CS224911B1 - The enzyme degradation method of the pectin substances - Google Patents

The enzyme degradation method of the pectin substances Download PDF

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CS224911B1
CS224911B1 CS434682A CS434682A CS224911B1 CS 224911 B1 CS224911 B1 CS 224911B1 CS 434682 A CS434682 A CS 434682A CS 434682 A CS434682 A CS 434682A CS 224911 B1 CS224911 B1 CS 224911B1
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enzyme
pectin substances
pectin
degradation method
enzyme degradation
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CS434682A
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Czech (cs)
Slovak (sk)
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Benkova Lubomira Rndr C Rexova
Jirina Ing Omelkova
Vladimir Doc Ing Csc Kubanek
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Benkova Lubomira Rndr C Rexova
Jirina Ing Omelkova
Kubanek Vladimir
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Priority to CS434682A priority Critical patent/CS224911B1/en
Publication of CS224911B1 publication Critical patent/CS224911B1/en

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Vynález sa týká. sposobu enzýmovej degradácie pektínových látok.The invention relates. method of enzyme degradation of pectin substances.

Enzýmové degradácie pektínoyých látok spočívajú v štiepení Qú -1,4 glykozidických vazieb medzi jednotkami D-galaktopyranurónovej kyseliny. Enzýmami, ktoré katalýzujú túto reakciu sú pektolytické enzýmy endopolygalakturonáza, exopolygalakturonáza a pektínlyáza, produkované mikroorganizmami alebo přítomné v rastlinách. Enzýmová degradácia pektínových látok sa v praxi uplatňuje napr. pri výrobě ovocných štiav a vín, kde jej účinnost* spočívá v zní žení viskozity· zlepšení klarifikácie a filtrácie a zvýšení stability finálnych produktov. Doteraz známe spósoby enzýmovej degradácie pektínových látok přítomných v týchto materiáloch sa zakladajú na ich inkubácii a uvedenými pektolytickými enzýmami, najmá endopolygalakturonázou, přidanými vo formě roztokov· alebo po ich rozpuštění v inkubačnom prostředí. Obecnou nevýhodou týchto postupov je, že po skončeni enzýmovej reakcie enzým zostává spolu ao sprievodnými látkami· pochádzajúcimi z povodného enzýmového preparátu, vo finálnom roztoku a spósobuje tak často nežiadúcu kontamináciu produktov. Ďaldím nedostat kom je, že enzým je použitelný len pre obmedzené množstvo substrátu a nie je možná jeho reutilizácia, ako aj skutočnosť, že zastavenie enzýmovej reakcie vžiadanom stupni je možné dosiahnuC lem inaktiváciou enzýmu jeho zahriatím.The enzymatic degradation of the pectic substances is based on the cleavage of Qu -1.4 glycosidic bonds between D-galactopyranuronic acid units. The enzymes that catalyze this reaction are the pectolytic enzymes endopolygalacturonase, exopolygalacturonase and pectin lyase, produced by microorganisms or present in plants. The enzymatic degradation of pectin substances is applied in practice e.g. in the production of fruit juices and wines, where its effectiveness * consists in reducing viscosity · improving clarification and filtration and increasing the stability of the final products. Hitherto known methods of enzymatic degradation of the pectin substances present in these materials are based on their incubation and by said pectolytic enzymes, in particular endopolygalacturonase, added in the form of solutions or after their dissolution in the incubation medium. A general disadvantage of these processes is that, after the enzyme reaction has ended, the enzyme remains together with the accompanying substances derived from the original enzyme preparation, in the final solution, and often causes undesirable product contamination. Another drawback is that the enzyme is only applicable to a limited amount of substrate and its reutilization is not possible, as is the fact that stopping the enzyme reaction at the desired stage can be achieved by inactivating the enzyme by heating it.

Uvedené nedostatky sa odstraňujú postupom podfa vynálezu, ktorého podstatou je, že enzýmová degradácia sa uskutočňuje pomocou pektolytického enzýmového preparátu, ireverzibilne zakotveného na nerozpustnú polymérnu látku typu polykondenzátu na báze diolov a dikarboxylových kyselin, s výhodou na polyetylóntereftalát.These drawbacks are overcome by the process according to the invention, wherein the enzymatic degradation is carried out by means of a pectolytic enzyme preparation irreversibly anchored to an insoluble polymer of the polyol type based on diols and dicarboxylic acids, preferably polyethylene terephthalate.

