee pollen

The Haematocoocus Pluvialls bee pollen inviting 24 personnel to prepare the embodiment of the present invention 2 is judged with the Haematocoocus Pluvialls bee pollen of commercially available two kinds of similar identical dates of manufacture, 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 includes outward appearance (20 points), quality (25 points), local flavor (30 points), four aspects of mouthfeel (25 points), and marking personnel independently carry out, are independent of each other, and judges result with guarantee accurate.Being added up judging result, equal score value takes approximation, retains integer, is specifically shown in Table 7:

Table 7: sensory evaluation statistical result

Note: show significant difference (P ＜ 0.05) with a line internal standard difference lowercase alphabet, the different capitalization of mark represents that difference is extremely notable (P ＜ 0.01), indicates same letter and represents that difference is not notable (P ＞ 0.05).

Result above shows, any one will be substantially better than commercially available Haematocoocus Pluvialls bee pollen to Haematocoocus Pluvialls bee pollen prepared by the present invention from outward appearance, quality, local flavor and mouthfeel, particularly outward appearance, local flavor and mouthfeel is fabulous, also is adapted for different age group, the consumer of different hierarchy of consumption eats simultaneously.

It should be understood that the bee pollen of reduced cholesterol 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 diversity.

The embodiment ten Haematocoocus Pluvialls of the present invention probiotic test of bee pollen

The Haematocoocus Pluvialls bee pollen embodiment of the present invention 2 prepared, being prepared as viable count with sterilized water is 2 × 10¹⁰The 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, uses ten times of stepwise dilutions to 10^-8, take 1mL diluent on flat board, the MRS agar culture medium being cooled to 45 DEG C poured on flat board (sterilizing), shake up rapidly after sterilizing.The test tube 2 that will be equipped with 10mL Lactobacillus plantarum solution again is placed in 80-90 DEG C of water-bath heating 15-25min, takes the Lactobacillus plantarum solution after heating and carries out ten times of stepwise dilutions to 10^-8, take 1mL diluent on flat board, the MRS agar culture medium being cooled to 45 DEG C poured into upper at flat board (sterilizing) and shake up rapidly after sterilizing.Finally the flat board before heating and after heating is all cultivated under the conditions of 35 DEG C 24h, calculates the quantity before and after heating.

Result shows, Viable detection has reached more than 88%.

2, simulated gastric fluid and the resistance test of intestinal juice: the hydrochloric acid 16.4mL taking 100g/L adds distilled water diluting, pH value is made to be respectively 1.5,2.5 and 3.5, take 100mL dilute hydrochloric acid solution, it is separately added into 1g pepsin, it is made fully to dissolve, obtaining simulated gastric fluid, microporous filter membrane degerming (0.22 μm) is standby.Taking potassium dihydrogen phosphate 6.8g, the 500mL that adds water makes dissolving, with 0.1moL/L sodium hydroxide solution regulation pH value to 6.8；Separately taking trypsin 10g, the 100mL that adds water makes dissolving, after two liquid mixing, is diluted with water to 1000ml, obtains simulated intestinal fluid, and microporous filter membrane degerming (0.22 μm) is standby.Take the Lactobacillus plantarum solution that 1mL keeps to join in the simulated gastric fluid of 9mL (i.e. ten times stepwise dilutions), and the most fully mix rapidly, be subsequently placed in 30-45 DEG C of quiescent culture 2-4h.Taking out culture fluid respectively 1h, 2h, 3h, 4h when and count remaining viable count immediately, comparing with former viable count, result shows, Viable detection is 98%.Then each 1mL of culture fluid digesting different time it is taken in simulated gastric fluid, it is inoculated in respectively in the simulated intestinal fluid that 9mL pH value is 6.8, it is placed in 30-45 DEG C of quiescent culture 2-4h, and respectively 0,3,6,24h sampling, measure its viable count, comparing with former viable count, result shows that Viable detection is 99%.

