CN107937380A - A kind of STEVIA REBAUDIANA extracts the carrier immobilized Trichoderma viride of steviol glycoside - Google Patents
A kind of STEVIA REBAUDIANA extracts the carrier immobilized Trichoderma viride of steviol glycoside Download PDFInfo
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- CN107937380A CN107937380A CN201711221337.5A CN201711221337A CN107937380A CN 107937380 A CN107937380 A CN 107937380A CN 201711221337 A CN201711221337 A CN 201711221337A CN 107937380 A CN107937380 A CN 107937380A
- Authority
- CN
- China
- Prior art keywords
- trichoderma viride
- carrier
- steviol glycoside
- stevia rebaudiana
- polyglutamic acid
- 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.)
- Granted
Links
- 241000223261 Trichoderma viride Species 0.000 title claims abstract description 56
- 235000019202 steviosides Nutrition 0.000 title claims abstract description 29
- 239000004383 Steviol glycoside Substances 0.000 title claims abstract description 28
- 235000019411 steviol glycoside Nutrition 0.000 title claims abstract description 28
- 229930182488 steviol glycoside Natural products 0.000 title claims abstract description 28
- 150000008144 steviol glycosides Chemical class 0.000 title claims abstract description 28
- 244000228451 Stevia rebaudiana Species 0.000 title claims abstract description 23
- 235000006092 Stevia rebaudiana Nutrition 0.000 title claims abstract description 23
- 239000000284 extract Substances 0.000 title claims description 11
- 229920002643 polyglutamic acid Polymers 0.000 claims abstract description 12
- 108010020346 Polyglutamic Acid Proteins 0.000 claims abstract description 9
- SXRSQZLOMIGNAQ-UHFFFAOYSA-N Glutaraldehyde Chemical compound O=CCCCC=O SXRSQZLOMIGNAQ-UHFFFAOYSA-N 0.000 claims abstract description 7
- 230000032050 esterification Effects 0.000 claims abstract description 7
- 238000005886 esterification reaction Methods 0.000 claims abstract description 7
- 238000004132 cross linking Methods 0.000 claims abstract description 5
- 239000003431 cross linking reagent Substances 0.000 claims abstract description 5
- 239000000243 solution Substances 0.000 claims description 29
- 239000007853 buffer solution Substances 0.000 claims description 28
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 15
- 239000006210 lotion Substances 0.000 claims description 13
- 238000003756 stirring Methods 0.000 claims description 10
- 238000002360 preparation method Methods 0.000 claims description 9
- QAOWNCQODCNURD-UHFFFAOYSA-N Sulfuric acid Chemical compound OS(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 claims description 8
- 239000007787 solid Substances 0.000 claims description 8
- 238000006243 chemical reaction Methods 0.000 claims description 7
- BHTJEPVNHUUIPV-UHFFFAOYSA-N pentanedial;hydrate Chemical compound O.O=CCCCC=O BHTJEPVNHUUIPV-UHFFFAOYSA-N 0.000 claims description 7
- XPFJYKARVSSRHE-UHFFFAOYSA-K trisodium;2-hydroxypropane-1,2,3-tricarboxylate;2-hydroxypropane-1,2,3-tricarboxylic acid Chemical compound [Na+].[Na+].[Na+].OC(=O)CC(O)(C(O)=O)CC(O)=O.[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O XPFJYKARVSSRHE-UHFFFAOYSA-K 0.000 claims description 6
- 239000000872 buffer Substances 0.000 claims description 5
- 210000004700 fetal blood Anatomy 0.000 claims description 5
- 238000005406 washing Methods 0.000 claims description 5
- 239000003595 mist Substances 0.000 claims description 4
- 239000000203 mixture Substances 0.000 claims description 4
- 239000012047 saturated solution Substances 0.000 claims description 4
- 230000008859 change Effects 0.000 claims description 3
- 239000007788 liquid Substances 0.000 claims description 3
- 238000010992 reflux Methods 0.000 claims description 3
- MWWQKLKFKPWECK-UHFFFAOYSA-N aminomethanetriol;hydrochloride Chemical compound Cl.NC(O)(O)O MWWQKLKFKPWECK-UHFFFAOYSA-N 0.000 claims description 2
- 239000007974 sodium acetate buffer Substances 0.000 claims description 2
- BHZOKUMUHVTPBX-UHFFFAOYSA-M sodium acetic acid acetate Chemical compound [Na+].CC(O)=O.CC([O-])=O BHZOKUMUHVTPBX-UHFFFAOYSA-M 0.000 claims description 2
