CN106366206A - Method for preparing radix-psuedostellariae-derived polysaccharides by steam explosion-bioenzyme combined technique - Google Patents
Method for preparing radix-psuedostellariae-derived polysaccharides by steam explosion-bioenzyme combined technique Download PDFInfo
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Abstract
The invention belongs to the field of preparation of radix psuedostellariae polysaccharides, and particularly discloses a method for preparing radix-psuedostellariae-derived polysaccharides by a steam explosion-bioenzyme combined technique. The method comprises the following eight steps: radix psuedostellariae pretreatment, double-stage steam explosion treatment, bioenzyme enzymolysis, hot water extraction, extract CO2 treatment, three-stage filtration, detoxification and alcohol precipitation. The double-stage steam explosion technique well achieves the effect of radix psuedostellariae extraction pretreatment, and can reduce and even remove the toxic substances generated by steam explosion. The nonionic surfactant introduced into the bioenzyme enzymolysis step can enhance the enzymolysis efficiency and reduce the addition of the external enzyme, thereby relieving the work load of subsequent treatment. The preparation method is suitable for preparing and producing radix-psuedostellariae-derived polysaccharide substances, and has important popularization meanings.
Description
Technical field
The invention belongs to Pseudostellaria Polysaccharide preparation field is and in particular to a kind of steam explosion combination biological enzyme technology prepares Radix Pseudostellariae
The method of source polysaccharide.
Background technology
Radix Pseudostellariae begin be loaded in " Bencao Congxin ", be the tuber of pinkwort Radix Ginseng, main product in Fujian, Jiangsu, Anhui,
Shandong etc. saves.2010 editions " Chinese Pharmacopoeia " description Radix Pseudostellariae sweet in the mouth, slight bitter, mild-natured；Returns spleen, lung meridian；There is replenishing QI to invigorate the spleen, promote the production of body fluid
The effect of lung moistening；Tired, inappetence, eak after being ill, deficiency of both vital energy and Yin for insufficiency of the spleen body, spontaneous perspiration is thirsty, the dryness of the lung.Radix Pseudostellariae system health
The kind of " can be used for the article list of health food " that portion promulgates, the research to Radix Pseudostellariae chemical composition shows, Radix Pseudostellariae contains
There are aminoacid, saccharide, glycoside, phospholipid, cyclic peptide, fatty acid, oils, volatile ingredient, sterols and micro unit
Element etc., wherein Pseudostellaria Polysaccharide are important component of Radix Pseudostellariae biological active agents.Modern pharmacology research proves: too
Sub- gracilis polysaccharide has enhancing immunologic function, improves the effect such as ability of learning and memory, resisting fatigue, slow down aging, antiinflammatory.
At present people to the utilization of Radix Pseudostellariae also in the starting exploratory stage, also constantly deep to the pharmacological research of Radix Pseudostellariae
Enter.If developing a kind of preparation method of Pseudostellaria Polysaccharide possess notable meaning.
Content of the invention
For overcoming above-mentioned weak point present in prior art, the purpose of the present invention aims to provide a kind of steam explosion combination life
The method that thing zymotechnic prepares Radix Pseudostellariae source polysaccharide.
For achieving the above object, the technical scheme that the present invention takes is as follows:
A kind of steam explosion is combined the method that biological enzyme technology prepares Radix Pseudostellariae source polysaccharide, and step is as follows:
S1, Radix Pseudostellariae pretreatment: five operations must be baked, cut into slices and steamed to drying Radix Pseudostellariae, after selecting, going successively；
Wherein, selecting is to be screened Radix Pseudostellariae drying then, and the Radix Pseudostellariae going mouldy or have impurity is rejected；Go must refer to adopt
Radix Pseudostellariae cercus is removed and separates by plant equipment；Baking refers to remove crown prince again using low temperature evacuation baking technology
Ginseng surface moisture is easy to follow-up section, 50 ~ 60 DEG C of baked temperature, baking time 10min ~ 20mim, and vacuum pressure 0.03 ~
0.05mpa；Section refers to using Chinese herbal medicine microtome, the Radix Pseudostellariae after baking be carried out the section that Radix Pseudostellariae joins body, after section
Radix Pseudostellariae piece thickness is 0.5 ~ 1.0mm；Steaming is Radix Pseudostellariae piece to be placed in food steamer process, and steams using vapor heating, steams
Time processed is 20 ~ 30min；
S2, double sections of Steam explosion treatment: the Radix Pseudostellariae piece after to steam as blasting substrates, successively pass through Steam explosion treatment i and
Steam explosion treatment ii, double sections of Steam explosion treatment interval times are 2 ~ 5min；The condition of Steam explosion treatment i is: explosion chamber
Coefficient is 0.7 ~ 0.9, and explosion steam pressure reaches 0.5 ~ 0.8mpa, and explosion pressurize process time is 10 ~ 15min, during explosion
Between not higher than 0.00875s；The condition of Steam explosion treatment ii is: the same cavity explosion treatment of blasting gear on the same stage, explosion steam
Pressure reaches 1.5 ~ 2.2mpa, and explosion pressurize process time is 30 ~ 50s, and blasting time is not higher than 0.00875s；Double sections of steam explosions
Entrainer is all added, entrainer is detailed catalogue corundum and ca (oh) in processing procedure2The complex of powder, corundum: ca (oh)2Powder
Mass values be 4:1 ~ 1:1；By percentage to the quality, entrainer total addition level accounts for the 1.5 ~ 3% of blasting substrates, and adds in batches
Plus, the 1/3 ~ 2/5 of Steam explosion treatment i total amount of adding, Steam explosion treatment ii add the entrainer of surplus；
S3, enzyme enzymolysis: by after explosion treatment and temperature is down to Radix Pseudostellariae material substrate during room temperature and water with solid-to-liquid ratio