3, simulating the resistance test of cholate: make the solution of 1g/L with pancreatin, and add the Fel Sus domestica salt of 0.8% in the solution, the NaOH adjustment pH with 10% is 8.0, then with 0.45 μm micro-filtrate membrane filtration degerming.The Lactobacillus plantarum solution inoculum kept by 0.5mL is simulated in cholate to 4.5mL, obtains culture fluid, the viable count of counting remaining after cultivating 24h.By culture fluid in sterile saline ten times of stepwise dilutions to 10^-8, and pour into MRS, it is subsequently placed in 35 DEG C of quiescent culture 24h.Result shows that Viable detection is 99%.

Above result of the test shows, Lactobacillus plantarum probiotic (thermostability, resistance to pH, bile tolerance) in Haematocoocus Pluvialls bee pollen of the present invention is stronger, being especially suitable for human intestines and stomach's environment, in simulated gastrointestinal environments, survival rate is big, can be effectively improved gastrointestinal function.

Embodiment 11 Haematocoocus Pluvialls of the present invention bee pollen mouse intestinal performance test

Choosing common Kunming white mice 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, prepare the mouse model of alteration of intestinal flora.Model group mouse diet declines, and dead and phenomenon of significantly suffering from diarrhoea does not occurs, arranges soft excrement, and profile normal aqueous divides more, and bedding and padding are moist.By 40 alteration of intestinal flora mices, being randomly divided into 2 groups, one group 20 is only used as treatment group, the Lactobacillus plantarum solution 0.5ml (2 × 10 that every day, gavage was kept¹⁰Cfu/ml)/only, another 20 are only used 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 white mice every day, weigh in the 8th, 21 days mices to Haematocoocus Pluvialls bee pollen treatment group and natural recovering group, calculate each group of weight average rate of increase, result such as table 8；Within every 5 days, survey each group of stool in mice escherichia coli quantity, calculate average, result such as table 9.Take stool in mice about 0.1g, in aseptic operating platform add 3 beades (adding 0.5mL diluent with 0.1g excrement sample), dilute and inoculate maconkey agar culture medium, calculate every gram wet just in coliform count.

Table 8: mice Gain weight

Packet	Average starting weight (g/ is only)	Average end weight (g/ is only)	Average rate of increase (%)
				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 9: the situation of coliform count in stool in mice

Haematocoocus Pluvialls bee pollen treatment group Mouse Weight average rate of increase (67.96%) is significantly higher than natural recovering group (32.14%)；Feed solution rear intestinal escherichia coli quantity to be remarkably decreased, reduce by 97.07%, it is substantially less than natural recovering group (24.78%), show the rapid field planting in white mice intestinal of the Lactobacillus plantarum in Haematocoocus Pluvialls bee pollen of the present invention, form dominant microflora, and the growth and breeding of effective suppression pathogen such as escherichia coli, and resident time is long, continues, effectively improves intestinal performance.

Embodiment 12 present invention can reduce the bee pollen impact on immunity of organisms of cholesterol

1 experiment purpose

Testing (mouse forced swimming) by exercise tolerance, the checking present invention can reduce the raising immunity of bee pollen of cholesterol, antifatigue effect.

2 experiment materials and reagent

2.1 for reagent thing:

The commercially available bee pollen (G1) reducing cholesterol；The commercially available bee pollen (G2) reducing cholesterol；The bee pollen (G3-G7) of reduced cholesterol 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；Concentrated sulphuric acid (AR), Nanjing Chemistry Reagent Co., Ltd.；Normal saline, Shangdong Changfu Jiejing Pharmaceutical Industry Co., Ltd..

3. laboratory animal

ICR mice, ♂, cleaning grade, body weight 18-22g, Ningxia Medical University's comparative medicine center provide, the free diet of mice during experiment.

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, upper Nereid grand experimental facilities company limited；Electronic scale: BS224S type, Sartorius company；Stopwatch, thermometer

5. experiment packet

5.1 dosage packets and given the test agent give the time and at random mice are divided into 8 groups, often group 10,1st group to the 7th group medicine giving G1～G7 respectively, 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 are to the 7th group: weigh 2.25g drug sample, be assigned to 150ml with distilled water；Blank group: distilled water 150ml.

