CS206992B1 - Method of separation of the glucoseoxidasis - Google Patents
Method of separation of the glucoseoxidasis Download PDFInfo
- Publication number
- CS206992B1 CS206992B1 CS725579A CS725579A CS206992B1 CS 206992 B1 CS206992 B1 CS 206992B1 CS 725579 A CS725579 A CS 725579A CS 725579 A CS725579 A CS 725579A CS 206992 B1 CS206992 B1 CS 206992B1
- Authority
- CS
- Czechoslovakia
- Prior art keywords
- cellulose
- glucose oxidase
- solution
- sorption
- separation
- Prior art date
Links
- 238000000034 method Methods 0.000 title claims description 10
- 238000000926 separation method Methods 0.000 title description 2
- 239000001913 cellulose Substances 0.000 claims description 16
- 229920002678 cellulose Polymers 0.000 claims description 16
- 108010015776 Glucose oxidase Proteins 0.000 claims description 15
- 239000004366 Glucose oxidase Substances 0.000 claims description 15
- 229940116332 glucose oxidase Drugs 0.000 claims description 15
- 235000019420 glucose oxidase Nutrition 0.000 claims description 15
- 238000001179 sorption measurement Methods 0.000 claims description 5
- 230000002538 fungal effect Effects 0.000 claims description 2
- 235000010980 cellulose Nutrition 0.000 description 12
- 230000000694 effects Effects 0.000 description 7
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 6
- 239000002594 sorbent Substances 0.000 description 5
- 239000011324 bead Substances 0.000 description 4
- 108090000790 Enzymes Proteins 0.000 description 3
- 102000004190 Enzymes Human genes 0.000 description 3
- 229940088598 enzyme Drugs 0.000 description 3
- 239000011780 sodium chloride Substances 0.000 description 3
- MZVQCMJNVPIDEA-UHFFFAOYSA-N [CH2]CN(CC)CC Chemical group [CH2]CN(CC)CC MZVQCMJNVPIDEA-UHFFFAOYSA-N 0.000 description 2
- -1 diethylaminoethyl groups Chemical group 0.000 description 2
- 229920000168 Microcrystalline cellulose Polymers 0.000 description 1
- 230000001419 dependent effect Effects 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 239000000706 filtrate Substances 0.000 description 1
- 238000005342 ion exchange Methods 0.000 description 1
- 235000019813 microcrystalline cellulose Nutrition 0.000 description 1
- 239000008108 microcrystalline cellulose Substances 0.000 description 1
- 229940016286 microcrystalline cellulose Drugs 0.000 description 1
- 239000002245 particle Substances 0.000 description 1
- 239000000725 suspension Substances 0.000 description 1
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 1
Landscapes
- Treatment Of Liquids With Adsorbents In General (AREA)
Description
Vynález se týká způsobu separace glukózooxidázy z roztoků získaných extrakci plísňových buněk.The invention relates to a process for separating glucose oxidase from solutions obtained by fungal cell extraction.
K separaci glukózooxidázy z roztoků se dosud užívá způsob, při kterém se glukózooxidáza sorbuje na mikrokrystalické fibrilárni celulóze s dietylaminoetylovymi nebo distylaminohydroxipropylovými výměnnými skupinami, ionexová celulóza se zachycenou glukózooxidázou se odfiltruje a čistá glukózooxidáza se izoluje eluci.The separation of glucose oxidase from solutions has hitherto used a method in which glucose oxidase is sorbed onto microcrystalline fibrillar cellulose with diethylaminoethyl or distylaminohydroxipropyl exchange groups, the ion exchange cellulose with the captured glucose oxidase is filtered off and the pure glucose oxidase is isolated.
Nevýhody tohoto způsobu tkvi především v tom,.že vláknitá struktura mikrokrystalické celulózy je příčinou špatné filtrovatelnosti a sorpci nelze provádět kontinuálně v kolonách, ale pouze násadou celulózy do zpracovávaného roztoku. Dalši nevýhodou tohoto způsobu je jeho nepřesnost a špatná reprodukovatelnost.The disadvantages of this method are, in particular, that the fibrous structure of microcrystalline cellulose is the cause of poor filterability and sorption cannot be carried out continuously in columns, but only by feeding the cellulose into the solution to be treated. Another disadvantage of this method is its inaccuracy and poor reproducibility.
Uvedené nevýhody odstraňuje způsob separace glukózooxidázy podle vynálezu, jehož podstata spočívá v tom, že se roztok glukózooxidázy uvádi do styku s celulózou v perlové formě s dietylaminoetylovými nebo dietylaminohydroxipropylovými výměnnými skupinami o velikosti zrn 30 až 800 fum a zachycená glukózooxidáza se izoluje ze sorbentu eluci.The above-mentioned disadvantages are eliminated by the process for separating the glucose oxidase according to the invention, which comprises contacting the glucose oxidase solution with cellulose in pearl form with diethylaminoethyl or diethylaminohydroxipropyl grain size groups of 30-800 [mu] m and recovering the glucose oxidase collected from the sorbent.