1kg:1l ~ 1kg:3l mixes, and the ph value of solidliquid mixture is adjusted to 4.5 ~ 5.0, then disposably adds non-ionic surface to live
Property agent, adding cellulase, xylanase and pectase is digested, 50 ~ 55 DEG C of hydrolysis temperature, enzymolysis time 30 ~
45min, the stage digesting front 20 ~ 25min is at the uniform velocity stirred (rotating speed 100 ~ 120rpm/min), adjusts mixed at the end of waiting to digest
The ph value of compound is to neutrality；Wherein, in terms of every g drying Radix Pseudostellariae piece (drying Radix Pseudostellariae piece be s1 section after, steam before crown prince
Ginseng piece), the addition of cellulase, xylanase, pectase and nonionic surfactant be respectively as follows: 10 ~ 15fpu/g, 100
~150u/g、80~110u/g、0.003~0.05g/g；Described nonionic surfactant is berol, peg, tween-80 and bsa
One or more of, ratio during multiple use preferably waits mass mixing；
S4, water hot extraction: by after explosion treatment Radix Pseudostellariae substrate collect after be dosed in water hot extraction's tank, in tank hot water amount with
The mass values of Radix Pseudostellariae substrate are 15:1 ~ 20:1, extract 90 ~ 100 DEG C of hot water temperature, extraction time 2 ~ 3h, extraction times 1
Secondary；Phase is at the uniform velocity stirred before extraction, to be extracted carry out to during 3/5 extraction time stop stirring；
S5, extract co2Process: after water hot extraction terminates, first with heat exchanger, the extract obtaining is cooled to room temperature,
Then move in the rustless steel container of cleaning, then in the extract after cooling, be passed through aseptic co2Gas, co2Flow velocity be 10 ~
15m3/ min, is passed through co2Gas time 5 ~ 15min；It is passed through co2During gas, co2Gas is by pipeline through rustless steel container bottom
Gas distributor enters inside extract；
S6, three-stage filtration: extract co2Respectively through the grade one filter of tandem compound, secondary filter and three-level after process
Filter is realizing filtration operation；Wherein, grade one filter and secondary filter are conventional cartridge filter, grade one filter
It is respectively 4.5 ~ 5 μm, 0.2 ~ 0.25 μm with the filter element aperture of secondary filter；Tertiary filter is hollow fiber filter, in
The molecular cut off of hollow fiber is 10000 ~ 30000 dalton；
S7, detoxification: the Radix Pseudostellariae extract priority after three-stage filtration is passed through d301 resin packed column and the filling of nkaii resin
Post, extract enters post pressure and is 0.15 ~ 0.2mpa, collects extract；
S8, precipitate with ethanol: first the Radix Pseudostellariae extract after detoxification is carried out decompression half and concentrate, concentration pressure is 0.01 ~ 0.05mpa,
50 ~ 70 DEG C of thickening temperature, concentration time 2 ~ 3h；It is followed by half concentrated solution that reduces pressure being cooled to 10 ~ 25 DEG C and being added thereto to 2 ~ 3
95% edible ethanol liquid of times volume, stands 8 ~ 10h, and finally centrifugation obtains precipitate and is Radix Pseudostellariae source polysaccharide；Wherein, it is centrifuged
Rotating speed be 10000 ~ 15000 rpm/min, centrifuging temperature temperature control at 10 ~ 15 DEG C, centrifugation time 15 ~ 25min.
Preferably, in s6 using self-control three-level filter system carry out three-stage filtration, this three-level filter system include tubular type from
Scheming, centrifugation liquid storage tank (in tank, temperature controlling range is 5～10 DEG C), power transmitting assembly i, power transmitting assembly ii, power
Transmission assembly iii, grade one filter, secondary filter, tertiary filter and its extraneous subsystem；Power transmitting assembly i by
Kinetic pump i and check-valves i is connected in series；Power transmitting assembly ii is connected in series by kinetic pump ii and check-valves ii；Power transmits
Assembly iii is connected in series by kinetic pump iii and check-valves iii；Grade one filter and secondary filter are conventional cartridge filter；
Tertiary filter is self-control filter, and its structure includes cylinder, doughnut tread assembly, pedestal；Cylinder body bottom and top are respectively
It is provided with inlet and liquid outlet, outside cylinder, is provided with interlayer, the upper and lower ends of interlayer are respectively equipped with interlayer import and interlayer outlet；
Doughnut tread assembly includes being evenly equipped with the package shell of porous, some doughnuts (preferably 300 ~ 600), these hollows
Fiber merges bunchy and is placed in inside package shell；Pedestal is cylinder, and cylinder epimere is cylindrical in shape for hollow, middle hypomere is solid,
The middle hypomere of pedestal is solid to be provided with the through hole being adapted with doughnut quantity and its size；Pedestal be located at liquid outlet below,
It is close to cylinder roof and be secured to connect；The upper end of doughnut is uncovered and is located in the through hole of pedestal, and lower end closed is simultaneously
It is fixedly connected with package shell diapire；The extraneous subsystem of tertiary filter includes kinetic pump iv, check-valves iv, constant temperature refrigeration
Water tank (controllable temperature scope is: 5～15 DEG C), interlayer outlet contacts check-valves iv, constant temperature refrigeration water tank and kinetic pump iv extremely successively
Interlayer import forms a closed cycle system；Tube centrifuge is contacted successively by pipeline and is centrifuged liquid storage tank, kinetic pump i, non-return
Valve i, grade one filter, kinetic pump ii, check-valves ii, secondary filter, kinetic pump iii, check-valves iii are to tertiary filter
Inlet；Grade one filter and the filter element aperture with secondary filter are respectively 4.5 ~ 5 μm, 0.2 ~ 0.22 μm；Doughnut
Molecular cut off is 10000 ~ 30000 dalton；
Three-stage filtration step is:
Extract a s5 is processed by () first after inputs in tube centrifuge inner chamber, starts this equipment and carries out centrifugal work, centrifugation