- 239000004359 castor oil Substances 0.000 claims 2
- 235000019438 castor oil Nutrition 0.000 claims 2
- ZEMPKEQAKRGZGQ-XOQCFJPHSA-N glycerol triricinoleate Natural products CCCCCC[C@@H](O)CC=CCCCCCCCC(=O)OC[C@@H](COC(=O)CCCCCCCC=CC[C@@H](O)CCCCCC)OC(=O)CCCCCCCC=CC[C@H](O)CCCCCC ZEMPKEQAKRGZGQ-XOQCFJPHSA-N 0.000 claims 2
- 241000223259 Trichoderma Species 0.000 claims 1
- 230000003139 buffering effect Effects 0.000 claims 1
- 238000001914 filtration Methods 0.000 claims 1
- 239000001257 hydrogen Substances 0.000 claims 1
- 229910052739 hydrogen Inorganic materials 0.000 claims 1
- 238000005984 hydrogenation reaction Methods 0.000 claims 1
- 238000009413 insulation Methods 0.000 claims 1
- 238000011084 recovery Methods 0.000 abstract description 7
- 241000196324 Embryophyta Species 0.000 abstract description 3
- 210000002421 cell wall Anatomy 0.000 abstract description 3
- 238000002386 leaching Methods 0.000 abstract description 3
- 238000000034 method Methods 0.000 abstract description 2
- IJGRMHOSHXDMSA-UHFFFAOYSA-N nitrogen Substances N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 abstract description 2
- 229910052757 nitrogen Inorganic materials 0.000 abstract description 2
- 125000002924 primary amino group Chemical group [H]N([H])* 0.000 abstract description 2
- 238000000605 extraction Methods 0.000 description 7
- 239000000969 carrier Substances 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- 239000002904 solvent Substances 0.000 description 2
- 229920001661 Chitosan Polymers 0.000 description 1
- 235000007516 Chrysanthemum Nutrition 0.000 description 1
- 240000005250 Chrysanthemum indicum Species 0.000 description 1
- 241001573881 Corolla Species 0.000 description 1
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 1
- DGAQECJNVWCQMB-PUAWFVPOSA-M Ilexoside XXIX Chemical compound C[C@@H]1CC[C@@]2(CC[C@@]3(C(=CC[C@H]4[C@]3(CC[C@@H]5[C@@]4(CC[C@@H](C5(C)C)OS(=O)(=O)[O-])C)C)[C@@H]2[C@]1(C)O)C)C(=O)O[C@H]6[C@@H]([C@H]([C@@H]([C@H](O6)CO)O)O)O.[Na+] DGAQECJNVWCQMB-PUAWFVPOSA-M 0.000 description 1
- 241000904014 Pappus Species 0.000 description 1
- 241000544066 Stevia Species 0.000 description 1
- UEDUENGHJMELGK-HYDKPPNVSA-N Stevioside Chemical compound O([C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@H]1O[C@]12C(=C)C[C@@]3(C1)CC[C@@H]1[C@@](C)(CCC[C@]1([C@@H]3CC2)C)C(=O)O[C@H]1[C@@H]([C@@H](O)[C@H](O)[C@@H](CO)O1)O)[C@@H]1O[C@H](CO)[C@@H](O)[C@H](O)[C@H]1O UEDUENGHJMELGK-HYDKPPNVSA-N 0.000 description 1
- 244000269722 Thea sinensis Species 0.000 description 1
- 238000011001 backwashing Methods 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 239000002131 composite material Substances 0.000 description 1
- 235000009508 confectionery Nutrition 0.000 description 1
- 230000002255 enzymatic effect Effects 0.000 description 1
- 235000013305 food Nutrition 0.000 description 1
- 229930182478 glucoside Natural products 0.000 description 1
- 150000008131 glucosides Chemical class 0.000 description 1
- 238000007654 immersion Methods 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 238000002955 isolation Methods 0.000 description 1
- 238000011031 large-scale manufacturing process Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 235000021096 natural sweeteners Nutrition 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
- HELXLJCILKEWJH-NCGAPWICSA-N rebaudioside A Chemical compound O([C@H]1[C@H](O)[C@@H](CO)O[C@H]([C@@H]1O[C@H]1[C@@H]([C@@H](O)[C@H](O)[C@@H](CO)O1)O)O[C@]12C(=C)C[C@@]3(C1)CC[C@@H]1[C@@](C)(CCC[C@]1([C@@H]3CC2)C)C(=O)O[C@H]1[C@@H]([C@@H](O)[C@H](O)[C@@H](CO)O1)O)[C@@H]1O[C@H](CO)[C@@H](O)[C@H](O)[C@H]1O HELXLJCILKEWJH-NCGAPWICSA-N 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 229910052708 sodium Inorganic materials 0.000 description 1
- 239000011734 sodium Substances 0.000 description 1
- 229940013618 stevioside Drugs 0.000 description 1
- OHHNJQXIOPOJSC-UHFFFAOYSA-N stevioside Natural products CC1(CCCC2(C)C3(C)CCC4(CC3(CCC12C)CC4=C)OC5OC(CO)C(O)C(O)C5OC6OC(CO)C(O)C(O)C6O)C(=O)OC7OC(CO)C(O)C(O)C7O OHHNJQXIOPOJSC-UHFFFAOYSA-N 0.000 description 1
