CN101629201B - Method for producing acteoside by biocatalysis and biotransformation of fresh herba cistanches - Google Patents
Method for producing acteoside by biocatalysis and biotransformation of fresh herba cistanches Download PDFInfo
- Publication number
- CN101629201B CN101629201B CN2009100416497A CN200910041649A CN101629201B CN 101629201 B CN101629201 B CN 101629201B CN 2009100416497 A CN2009100416497 A CN 2009100416497A CN 200910041649 A CN200910041649 A CN 200910041649A CN 101629201 B CN101629201 B CN 101629201B
- Authority
- CN
- China
- Prior art keywords
- industrial
- herba cistanches
- temperature
- enzyme
- pure water
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Expired - Fee Related
Links
- 241000005787 Cistanche Species 0.000 title claims abstract description 114
- QFRYQWYZSQDFOS-UHFFFAOYSA-N verbascoside Natural products CC1OC(COC2C(O)C(COC3OC(C(O)C(O)C3O)C(=O)O)OC(Oc4cc(O)cc5OC(=CC(=O)c45)c6ccc(O)c(O)c6)C2O)C(O)C(O)C1O QFRYQWYZSQDFOS-UHFFFAOYSA-N 0.000 title claims abstract description 12
- 229930185474 acteoside Natural products 0.000 title claims abstract description 10
- FBSKJMQYURKNSU-ZLSOWSIRSA-N acteoside Chemical compound O[C@@H]1[C@H](O)[C@@H](O)[C@H](C)O[C@H]1O[C@H]1[C@H](OC(=O)\C=C\C=2C=C(O)C(O)=CC=2)[C@@H](CO)O[C@@H](OCCC=2C=C(O)C(O)=CC=2)[C@@H]1O FBSKJMQYURKNSU-ZLSOWSIRSA-N 0.000 title claims abstract description 10
- FBSKJMQYURKNSU-UKQWSTALSA-N acteoside I Natural products C[C@@H]1O[C@H](O[C@@H]2[C@@H](O)[C@H](OCCc3ccc(O)c(O)c3)O[C@H](CO)[C@H]2OC(=O)C=Cc4ccc(O)c(O)c4)[C@H](O)[C@H](O)[C@H]1O FBSKJMQYURKNSU-UKQWSTALSA-N 0.000 title claims abstract description 10
- 230000036983 biotransformation Effects 0.000 title claims abstract description 5
- 238000004519 manufacturing process Methods 0.000 title abstract description 3
- 150000001875 compounds Chemical class 0.000 claims abstract description 104
- 238000000034 method Methods 0.000 claims abstract description 96
- 238000006555 catalytic reaction Methods 0.000 claims abstract description 85
- 102000004190 Enzymes Human genes 0.000 claims abstract description 51
- 108090000790 Enzymes Proteins 0.000 claims abstract description 51
- 238000000746 purification Methods 0.000 claims abstract description 42
- 239000000758 substrate Substances 0.000 claims abstract description 38
- 238000001035 drying Methods 0.000 claims abstract description 36
- FSBUXLDOLNLABB-ISAKITKMSA-N echinacoside Chemical compound O[C@@H]1[C@H](O)[C@@H](O)[C@H](C)O[C@H]1O[C@H]1[C@H](OC(=O)\C=C\C=2C=C(O)C(O)=CC=2)[C@@H](CO[C@H]2[C@@H]([C@@H](O)[C@H](O)[C@@H](CO)O2)O)O[C@@H](OCCC=2C=C(O)C(O)=CC=2)[C@@H]1O FSBUXLDOLNLABB-ISAKITKMSA-N 0.000 claims abstract description 30
- NJYVDFDTLLZVMG-UHFFFAOYSA-N echinacoside Natural products CC1OC(OC2C(O)C(OCCc3ccc(O)c(O)c3)OC(COC4OC(CO)C(O)C(O)C4O)C2OC(=O)C=Cc5cc(O)cc(O)c5)C(O)C(O)C1O NJYVDFDTLLZVMG-UHFFFAOYSA-N 0.000 claims abstract description 30
- 108010093096 Immobilized Enzymes Proteins 0.000 claims abstract description 20
- 108010047754 beta-Glucosidase Proteins 0.000 claims abstract description 19
- 102000006995 beta-Glucosidase Human genes 0.000 claims abstract description 19
- 230000000694 effects Effects 0.000 claims abstract description 19
- 238000000926 separation method Methods 0.000 claims abstract description 16
- 230000007062 hydrolysis Effects 0.000 claims abstract description 14
- 238000006460 hydrolysis reaction Methods 0.000 claims abstract description 14
- 238000010521 absorption reaction Methods 0.000 claims abstract description 7
- 239000002994 raw material Substances 0.000 claims abstract description 7
- 210000004027 cell Anatomy 0.000 claims description 94
- 241001530097 Verbascum Species 0.000 claims description 66
- 235000010599 Verbascum thapsus Nutrition 0.000 claims description 66
- 229930182478 glucoside Natural products 0.000 claims description 66
- 150000008131 glucosides Chemical class 0.000 claims description 66
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 63
- 229940088598 enzyme Drugs 0.000 claims description 48
- 239000002002 slurry Substances 0.000 claims description 45
- 238000005516 engineering process Methods 0.000 claims description 42
- 239000012528 membrane Substances 0.000 claims description 33
- 235000014347 soups Nutrition 0.000 claims description 31
- 238000005406 washing Methods 0.000 claims description 26
- 238000013467 fragmentation Methods 0.000 claims description 24
- 238000006062 fragmentation reaction Methods 0.000 claims description 24
- 238000003801 milling Methods 0.000 claims description 19
- 238000001179 sorption measurement Methods 0.000 claims description 17
- 239000000376 reactant Substances 0.000 claims description 15
- 238000000108 ultra-filtration Methods 0.000 claims description 15
- 238000004587 chromatography analysis Methods 0.000 claims description 14
- 239000000463 material Substances 0.000 claims description 14
- 239000003054 catalyst Substances 0.000 claims description 13
- 230000003197 catalytic effect Effects 0.000 claims description 13
- 238000006243 chemical reaction Methods 0.000 claims description 13
- 238000001914 filtration Methods 0.000 claims description 13
- 238000009776 industrial production Methods 0.000 claims description 13
- 150000008495 β-glucosides Chemical class 0.000 claims description 13
- 210000000805 cytoplasm Anatomy 0.000 claims description 12
- 238000001728 nano-filtration Methods 0.000 claims description 12
- 229920005989 resin Polymers 0.000 claims description 12
- 239000011347 resin Substances 0.000 claims description 12
- 239000007787 solid Substances 0.000 claims description 12
- JEGUKCSWCFPDGT-UHFFFAOYSA-N h2o hydrate Chemical compound O.O JEGUKCSWCFPDGT-UHFFFAOYSA-N 0.000 claims description 10
- 238000010438 heat treatment Methods 0.000 claims description 10
- 238000001704 evaporation Methods 0.000 claims description 9
- 235000012054 meals Nutrition 0.000 claims description 7
- 239000012535 impurity Substances 0.000 claims description 6
- 239000003262 industrial enzyme Substances 0.000 claims description 6
- 238000002360 preparation method Methods 0.000 claims description 6
- 108010059892 Cellulase Proteins 0.000 claims description 3
- FBPFZTCFMRRESA-KVTDHHQDSA-N D-Mannitol Chemical compound OC[C@@H](O)[C@@H](O)[C@H](O)[C@H](O)CO FBPFZTCFMRRESA-KVTDHHQDSA-N 0.000 claims description 3
- 241000196324 Embryophyta Species 0.000 claims description 3
- 229930195725 Mannitol Natural products 0.000 claims description 3
- 229940106157 cellulase Drugs 0.000 claims description 3
- KWIUHFFTVRNATP-UHFFFAOYSA-N glycine betaine Chemical compound C[N+](C)(C)CC([O-])=O KWIUHFFTVRNATP-UHFFFAOYSA-N 0.000 claims description 3
- 239000000594 mannitol Substances 0.000 claims description 3
- 235000010355 mannitol Nutrition 0.000 claims description 3