6. experimental technique

6.1 swimmings with a load attached to the body experiment lasts are administered after 30min, put mice in swimming trunk, and the depth of water is no less than 30cm, water temperature 25 ± 1 DEG C, the sheet lead of rat-tail root load 5% body weight, and record mice swimming starts to the dead time, as mice swimming time.

After 6.2 mice serum carbamide measure last administration 30min, not swimming with a load attached to the body 90min in the water that temperature is 30 DEG C, eyeball blood sampling 0.5mL (being not added with anticoagulant) is plucked after rest 60min, put 4 DEG C of refrigerator 3h, after hemopexis, 2000r/min is centrifuged 15min, takes serum and send clinical laboratory of Affiliated Hospital of Ningxia Medical University to detect.

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

The mensuration last of 6.4 blood lactase acid is taken a blood sample after being administered 30min, does not the most bear a heavy burden and stops after the water went swimming 10min that temperature is 30 DEG C.Lactic acid instrument assay method: respectively before swimming, each blood sampling 20 μ L add in 40 μ L rupture of membranes liquid after rest 20min after swimming, after swimming, the most fully vibration smudge cells lactic acid instrument measures.(blood lactase acid area under curve=5 × (the blood lactase acid value of 20min after blood lactase acid value+2 × swimming of 0min after swimming front blood lactase acid value+3 × swimming)

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

8. statistical method experimental data is usedRepresent, use t inspection to compare between organizing

9. experimental result

9.1 present invention can reduce the bee pollen impact on Mouse Weight of cholesterol

Each group mice is after giving G1～G9 medicine, before, in, shown in post-weight see table respectively, original body mass and the weightening finish body weight of each group mice compare equal no difference of science of statistics (P ＞ 0.05) with matched group, show that G1～G9 medicine is all without obvious toxicity.Experimental result refers to table 10.

The original body mass of table 10 swimming with a load attached to the body experiment mice, body weight in mid-term and end body weight

9.2 present invention can reduce the impact on the mice burden swimming time of the bee pollen of cholesterol

After per os gives mice G1～G7 medicine, G1～G2 medicine compares with blank group, the mice burden swimming time can be obviously prolonged, there is significant difference (P ＜ 0.05), the present invention can reduce the bee pollen G3 of cholesterol～G7 medicine compares with blank group, can significantly extend the mice burden swimming time, there is pole significant difference (P ＜ 0.01), and be substantially better than G1～G2 medicine.The results detailed in Table 11.

Table 11 can reduce the impact on the mice burden swimming time of the bee pollen of cholesterol

" * " p < 0.05vs blank；

" * * " p < 0.01vs blank；

9.3 present invention can reduce the bee pollen of cholesterol to the impact of blood lactase acid before and after mouse movement

After the bee pollen of the reduced cholesterol that per os gives the mice present invention, the present invention can reduce the bee pollen G3 of cholesterol～G7 medicine and compare blood lactase acid area under curve after mouse movement with matched group and have significant difference (P ＜ 0.05), decrease though G1～G2 medicine group Mouse Blood lactic acid area under curve compares with matched group, but and no difference of science of statistics (P ＞ 0.05).The results are shown in Table 12.

Table 12 present invention can reduce the bee pollen of cholesterol to the impact of blood lactase acid level before and after mouse movement

" * " p < 0.05vs blank；

9.4 the present invention can reduce the impact on Mouse Liver glycogen of the bee pollen of cholesterol

After per os gives mice G1～G7 medicine, G1～G2 medicine compares with blank group, Mouse Liver glycogen content all has significantly rising, there is significant difference (P ＜ 0.05), the present invention can reduce the bee pollen G3 of cholesterol～G7 medicine compares with blank group, Mouse Liver glycogen content all has significantly rising, has pole significant difference (P ＜ 0.01), and is substantially better than G1～G2 medicine.The results detailed in Table 13.

Table 13 present invention can reduce the impact on Mouse Liver glycogen content of the bee pollen of cholesterol

" * " p < 0.05vs blank；

" * * " p < 0.01vs blank；

9.5 present invention can reduce the impact on mice serum carbamide of the bee pollen of cholesterol

After per os gives mice G1～G7 medicine, G1～G2 medicine group compares with blank group, after mouse movement, serum urea content all has significantly reduction, there is significant difference (P ＜ 0.05), the present invention can reduce the bee pollen G3 of cholesterol～G7 medicine compares with blank group, after mouse movement, serum urea content all has significantly reduction, has pole significant difference (P ＜ 0.01), and is substantially better than G1～G2 medicine.The results detailed in Table 14.