- 238000003809 water extraction Methods 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N11/00—Carrier-bound or immobilised enzymes; Carrier-bound or immobilised microbial cells; Preparation thereof
- C12N11/02—Enzymes or microbial cells immobilised on or in an organic carrier
- C12N11/08—Enzymes or microbial cells immobilised on or in an organic carrier the carrier being a synthetic polymer
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N11/00—Carrier-bound or immobilised enzymes; Carrier-bound or immobilised microbial cells; Preparation thereof
- C12N11/02—Enzymes or microbial cells immobilised on or in an organic carrier
- C12N11/06—Enzymes or microbial cells immobilised on or in an organic carrier attached to the carrier via a bridging agent
Abstract
The invention discloses a kind of STEVIA REBAUDIANA to extract the carrier immobilized Trichoderma viride of steviol glycoside, it is related to STEVIA REBAUDIANA extractive technique field, using polyglutamic acid rilanit special as carrier, by crosslinking agent glutaraldehyde by Trichoderma viride covalent cross-linking on carrier, obtain carrier immobilized Trichoderma viride；The polyglutamic acid rilanit special is made by food-grade polyglutamic acid with rilanit special through esterification.The present invention is used as crosslinking agent using glutaraldehyde, respectively react to form carbon-to-nitrogen double bon with the amino on carrier and Trichoderma viride using the aldehyde radical at glutaraldehyde both ends, make Trichoderma viride effectively covalent cross-linking on carrier, so as to destroy plant cell wall using Trichoderma viride, in favor of the Rapid Leaching of contained steviol glycoside in STEVIA REBAUDIANA, and then improve the recovery rate of steviol glycoside.
Description
Technical field：
The present invention relates to STEVIA REBAUDIANA extractive technique field, and in particular to a kind of STEVIA REBAUDIANA extraction steviol glycoside is fixed with carrier
Change Trichoderma viride.
Background technology：
STEVIA REBAUDIANA, composite family, sweetleaf Chrysanthemum herbaceos perennial.Stem is upright, and base portion tip lignifying, top is tender, dense
Short down, corolla base portion light violet magenta or white, top white.Achene is linear, slightly flat, brown, has pappus.STEVIA REBAUDIANA is liked in temperature
Grown in the environment of warm moistening, it is light sensitive.Steviol glycoside, also known as stevioside, are one extracted from Stevia leaves
Kind glucosides.Steviol glycoside is a kind of natural sweetener of sugariness low in calories, high, is one of raw material of food and pharmaceutical industries.
At present, the general extraction methods of steviol glycoside be by by STEVIA REBAUDIANA cured leaf immersion in water, filter by liquid with
Leaf, stem separation, are further purified using water or food-grade alcohol etc..Although this water extraction is simple to equipment requirement, with
For water as Extraction solvent, cost is low and environmentally friendly, but extraction time is grown, and the recovery rate of steviol glycoside is low, it is difficult to full
The large-scale production of sufficient steviol glycoside.
The content of the invention：
The technical problems to be solved by the invention are to provide a kind of sweet tea for improving steviol glycoside recovery rate using enzymatic isolation method
Ye Ju extracts the carrier immobilized Trichoderma viride of steviol glycoside.
The technical problems to be solved by the invention are realized using following technical solution：
A kind of STEVIA REBAUDIANA extracts the carrier immobilized Trichoderma viride of steviol glycoside, is made with polyglutamic acid-rilanit special
For carrier, by crosslinking agent glutaraldehyde by Trichoderma viride covalent cross-linking on carrier, obtain carrier immobilized Trichoderma viride；It is described
Polyglutamic acid-rilanit special is made by food-grade polyglutamic acid with rilanit special through esterification.