- 150000001413 amino acids Chemical class 0.000 claims description 2
- 150000004676 glycans Chemical class 0.000 claims description 2
- 239000000419 plant extract Substances 0.000 claims description 2
- 229920001282 polysaccharide Polymers 0.000 claims description 2
- 239000005017 polysaccharide Substances 0.000 claims description 2
- 102000004169 proteins and genes Human genes 0.000 claims description 2
- 108090000623 proteins and genes Proteins 0.000 claims description 2
- 229920002521 macromolecule Polymers 0.000 claims 1
- 239000003814 drug Substances 0.000 abstract description 4
- 229940079593 drug Drugs 0.000 abstract description 2
- 238000000605 extraction Methods 0.000 abstract description 2
- 150000003384 small molecules Chemical class 0.000 abstract description 2
- 230000026676 system process Effects 0.000 abstract description 2
- 230000009849 deactivation Effects 0.000 abstract 2
- 239000011942 biocatalyst Substances 0.000 abstract 1
- 125000002791 glucosyl group Chemical group C1([C@H](O)[C@@H](O)[C@H](O)[C@H](O1)CO)* 0.000 abstract 1
- 239000003446 ligand Substances 0.000 abstract 1
- 239000007788 liquid Substances 0.000 description 23
- 239000002594 sorbent Substances 0.000 description 14
- 230000009466 transformation Effects 0.000 description 11
- 238000003556 assay Methods 0.000 description 10
- 230000035484 reaction time Effects 0.000 description 10
- 239000003463 adsorbent Substances 0.000 description 9
- 238000005194 fractionation Methods 0.000 description 9
- 238000005057 refrigeration Methods 0.000 description 7
- 229930182470 glycoside Natural products 0.000 description 6
- -1 phenylethyl alcohol glycoside Chemical class 0.000 description 5
- 238000001291 vacuum drying Methods 0.000 description 5
- GMWTXQKKRDUVQG-WOPPDYDQSA-N 4-amino-5-bromo-1-[(2r,3s,4s,5r)-4-hydroxy-5-(hydroxymethyl)-3-methyloxolan-2-yl]pyrimidin-2-one Chemical compound C[C@H]1[C@H](O)[C@@H](CO)O[C@H]1N1C(=O)N=C(N)C(Br)=C1 GMWTXQKKRDUVQG-WOPPDYDQSA-N 0.000 description 3
- XYJRXVWERLGGKC-UHFFFAOYSA-D pentacalcium;hydroxide;triphosphate Chemical compound [OH-].[Ca+2].[Ca+2].[Ca+2].[Ca+2].[Ca+2].[O-]P([O-])([O-])=O.[O-]P([O-])([O-])=O.[O-]P([O-])([O-])=O XYJRXVWERLGGKC-UHFFFAOYSA-D 0.000 description 3
- WRMNZCZEMHIOCP-UHFFFAOYSA-N 2-Phenylethanol Natural products OCCC1=CC=CC=C1 WRMNZCZEMHIOCP-UHFFFAOYSA-N 0.000 description 2
- 229920000298 Cellophane Polymers 0.000 description 2
- 229920002678 cellulose Polymers 0.000 description 2
- 239000001913 cellulose Substances 0.000 description 2
- 229920001429 chelating resin Polymers 0.000 description 2
- 150000002338 glycosides Chemical class 0.000 description 2
- 230000004060 metabolic process Effects 0.000 description 2
- 229940067107 phenylethyl alcohol Drugs 0.000 description 2
- 229920003002 synthetic resin Polymers 0.000 description 2
- 239000000057 synthetic resin Substances 0.000 description 2
- 238000002137 ultrasound extraction Methods 0.000 description 2
- KDSWDGKIENPKLB-QJDQKFITSA-N verbascoside Chemical compound O[C@@H]1[C@H](O)[C@@H](O)[C@H](C)O[C@H]1O[C@H]1[C@H](OC(=O)CCC=2C=C(O)C(O)=CC=2)[C@@H](CO)O[C@@H](OCCC=2C=C(O)C(O)=CC=2)[C@@H]1O KDSWDGKIENPKLB-QJDQKFITSA-N 0.000 description 2
- 241000894006 Bacteria Species 0.000 description 1
- 241000336291 Cistanche deserticola Species 0.000 description 1
- 241000336316 Cistanche tubulosa Species 0.000 description 1
- 239000002033 PVDF binder Substances 0.000 description 1
- 239000004480 active ingredient Substances 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 229960004217 benzyl alcohol Drugs 0.000 description 1
- 235000019445 benzyl alcohol Nutrition 0.000 description 1
- 239000002775 capsule Substances 0.000 description 1
- 239000002537 cosmetic Substances 0.000 description 1
- 230000002255 enzymatic effect Effects 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 239000000835 fiber Substances 0.000 description 1
- 235000013305 food Nutrition 0.000 description 1
- 239000011521 glass Substances 0.000 description 1
- 239000008187 granular material Substances 0.000 description 1
- 239000012510 hollow fiber Substances 0.000 description 1
- 210000000936 intestine Anatomy 0.000 description 1
- 229930182489 iridoid glycoside Natural products 0.000 description 1
- 150000008145 iridoid glycosides Chemical class 0.000 description 1
- 210000003734 kidney Anatomy 0.000 description 1
- 210000002429 large intestine Anatomy 0.000 description 1
- 239000012567 medical material Substances 0.000 description 1
- 229920001542 oligosaccharide Polymers 0.000 description 1
- 150000002482 oligosaccharides Chemical class 0.000 description 1
- 230000003071 parasitic effect Effects 0.000 description 1
- 239000002245 particle Substances 0.000 description 1
- 239000006187 pill Substances 0.000 description 1
- 238000011020 pilot scale process Methods 0.000 description 1
- 229920002981 polyvinylidene fluoride Polymers 0.000 description 1
- 238000002953 preparative HPLC Methods 0.000 description 1
- 238000004321 preservation Methods 0.000 description 1
- 239000003826 tablet Substances 0.000 description 1
- 230000001256 tonic effect Effects 0.000 description 1
Landscapes
- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Saccharide Compounds (AREA)
Abstract
The invention provides a method for producing acteoside by biocatalysis and biotransformation of fresh herba cistanches, comprising the following steps: taking the fresh herba cistanches as raw materials to undergo such system processes as cell disruption and size extraction as well as enzyme deactivation at high temperature, taking herba cistanches cell size undergoing enzyme deactivation as substrates for catalytic reaction and beta-glucosidase, an industrial immobilized enzyme, as a biocatalyst for discontinuous or continuous catalytic reaction to catalyze echinacoside in hydrolysis substrates to enable the ends of glucosyl ligands of the echinacoside to be broken and the echinacoside to be directionally converted into high-activity monomeric compounds of the acteoside, and obtaining the monomeric compounds of the acteoside by the processes of separation and purification, concentration and drying. By adjusting the conditions of the separation and purification processes, the purity percentage content of the obtained monomeric compounds of the acteoside can reach 50-90%. The monomeric compounds of the acteoside have the characteristics of small molecules and low polarity and show good absorption rate and particular pharmacology activity in drug absorption.
Description
Technical field
The present invention relates to a kind of method of utilizing the producing acteoside by biocatalysis and biotransformation of fresh herba cistanches monomeric compound.