Turn up 4000 ~ 6000rpm/min, is centrifuged 20 ~ 30min；Centrifugation proceeds to the centrifugal clear liquid of acquisition to centrifugation after terminating in time
In liquid storage tank, and in this tank, temperature controlling range is 5 ~ 10 DEG C；
B the cooling centrifugal liquid that upper operation obtains is crossed check-valves i and is delivered in grade one filter by () startup kinetic pump i by the road,
Input pressure 0.2 ~ 0.6 mpa, realizes first time coarse filtration；
C the single filter liquid obtaining after grade one filter is crossed check-valves ii and is delivered to two grades by () startup kinetic pump ii by the road
In filter, input pressure 0.2 ~ 0.6 mpa, realizes second coarse filtration；
D () starts kinetic pump iv, the cold water that obtained temperature in constant temperature refrigeration water tank is 5 ~ 15 DEG C enters from interlayer by the road
Mouthful enter interlayer in, and by interlayer export by the road cross check-valves iv be recycled in constant temperature refrigeration water tank；Treat cylinder lumen temperature
Degree is restarted kinetic pump iii and the cascade filtration liquid obtaining after secondary filter is crossed check-valves by the road when reaching 5 ~ 15 DEG C
Iii is delivered in tertiary filter by inlet, input pressure 0.2 ~ 0.6 mpa, and cascade filtration liquid is first from package shell
Porous enters inside package shell, and due to the crown_interception of doughnut, the impurity in cascade filtration liquid is blocked in doughnut
Outside, and extract then pass through doughnut and from doughnut top opening enter pedestal, then from liquid outlet out.
Further, s6 also includes the anti-cleaning of step (e): during end-of-job to be filtered, can carry out the reverse cleaning of this system
Operation, manual 180 ° of integrated regulation power transmitting assembly i, power transmitting assembly ii, the direction of power transmitting assembly iii are original
Import be changed into export, reconnect corresponding pipeline, start above-mentioned kinetic pump i, kinetic pump ii, kinetic pump iii simultaneously, will clean
Purified water through barrel liquid discharge mouth enter cylinder in, then rinse water finally from centrifugation liquid storage tank discharge, each filter pressure in operation
Power list index must not be higher than 0.3 mpa, repeat 2 ~ 3 times.
Preferably, between centrifugation liquid storage tank and kinetic pump i, between check-valves i and grade one filter, grade one filter with dynamic
Between power pump ii, between check-valves ii and secondary filter, between secondary filter and kinetic pump iii, check-valves iii and three-level
Pipeline between filter takes movable connection method.
Further, described movable connection method is clamp connection.
Preferably, tube centrifuge is gf- divergence type.
Steam explosion technology can finish infiltering the sudden release of inside plant tissues gas compressed short time, purpose
It is with less energy, raw material to be decomposed on request.Steam explosion by the whole raw materials in container in Millisecond time range,
It is suspended in airspace in the form of fried dissipating.Because its action time is short, energy density is high and concentrates, and gas molecule can permeate
To between the macromole such as cellulose and lignin, sufficiently on macromolecular level, material is decomposed.Therefore, steam explosion process
Can by the copolymer of lignin, hemicellulose and cellulose that plant cell wall pectin is inlayed break up for lignin, pectin, half
Cellulose and the dispersion of cellulose.The present invention is just allowing for steam explosion technology and can destroy the plant cell wall natural cover for defense
System, thus using steam explosion technology using the pretreatment process before extracting as Radix Pseudostellariae, thus being conducive to Radix Pseudostellariae effective
The extraction of component and separation.But steam explosion technology pretreatment Radix Pseudostellariae still suffers from subproblem, such as in the height of steam explosion
In warm high-pressure process, the lignocellulose of plant cell wall can produce certain noxious substance, such as formic acid, furfural, phenolic compound
Deng.Therefore, with the addition of entrainer in double section steam explosion stages of the present invention, corundum presented in the moment of steam explosion again
Stronger shear action, strengthens breaking the cell natural cover for defense system of Radix Pseudostellariae, ca (oh) further2Interpolation then quick-fried in steam
It is unfavorable for the generation of noxious substance in broken process environment, reduce the conversion of noxious substance.It is passed through co in extract2Process, co2With
ca(oh)2Reaction generates caco3Precipitation, is conducive to removing together with corundum in three-stage filtration operation, adjusts extract simultaneously
Ph value, so that d301 resin packed column and nkaii resin packed column slant acidity, improves detoxification efficiency.In addition, the present invention is steamed
Steam explosion adds enzyme preparation enzymolysis and is remarkably improved the extraction ratio of effective ingredient after breaking, using having arrived cellulase, wood in the present invention
Dextranase and the mixing enzyme system of three kinds of enzymes of pectase, but Cellulase Unit enzyme activity is relatively low, enzyme dosage is higher and enzyme itself is easy
Inactivation, simultaneously during ligocellulose degradation, a large amount of hydrophobic group on lignin can be with the carbon of molecule on cellulase
There is hydrophobic interaction in chain, it is adsorbed, and reduces the effective enzyme amount in enzymolysis process, hinders the enzymolysis of cellulose.Cause
This, add nonionic surfactant to reduce cellulase consumption, and utilize these in lignocellulose Matrix lysis
Nonionic surfactant promotes enzymolysis efficiency to improve.Therefore, the Radix Pseudostellariae pretreatment of preparation method of the present invention, double sections of steam
Explosion treatment, enzyme enzymolysis, water hot extraction, extract co2Process, three-stage filtration, detoxification and precipitate with ethanol are between totally eight steps
It is complementary, assume positive synergism.