The preparation method of the carrier immobilized Trichoderma viride is：First carrier polyglutamic acid-rilanit special is dissolved in
Saturated solution is made in 25-30 DEG C of water, and adds 25% glutaraldehyde water solution and Trichoderma viride, then at insulated and stirred at 25-30 DEG C
Reaction, is adjusted pH value to 3-4, carrier immobilized Trichoderma viride is separated out with gel form, filter, gained gel after reaction
Washed repeatedly into washing lotion without free Trichoderma viride vigor with buffer solution I, then the stirring of gained gel is dissolved in buffer solution II
In, it is spare up to carrier immobilized Trichoderma viride solution, 4 DEG C of Cord bloods.
Polyglutamic acid-the rilanit special, 25% glutaraldehyde water solution, the mass ratio of Trichoderma viride are 50:5-10:
0.5-5。
The buffer solution I is selected from glycine-HCI buffer solution, acetic acid-sodium acetate buffer solution, citric acid-lemon
One kind in sour sodium buffer solution, solution ph is in 3-4.
The buffer solution II is selected from citric acid-sodium citrate buffer, trihydroxy aminomethane-hydrochloric acid buffer solution
In one kind, solution ph is in 6-7.
The preparation process of the polyglutamic acid-rilanit special is：It is added dropwise under stirring into food-grade polyglutamic acid
45-50 DEG C of water is until be completely dissolved, and added rilanit special to 4-5 using dilute sulfuric acid adjusting solution ph, be heated to back
Stream mode insulated and stirred is reacted, and gained mixture, which is concentrated under reduced pressure, after esterification is made the lotion of solid content 65-70%, treats
It is sent into after lotion cooled to room temperature in freeze drier, micro mist is made through micronizer in dry obtained solid, i.e., much
Polyglutamic acid-rilanit special.
The mass ratio of the food-grade polyglutamic acid and rilanit special is 40:5-10.
The beneficial effects of the invention are as follows：
(1) present invention using glutaraldehyde as crosslinking agent, using glutaraldehyde both ends aldehyde radical respectively with carrier and Trichoderma viride
On amino react to form carbon-to-nitrogen double bon, making Trichoderma viride, effectively covalent cross-linking is on carrier, so as to be broken using Trichoderma viride
Bad plant cell wall, in favor of the Rapid Leaching of contained steviol glycoside in STEVIA REBAUDIANA, and then improves the recovery rate of steviol glycoside；
(2) present invention belongs to water-solubility carrier, utilizes its spy to make polyglutamic acid-rilanit special by oneself as carrier
Some water-soluble and adsorptivities make Trichoderma viride effectively be attached in STEVIA REBAUDIANA, pass through the plant cell wall of rapid damage STEVIA REBAUDIANA
To further speed up the leaching velocity of steviol glycoside, make steviol glycoside extraction rate reached to more than 90%.
Embodiment：
In order to make the technical means, the creative features, the aims and the efficiencies achieved by the present invention easy to understand, tie below
Specific embodiment is closed, the present invention is further explained.
Embodiment 1
The preparation of carrier immobilized Trichoderma viride：50g carriers polyglutamic acid-rilanit special is first dissolved in 25-30 DEG C
Saturated solution is made in water, and adds 25% glutaraldehyde water solutions of 5g and 2g Trichoderma virides, anti-then at insulated and stirred at 25-30 DEG C
Should, pH value is adjusted after reaction to 3-4, carrier immobilized Trichoderma viride is separated out with gel form, is filtered, gained gel is used
Buffer solution I is washed into washing lotion without free Trichoderma viride vigor repeatedly, then the stirring of gained gel is dissolved in buffer solution II,
It is spare up to carrier immobilized Trichoderma viride solution, 4 DEG C of Cord bloods.
Buffer solution I is selected from glycine-HCI buffer solution, and solution ph is in 3-4；Buffer solution II be selected from citric acid-
Sodium citrate buffer, solution ph is in 6-7.
The preparation of polyglutamic acid-rilanit special：45-50 DEG C is added dropwise under stirring into 40g food-grade polyglutamic acids
Water is until be completely dissolved, and using dilute sulfuric acid adjusting solution ph to 4-5, add 5g rilanit specials, be heated to reflux shape
State insulated and stirred is reacted, and gained mixture, which is concentrated under reduced pressure, after esterification is made the lotion of solid content 65-70%, treats lotion
It is sent into after cooled to room temperature in freeze drier, micro mist is made through micronizer in dry obtained solid, up to poly paddy
Propylhomoserin-rilanit special.