Background technology
Herba Cistanches (Herba Cistanches) is parasitic for many years herbage, is famous traditional tonic medicine, effects such as having beneficial vital essence, tonify the kidney and support yang and delay senility.Modern analysis shows that the main active ingredient of Herba Cistanches is phenylethyl alcohol glycoside, phenylcarbinol glycoside, iridoid glycosides, lignanoid's glycoside, oligosaccharides verivate, D-N.F,USP MANNITOL and trimethyl-glycine etc.Phenylethyl alcohol glycoside (Phenylethanoid Glycosides; Ph Gs) comprises echinacoside (Echinacoside), verbascoside (Acteoside) isoreactivity material; Discover through bacterium metabolism in the intestines; Benzyl carbinol glycosides staple echinacoside moving process need micro bioenzyme catalysis in large intestine in gi tract is converted into the metabolism passage of verbascoside, is to cause oral artifact availability and curative effect to have the principal element of significant difference.Mullein glucoside monomeric compound has small molecules and low polar characteristic, in drug absorption, shows good specific absorption and special pharmacologically active.
Summary of the invention
The purpose of this invention is to provide a kind of is raw material with the fresh herba cistanches, carries out the method that catalyzed conversion is produced mullein glucoside monomeric compound through system process processing and industrial immobilized enzyme reactor.
Method of the present invention is; Utilize fresh herba cistanches to be raw material; Handle through cytoclasis, slurries extraction process and the high temperature enzymatic process that goes out, the Herba Cistanches cell slurries behind the enzyme that will go out are as the catalyzed reaction substrate, through beta-glucosidase industrial enzyme preparation immobilized enzyme as biological catalyst; Echinacoside makes the terminal fracture of glucone part, orientation be converted into highly active mullein glucoside monomeric compound in the catalytic hydrolysis substrate; Through process for separating and purifying, reach concentrated, drying process again, the results mullein glucoside monomeric compound; The catalytic reaction process temperature is 38 ℃～60 ℃, and pH is 4.0～5.8, and the time is 4h～8h, and the beta-glucoside enzyme activity in the catalytic reaction process reactant is 2U/g～1000U/g; Through adjustment process for separating and purifying condition, the mullein glucoside monomeric compound purity degree of results can reach 50%～90%.
The concrete steps of the inventive method comprise:
1, selects the fresh herba cistanches chylocaulous, place washing bath with cold water or technical pure water rinse; After picking up, place the broken milling device of common industry that the fragmentation of Herba Cistanches chylocaulous is milled, add the technical pure water purification or become Herba Cistanches smudge cells soup compound through the technical pure water washing fragmented cell of precooling; Adopt common industrial pressure filter press filtration or industrial centrifugal machine to separate then, obtain Herba Cistanches cell slurries; Can be again with filter residue through once adding the technical pure water purification respectively or through the technical pure water purification of precooling to (being generally 1～3 time) for several times; Once to further being milled into soup compound through common industrial broken runner milling for several times; Also can adopt common industrial ultrasonic extraction device that smudge cells is extracted simultaneously, further obtain Herba Cistanches cell slurries through industrial pressure filter press filtration or industrial centrifugal machine separation again; With fragmentation mill with separating technology process Herba Cistanches smudge cells soup compound and cytoplasm hydraulic control built in the low temperature environment that (is generally 4～20 ℃) below 20 ℃;
2, the Herba Cistanches cell slurries that through common industrial heating unit step 1 obtained carry out the high temperature enzyme that goes out, or carry out the instantaneous enzyme that goes out of industrial ultrahigh-temperature; To go out Herba Cistanches cell slurries behind the enzyme as the catalyzed reaction substrate; The beta-glucosidase industrial enzyme preparation immobilized enzyme that adopts the absorption method fixed form acquisition in the common immobilized enzyme industrial production is as biological catalyst; Carry out discontinuous or continous way catalyzed reaction through common industrial immobilized enzyme reactor, catalytic hydrolysis removes substrate echinacoside glucone part terminal glucone, an orientation and is converted into mullein glucoside monomeric compound; The catalytic reaction process temperature is 38 ℃～60 ℃, and optimum temps is 42 ℃～48 ℃; PH is 4.0～5.8, and best pH is 4.5～5.3; Time is 4h～8h, and Best Times is 5h～6h, and the beta-glucoside enzyme activity in the catalytic reaction process reactant is 2U/g～1000U/g, and best enzyme activity is 25U/g～150U/g; Through measuring, the transformation efficiency that the echinacoside in the catalytic reaction process reactant is converted into mullein glucoside monomeric compound reaches 78%～95%
3, will adopt common industrial ultra-filtration membrane to separate through the reacted reaction solution of step 2, macromole impurity such as removing polysaccharide, protein and impurity such as particulate and submicron are sieved and held back to selectivity; Adopt common industrial nanofiltration membrane separation again, small molecular weight impurities such as removing amino acid, D-N.F,USP MANNITOL, trimethyl-glycine is sieved and held back to selectivity; Also can adopt common industrial macroporous resin adsorption separating technology again, or/and adopt common industrial chromatography column separating purification technology, or/and adopt common separation system of simulated moving bed chromatography technology or the further separation and purification of common multi-functional chromatogram separating technology; Through adjusting above process for separating and purifying condition, can obtain the mullein glucoside monomeric compound extracting solution of different purity;
4, the mullein glucoside monomeric compound extracting solution that step 3 is obtained, through common industrial vacuum concentration technology or industrial film evaporating and concentrating process, the acquisition solid content is 30%～50% enriched material; Pass through industrial drying process with atomizing again, or other common industrially drying method, the meal or the dry thing of acquisition mullein glucoside monomeric compound.The mullein glucoside monomeric compound purity degree of results can reach 50%～90%.
The fresh herba cistanches raw material that the inventive method relates to; It can be the fresh herba cistanches chylocaulous of new results (without refrigeration or freeze preservation); Or place the fresh-keeping fresh herba cistanches chylocaulous of industrial refrigerator chamber, or place the fresh herba cistanches chylocaulous of industrial freezer refrigeration; Or the section of above-mentioned fresh herba cistanches chylocaulous or particle.
Herba Cistanches of the present invention is to meet orobanchaceae plant cistanche Cistanchedeserticola Y.C.Ma or Cistanche Tubulosa Cistanche tubulosa (Schrenk) Wight that Pharmacopoeia of the People's Republic of China version in 2005 is recorded.
The fresh-keeping Ku Wen of employing industry refrigerator chamber that the inventive method relates to is generally 1 ℃～13 ℃, and best refrigerator chamber fresh-keeping warehouse temperature is 4 ℃～10 ℃; Adopt the Ku Wen of industrial freezer refrigeration to be generally-52 ℃～-10 ℃, the storehouse temperature of best freezer refrigeration is-36 ℃～-18 ℃; The cold water water temperature that places tank that adopts is 1 ℃～20 ℃, and best cold water water temperature is 4 ℃～10 ℃; Each add the technical pure water purification or be 100%～400% (weight ratio) of Herba Cistanches smudge cells or filter residue through the addition of the technical pure water purification of precooling, optimum addition is 200%～300% (weight ratio); Technical pure water purification water temperature through precooling is 1 ℃～20 ℃, and optimum water temperature is 4 ℃～10 ℃.
Absorption method fixed form in the industrial production that the inventive method relates to obtains immobilized enzyme, normally beta-glucosidase industrial enzyme preparation enzyme liquid is contacted with sorbent material, removes non-adsorbable enzyme liquid through washing again and just can make immobilized enzyme; Comprise physisorphtion, ionic adsorption method; Sorbent material comprises micropore glass, Win 40350, cellophane, macroporous type synthetic resins, special cermacis, DEAE-Mierocrystalline cellulose, DEAE-polydextran gel, Amberlite IRA-93, IRA-410, IRA-900, Dowex-50, Amberlite CG-50, IRC-50, IR-120, can select usually external and domestic goods for use.
The beta-glucosidase industrial enzyme preparation that the inventive method relates to comprises beta-glucosidase and is rich in cellulase, plant extract prozyme, the plant hydrolyzed prozyme of beta-glucosidase.