Výhodou způsobu podle vynálezu je, že jej lze provádět jak šaržovitě násadou celulózy do roztoku glukózooxidázy, tak i kontinuálně průtokem roztoku kolonou naplněnou perlovou celulózou. Oe to umožněno vlastnosti perlové celulózy a jejich derivátů, které jsou v potřebném rozsahu tlaků nestlačitelné a nezpůsobuji pokles průtoku v závislosti na tlaku. Dalši výhodou tohoto způsobu je jeho přesnost a reprodukovatelnost. Makroporézni charakter perlové celulózy umožňuje, na rozdil od fibrilárni celulózy,uplatnit při sorpci nejen vnější, ale i vnitřní povrch a perlová celulóza obsahuje vyšší množství sorpčně aktivních skupin, takže dochází ke zvýšeni sorpčni kapacity vůči glukózooxidáze.An advantage of the process according to the invention is that it can be carried out both batchwise by loading the cellulose into the glucose oxidase solution and continuously by flowing the solution through a column packed with pearl cellulose. This has enabled the properties of the bead cellulose and its derivatives, which are incompressible to the required pressure range and do not cause a pressure-dependent drop in flow. Another advantage of this method is its accuracy and reproducibility. The macroporous nature of the bead cellulose makes it possible, in contrast to the fibrillar cellulose, to apply not only the outer but also the inner surface of the sorption and the bead cellulose contains higher amounts of sorption active groups, thus increasing the sorption capacity towards glucose oxidase.
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Přiklad 1 t v 1 000 ml roztoku glukózooxidázy o aktivitě 4.500/ul Og ml“^ h“1- bylo suspendováno 2,5 g OEAE-perlové celulózy obsahujici 2,0 milimolů dietylaminoetylových skupin v 1 g při velikosti zrna 30 <um. Suspenze byla utichána 30 minut při 20 °C a potom byl sorbent odfiltrován. Ve filtrátu bylo zjištěno 7,75 % původni aktivity. Veškerá sorbované enzymová aktivita byla ze sorbentu eluována 1 M roztokem chloridu sodného při teplotě 15 až 20 °C. Přiklad 2 t v 1 000 ml roztoku glukozooxidázy o aktivitě 4.500 ^ul 0gml*^h“^ bylo suspendováno 10 g DEAHP-cslulózy obsahujici 2,5 millmolu distylaminohydroxipropylových skupin v 1 g při velikosti zrna 30 (um. Suspenze byla michána 30 minut při 20 °C a potom byl sorbent odfiltrován. Ve filtrátu bylo zjištěno 15 % původni enzymové aktivity. Veškerá sorbované enzymová aktivita byla ze sorbentu eluována 1 M roztokem chloridu sodného při teplotě 15 až 20 °C.Example 1 tv 1000 ml solution of glucose oxidase activity of 4500 / ul Og ml "^ h" 1 were suspended 2.5 g DEAE-bead cellulose containing 2.0 millimoles diethylaminoethyl groups in 1 g at a particle size of 30 <m. The suspension was quenched for 30 minutes at 20 ° C and then the sorbent was filtered off. 7.75% of the original activity was found in the filtrate. All sorbed enzyme activity was eluted from the sorbent with 1 M sodium chloride solution at 15-20 ° C. Example 2 t 1000 ml of a glucosoxidase activity of 4.500 µl of 0 gml * 4 h -1 was suspended in 10 g of DEAHP-cellulose containing 2.5 ml of distylamino-hydroxypropyl groups per g at a grain size of 30 (µm). The sorbent was eluted from the sorbent with 1 M sodium chloride solution at 15-20 ° C.
Přiklad 3 » □EAE - celulóza perlová o koncentraci dietylaminoetylových skupin 2,0 milimolů / g byla naplněna do sloupce rozměrů 0,9 x 15 cm. Sloupcem byl propouštěn roztok glukozooxidázy o aktivitě 4.500 ^um Og ml“^ h“1, rychlostí 1 mí cm“2' min.“'1'. Při zjištěné aktivitě 340 ml Og ml“ h“1 ve vytékajicim roztoku z kolony, tj. po průtoku 500 ml roztoku kolonou, bylo propouštěni zastaveno. Potom byl sloupec promyt vodou a enzym byl eluován 1 M roztokem chloridu sodného. V eluátu bylo získáno 92,25 % veškeré vložené enzymově aktivity.EXAMPLE 3 EAE pearl cellulose having a diethylaminoethyl group concentration of 2.0 millimoles / g was filled into a 0.9 x 15 cm column. Column redundancy solution was glucose oxidase activity of 4500 microns Og ml "^ h" 1, a rate of 1 mu cm "2" min. "'1'. At the detected activity of 340 ml Og ml " h " 1 in the effluent from the column, i.e. after 500 ml of solution flowing through the column, the discharge was stopped. The column was washed with water and the enzyme eluted with 1 M sodium chloride solution. 92.25% of the total enzyme activity loaded was recovered in the eluate.
Claims (2)
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CS725579A CS206992B1 (en) | 1979-10-25 | 1979-10-25 | Method of separation of the glucoseoxidasis |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CS725579A CS206992B1 (en) | 1979-10-25 | 1979-10-25 | Method of separation of the glucoseoxidasis |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CS206992B1 true CS206992B1 (en) | 1981-07-31 |
Family
ID=5421370
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CS725579A CS206992B1 (en) | 1979-10-25 | 1979-10-25 | Method of separation of the glucoseoxidasis |
Country Status (1)
| Country | Link |
|---|---|
| CS (1) | CS206992B1 (en) |
-
1979
- 1979-10-25 CS CS725579A patent/CS206992B1/en unknown
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