Základný účinok spósobu degradácie pektínových látok podfa vynálezu spočívá v možnosti oddelenia enzýmu od produktov reakcie a to v ktoromkofvek stádiu reakcie, ako aj v možnosti mnohonásobného kontinuálneho alebo diskoxt&inuálneho použitia ireverzibilne zakotveného enzýmu, vyplývajúcej z jeho neobyčajme vysokej operačnej stability. ĎalšouThe basic effect of the method of degrading the pectin substances of the invention lies in the possibility of separating the enzyme from the reaction products at any stage of the reaction as well as in the possibility of multiple continuous or discoxual use of the irreversibly anchored enzyme resulting from its unusually high operational stability. Another

-h 224 911 výhodou spósobu podlá vynálezu je, že pektolytické enzýiay zakotvené na polyetyléntereftalát sa svojimi vlastnosťamž /závialosť aktivity na pH a teplote/ nelíšia od rozpustného enzymu, takže enzýmová degradácia sa móže uskutočňovať za rovnakých podmienok ako pri použití rozpustnéj pektinázy.It is an advantage of the method according to the invention that the pectolytic enzymes anchored to polyethylene terephthalate do not differ from the soluble enzyme in their properties (depending on the pH and temperature activity), so that the enzyme degradation can be carried out under the same conditions as using soluble pectinase.

Příklad 1 g pektolytického enzýmového preparátu zmobilizovaného na polyetyléntereftalát /specifický povrch nosiča 50 až 120 m /g; objem porov 2 až 3 cm /g/ sa suspenduje vil roztoku obsahujúceho pektín. Po úpravě pH na 3,8 až 4,8 sa zmes inkubuje ža miešania pri teplote nepresahujúcej 45 °C dovtedy, kým sa nedosiahne žiadaný stupeň: degradácie pektínových látok prejavujúci sa příslušnou hodnotou viskozity a obsahu redukujúcich skupím v roztoku. Po skončeni reakcie sa zmobilizovaný enzým oddeli filtráciou alebo ceotrifugácíou & móže sa použit? k degradácii ďalšej dávky substrátu. Dosiahnutie maximálněj degradácie pektínu sa prejaví nemeniacim. sa pomerom viskozity a obsahu redukujúcich skupin produktu.Example 1 g of a pectolytic enzyme preparation mobilized to polyethylene terephthalate / specific surface area of 50-120 m / g; a pore volume of 2-3 cm (g) is suspended in 1 pectin-containing solution. After adjusting the pH to 3.8 to 4.8, the mixture is incubated with stirring at a temperature not exceeding 45 ° C until the desired degree of degradation of pectin substances is achieved, exhibiting the appropriate viscosity and reducing groups content in the solution. When the reaction is complete, the enzyme is mobilized by filtration or by centrifugation. to degrade the next substrate dose. Achieving maximum degradation of pectin is manifested as unchanged. by the ratio of viscosity and content of reducing product groups.

Příklad 2Example 2

Pektolytický enzýmový preparát imobilizovaný na polyetyléntereftalát /poměr enzýmu k nosičů = 1 : 100/ sa naplní do kolony. Cez vzniklý štipec enzýmového gelu sa filtruje roztok obsahujúci pektínové látky, upravený na pH 3,8 až 4,8, Rýchlosť prietoku kolonou a výška stípca zmobilizovaného enzýrnu sa volia tak, ahy sa počas prietoku pektínovej látky cez vrstvu zmobilizovaného enzýmu dosiahol žiadaný stupem jej degradácie» prejavujúci sa příslušnou hodnotou viskozity a obsahu redukujúcich skupím.The pectolytic enzyme preparation immobilized to polyethylene terephthalate (enzyme to carrier ratio = 1: 100) is loaded onto the column. The pectin-containing solution, adjusted to a pH of 3.8 to 4.8, is filtered through the resulting gel gel column. The column flow rate and column height of the mobilized enzyme are selected such that the desired degree of degradation is achieved during the flow of pectin substance through the layer of mobilized enzyme. Exhibiting an appropriate value of viscosity and content of reducing groups.

Pri maximálnej degradácii sa v obidvoch prípadoch podlá příkladu X a 2 zfska produkt nekoutaminováný enzýmom alebo jeho sprievodnými látkami, vyznačujúci sa nízkou viskozitou· ktorý obsahuje D-galaktopyramurónovú kyselinu a jej 3 až 4 najnižšie oligomery.At maximum degradation, in both cases according to Examples X and 2, the product is non-contaminated by the enzyme or its accompanying substances, characterized by a low viscosity containing D-galactopyramuronic acid and its 3 to 4 lowest oligomers.

Vynález je možné využiť v konzervárenském priemysle, pri výrob» ovocných štiav a vo vinárskom priemysle při stabili zácii vín.The invention can be used in the canning industry, in the production of fruit juices and in the wine industry for the stabilization of wines.

Claims (1)

PREDMET VYNÁLEZUOBJECT OF THE INVENTION 224 911224 911 Spósob enzýmovej degradácie pektínových látok vyznačený tým, že sa na pektínové látky pósobí pektolytickým enzýmovým preparát om ireverzibilne imobilizováným na polymému látku typu polykondenzátu na báze diolov a dikarboxylových kyselin, s výhodou na polyetyléntereftalát·Method of enzymatic degradation of pectin substances characterized in that pectin substances are treated with a pectolytic enzyme preparation irreversibly immobilized on a polyol type of polycondensate based on diols and dicarboxylic acids, preferably polyethylene terephthalate.
CS434682A 1982-06-11 1982-06-11 The enzyme degradation method of the pectin substances CS224911B1 (en)

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