The go out temperature of enzyme of the high temperature that the inventive method relates to is generally 75 ℃～100 ℃, and the time is generally 3min～10min; The temperature of the instantaneous enzyme that goes out of ultrahigh-temperature is generally 135 ℃～141 ℃.
The industrial immobilized enzyme reactor that the inventive method relates to can be common industrial stirred tank, immobilization bed bioreactor or fluidized-bed reactor.
The relative molecular mass of the ultra-filtration membrane that the inventive method relates to is 1000～4500, and best relative molecular mass is 1500～3500; The nf membrane relative molecular mass is 200～300, and best relative molecular mass is 250～280; Ultra-filtration membrane is selected the series membranes model of CA or CTA, PAN, PS, PSA, PES, PVDF, PEK, SPS for use; Nf membrane is selected the series membranes model of 4040-UHT-ESNA or 8540-UHY-ESNA, ESNA-FREE650, ESNA-FREE1700, NTR7450HG, NTR729HG, NF-CA film, NF-CA wound element and tubular fibre spare for use; Macroporous adsorbent resin is selected XAD for use; Diaion; SP; Posapak and Chromosorb and non-ionic type polymeric sorbent AB-8; CHA-III; CAD-40; D101; D301; D296; D396R; D4006; D4020; D3520; DA201; DM301; D130; GDX104; HPD100; HPD450; HPD500; HPD600; HPD8; H107; JD-KW; LD601; LD605; ME-1; ME-2; ME-3; NKA-2; NKA-9; R-A; S8; SIP; WLD and X-5 type serial model No.; Separation system of simulated moving bed chromatography and multi-functional chromatogram separation system can select for use MB, MD, ME, MF preparative scale chromatography to separate the chromatographic fractionation system of chromatograph, SMBC experiment type, SMBC pilot scale type, XZ12E-4L type, XZ20Z-2L type and the continuous preparative hplc type of SMBC industriallization.The ultra-filtration membrane that the inventive method relates to, nf membrane, macroporous adsorbent resin, non-ionic type polymeric sorbent, industrial chromatography column, moving bed chromatographic fractionation system, multi-functional chromatogram separation system can be selected usually external and domestic goods for use.
The industrial vacuum concentration technology that the inventive method relates to or the temperature of industrial film evaporating and concentrating process are generally 42 ℃～80 ℃, and optimum temps is 45 ℃～65 ℃; The drying tower EAT of the industrial drying process with atomizing that adopts is generally 125 ℃～285 ℃, and best EAT is 135 ℃～185 ℃.
The method of other industrially drying that the inventive method relates to comprises common industrial roller drying or air stream drying, fluidised bed drying, microwave vacuum drying, microwave drying, vacuum-drying or heat-wind circulate drying; Drying temperature is generally 50～100 ℃, and the best is 58～80 ℃.
The meal or the dry thing of the mullein glucoside monomeric compound that the inventive method is produced; Can directly be used for medical material; Or, also can be used as raw material or functional health-care foods such as compound raw material production oral liquid, capsule, tablet, granule, electuary, pill or teabag as cosmetic material.The promotion and implementation of the inventive method are with the health that this famous and precious Chinese medicine of Herba Cistanches is brought benefit to the mankind better.
The per-cent of the various amounts that relate in the inventive method (%) except other has explanation, all is weight percentage.
Embodiment
Below in conjunction with embodiment the present invention is done further explain.
Embodiment one:
1, selects that to place the storehouse temperature be 4 ℃ the fresh-keeping fresh herba cistanches chylocaulous of industrial refrigerator chamber; Place washing bath to use cold rinse; The cold water water temperature is 18 ℃, after picking up, places industrial broken milling device that the fragmentation of Herba Cistanches chylocaulous is milled; Adding technical pure water washing smudge cells becomes Herba Cistanches smudge cells soup compound, and the add-on of water is 300% of a smudge cells weight; Adopt industrial pressure filter press filtration then, obtain Herba Cistanches cell slurries; Through detecting, fragmentation is milled and separating technology process Herba Cistanches smudge cells soup compound and cytoplasm liquid temp are 20 ℃;
2, through the industrial microwave heating unit Herba Cistanches cell slurries are carried out the high temperature enzyme that goes out, temperature is 100 ℃, and the time is 3min; To go out Herba Cistanches cell slurries behind the enzyme as the catalyzed reaction substrate; Adopt physisorphtion Win 40350 in the common industrial production as the fixed form of sorbent material; Obtain the beta-glucosidase immobilized enzyme as biological catalyst; Carry out the continous way catalyzed reaction through common immobilization bed bioreactor, catalytic hydrolysis removes substrate echinacoside glucone part terminal glucone, an orientation and is converted into mullein glucoside monomeric compound; The catalytic reaction process temperature is 46 ℃, and pH is 4.9, and every batch of catalyzed reaction time is 5.5h, carries out 10 batches of catalyzed reactions continuously; Beta-glucoside enzyme activity in the catalytic reaction process reactant is 10U/g; Through detecting, the transformation efficiency that echinacoside is converted into mullein glucoside monomeric compound in the substrate reaches 94%;
3, the reaction solution after the above-mentioned catalyzed reaction being adopted relative molecular mass is that 4000 ultra-filtration membrane separates; Adopting relative molecular mass again is that 250 nf membrane is carried out nanofiltration; Adopt the industrial macroporous adsorbent resin of AB-8 type to carry out fractionation by adsorption then; And, adopt the multi-functional chromatogram separation system of XZ12E-4L type further to separate purification again through industrial chromatography column separating purification technology separation purification, obtain the mullein glucoside monomeric compound extracting solution of higher degree;
4, said extracted liquid is passed through the industrial vacuum concentration technology, temperature is 60 ℃, and the acquisition solid content is 42% enriched material; Further through the industrial vacuum drying process, drying temperature is 58 ℃, obtains the dry thing of mullein glucoside monomeric compound; Through assay determination, in the dry thing of being gathered in the crops (in dry-matter), mullein glucoside monomeric compound purity degree is 87.5%.
Embodiment two:
1, selects that to place the storehouse temperature be 10 ℃ the fresh-keeping fresh herba cistanches chylocaulous of industrial refrigerator chamber; Place washing bath to use cold rinse; The cold water water temperature is 4 ℃, after picking up, places industrial broken milling device that the fragmentation of Herba Cistanches chylocaulous is milled; Adding is that 4 ℃ technical pure water washing smudge cells becomes Herba Cistanches smudge cells soup compound through precooling to water temperature, and the add-on of water is 200% of a smudge cells weight; Adopt industrial centrifugal machine to separate then, obtain Herba Cistanches cell slurries; And again filter residue is added the technical pure water purification; The add-on of water is 200% of a filter residue weight; Further be milled into soup compound through common industrial broken runner milling, and adopt the ultrasonic extraction device that smudge cells is extracted, separate through industrial centrifugal machine again and further obtain Herba Cistanches cell slurries; Through detecting, fragmentation is milled and separating technology process Herba Cistanches smudge cells soup compound and cytoplasm liquid temp are 18 ℃;
2, through industrial heating unit Herba Cistanches cell slurries are carried out the high temperature enzyme that goes out, temperature is 100 ℃, and the time is 5min; To go out Herba Cistanches cell slurries behind the enzyme as the catalyzed reaction substrate; Adopt physisorphtion cellophane in the common industrial production as the fixed form of sorbent material; Obtain the beta-glucosidase immobilized enzyme as biological catalyst; Carry out the continous way catalyzed reaction through common industrial immobilization reactor, catalytic hydrolysis removes substrate echinacoside glucone part terminal glucone, an orientation and is converted into mullein glucoside monomeric compound; The catalytic reaction process temperature is 45 ℃, and pH is 4.5, and every batch of catalyzed reaction time is 6.0h, carries out 12 batches of catalyzed reactions continuously, and the beta-glucoside enzyme activity in the catalytic reaction process reactant is 29U/g; Through detecting, the transformation efficiency that echinacoside is converted into mullein glucoside monomeric compound in the substrate reaches 95%;
3, the reaction solution after the above-mentioned catalyzed reaction being adopted relative molecular mass is that 300 nf membrane is carried out nanofiltration; Adopt the industrial macroporous adsorbent resin of XAD type to carry out fractionation by adsorption then; And through industrial chromatography column separating purification technology purification; Adopt the multi-functional chromatogram separation system of XZ20Z-ZL type further to separate purification again, obtain the mullein glucoside monomeric compound extracting solution of higher degree;
4, said extracted liquid is passed through the industrial film evaporating and concentrating process, temperature is 55 ℃, and the acquisition solid content is 45% enriched material; Further through industrial roller drying technology, drying temperature is 55 ℃, obtains the dry thing of mullein glucoside monomeric compound; Through assay determination, in the dry thing of being gathered in the crops (in dry-matter), mullein glucoside monomeric compound purity degree is 89.9%.