Beneficial effect:
1st, two sections of steam explosion technology in the present invention have not only admirably achieved the effect that Radix Pseudostellariae extracts pretreatment, should simultaneously
Technique also can reduce or even remove the noxious substance producing in steam explosion；
2nd, it is passed through co in extract2Process, co2With ca (oh)2Reaction generates caco3Precipitation, not only effectively remove the calcium introduced from
Son may also be advantageous for removing together with corundum in three-stage filtration operation, adjusts extract ph value so that d301 resin simultaneously
Packed column and nkaii resin packed column slant acidity, improve detoxification efficiency；
3rd, the present invention have that technique is accurate, the scale process standard of being easy to operates with, can continuous operation, significantly improve crown prince
The extraction efficiency of ginseng source polysaccharide, reduces the feature of labor intensity；
4th, introduce nonionic surfactant in the operation of enzyme enzymolysis to improve enzymolysis efficiency and reduce adding of exogenous enzyme
Add and mitigate the work load of subsequent treatment；The preparation method of the present invention is suitable to preparation and the life of Radix Pseudostellariae source polysaccharide material
Produce, there is important dissemination.
Brief description
Fig. 1: the Radix Pseudostellariae substrate photo after explosion treatment；
Fig. 2: make the structural representation of three-level filter system in the present invention by oneself；
Fig. 3: in the present invention, self-control three-level filter system is in structural representation during anti-cleaning state；
Description of reference numerals: 1- tube centrifuge, 2- centrifugation liquid storage tank, 31- kinetic pump i, 41- check-valves i, 32- kinetic pump ii,
42- check-valves ii, 33- kinetic pump iii, 43- check-valves iii, 34- kinetic pump iv, 44- check-valves iv, 5- grade one filter, 6-
Secondary filter, 7- cylinder, 8- inlet, 9- liquid outlet, 10- interlayer, 11- interlayer import, 12- interlayer exports, 13- pedestal,
14- package shell, 15- doughnut, 16- constant temperature refrigeration water tank.
Specific embodiment
With reference to the accompanying drawings and detailed description present disclosure is described in further detail:
Embodiment 1
A kind of steam explosion is combined the method that biological enzyme technology prepares Radix Pseudostellariae source polysaccharide, and step is as follows:
S1, Radix Pseudostellariae pretreatment: five operations must be baked, cut into slices and steamed to drying Radix Pseudostellariae, after selecting, going successively；
Wherein, selecting is to be screened Radix Pseudostellariae drying then, and the Radix Pseudostellariae going mouldy or have impurity is rejected；Go must refer to adopt
Radix Pseudostellariae cercus is removed and separates by plant equipment；Baking refers to remove crown prince again using low temperature evacuation baking technology
Ginseng surface moisture is easy to follow-up section, 50 DEG C of baked temperature, baking time 15mim, vacuum pressure 0.04mpa；Section refers to
Radix Pseudostellariae after baking is carried out, using Chinese herbal medicine microtome, the section that Radix Pseudostellariae joins body, the Radix Pseudostellariae piece thickness after section is
0.8mm；Steaming is Radix Pseudostellariae piece to be placed in food steamer process, and steams using vapor heating, steaming time is 25min；
S2, double sections of Steam explosion treatment: the Radix Pseudostellariae after to cut off, as blasting substrates, successively passes through Steam explosion treatment i and steaming
Vapour explosion treatment ii, double sections of Steam explosion treatment interval times are 3min；The condition of Steam explosion treatment i is: the charging of explosion chamber
Coefficient is 0.8, and explosion steam pressure reaches 0.6mpa, and explosion pressurize process time is 12min, and blasting time is not higher than
0.00875s；The condition of Steam explosion treatment ii is: the same cavity explosion treatment of blasting gear on the same stage, and explosion steam pressure reaches
2.0mpa, explosion pressurize process time is 40s, and blasting time is not higher than 0.00875s；During double sections of Steam explosion treatment all
Add entrainer, entrainer is detailed catalogue corundum and ca (oh)2The complex of powder, corundum: ca (oh)2The mass values of powder are
1:1；By percentage to the quality, entrainer total addition level accounts for the 2% of blasting substrates, and is added batch-wise, and Steam explosion treatment i adds
The 1/3 of total amount, Steam explosion treatment ii adds the entrainer of surplus；Fig. 1 is shown in by Radix Pseudostellariae substrate photo after explosion treatment；
S3, enzyme enzymolysis: by after explosion treatment and temperature is down to Radix Pseudostellariae material substrate during room temperature and water with solid-to-liquid ratio
1kg:2l mixes, and the ph value of solidliquid mixture is adjusted to 5.0, then disposably adds nonionic surfactant, add
Cellulase, xylanase and pectase are digested, 50 DEG C of hydrolysis temperature, enzymolysis time 45min, the rank of 25min before enzymolysis
Duan Jinhang at the uniform velocity stirs (rotating speed 110rpm/min), adjusts the ph value of mixture to neutrality at the end of waiting to digest；Wherein, with every g
Drying Radix Pseudostellariae piece meter (drying Radix Pseudostellariae piece be s1 section after, steam before Radix Pseudostellariae piece), cellulase, xylanase, really
The addition of glue enzyme and nonionic surfactant is respectively as follows: 12fpu/g, 130u/g, 100u/g, 0.02g/g；Described nonionic
Surfactant is the mixture of the quality such as berol, tween-80 and bsa；
