SU436854A1 - The method of obtaining o-diphenol oxidase - Google Patents
The method of obtaining o-diphenol oxidaseInfo
- Publication number
- SU436854A1 SU436854A1 SU1768525A SU1768525A SU436854A1 SU 436854 A1 SU436854 A1 SU 436854A1 SU 1768525 A SU1768525 A SU 1768525A SU 1768525 A SU1768525 A SU 1768525A SU 436854 A1 SU436854 A1 SU 436854A1
- Authority
- SU
- USSR - Soviet Union
- Prior art keywords
- enzyme
- obtaining
- diphenol oxidase
- buffer solution
- precipitate
- Prior art date
Links
- 102000004316 Oxidoreductases Human genes 0.000 title description 5
- 108090000854 Oxidoreductases Proteins 0.000 title description 5
- 238000000034 method Methods 0.000 title description 2
- 108090000790 Enzymes Proteins 0.000 description 9
- 102000004190 Enzymes Human genes 0.000 description 9
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 description 6
- 239000007853 buffer solution Substances 0.000 description 6
- 244000269722 Thea sinensis Species 0.000 description 5
- 239000002244 precipitate Substances 0.000 description 5
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 4
- 239000002994 raw material Substances 0.000 description 4
- ISWSIDIOOBJBQZ-UHFFFAOYSA-N Phenol Chemical compound OC1=CC=CC=C1 ISWSIDIOOBJBQZ-UHFFFAOYSA-N 0.000 description 3
- 150000002500 ions Chemical class 0.000 description 3
- 239000000463 material Substances 0.000 description 3
- 235000010378 sodium ascorbate Nutrition 0.000 description 3
- 229960005055 sodium ascorbate Drugs 0.000 description 3
- PPASLZSBLFJQEF-RKJRWTFHSA-M sodium ascorbate Substances [Na+].OC[C@@H](O)[C@H]1OC(=O)C(O)=C1[O-] PPASLZSBLFJQEF-RKJRWTFHSA-M 0.000 description 3
- PPASLZSBLFJQEF-RXSVEWSESA-M sodium-L-ascorbate Chemical compound [Na+].OC[C@H](O)[C@H]1OC(=O)C(O)=C1[O-] PPASLZSBLFJQEF-RXSVEWSESA-M 0.000 description 3
- 239000000126 substance Substances 0.000 description 3
- 108010044467 Isoenzymes Proteins 0.000 description 2
- 235000006468 Thea sinensis Nutrition 0.000 description 2
- BFNBIHQBYMNNAN-UHFFFAOYSA-N ammonium sulfate Chemical compound N.N.OS(O)(=O)=O BFNBIHQBYMNNAN-UHFFFAOYSA-N 0.000 description 2
- 229910052921 ammonium sulfate Inorganic materials 0.000 description 2
- 235000011130 ammonium sulphate Nutrition 0.000 description 2
- 238000004519 manufacturing process Methods 0.000 description 2
- 230000001376 precipitating effect Effects 0.000 description 2
- 239000011780 sodium chloride Substances 0.000 description 2
- DHMQDGOQFOQNFH-UHFFFAOYSA-M Aminoacetate Chemical compound NCC([O-])=O DHMQDGOQFOQNFH-UHFFFAOYSA-M 0.000 description 1
- 229920006051 Capron® Polymers 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
- 229920002292 Nylon 6 Polymers 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 229910021538 borax Inorganic materials 0.000 description 1
- 239000000872 buffer Substances 0.000 description 1
- 229920001429 chelating resin Polymers 0.000 description 1
- 239000007979 citrate buffer Substances 0.000 description 1
- UQGFMSUEHSUPRD-UHFFFAOYSA-N disodium;3,7-dioxido-2,4,6,8,9-pentaoxa-1,3,5,7-tetraborabicyclo[3.3.1]nonane Chemical compound [Na+].[Na+].O1B([O-])OB2OB([O-])OB1O2 UQGFMSUEHSUPRD-UHFFFAOYSA-N 0.000 description 1
- 238000010828 elution Methods 0.000 description 1
- JBKVHLHDHHXQEQ-UHFFFAOYSA-N epsilon-caprolactam Chemical compound O=C1CCCCCN1 JBKVHLHDHHXQEQ-UHFFFAOYSA-N 0.000 description 1
- 238000001914 filtration Methods 0.000 description 1
- 239000007791 liquid phase Substances 0.000 description 1
- 230000004060 metabolic process Effects 0.000 description 1
- 239000002245 particle Substances 0.000 description 1
- 239000000049 pigment Substances 0.000 description 1
- 239000000843 powder Substances 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 229920005989 resin Polymers 0.000 description 1
- 239000011347 resin Substances 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 239000011734 sodium Substances 0.000 description 1
- 229910052708 sodium Inorganic materials 0.000 description 1
- 235000010339 sodium tetraborate Nutrition 0.000 description 1
- 239000004328 sodium tetraborate Substances 0.000 description 1
- 239000000243 solution Substances 0.000 description 1
Landscapes
- Enzymes And Modification Thereof (AREA)
Description
1one
Изобретение относитс к ферментной промышленности , точнее к способу получени о-дифенолоксидазы, играющей важную роль в метаболизме клетки чайного растени , а также в формировании таких качественных показателей ча , как вкус, цвет насто и частично аромат.The invention relates to the enzyme industry, more precisely to a method for producing o-diphenol oxidase, which plays an important role in the metabolism of a tea plant cell, as well as in the formation of tea quality indicators such as taste, color, and partly aroma.