Embodiment three:
1, selects and place the fresh herba cistanches chylocaulous of storehouse temperature for-52 ℃ industrial freezer refrigeration; Place washing bath with industrial pure water rinsing; After picking up; Place industrial broken milling device that the fragmentation of Herba Cistanches chylocaulous is milled, adding technical pure water washing smudge cells becomes Herba Cistanches smudge cells soup compound, and the add-on of water is 400% of a smudge cells weight; Adopt industrial pressure filter press filtration then, obtain Herba Cistanches cell slurries; Through detecting, fragmentation is milled and separating technology process Herba Cistanches smudge cells soup compound and cytoplasm liquid temp are 18 ℃;
2, through industrial heating unit Herba Cistanches cell slurries are carried out the high temperature enzyme that goes out, temperature is 95 ℃, and the time is 10min; To go out Herba Cistanches cell slurries behind the enzyme as the catalyzed reaction substrate; Adopt physisorphtion macroporous type synthetic resins in the common industrial production as the fixed form of sorbent material; Obtain the beta-glucosidase immobilized enzyme as biological catalyst; Carry out the continous way catalyzed reaction through common industrial fluidized bed reactor drum, catalytic hydrolysis removes substrate echinacoside glucone part terminal glucone, an orientation and is converted into mullein glucoside monomeric compound; The catalytic reaction process temperature is 42 ℃, and pH is 4.9, and every batch of catalyzed reaction time is 5h, carries out 8 batches of catalyzed reactions continuously, and through detecting, the beta-glucosidase enzyme activity in the catalytic reaction process reactant is 2U/g; Through detecting, the transformation efficiency that echinacoside is converted into mullein glucoside monomeric compound in the substrate reaches 81.2%;
3, the reaction solution after the above-mentioned catalyzed reaction being adopted relative molecular mass is that 4500 ultra-filtration membrane separates; Adopting relative molecular mass again is that 250 nf membrane is carried out nanofiltration; Adopt the industrial macroporous adsorbent resin of NKA-2 type to carry out fractionation by adsorption then; And further separate purification through industrial chromatography column separating purification technology, obtain the mullein glucoside monomeric compound extracting solution of higher degree;
4, said extracted liquid is passed through the industrial vacuum concentration technology, temperature is 60 ℃, and the acquisition solid content is 41% enriched material; Further through industrial drying process with atomizing, the drying tower EAT is 185 ℃, obtains the meal of mullein glucoside monomeric compound; Through assay determination, in the meal of being gathered in the crops (in dry-matter), mullein glucoside monomeric compound purity degree is 76.3%.
Embodiment four:
1, selects that to place the storehouse temperature be 13 ℃ the fresh-keeping fresh herba cistanches chylocaulous of industrial refrigerator chamber; Place washing bath to use cold rinse; The cold water water temperature is 13 ℃, after picking up, places industrial broken milling device that the fragmentation of Herba Cistanches chylocaulous is milled; Adding is that 2 ℃ technical pure water washing smudge cells becomes Herba Cistanches smudge cells soup compound through precooling to water temperature, and the add-on of water is 100% of a smudge cells weight; Adopt industrial pressure filter press filtration to separate then, obtain Herba Cistanches cell slurries; And filter residue added the technical pure water purification respectively through 2 times, the add-on of water is 300% of a filter residue weight, further is milled into soup compound through industrial broken runner milling usually again, separates through industrial pressure filter press filtration and further obtains Herba Cistanches cell slurries; Through detecting, fragmentation is milled and separating technology process Herba Cistanches smudge cells soup compound and cytoplasm liquid temp are 18 ℃;
2, the Herba Cistanches cell slurries that above-mentioned acquisition obtained carry out the instantaneous enzyme that goes out of industrial ultrahigh-temperature, and temperature is 138 ℃; To go out Herba Cistanches cell slurries behind the enzyme as the catalyzed reaction substrate; Adopt ionic adsorption method DEAE-polydextran gel in the common industrial production as the fixed form of sorbent material; Obtain the beta-glucosidase immobilized enzyme as biological catalyst; Carry out the continous way catalyzed reaction through common industrial stirred tank, catalytic hydrolysis removes substrate echinacoside glucone part terminal glucone, an orientation and is converted into mullein glucoside monomeric compound; The catalytic reaction process temperature is 47 ℃, and pH is 5.0, and every batch of catalyzed reaction time is 5.0h, carries out 16 batches of catalyzed reactions continuously, and the beta-glucoside enzyme activity in the catalytic reaction process reactant is 3.5U/g; Through detecting, the transformation efficiency that echinacoside is converted into mullein glucoside monomeric compound in the substrate reaches 88.1%;
3, the reaction solution after the above-mentioned catalyzed reaction being adopted relative molecular mass is that 1000 ultra-filtration membrane separates; Adopting relative molecular mass again is that 300 nf membrane is carried out nanofiltration; Adopt the industrial macroporous adsorbent resin of CAD-40 type to carry out fractionation by adsorption then; And further separate purification through industrial chromatography column separating purification technology, obtain the mullein glucoside monomeric compound extracting solution of higher degree;
4, said extracted liquid is passed through the industrial film evaporating and concentrating process, temperature is 60 ℃, and the acquisition solid content is 48% enriched material; Further through industrial roller drying technology, drying temperature is 55 ℃, obtains the dry thing of mullein glucoside monomeric compound; Through assay determination, in the dry thing of being gathered in the crops (in dry-matter), mullein glucoside monomeric compound purity degree is 87.6%.
Embodiment five:
1, selects and place the fresh herba cistanches chylocaulous of storehouse temperature for-18 ℃ industrial freezer refrigeration; Place washing bath with industrial pure water rinsing; After picking up; Place industrial broken milling device that the fragmentation of Herba Cistanches chylocaulous is milled, adding technical pure water washing smudge cells becomes Herba Cistanches smudge cells soup compound, and the add-on of water is 250% of a smudge cells weight; Adopt industrial pressure filter press filtration then, obtain Herba Cistanches cell slurries; Through detecting, fragmentation is milled and separating technology process Herba Cistanches smudge cells soup compound and cytoplasm liquid temp are 15 ℃;
2, through industrial heating unit Herba Cistanches cell slurries are carried out the high temperature enzyme that goes out, temperature is 85 ℃, and the time is 10min; To go out Herba Cistanches cell slurries behind the enzyme as the catalyzed reaction substrate; Adopt ionic adsorption method IRC-50 in the common industrial production as the fixed form of sorbent material; Obtain the beta-glucosidase immobilized enzyme as biological catalyst; Carry out the continous way catalyzed reaction through common industrial stirred tank, catalytic hydrolysis removes substrate echinacoside glucone part terminal glucone, an orientation and is converted into mullein glucoside monomeric compound; The catalytic reaction process temperature is 45 ℃, and pH is 4.1, and every batch of catalyzed reaction time is 6h, carries out 4 batches of catalyzed reactions continuously, and the beta-glucoside enzyme activity in the catalytic reaction process reactant is 70U/g; Through detecting, the transformation efficiency that echinacoside is converted into mullein glucoside monomeric compound in the substrate reaches 93.3%;
3, the reaction solution after the above-mentioned catalyzed reaction being adopted relative molecular mass is that 3500 ultra-filtration membrane separates; Adopting relative molecular mass again is that 250 nf membrane is carried out nanofiltration; Adopt the industrial macroporous adsorbent resin of DM301 type to carry out fractionation by adsorption then; And further separate purification through industrial chromatography column separating purification technology, obtain the mullein glucoside monomeric compound extracting solution of higher degree;
4, said extracted liquid is passed through the industrial vacuum concentration technology, temperature is 56 ℃, and the acquisition solid content is 30% enriched material; Further through the industrial vacuum drying process, drying temperature is 56 ℃, obtains the dry thing of mullein glucoside monomeric compound; Through assay determination, in the dry thing of being gathered in the crops (in dry-matter), mullein glucoside monomeric compound purity degree is 87.2%.