Table 14 present invention can reduce the impact on mice serum urea content of the bee pollen of cholesterol

" * " p < 0.05vs blank；

" * * " p < 0.01vs blank；

10. experiment conclusion

This experiment is mainly tested by mice burden swimming, simultaneously detection Mouse Liver glycogen deposit observe the present invention can reduce cholesterol bee pollen improve immunity, the effect of resisting fatigue.Preliminary Results shows as follows:

1, G3～G7 of the present invention can reduce the bee pollen of cholesterol and all can extend the mice burden swimming time (P ＜ 0.01), and effect is substantially better than the bee pollen of reduced cholesterol of other G1～G2.

2, biochemistry detection aspect shows, the bee pollen each dosage group of cholesterol of can reducing G3～G7 of the present invention all can reduce move after lactic acid content produced by glucose anerobic glycolysis in mice serum, compare with matched group and have significant difference (P ＜ 0.05), and although the bee pollen of the reduced cholesterol of other G1～G2 also can reduce after motion lactic acid content produced by glucose anerobic glycolysis in mice serum, but compare with matched group, no difference of science of statistics (P ＞ 0.05)；

3, G3～G7 of the present invention can reduce bee pollen each dosage group of cholesterol and all can significantly improve the deposit (P ＜ 0.01) of glycogen in mouse liver, and effect is substantially better than the bee pollen of reduced cholesterol of other G1～G2；

4, metabolic arthritis model finds, G3～G7 of the present invention can reduce the bee pollen of cholesterol can significantly reduce the content (P ＜ 0.01) of urea in serum after mice is swum, and effect is substantially better than other G1～G2 and can reduce the bee pollen of cholesterol；

11. conclusions

The above-mentioned experiment proof present invention can reduce the bee pollen of cholesterol can significantly improve immunity of organisms, improve muscle power and the endurance of mice, reduce urea in serum and the content of lactic acid after mouse movement, and the deposit of glycogen in mouse liver can be significantly improved, contribute to alleviating the fatigue that sports load causes；The time that mice burden swimming to power exhausts can be extended.

Claims

1. can reduce a bee pollen for cholesterol, mainly have the raw material of following parts by weight to prepare: bee pollen 150-180 part, plant Extract 10-30 part, pupa worm intestinal extract 10-20 part, modified dietary fiber 10-20 part, oligosaccharide 20-30 part, plant Thing lactobacillus powder 6-10 part, hydroxyl isomaltulose 2-6 part, mango powder 2-6 part；

The described bee pollen reducing cholesterol also includes one or more adjuvants 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, plant sterol 0.2-0.8 part, phosphatidyl Serine 0.1-0.7 part, oligochitosan 0.1-0.7 part, marine fishbone collagen oligopeptide powder 0.1-0.7 part, Chinese herbal medicine 0.1-0.6 Part, Cordyceps militaris (L.) Link. 0.1-0.5 part；Soybean protein isolate 0.1-0.5 part；

Described Lactobacillus plantarum powder is to prepare according to a conventional method with Lactobacillus plantarum CGMCC NO.11763 for starting strain.

The bee pollen of cholesterol can be reduced the most as claimed in claim 1, it is characterised in that the preparation side of described pupa worm intestinal extract Method, comprises the steps: the drone pupa intestinal 3-5:1-3 in mass ratio of the queen bee nit intestinal by 2-3 age in days and 11-12 age in days Mixing, adds the sericin peptide taken solution that mass percent is 8-12% of mixture quality 1-2 times, stirs, put-18-22 DEG C 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, under the conditions of 4 DEG C, centrifugal 20-30min, collects the middle level liquid after being centrifuged for the first time, then exists 18000-25000g, centrifugal 20-30min under the conditions of 4 DEG C, collect the middle level liquid after second time is centrifuged, obtain pupa worm intestinal and carry Take thing.