S4, water hot extraction: by after explosion treatment Radix Pseudostellariae substrate collect after be dosed in water hot extraction's tank, in tank hot water amount with
The mass values of Radix Pseudostellariae substrate are 18:1, extract 95 DEG C of hot water temperature, extraction time 2.5h, extraction times 1 time；Before extraction
Phase is at the uniform velocity stirred (rotating speed be 100rpm/min), to be extracted carries out to stopping stirring during 3/5 extraction time；
S5, extract co2Process: after water hot extraction terminates, first with heat exchanger, the extract obtaining is cooled to room temperature,
Then move in the rustless steel container of cleaning, then in the extract after cooling, be passed through aseptic co2Gas, co2Flow velocity is 13m3/
Min, is passed through co2Gas time 10min；It is passed through co2During gas, co2By pipeline, the gas through rustless steel container bottom is distributed gas
Device enters inside extract；
S6, three-stage filtration: carry out three-stage filtration using self-control three-level filter system, as shown in Fig. 2 this three-level filter system includes
In gf- divergence type tube centrifuge 1, centrifugation liquid storage tank 2(tank, temperature controlling range is 5～10 DEG C), power transmitting assembly i, dynamic
Power transmission assembly ii, power transmitting assembly iii, grade one filter 5, secondary filter 6, tertiary filter and its extraneous attached system
System；
Power transmitting assembly i is connected in series by kinetic pump i 31 and check-valves i 41；Power transmitting assembly ii is by kinetic pump ii 32
It is connected in series with check-valves ii 42；Power transmitting assembly iii is connected in series by kinetic pump iii 33 and check-valves iii 43；
Grade one filter 5 and secondary filter 6 are conventional cartridge filter；Tertiary filter is self-control filter, its structure bag
Include cylinder 7, doughnut tread assembly, pedestal 13；Cylinder 7 bottom and top are respectively equipped with inlet 8 and liquid outlet 9, outside cylinder 7
Portion is provided with interlayer 10, and the upper and lower ends of interlayer 10 are respectively equipped with interlayer import 11 and interlayer outlet 12；Doughnut tread assembly bag
Include and be evenly equipped with the package shell 14 of porous, quantity 600, molecular cut off be 10000 dalton doughnut 15, in these
Hollow fiber 15 merges bunchy and is placed in inside package shell 14；Pedestal 13 is cylinder, cylinder epimere is cylindrical in shape for hollow, under
Duan Shixin, the middle hypomere of pedestal 13 is solid to be provided with the through hole being adapted with doughnut 15 quantity and its size；Pedestal 13
In liquid outlet 9 lower section, it is close to cylinder 7 roof and is secured to connect；The upper end of doughnut 15 is uncovered and is located in pedestal 13
Through hole in, lower end closed is simultaneously fixedly connected with package shell 14 diapire；
The extraneous subsystem of tertiary filter includes kinetic pump iv 34, check-valves iv 44, constant temperature refrigeration water tank 16, and interlayer goes out
Mouth 12 contacts check-valves iv 44, constant temperature refrigeration water tank 16 (controllable temperature scope is: 5～15 DEG C) and kinetic pump iv 34 successively to folder
Layer import 11 forms a closed cycle system；
Tube centrifuge 1 pass through pipeline successively contact centrifugation liquid storage tank 2, kinetic pump i 31, check-valves i 41, grade one filter 5,
Kinetic pump ii 32, check-valves ii 42, secondary filter 6, kinetic pump iii 33, the entering of check-valves iii 43 to tertiary filter
Liquid mouth 8；Wherein, between centrifugation liquid storage tank 2 and kinetic pump i 31, between check-valves i 41 and grade one filter 5, grade one filter
Between 5 and kinetic pump ii 32, between check-valves ii 42 and secondary filter 6, secondary filter 6 and kinetic pump iii 33 it
Between, the pipeline between check-valves iii 43 and tertiary filter take clamp connection；
Aforementioned grade one filter 5 and secondary filter 6 pass through city available from the filter element of grade one filter 5 and secondary filter 6 is equal
For folding type filter element, grade one filter 5 and the filter element number respectively 5 and 7 with secondary filter 6, grade one filter 5 He
The filter element aperture of secondary filter 6 is respectively 5 μm, 0.22 μm；The filter element material of grade one filter 5 and secondary filter 6 is poly-
Propylene, frame of filter element (bracing frame) is rigid plastics, and filter element can compression resistance be 0.4 mpa；
Three-stage filtration step is:
A () inputs the extract that s5 obtains in tube centrifuge 1 inner chamber first, start this equipment and carry out centrifugal work, centrifugation
Turn up 5000rpm/min, is centrifuged 30min；Centrifugation proceeds to the centrifugal clear liquid of acquisition to centrifugation liquid storage tank 2 after terminating in time
Interior, and in this tank, temperature controlling range is 10 DEG C；
B the cooling centrifugal liquid that upper operation obtains is crossed check-valves i 41 through pipeline and is delivered to single filter by () startup kinetic pump i 31
In device 5, input pressure 0.2 mpa, realizes first time coarse filtration；
C the single filter liquid obtaining after grade one filter 5 is crossed check-valves ii 42 through pipeline by () startup kinetic pump ii 32 defeated
Deliver in secondary filter 6, input pressure 0.2 mpa, realizes second coarse filtration；
(d) start kinetic pump iv 34, by obtained temperature in constant temperature refrigeration water tank 16 be 5 DEG C cold water through pipeline from interlayer
Import 11 enters in interlayer 10, and is crossed check-valves iv 44 through pipeline and be recycled in constant temperature refrigeration water tank 16 by interlayer outlet 12；