Embodiment six:
1, selects that to place the storehouse temperature be 10 ℃ the fresh-keeping fresh herba cistanches chylocaulous of industrial refrigerator chamber; Place washing bath to use cold rinse; The cold water water temperature is 8 ℃, after picking up, places industrial broken milling device that the fragmentation of Herba Cistanches chylocaulous is milled; Adding is that 10 ℃ technical pure water washing smudge cells becomes Herba Cistanches smudge cells soup compound through precooling to water temperature, and the add-on of water is 200% of a smudge cells weight; Adopt industrial pressure filter press filtration then, obtain Herba Cistanches cell slurries; And filter residue is added through precooling to water temperature again is 10 ℃ technical pure water purification; The add-on of water is 100% of a filter residue weight; Further be milled into soup compound through common industrial broken runner milling, separate through industrial pressure filter press filtration again and further obtain Herba Cistanches cell slurries; Through detecting, fragmentation is milled and separating technology process Herba Cistanches smudge cells soup compound and cytoplasm liquid temp are 12 ℃;
2, through industrial heating unit Herba Cistanches cell slurries are carried out the high temperature enzyme that goes out, temperature is 100 ℃, and the time is 4min; To go out Herba Cistanches cell slurries behind the enzyme as the catalyzed reaction substrate; Adopt ionic adsorption method IRC-120 in the common industrial production as the fixed form of sorbent material; Obtain plant hydrolyzed complex enzyme immobilization enzyme as biological catalyst; Carry out the discontinuous catalyzed reaction through common industrial stirred tank, catalytic hydrolysis removes substrate echinacoside glucone part terminal glucone, an orientation and is converted into mullein glucoside monomeric compound; The catalytic reaction process temperature is 48 ℃; PH is 4.9; The catalyzed reaction time is 6h, and the beta-glucoside enzyme activity in the catalytic reaction process reactant is 100U/g through detecting, and the transformation efficiency that echinacoside is converted into mullein glucoside monomeric compound in the substrate reaches 84.2%;
3, the reaction solution after the above-mentioned catalyzed reaction being adopted relative molecular mass is that 4000 hollow fiber ultrafiltration membrane is separated; Adopting relative molecular mass again is that 200 nf membrane is carried out nanofiltration; Adopt the industrial macroporous adsorbent resin of X-5 type to carry out fractionation by adsorption then; And further separate purification through industrial chromatography column separating purification technology, obtain the mullein glucoside monomeric compound extracting solution of higher degree;
4, said extracted liquid is passed through the industrial film evaporating and concentrating process, temperature is 70 ℃, and the acquisition solid content is 38% enriched material; Further through industrial roller drying technology, drying temperature is 60 ℃, obtains the dry thing of mullein glucoside monomeric compound; Through assay determination, in the dry thing of being gathered in the crops (in dry-matter), mullein glucoside monomeric compound purity degree is 76.2%.
Embodiment seven:
1, selects that to place the storehouse temperature be 4 ℃ the fresh-keeping fresh herba cistanches chylocaulous of industrial refrigerator chamber; Place washing bath to use cold rinse; The cold water water temperature is 15 ℃, after picking up, places industrial broken milling device that the fragmentation of Herba Cistanches chylocaulous is milled; Adding technical pure water washing smudge cells becomes Herba Cistanches smudge cells soup compound, and the add-on of water is 100% of a smudge cells weight; Adopt industrial pressure filter press filtration then, obtain Herba Cistanches cell slurries; Through detecting, fragmentation is milled and separating technology process Herba Cistanches smudge cells soup compound and cytoplasm liquid temp are 19 ℃;
2, through the industrial microwave heating unit Herba Cistanches cell slurries are carried out the high temperature enzyme that goes out, temperature is 95 ℃, and the time is 4min; To go out Herba Cistanches cell slurries behind the enzyme as the catalyzed reaction substrate; Adopt physisorphtion Win 40350 in the common industrial production as the fixed form of sorbent material; Obtain the beta-glucosidase immobilized enzyme as biological catalyst; Carry out the continous way catalyzed reaction through common immobilization bed bioreactor, catalytic hydrolysis removes substrate echinacoside glucone part terminal glucone, an orientation and is converted into mullein glucoside monomeric compound; The catalytic reaction process temperature is 52 ℃, and pH is 4.8, and every batch of catalyzed reaction time is 7h, carries out 11 batches of catalyzed reactions continuously, and the beta-glucoside enzyme activity that catalyzed reaction is crossed in the reactant is 900U/g; Through detecting, the transformation efficiency that echinacoside is converted into mullein glucoside monomeric compound in the substrate reaches 83.4%;
3, the reaction solution after the above-mentioned catalyzed reaction being adopted relative molecular mass is that 2000 ultra-filtration membrane separates, and adopting relative molecular mass again is that 250 nf membrane is carried out nanofiltration, and further separates purification through industrial chromatography column separating purification technology; Obtain the mullein glucoside monomeric compound extracting solution of higher degree;
4, said extracted liquid is passed through the industrial vacuum concentration technology, temperature is 60 ℃, and the acquisition solid content is 40% enriched material; Further through the industrial vacuum drying process, drying temperature is 58 ℃, obtains the dry thing of mullein glucoside monomeric compound; Through assay determination, in the dry thing of being gathered in the crops (in dry-matter), mullein glucoside monomeric compound purity degree is 65.4%.