The bee pollen of cholesterol can be reduced the most as claimed in claim 2, it is characterised in that the pH value of described sericin peptide taken solution is 6-7.

The bee pollen of cholesterol can be reduced the most as claimed in claim 2, it is characterised in that the quality group of described biological demulsifying agent becomes: Glycolipid class: lipopeptid class: cell wall combines class=5-7:4-6:2-4.

The bee pollen of cholesterol can be reduced the most as claimed in claim 4, it is characterised in that the quality of described glycolipid class biological demulsifying agent Consist of: rhamnolipid: alkyl polyglucoside=3-5:1-2.

The bee pollen of cholesterol can be reduced the most as claimed in claim 1, it is characterised in that the preparation method of described plant extract is: Fructus Hordei Germinatus and Fructus Tritici aestivi 8-10:1-3 in mass ratio are uniformly mixed, is crushed to granularity 0.5-1mm, obtains pulverizing Fructus Hordei Germinatus；Then will Fructus Chaenomelis, Fructus Ananadis comosi, Fructus Fici in ultrasonic washing unit under the conditions of power 200W, frequency 30KHz ultrasonic cleaning 5-10min, Drain, be crushed to granularity 0.5-1mm under room temperature, and 7-9:1-3:1-2 uniformly mixes in mass ratio, add mixture quality 3-5 Pulverizing Fructus Hordei Germinatus again obtains raw mixture, adds the water of raw mixture quality 1-3 times, is 3-4 with Fructus Citri Limoniae acid for adjusting pH value, Microwave extraction, wherein, each microwave exposure total time 60-80s is carried out under the conditions of power 150-300W, frequency 2000Hz, Carry out compartment irradiation: irradiation 10s, be spaced 10s, control temperature 20-35 DEG C, such irradiation 10 times, simultaneously at power 200-300W, Ultrasonic assistant extraction is carried out under the conditions of frequency 30-40KHz；Insulation 1-3h, then, in power 200-400W, frequency 2000Hz Under the conditions of carry out microwave extraction, wherein, each microwave exposure total time 90-105s, carry out compartment irradiation: irradiation 15s, Every 10s, controlling temperature 40-60 DEG C, such irradiation 10 times, simultaneously at power 300-500W, under the conditions of frequency 40-50KHz Carry out ultrasonic assistant extraction, be finally naturally cooling to room temperature, in electric field intensity 25-35kV/cm, burst length 400-600 μ s, Carry out high-pressure pulse electric under the conditions of pulse frequency 200-300Hz and extract 15-20min；Extracting solution filters to obtain the first filtrate, adds filter The water that slag is 4 times rinses, filters to obtain the second filtrate, the first filtrate and the second filtrate 1:3-5 is in mass ratio uniformly mixed, decompression Being concentrated into solid content is more than 20%, obtains plant extract.

The bee pollen of cholesterol can be reduced the most as claimed in claim 1, it is characterised in that the preparation method of described modified dietary fiber, Comprise the following steps: inulin, Semen Tritici aestivi fiber, Herba avenae fatuae fiber 5-7:2-4:1-3 in mass ratio is uniformly mixed, adds its quality The water of 3-7 times, room temperature 100-300W, 35-40KHz condition supersound extraction 10-15min, then in electric field intensity 20-40kV/cm, Burst length 300-500 μ s, carries out high voltage pulse electric field processing 10-15min under the conditions of pulse frequency 200-400Hz；Adjust with lactic acid Joint pH value is 4.5-6.5, adds the enzyme of mixture quality 0.1-0.3%, in 45-55 DEG C of enzymolysis 20-48min；Enzymolysis solution Filtering, filtrate reduced in volume, lyophilization to moisture are that 5-8% i.e. obtains modified dietary fiber；

The bee pollen of cholesterol can be reduced the most as claimed in claim 1, it is characterised in that described Chinese herbal medicine be with Flos Chrysanthemi, Flos Sophorae, Radix Ginseng, Bulbus Lilii, Radix Puerariae, Fructus Lycii, Herba Taraxaci, Rhizoma Dioscoreae, Poria, Fructus Hippophae, Flos Rosae Rugosae, Flos Lonicerae, Herba Menthae, Fructus Momordicae, Ah One or more in glue, Fructus Jujubae, Folium Mori, Fructus Momordicae charantiae, Agaricus blazei Murrill are prepared through micronizing.