Embodiment 2
The preparation of carrier immobilized Trichoderma viride：50g carriers polyglutamic acid-rilanit special is first dissolved in 25-30 DEG C
Saturated solution is made in water, and adds 25% glutaraldehyde water solutions of 5g and 2.5g Trichoderma virides, then at insulated and stirred at 25-30 DEG C
Reaction, is adjusted pH value to 3-4, carrier immobilized Trichoderma viride is separated out with gel form, filter, gained gel after reaction
Washed repeatedly into washing lotion without free Trichoderma viride vigor with buffer solution I, then the stirring of gained gel is dissolved in buffer solution II
In, it is spare up to carrier immobilized Trichoderma viride solution, 4 DEG C of Cord bloods.
Buffer solution I is selected from glycine-HCI buffer solution, and solution ph is in 3-4；Buffer solution II be selected from citric acid-
Sodium citrate buffer, solution ph is in 6-7.
The preparation of polyglutamic acid-rilanit special：45-50 DEG C is added dropwise under stirring into 40g food-grade polyglutamic acids
Water is until be completely dissolved, and using dilute sulfuric acid adjusting solution ph to 4-5, add 8g rilanit specials, be heated to reflux shape
State insulated and stirred is reacted, and gained mixture, which is concentrated under reduced pressure, after esterification is made the lotion of solid content 65-70%, treats lotion
It is sent into after cooled to room temperature in freeze drier, micro mist is made through micronizer in dry obtained solid, up to poly paddy
Propylhomoserin-rilanit special.
Reference examples 1
With the carrier in N- succinyl-chitosans alternate embodiment 2.
Reference examples 2
The preparation of carrier immobilized Trichoderma viride：50g carrier polyglutamic acids first are dissolved in 25-30 DEG C of water, and that saturation is made is molten
Liquid, and 25% glutaraldehyde water solutions of 5g and 2.5g Trichoderma virides are added, reacted then at insulated and stirred at 25-30 DEG C, reaction terminates
PH value is adjusted afterwards to 3-4, carrier immobilized Trichoderma viride is separated out with gel form, is filtered, gained gel is anti-with buffer solution I
After backwashing is washed into washing lotion without free Trichoderma viride vigor, then the stirring of gained gel is dissolved in buffer solution II, is consolidated up to carrier
Surely Trichoderma viride solution is changed, 4 DEG C of Cord bloods are spare.
Buffer solution I is selected from glycine-HCI buffer solution, and solution ph is in 3-4；Buffer solution II be selected from citric acid-
Sodium citrate buffer, solution ph is in 6-7.
Embodiment 3
Embodiment 1, embodiment 2, reference examples 1, the 2 carrier immobilized Trichoderma virides of made 5g of reference examples are utilized respectively to same batch
100g STEVIA REBAUDIANAs carry out the extraction of steviol glycoside, and using 500g water as Extraction solvent, 45-50 DEG C of water temperature raising degree, water carry the time
90min, measures steviol glycoside recovery rate, the results are shown in Table 1.Addition and the carrier immobilized Trichoderma viride vigor of 5g are set at the same time
Suitable free Trichoderma viride is as reference examples 3 and does not add the sky of carrier immobilized Trichoderma viride and free Trichoderma viride
White reference examples 4.
The recovery rate of 1 steviol glycoside of the present invention of table
| Group | Recovery rate/% |
| Embodiment 1 | 93.8 |
| Embodiment 2 | 94.2 |
| Reference examples 1 | 88.7 |
| Reference examples 2 | 79.4 |
| Reference examples 3 | 75.6 |
| Reference examples 4 | 58.3 |
The basic principles, main features and the advantages of the invention have been shown and described above.The technology of the industry
Personnel are it should be appreciated that the present invention is not limited to the above embodiments, and the above embodiments and description only describe this
The principle of invention, without departing from the spirit and scope of the present invention, various changes and modifications of the present invention are possible, these changes
Change and improvement all fall within the protetion scope of the claimed invention.The claimed scope of the invention by appended claims and its
Equivalent thereof.
Claims (7)
1. a kind of STEVIA REBAUDIANA extracts the carrier immobilized Trichoderma viride of steviol glycoside, it is characterised in that：With polyglutamic acid-hydrogenation
Castor oil as carrier, by crosslinking agent glutaraldehyde by Trichoderma viride covalent cross-linking on carrier, obtain carrier immobilized green
Trichoderma；Polyglutamic acid-the rilanit special is made by food-grade polyglutamic acid with rilanit special through esterification.