Embodiment eight:
1, selects that to place the storehouse temperature be 8 ℃ the fresh-keeping fresh herba cistanches chylocaulous of industrial refrigerator chamber; Place washing bath to use cold rinse; The cold water water temperature is 14 ℃, after picking up, places industrial broken milling device that the fragmentation of Herba Cistanches chylocaulous is milled; Adding is that 6 ℃ technical pure water washing smudge cells becomes Herba Cistanches smudge cells soup compound through precooling to water temperature, and the add-on of water is 300% of a smudge cells weight; Adopt industrial centrifugal machine to separate then, obtain Herba Cistanches cell slurries; And again filter residue is added the technical pure water purification through 2 times respectively, and further be milled into soup compound through industrial broken runner milling usually, separate through industrial centrifugal machine again and further obtain Herba Cistanches cell slurries; Through detecting, fragmentation is milled and separating technology process Herba Cistanches smudge cells soup compound and cytoplasm liquid temp are 14 ℃;
2, through industrial heating unit Herba Cistanches cell slurries are carried out the high temperature enzyme that goes out, temperature is 100 ℃, and the time is 3min; To go out Herba Cistanches cell slurries behind the enzyme as the catalyzed reaction substrate; Adopt ionic adsorption method DEAE-Mierocrystalline cellulose in the common industrial production as the fixed form of sorbent material; Obtain the cellulase immobilization enzyme as biological catalyst; Carry out the continous way catalyzed reaction through common fluidized-bed reactor, catalytic hydrolysis removes substrate echinacoside glucone part terminal glucone, an orientation and is converted into mullein glucoside monomeric compound; The catalytic reaction process temperature is 53 ℃, and pH is 5.2, and every batch of catalyzed reaction time is 8h, carries out 7 batches of catalyzed reactions continuously, and the beta-glucoside enzyme activity in the catalytic reaction process reactant is 22U/g; Through detecting, the transformation efficiency that echinacoside is converted into mullein glucoside monomeric compound in the substrate reaches 69.3%;
3, the reaction solution after the above-mentioned catalyzed reaction being adopted relative molecular mass is that 4000 ultra-filtration membrane separates, and adopting relative molecular mass again is that 300 nf membrane is carried out nanofiltration, adopts the industrial macroporous adsorbent resin of R-A type to carry out fractionation by adsorption then; Obtain the mullein glucoside monomeric compound extracting solution of higher degree;
4, said extracted liquid is passed through the industrial film evaporating and concentrating process, temperature is 60 ℃, and the acquisition solid content is 40% enriched material; Further through industrial roller drying technology, drying temperature is 65 ℃, obtains the dry thing of mullein glucoside monomeric compound; Through assay determination, in the dry thing of being gathered in the crops (in dry-matter), mullein glucoside monomeric compound purity degree is 52.2%.
Embodiment nine:
1, selects and place the fresh herba cistanches chylocaulous of storehouse temperature for-36 ℃ industrial freezer refrigeration; Place washing bath with industrial pure water rinsing; After picking up; Place industrial broken milling device that the fragmentation of Herba Cistanches chylocaulous is milled, adding technical pure water washing smudge cells becomes Herba Cistanches smudge cells soup compound, and the add-on of water is 200% of a smudge cells weight; Adopt industrial pressure filter press filtration then, obtain Herba Cistanches cell slurries; Through detecting, fragmentation is milled and separating technology process Herba Cistanches smudge cells soup compound and cytoplasm liquid temp are 11 ℃;
2, through industrial heating unit Herba Cistanches cell slurries are carried out the high temperature enzyme that goes out, temperature is 100 ℃, and the time is 8min; To go out Herba Cistanches cell slurries behind the enzyme as the catalyzed reaction substrate; Adopt ionic adsorption method DEAE-polydextran gel in the common industrial production as the fixed form of sorbent material; Obtain the beta-glucosidase immobilized enzyme as biological catalyst; Carry out the continous way catalyzed reaction through common immobilization bed bioreactor, catalytic hydrolysis removes substrate echinacoside glucone part terminal glucone, an orientation and is converted into mullein glucoside monomeric compound; The catalytic reaction process temperature is 40 ℃, and pH is 4.6, and every batch of catalyzed reaction time is 4.5h, carries out 13 batches of catalyzed reactions continuously, and the beta-glucoside enzyme activity in the catalytic reaction process reactant is 21U/g; Through detecting, the transformation efficiency that echinacoside is converted into mullein glucoside monomeric compound in the substrate reaches 65.6%;
3, the reaction solution after the above-mentioned catalyzed reaction being adopted relative molecular mass is that 3500 ultra-filtration membrane separates, and adopting relative molecular mass again is that 250 nf membrane is carried out nanofiltration; Obtain the mullein glucoside monomeric compound extracting solution;
4, said extracted liquid is passed through the industrial vacuum concentration technology, temperature is 60 ℃, and the acquisition solid content is 38% enriched material; Further through industrial drying process with atomizing, the drying tower EAT is 185 ℃, obtains the meal of mullein glucoside monomeric compound; Through assay determination, in the meal of being gathered in the crops (in dry-matter), mullein glucoside monomeric compound purity degree is 50%.
Embodiment ten:
1, selects that to place the storehouse temperature be 13 ℃ the fresh-keeping fresh herba cistanches chylocaulous of industrial refrigerator chamber; Place washing bath to use cold rinse; The cold water water temperature is 6 ℃, after picking up, places industrial broken milling device that the fragmentation of Herba Cistanches chylocaulous is milled; Adding is that 4 ℃ technical pure water washing smudge cells becomes Herba Cistanches smudge cells soup compound through precooling to water temperature, and the add-on of water is 100% of a smudge cells weight; Adopt industrial centrifugal machine to separate then, obtain Herba Cistanches cell slurries; And again filter residue being added the technical pure water purification, the add-on of water is 100% of a filter residue weight, further is milled into soup compound through common industrial broken runner milling, separates through industrial centrifugal machine and further obtains Herba Cistanches cell slurries; Through detecting, fragmentation is milled and separating technology process Herba Cistanches smudge cells soup compound and cytoplasm liquid temp are 18 ℃;
2, the Herba Cistanches cell slurries with above-mentioned acquisition carry out the instantaneous enzyme that goes out of industrial ultrahigh-temperature, and temperature is 138 ℃; To go out Herba Cistanches cell slurries behind the enzyme as the catalyzed reaction substrate; Adopt ionic adsorption method IR-120 in the common industrial production as the fixed form of sorbent material; Obtain the beta-glucosidase immobilized enzyme as biological catalyst; Carry out the discontinuous catalyzed reaction through common industrial stirred tank, catalytic hydrolysis removes substrate echinacoside glucone part terminal glucone, an orientation and is converted into mullein glucoside monomeric compound; The catalytic reaction process temperature is 60 ℃, and pH is 4.8, and the catalyzed reaction time is 4h, and the beta-glucoside enzyme activity in the catalytic reaction process reactant is 500U/g; Through detecting, the transformation efficiency that echinacoside is converted into mullein glucoside monomeric compound in the substrate reaches 78%;
3, the reaction solution after the above-mentioned catalyzed reaction being adopted relative molecular mass is that 1500 ultra-filtration membrane separates; Adopting the SP macroporous resin adsorption to separate again purifies; And be that 300 nf membrane is carried out nanofiltration through relative molecular mass, obtain the mullein glucoside monomeric compound extracting solution of higher degree;
4, said extracted liquid is passed through the industrial film evaporating and concentrating process, temperature is 55 ℃, and the acquisition solid content is 42% enriched material; Further through industrial roller drying technology, drying temperature is 59 ℃, obtains the dry thing of mullein glucoside monomeric compound; Through assay determination, in the dry thing of being gathered in the crops (in dry-matter), mullein glucoside monomeric compound purity degree is 50.2%.