Известен способ получени о-дифенолоксидазы , предусматривающий измельчение сырь , экстракцию фермента, удаление из экстракта нерастворимых частиц, например, фильтрацией , осаждение фермента, например сульфатом аммони , отделение полученного осадка, например , центрифугированием, растворение осадка в буферном растворе, пропуск раствора через ионообменники и элюцию фермента с ионообменника.A known method for producing o-diphenol oxidase involves grinding the raw material, extracting the enzyme, removing insoluble particles from the extract, for example, by filtration, precipitating the enzyme, for example ammonium sulfate, separating the precipitate obtained, for example, by centrifuging, dissolving the precipitate in a buffer solution, passing the solution through ion exchangers and elution of the enzyme from the ion exchanger.
Предлагаемый способ позвол ет получить изоферменты о-дифенолоксидазы из чайного растени .The proposed method allows to obtain o-diphenol oxidase isoenzymes from a tea plant.
Достигаетс это тем, что из измельченного сырь удал ют фенольные вещества, экстракцию фермента осуществл ют буферным раствором , содержащим аскорбат натри , теграборат натри и хлористый натрий, рН не ниже 6,0.This is achieved by removing phenol substances from the crushed raw material, extracting the enzyme with a buffer solution containing sodium ascorbate, sodium tegraborate and sodium chloride, pH not lower than 6.0.
Дл экстракции фермента в качестве буферного раствора используют глицинатный буфер при значени х рН 9,0, в котором содержание аскорбата натри , тетрабората натри To extract the enzyme, glycinate buffer is used as a buffer solution at pH values of 9.0, in which the content of sodium ascorbate, sodium tetraborate
и хлористого натри соответственно составл ют (в мол х): 0,03; 0,05 и 1,0, а дл растворени осадка в качестве буферного раствора используют 0,1 М цитратный буфер при рНand sodium chloride, respectively, are (in moles): 0.03; 0.05 and 1.0, and to dissolve the precipitate as a buffer solution, use 0.1 M citrate buffer at pH
не ниже 5,0. При этом элюцию фермента с ионообменника целесообразно осуществл ть буферным раствором с ионной силой 0,5, а осаждение фермента - из экстракта или сульфатом аммони при 100%-ном насыщении,not lower than 5.0. In this case, it is advisable to elute the enzyme from the ion exchanger with a buffer solution with an ionic strength of 0.5, and precipitate the enzyme from the extract or ammonium sulfate at 100% saturation,
или ацетоном при соотнощении 1 : 4.or acetone at a ratio of 1: 4.
Дл получени гомогенных изоферментов о-дифенолоксидазы элюаты дополнительно хроматографируют на колонках с гидроксилаппатитом и амберлитом, а жидкую фазу, полученную после отделени осадка фермента, можно использовать дл выделени тапинокатехинового комплекса.To obtain homogeneous o-diphenol oxidase isoenzymes, the eluates are additionally chromatographed on columns with hydroxylappatite and amberlite, and the liquid phase obtained after separation of the enzyme precipitate can be used to isolate the tapene-catechol complex.
Пример 1. Сырье (чайпый лист) измельчают в замороженном виде до максимальногоExample 1. Raw materials (tea leaf) are crushed frozen to a maximum
разрушени клеток, а затем удал ют фенольные пигменты, смолы и другие вещества путем осаждени 80%-ным ацетоном (соотнощение ча к ацетону 1 :4) или экстракцией измельченного сырь поликапролактамовым порощком (капроном) в количестве 2 г на 1 г сырь в зависимости от содержани фенольных веществ в материале.destroying the cells, and then removing the phenolic pigments, resins and other substances by precipitating with 80% acetone (tea ratio to acetone 1: 4) or extracting the crushed material with a polycaprolactam powder (capron) in an amount of 2 g per 1 g of raw material, depending on content of phenolic substances in the material.
Белки из обработанного материала эксграгируют буферным раствором при значени хProteins from the treated material are extracted with a buffer solution at
рН 6,0 и выше, содержащим аскорбат натри ,pH 6.0 and above, containing sodium ascorbate,
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| SU1768525A SU436854A1 (en) | 1972-04-04 | 1972-04-04 | The method of obtaining o-diphenol oxidase |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| SU1768525A SU436854A1 (en) | 1972-04-04 | 1972-04-04 | The method of obtaining o-diphenol oxidase |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| SU436854A1 true SU436854A1 (en) | 1974-07-25 |
Family
ID=20509337
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| SU1768525A SU436854A1 (en) | 1972-04-04 | 1972-04-04 | The method of obtaining o-diphenol oxidase |
Country Status (1)
| Country | Link |
|---|---|
| SU (1) | SU436854A1 (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2010089385A1 (en) * | 2009-02-06 | 2010-08-12 | Novozymes Biopharma Dk A/S | Purification process |
-
1972
- 1972-04-04 SU SU1768525A patent/SU436854A1/en active
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2010089385A1 (en) * | 2009-02-06 | 2010-08-12 | Novozymes Biopharma Dk A/S | Purification process |
| EP2617733A1 (en) * | 2009-02-06 | 2013-07-24 | Novozymes Biopharma DK A/S | Purification process |
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