Restart the cascade filtration liquid warp that kinetic pump iii 33 will obtain after secondary filter 6 when cylinder 7 inner chamber temperature reaches 5 DEG C
Pipeline is crossed check-valves iii 43 and is delivered in tertiary filter by inlet 8, input pressure 0.2 mpa, cascade filtration liquid first from
Porous on package shell 14 enters inside package shell 14, due to the crown_interception of doughnut 15, in cascade filtration liquid
Impurity is blocked in the outside of doughnut 15, and extract then passes through doughnut 15 and enters pedestal from doughnut 15 top opening
13, then from liquid outlet 9 out；
E () device is counter to clean: during end-of-job to be filtered, can carry out the reverse cleaning operation of this equipment series, dismantle first
Between centrifugation liquid storage tank 2 and kinetic pump i 31, between check-valves i 41 and grade one filter 5, grade one filter 5 and kinetic pump ii
Between 32, between check-valves ii 42 and secondary filter 6, between secondary filter 6 and kinetic pump iii 33, check-valves iii
The clip on pipeline between 43 and tertiary filter, manual 180 ° of integrated regulation power transmitting assembly i, power transmitting assemblies
Ii, the direction (original import is changed into exporting) of power transmitting assembly iii, then pass through the corresponding pipeline of clamp connection, after adjustment
State as shown in figure 3, start above-mentioned kinetic pump i 31, kinetic pump ii 32, kinetic pump iii 33 simultaneously, will cleaning purification
Water enters in cylinder 7 through cylinder 7 liquid outlet 9, then rinse water is finally discharged from centrifugation liquid storage tank 2, each filter pressure in operation
List index must not be higher than 0.3 mpa, is repeated 2 times；
S7, detoxification: the Radix Pseudostellariae extract priority after three-stage filtration is passed through d301 resin packed column and the filling of nkaii resin
Post, extract enters post pressure and is 0.15mpa, collects extract；
S8, precipitate with ethanol: first the Radix Pseudostellariae extract after detoxification is carried out decompression half and concentrate, concentration pressure is 0.03mpa, concentrates temperature
60 DEG C of degree, concentration time 2.5h；It is followed by half concentrated solution that reduces pressure being cooled to 20 DEG C and being added thereto to the 95% of 2.5 times of volumes
Edible ethanol liquid, stands 9h, and finally centrifugation obtains precipitate and is Radix Pseudostellariae source polysaccharide；Wherein, centrifugal rotational speed is 13000rpm/
Min, centrifuging temperature temperature control at 10 DEG C, centrifugation time 20min.
Reference examples 1
It is s2 with the difference of embodiment 1: Steam explosion treatment i and ii is all without entrainer；Other steps are with enforcement
Example 1.
Reference examples 2
It is s2 with the difference of embodiment 1: only implement Steam explosion treatment i, and add the entrainer of full dose；Other steps
With embodiment 1.
Reference examples 3
It is s2 with the difference of embodiment 1: only implement Steam explosion treatment ii, and add the entrainer of full dose；Other steps
Suddenly with embodiment 1.
Reference examples 4
It is to omit s3 with the difference of embodiment 1, digested without enzyme；Other steps are with embodiment 1.
Reference examples 5
It is s3 with the difference of embodiment 1: only add cellulase by the addition of cellulase；Other steps are with enforcement
Example 1.
Reference examples 6
It is s3 with the difference of embodiment 1: only add xylanase by the addition of xylanase；Other steps are with enforcement
Example 1.
Reference examples 7
It is s3 with the difference of embodiment 1: only add pectase by the addition of pectase；Other steps are with embodiment 1.
With polysaccharide extraction yield=(quality of the quality of the Radix Pseudostellariae source polysaccharide preparing/drying Radix Pseudostellariae piece) × 100%
Meter, drying Radix Pseudostellariae piece therein be s1 section after, steam before Radix Pseudostellariae piece, to embodiment 1 and reference examples 1 ~ 7 gained crown prince
The extraction yield statistical computation of ginseng source polysaccharide, result is as follows:
As can be seen from the above table:
(1), compared with Example 1, the polysaccharide extraction yield of reference examples 1 is significantly lower than the polysaccharide extraction yield of embodiment 1, illustrates " to add
Plus entrainer " relatively " without entrainer " be remarkably improved the extraction yield of Radix Pseudostellariae source polysaccharide；
(2), compared with Example 1, the polysaccharide extraction yield of reference examples 2 and 3 is significantly lower than the polysaccharide extraction yield of embodiment 1, explanation
" two section type steam explosion " relatively " one-part form steam explosion " is remarkably improved the extraction yield of Radix Pseudostellariae source polysaccharide；
(3), compared with reference examples 4, the polysaccharide extraction yield of embodiment 1 and reference examples 5 ~ 7 is apparently higher than the polysaccharide extraction of reference examples 4
Take rate, after two section type steam explosion is described, " interpolation enzyme " relatively " without enzyme " can carry significantly high Radix Pseudostellariae source polysaccharide
Extraction yield；Meanwhile, compared with Example 1, the polysaccharide extraction yield of reference examples 5 ~ 7 is significantly lower than the polysaccharide extraction yield of embodiment 1 again,
The compound use of three kinds of enzymes is described, more single enzyme, more can improve the extraction yield of Radix Pseudostellariae source polysaccharide.