2. STEVIA REBAUDIANA according to claim 1 extracts the carrier immobilized Trichoderma viride of steviol glycoside, it is characterised in that its
Preparation method is：Carrier polyglutamic acid-rilanit special is first dissolved in 25-30 DEG C of water saturated solution is made, and add 25%
Glutaraldehyde water solution and Trichoderma viride, react then at insulated and stirred at 25-30 DEG C, adjust pH value after reaction to 3-4, make load
Body immobilization Trichoderma viride is separated out with gel form, and filtering, gained gel is washed into washing lotion without free repeatedly with buffer solution I
Trichoderma viride vigor, then the stirring of gained gel is dissolved in buffer solution II, up to carrier immobilized Trichoderma viride solution, 4 DEG C
Cord blood is spare.
3. STEVIA REBAUDIANA according to claim 2 extracts the carrier immobilized Trichoderma viride of steviol glycoside, it is characterised in that：Institute
It is 50 to state polyglutamic acid-rilanit special, 25% glutaraldehyde water solution, the mass ratio of Trichoderma viride:5-10:0.5-5.
4. STEVIA REBAUDIANA according to claim 2 extracts the carrier immobilized Trichoderma viride of steviol glycoside, it is characterised in that：Institute
It is molten selected from glycine-HCI buffer solution, acetic acid-sodium acetate buffer solution, citric acid-sodium citrate buffering to state buffer solution I
One kind in liquid, solution ph is in 3-4.
5. STEVIA REBAUDIANA according to claim 2 extracts the carrier immobilized Trichoderma viride of steviol glycoside, it is characterised in that：Institute
The one kind of buffer solution II in citric acid-sodium citrate buffer, trihydroxy aminomethane-hydrochloric acid buffer solution is stated,
Solution ph is in 6-7.
6. STEVIA REBAUDIANA according to claim 2 extracts the carrier immobilized Trichoderma viride of steviol glycoside, it is characterised in that：Institute
The preparation process for stating polyglutamic acid-rilanit special is：Into food-grade polyglutamic acid, 45-50 DEG C of water of dropwise addition is straight under stirring
Solution ph is adjusted to 4-5 to being completely dissolved, and using dilute sulfuric acid, is added rilanit special, is heated to reflux state insulation
Stirring is reacted, and gained mixture, which is concentrated under reduced pressure, after esterification is made the lotion of solid content 65-70%, treats that lotion is naturally cold
But into feeding freeze drier after room temperature, micro mist is made through micronizer in dry obtained solid, up to polyglutamic acid-hydrogen
Change castor oil.
7. STEVIA REBAUDIANA according to claim 6 extracts the carrier immobilized Trichoderma viride of steviol glycoside, it is characterised in that：Institute
The mass ratio for stating food-grade polyglutamic acid and rilanit special is 40:5-10.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201711221337.5A CN107937380B (en) | 2017-11-29 | 2017-11-29 | Carrier immobilized trichoderma viride for extracting stevioside from stevia rebaudiana |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201711221337.5A CN107937380B (en) | 2017-11-29 | 2017-11-29 | Carrier immobilized trichoderma viride for extracting stevioside from stevia rebaudiana |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN107937380A true CN107937380A (en) | 2018-04-20 |
| CN107937380B CN107937380B (en) | 2020-04-14 |
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| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201711221337.5A Active CN107937380B (en) | 2017-11-29 | 2017-11-29 | Carrier immobilized trichoderma viride for extracting stevioside from stevia rebaudiana |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109628321A (en) * | 2018-12-19 | 2019-04-16 | 南京农业大学 | It is a kind of to utilize stevia rebaudiana dregs solid fermentation trichoderma strain and preparation method thereof |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101550411A (en) * | 2009-02-09 | 2009-10-07 | 苏州大学 | Immobilized cellulose and a method for hydrolyzing plant material thereby for extracting active ingredients |
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| CN105541937A (en) * | 2016-01-19 | 2016-05-04 | 苏州赛分科技有限公司 | Extracting method for stevioside in stevia rebaudiana |
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| CN109628321A (en) * | 2018-12-19 | 2019-04-16 | 南京农业大学 | It is a kind of to utilize stevia rebaudiana dregs solid fermentation trichoderma strain and preparation method thereof |
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| CN107937380B (en) | 2020-04-14 |
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