The preparation method of the bee pollen of cholesterol can be reduced the most as claimed in claim 1, it is characterised in that comprise the steps:

1) each supplementary material is weighed according to formula；

2) bee pollen is put in the ultrasonic washing unit equipped with 0.3-0.5% sodium bicarbonate solution, in 200W, 40KHz under room temperature Clean 3-5min, rinsing, drain, the sericin peptide taken solution that mass percent is 13-18% soaks 5-7min, takes out, put -18-22 DEG C of freezing 5-10min, broken, immediately at power 3-5Kw, frequency 2450MHz, temperature 120-140 DEG C microwave heating 5-7s, adds the water of former bee pollen quality 3-5 times, is 4-6 with breast acid for adjusting pH value, in electric field intensity 20-40kV/cm, arteries and veins Rush time 400-600 μ s, under pulse frequency 200-400Hz room temperature condition, carry out high voltage pulse electric field processing 10-15min, then room Temperature 400W, 40KHz condition supersound process 10-15min obtains bee pollen pretreatment fluid；

3) bee pollen pretreatment fluid is warming up to 40-50 DEG C, is added thereto to plant extract and pupa worm intestinal extract, stirring Uniformly, insulation, enzymolysis 10-15min, obtain bee pollen enzymolysis solution；

4) in bee pollen enzymolysis solution, add oligosaccharide, the modified dietary fiber of 20-30%, fully dissolve 5-10min, naturally fall Temperature, to 40-45 DEG C, adds the 50-60% ferment at constant temperature 3-5h of Lactobacillus plantarum opaque amount, then with the speed of 0.8-1.0 DEG C/min Rate is warming up to 50-55 DEG C, add Lactobacillus plantarum opaque amount 40-50% ferment at constant temperature 1.5-3.5h, finally with The speed of 0.6-0.8 DEG C/min is cooled to 40-45 DEG C, continues fermentation 0.5-1h, and fermentation liquor 100-300 eye mesh screen filters, filter Liquid i.e. obtains broker wall bee pollen through ultrafiltration, concentrating under reduced pressure, lyophilization；

5) broker wall bee pollen is added V-type blending tank with hydroxyl isomaltulose, mango powder and adjuvant and uniformly mix 25-35min, stirring Rotating speed 20-40r/min, is subsequently adding residue modified dietary fiber and uniformly mixes 12-18min, speed of agitator 40-60r/min, aseptic Fill, seal, pack and i.e. obtain the bee pollen that can reduce cholesterol.

The preparation method of the bee pollen of cholesterol can be reduced the most as claimed in claim 9, it is characterised in that described Lactobacillus plantarum powder The preparation method of cryoprotective agent in preparation process, comprises the steps: winter rye, Caulis et Folium Ammopiptanthi Mongolici, shark Collagen protein of fish skin is respectively washed, drains, and 8-10:3-5:2-4 in mass ratio uniformly mixes, and adds mixed material quality 0.1-1 The lactic acid moistening 3-8h that pH value is 3.8-4.5 again, pulverizes after-18--22 DEG C of freezing 1-2h immediately, and the freezing bed of material is thick Degree 3-5cm, ground product particle diameter 0.5-3mm, be subsequently added into the water of ground product quality 10-20 times, with breast acid for adjusting pH value is 3.5-5.5, at room temperature in electric field intensity 25-35kV/cm, burst length 300-500 μ s, pulse frequency 200-300Hz condition Under carry out high voltage pulse electric field processing 20-30min；Then under the conditions of power 150-300W, microwave irradiation and extraction is carried out in room temperature 15-20min, simultaneously at power 200-300W, carries out ultrasonic assistant extraction under the conditions of frequency 30-40KHz；Add extracting solution The compound enzyme of quality 1-2%, in 45-55 DEG C of enzymolysis 30-50min；Enzymolysis solution filters, filtrate concentrates, low-temperature grinding to particle diameter is 0.1-0.3mm i.e. obtains protective agent；

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