To sum up, the present invention adopts two section type steam explosion, and adds entrainer during it, and two section type steam is quick-fried simultaneously
Add enzyme enzymolysis after broken, the polysaccharide extraction yield of Radix Pseudostellariae can be made farthest to be improved.
Claims (6)
1. a kind of steam explosion combination biological enzyme technology prepares the method for Radix Pseudostellariae source polysaccharide it is characterised in that step is as follows:
S1, Radix Pseudostellariae pretreatment: five operations must be baked, cut into slices and steamed to drying Radix Pseudostellariae, after selecting, going successively；
S2, double sections of Steam explosion treatment: the Radix Pseudostellariae piece after to steam as blasting substrates, successively pass through Steam explosion treatment i and
Steam explosion treatment ii, double sections of Steam explosion treatment interval times are 2 ~ 5min；The condition of Steam explosion treatment i is: explosion chamber
Coefficient is 0.7 ~ 0.9, and explosion steam pressure reaches 0.5 ~ 0.8mpa, and explosion pressurize process time is 10 ~ 15min, during explosion
Between not higher than 0.00875s；The condition of Steam explosion treatment ii is: the same cavity explosion treatment of blasting gear on the same stage, explosion steam
Pressure reaches 1.5 ~ 2.2mpa, and explosion pressurize process time is 30 ~ 50s, and blasting time is not higher than 0.00875s；Double sections of steam explosions
Entrainer is all added, entrainer is detailed catalogue corundum and ca (oh) in processing procedure2The complex of powder, corundum: ca (oh)2Powder
Mass values be 4:1 ~ 1:1；By percentage to the quality, entrainer total addition level accounts for the 1.5 ~ 3% of blasting substrates, and adds in batches
Plus, the 1/3 ~ 2/5 of Steam explosion treatment i total amount of adding, Steam explosion treatment ii add the entrainer of surplus；
S3, enzyme enzymolysis: by after explosion treatment and temperature is down to Radix Pseudostellariae material substrate during room temperature and water with solid-to-liquid ratio
1kg:1l ~ 1kg:3l mixes, and the ph value of solidliquid mixture is adjusted to 4.5 ~ 5.0, then disposably adds non-ionic surface to live
Property agent, adding cellulase, xylanase and pectase is digested, 50 ~ 55 DEG C of hydrolysis temperature, enzymolysis time 30 ~
45min, the stage digesting front 20 ~ 25min is at the uniform velocity stirred, and adjusts the ph value of mixture to neutrality at the end of waiting to digest；Its
In, in terms of every g drying Radix Pseudostellariae piece, the addition of cellulase, xylanase, pectase and nonionic surfactant is respectively
For: 10 ~ 15fpu/g, 100 ~ 150u/g, 80 ~ 110u/g, 0.003 ~ 0.05g/g；Described nonionic surfactant be berol,
One or more of peg, tween-80 and bsa；
S4, water hot extraction: by after explosion treatment Radix Pseudostellariae substrate collect after be dosed in water hot extraction's tank, in tank hot water amount with
The mass values of Radix Pseudostellariae substrate are 15:1 ~ 20:1, extract 90 ~ 100 DEG C of hot water temperature, extraction time 2 ~ 3h, extraction times 1
Secondary；Phase is at the uniform velocity stirred before extraction, to be extracted carry out to during 3/5 extraction time stop stirring；
S5, extract co2Process: after water hot extraction terminates, the extract obtaining is cooled to room temperature, then moves to cleaning not
In rust steel container, then in the extract after cooling, it is passed through aseptic co2Gas, co2Flow velocity is 10 ~ 15m3/ min, is passed through co2
Gas time 5 ~ 15min；
S6, three-stage filtration: extract co2Respectively through the grade one filter of tandem compound, secondary filter and three-level mistake after process
Filter is realizing filtration operation；Wherein, grade one filter and secondary filter are conventional cartridge filter, grade one filter and
The filter element aperture of secondary filter is respectively 4.5 ~ 5 μm, 0.2 ~ 0.25 μm；Tertiary filter is hollow fiber filter, hollow
The molecular cut off of fiber is 10000 ~ 30000 dalton；
S7, detoxification: the Radix Pseudostellariae extract priority after three-stage filtration is passed through d301 resin packed column and the filling of nkaii resin
Post, extract enters post pressure and is 0.15 ~ 0.2mpa, collects extract；
S8, precipitate with ethanol: first the Radix Pseudostellariae extract after detoxification is carried out decompression half and concentrate, concentration pressure is 0.01 ~ 0.05mpa,
50 ~ 70 DEG C of thickening temperature, concentration time 2 ~ 3h；It is followed by half concentrated solution that reduces pressure being cooled to 10 ~ 25 DEG C and being added thereto to 2 ~ 3
95% edible ethanol liquid of times volume, stands 8 ~ 10h, and finally centrifugation obtains precipitate and is Radix Pseudostellariae source polysaccharide.