Claims (6)
1. method of utilizing the fresh herba cistanches raw material through the producing acteoside by biocatalysis and biotransformation monomeric compound is characterized in that the concrete steps of this method comprise:
(1) select the fresh herba cistanches chylocaulous, place washing bath with cold water or technical pure water rinse, the cold water water temperature in the washing bath is 1 ℃～20 ℃; After picking up, place the broken milling device of common industry that the fragmentation of Herba Cistanches chylocaulous is milled, add the technical pure water purification or become Herba Cistanches smudge cells soup compound through the technical pure water washing smudge cells of precooling; Described adding technical pure water purification or at every turn be 100%～400% of Herba Cistanches smudge cells weight through the amount of the technical pure water purification of precooling is 1 ℃～20 ℃ through the technical pure water purification water temperature of precooling; Adopt common industrial pressure filter press filtration or industrial centrifugal machine to separate then, obtain Herba Cistanches cell slurries; Fragmentation is milled the Herba Cistanches cytoplasm hydraulic control of Herba Cistanches smudge cells soup compound and separating technology process of technological process built in 4～20 ℃ of low temperature environments;
(2) the Herba Cistanches cell slurries that through common industrial heating unit step (1) obtained carry out high temperature go out enzyme or the instantaneous enzyme that goes out of ultrahigh-temperature; Described high temperature enzyme-removal temperature is 75 ℃～100 ℃, and the time is 3min～10min; The instantaneous enzyme-removal temperature of ultrahigh-temperature is 135 ℃～141 ℃; To go out Herba Cistanches cell slurries behind the enzyme as the catalyzed reaction substrate; The beta-glucosidase industrial enzyme preparation immobilized enzyme that adopts the absorption method fixed form acquisition in the common immobilized enzyme industrial production is as biological catalyst; Carry out catalyzed reaction through common industrial immobilized enzyme reactor, catalytic hydrolysis removes substrate echinacoside glucone part terminal glucone, an orientation and is converted into mullein glucoside monomeric compound; The catalytic reaction process temperature is 38 ℃～60 ℃, and pH is 4.0～5.8, and the time is 4h～8h, and the beta-glucoside enzyme activity in the catalytic reaction process reactant is 2U/g～1000U/g;
(3) will adopt common industrial ultra-filtration membrane to separate through the reacted reaction solution of step (2), selectivity be sieved and is held back and remove polysaccharide, protein macromolecule impurity and particulate and submicron impurity; Adopt common industrial nanofiltration membrane separation again, selectivity is sieved and is held back and remove amino acid, D-N.F,USP MANNITOL, trimethyl-glycine small molecular weight impurity; Obtain the mullein glucoside monomeric compound extracting solution; The relative molecular mass of described ultra-filtration membrane is 1000～4500, and the nf membrane relative molecular mass is 200～300;
(4) the mullein glucoside monomeric compound extracting solution that step (3) is obtained, through common industrial vacuum concentration technology or industrial film evaporating and concentrating process, obtaining solid content is the enriched material of 30%～50% weight ratio; Pass through industrial drying process with atomizing again, or other common industrially drying method, the meal or the dry thing of acquisition mullein glucoside monomeric compound.
2. according to the described method of claim 1, it is characterized in that the described catalytic reaction process temperature of step (2) is 42 ℃～48 ℃; PH is 4.5～5.3; Time, the beta-glucoside enzyme activity in the catalytic reaction process reactant was 25U/g～150U/g in order to be 5h～6h.
3. according to the described method of claim 1; It is characterized in that in the step (1) again filter residue through once to adding for several times the technical pure water purification or through the technical pure water purification of precooling; Once, further obtain Herba Cistanches cell slurries through industrial pressure filter press filtration or industrial centrifugal machine separation again to further being milled into soup compound through common industrial broken runner milling for several times; Described adding technical pure water purification or at every turn be 100%～400% of filter residue weight through the amount of the technical pure water purification of precooling is 1 ℃～20 ℃ through the technical pure water purification water temperature of precooling;
4. according to the described method of claim 1, it is characterized in that described beta-glucosidase industrial enzyme preparation, comprise that beta-glucosidase, cellulase, plant extract prozyme are or/and plant hydrolyzed prozyme.
5. according to the described method of claim 1; It is characterized in that adopting again in the step (3) common industrial macroporous resin adsorption separating technology; Or/and adopt common industrial chromatography column separating purification technology, or/and adopt common separation system of simulated moving bed chromatography technology or the further separation and purification of common multi-functional chromatogram separating technology.
6. according to the described method of claim 1, the temperature that it is characterized in that industrial vacuum concentration technology described in the step (4) or industrial film evaporating and concentrating process is 42 ℃～80 ℃; The drying tower EAT of industry drying process with atomizing is 125 ℃～285 ℃; The drying temperature of the method for other industrially drying is 50 ℃～100 ℃.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2009100416497A CN101629201B (en) | 2009-08-04 | 2009-08-04 | Method for producing acteoside by biocatalysis and biotransformation of fresh herba cistanches |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2009100416497A CN101629201B (en) | 2009-08-04 | 2009-08-04 | Method for producing acteoside by biocatalysis and biotransformation of fresh herba cistanches |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN101629201A CN101629201A (en) | 2010-01-20 |
| CN101629201B true CN101629201B (en) | 2012-07-04 |
Family
ID=41574480
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN2009100416497A Expired - Fee Related CN101629201B (en) | 2009-08-04 | 2009-08-04 | Method for producing acteoside by biocatalysis and biotransformation of fresh herba cistanches |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN101629201B (en) |
Families Citing this family (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104957640B (en) * | 2015-06-26 | 2017-07-11 | 内蒙古中兴华冠生物工程股份有限公司 | A kind of saline cistanche dietary fiber and preparation method thereof |
| CN106692311A (en) * | 2016-12-23 | 2017-05-24 | 庆阳敦博科技发展有限公司 | Processing method of herba cistanche tablet |
-
2009
- 2009-08-04 CN CN2009100416497A patent/CN101629201B/en not_active Expired - Fee Related
Non-Patent Citations (3)
| Title |
|---|
| 王丽楠等.不同初加工温度对肉苁蓉有效成分含量的影响.《中国药房》.2007,第18卷(第21期),1620-1623. * |
| 王丽楠等.鲜品肉苁蓉渣和汁有效成分的比较.《中草药》.2008,第39卷(第12期),1887-1889. * |
| 陈敏等.管花肉苁蓉中松果菊苷和毛蕊花糖苷的含量测定.《中国中药杂志》.2005,第30卷(第11期),839-841. * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN101629201A (en) | 2010-01-20 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN106008645B (en) | A kind of method that momordica grosvenori glycoside V is extracted from Momordica grosvenori | |
| CN109593034B (en) | Method for preparing shikimic acid from ginkgo leaf extraction waste liquid | |
| KR20000062140A (en) | manufacture method for ginseng saponin | |
| CN101214281B (en) | Method for producing cistanche salsa extract | |
| CN109651480A (en) | A method of separation momordica glycoside V | |
| CN103113422B (en) | Method for separating and refining high-purity L-arabinose and D-xylose with simulated moving bed | |
| CN107501224A (en) | A kind of method of Zeolite molecular sieve catalysis hydrolysis ginkgo flavone glycosides production flavone aglycone | |
| CN104357332B (en) | Aspergillus niger JH-2 and application to biotransformation and synthesis of asiatic acid | |
| CN101194934B (en) | Method for abstracting active compound rich in echinacoside and feltwort glycoside | |
| CN101629201B (en) | Method for producing acteoside by biocatalysis and biotransformation of fresh herba cistanches | |
| US8173837B1 (en) | Process for the production of L-citrulline from watermelon flesh and rind | |
| CN101629200B (en) | Method for producing acteoside by fresh herba cistanches | |
| CN101629198B (en) | Method for producing monomeric compounds of acteoside | |
| CN101619337A (en) | Method for producing monomeric compounds of acteoside by biocatalysis and biotransformation of echinacoside | |
| CN102949609A (en) | Preparation method of appetite and digestion promoting particles | |
| CN101654692B (en) | Method for producing mullein glucoside monomeric compound | |
| CN100497366C (en) | Process for extracting cyclic adenosine monophosphate (cAMP) from Chinese date | |
| Moreno et al. | Fractionation of food bioactive oligosaccharides | |
| CN101629197B (en) | Method for increasing content of acteoside in herba cistanches | |
| CN101619336B (en) | Method for producing acteoside by biocatalysis and biotransformation | |
| CN101629199A (en) | Method for producing acteoside by immobilized enzyme | |
| CN108147964A (en) | A kind of method with Integrated Membrane Technology separating chlorogenic acid | |
| CN107893092A (en) | A kind of tea fruit shell xylo-oligosaccharide and preparation method thereof | |
| CN104744524B (en) | Method for preparation of D-fructose and D-mannose by simulated moving bed reactor | |
| CN103131643B (en) | Strain for producing mannitol and method for producing mannitol through fermentation of strain |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20120704 |
|
| CF01 | Termination of patent right due to non-payment of annual fee |