2. preparation method as claimed in claim 1 it is characterised in that: in s6 using self-control three-level filter system carry out three-level mistake
Filter, this three-level filter system includes tube centrifuge, centrifugation liquid storage tank, power transmitting assembly i, power transmitting assembly ii, power
Transmission assembly iii, grade one filter, secondary filter, tertiary filter and its extraneous subsystem；Power transmitting assembly i by
Kinetic pump i and check-valves i is connected in series；Power transmitting assembly ii is connected in series by kinetic pump ii and check-valves ii；Power transmits
Assembly iii is connected in series by kinetic pump iii and check-valves iii；Grade one filter and secondary filter are conventional cartridge filter；
Tertiary filter is self-control filter, and its structure includes cylinder, doughnut tread assembly, pedestal；Cylinder body bottom and top are respectively
It is provided with inlet and liquid outlet, outside cylinder, is provided with interlayer, the upper and lower ends of interlayer are respectively equipped with interlayer import and interlayer outlet；
Doughnut tread assembly includes being evenly equipped with the package shell of porous, some doughnuts, and these doughnuts merge bunchy and put
Inside package shell；Pedestal is cylinder, and cylinder epimere is cylindrical in shape for hollow, middle hypomere is solid, and the middle hypomere of pedestal is real
It is provided with the through hole being adapted with doughnut quantity and its size in the heart；Pedestal is located at below liquid outlet, is close to cylinder roof simultaneously
It is secured to connect；The upper end of doughnut is uncovered and is located in the through hole of pedestal, lower end closed and with package shell diapire
It is fixedly connected；The extraneous subsystem of tertiary filter includes kinetic pump iv, check-valves iv, constant temperature refrigeration water tank, and interlayer exports
Series winding check-valves iv, constant temperature refrigeration water tank and kinetic pump iv form a closed cycle system to interlayer import successively；Tubular type is centrifuged
Machine pass through pipeline successively contact centrifugation liquid storage tank, kinetic pump i, check-valves i, grade one filter, kinetic pump ii, check-valves ii, two
Level filter, kinetic pump iii, the inlet of check-valves iii to tertiary filter；Grade one filter and the filter with secondary filter
Core bore footpath is respectively 4.5 ~ 5 μm, 0.2 ~ 0.22 μm；The molecular cut off of doughnut is 10000 ~ 30000 dalton；
Three-stage filtration step is:
Extract a s5 is processed by () first after inputs in tube centrifuge inner chamber, starts this equipment and carries out centrifugal work, centrifugation
Turn up 4000 ~ 6000rpm/min, is centrifuged 20 ~ 30min；Centrifugation proceeds to the centrifugal clear liquid of acquisition to centrifugation after terminating in time
In liquid storage tank, and in this tank, temperature controlling range is 5 ~ 10 DEG C；
B the cooling centrifugal liquid that upper operation obtains is crossed check-valves i and is delivered in grade one filter by () startup kinetic pump i by the road,
Input pressure 0.2 ~ 0.6 mpa, realizes first time coarse filtration；
C the single filter liquid obtaining after grade one filter is crossed check-valves ii and is delivered to two grades by () startup kinetic pump ii by the road
In filter, input pressure 0.2 ~ 0.6 mpa, realizes second coarse filtration；
D () starts kinetic pump iv, the cold water that obtained temperature in constant temperature refrigeration water tank is 5 ~ 15 DEG C enters from interlayer by the road
Mouthful enter interlayer in, and by interlayer export by the road cross check-valves iv be recycled in constant temperature refrigeration water tank；Treat cylinder lumen temperature
Degree is restarted kinetic pump iii and the cascade filtration liquid obtaining after secondary filter is crossed check-valves by the road when reaching 5 ~ 15 DEG C
Iii is delivered in tertiary filter by inlet, input pressure 0.2 ~ 0.6 mpa, and cascade filtration liquid is first from package shell
Porous enters inside package shell, and due to the crown_interception of doughnut, the impurity in cascade filtration liquid is blocked in doughnut
Outside, and extract then pass through doughnut and from doughnut top opening enter pedestal, then from liquid outlet out.
3. it is characterised in that s6 also includes, step (e) is anti-to clean preparation method as claimed in claim 2: work knot to be filtered
Shu Shi, can carry out the reverse cleaning operation of this system, manual 180 ° of integrated regulation power transmitting assembly i, power transmitting assembly ii, dynamic
The import that the direction of power transmission assembly iii is original be changed into export, reconnect corresponding pipeline, start simultaneously above-mentioned kinetic pump i,
Kinetic pump ii, kinetic pump iii, the purified water of cleaning are entered in cylinder through barrel liquid discharge mouth, then rinse water is finally from centrifugal liquid
Storage tank is discharged, and in operation, each filter pressure list index must not be higher than 0.3 mpa, repeats 2 ~ 3 times.
4. preparation method as claimed in claim 2 it is characterised in that: centrifugation liquid storage tank and kinetic pump i between, check-valves i with
Between grade one filter, between grade one filter and kinetic pump ii, between check-valves ii and secondary filter, secondary filter with
Pipeline between kinetic pump iii, between check-valves iii and tertiary filter takes movable connection method.
5. preparation method as claimed in claim 4 it is characterised in that: described movable connection method be clamp connection.
6. preparation method as claimed in claim 2 it is characterised in that: tube centrifuge be gf- divergence type.
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