EP0513709A2 - Enzymatic method for reducing the amount of phosphorous-containing components in vegetable and animal oils - Google Patents

Enzymatic method for reducing the amount of phosphorous-containing components in vegetable and animal oils Download PDF

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EP0513709A2
EP0513709A2 EP92107888A EP92107888A EP0513709A2 EP 0513709 A2 EP0513709 A2 EP 0513709A2 EP 92107888 A EP92107888 A EP 92107888A EP 92107888 A EP92107888 A EP 92107888A EP 0513709 A2 EP0513709 A2 EP 0513709A2
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oil
phospholipase
degummed
ppm
content
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EP0513709B2 (en
EP0513709B1 (en
EP0513709A3 (en
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Erik Aalrust
Wolfgang Beyer
Hans Ottofrickenstein
Georg Penk
Hermann Dr. Plainer
Roland Dr. Reiner
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AB Enzymes GmbH
Roehm GmbH Darmstadt
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Metallgesellschaft AG
Roehm GmbH Darmstadt
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    • C—CHEMISTRY; METALLURGY
    • C11—ANIMAL OR VEGETABLE OILS, FATS, FATTY SUBSTANCES OR WAXES; FATTY ACIDS THEREFROM; DETERGENTS; CANDLES
    • C11B—PRODUCING, e.g. BY PRESSING RAW MATERIALS OR BY EXTRACTION FROM WASTE MATERIALS, REFINING OR PRESERVING FATS, FATTY SUBSTANCES, e.g. LANOLIN, FATTY OILS OR WAXES; ESSENTIAL OILS; PERFUMES
    • C11B3/00—Refining fats or fatty oils
    • C11B3/003—Refining fats or fatty oils by enzymes or microorganisms, living or dead

Definitions

  • the NHP arise from the action of the plant's own enzymes. In the "Alcon process", these are inactivated by steam treatment of the soy flakes, so that the formation of the NHP is prevented and the phosphatide content can be almost completely removed when the crude oil is degummed.
  • Phospholipase A2 and B are available as commercial products.
  • the use of purified enzymes is generally not necessary for technical applications.
  • a phospholipase preparation which is obtained from ground pancreatic pulp and contains above all phospholipase A2, is suitable for the process of the invention.
  • the enzyme is used in amounts of 0.001 to 1% by weight, based on oil.
  • a good distribution of the enzyme in the oil is ensured if it is dissolved in an amount of water of 0.5 to 5% by weight, based on oil, and emulsified in this form in the oil into droplets of less than 10 micrometers in diameter (weight average) becomes.
  • Turbulent stirring at radial speeds above 100 cm / s has proven itself. Instead, the oil can be circulated in the reactor using an external centrifugal pump. The enzymatic reaction can also be promoted by means of ultrasound.
  • emulsifying additives are helpful.
  • Water-soluble emulsifiers are suitable, in particular those with an HLB value above 9, such as sodium dodecyl sulfate. It is effective in an amount of, for example, 0.001% by weight based on oil if it is added to the enzyme solution prior to emulsification in the oil.
  • 1 l wet-degummed soybean oil with a residual phosphorus content of 110 ppm is heated to 75 o C in a round bottom flask. With vigorous stirring using a 5 cm diameter paddle stirrer at 700 rpm, 10 ml of water containing 1 g of citric acid are added and stirring is continued for 1 hour. Then it is cooled to 40 ° C. and a solution of 0.1 g of phospholipase A2 of the quality given in Example 1 and 50 mg of calcium chloride in 20 ml of 0.1 molar acetate buffer pH 5.5 are added. After a further 5 hours of intensive stirring, the water phase is centrifuged off. The oil obtained contains 2 ppm P and is suitable for physical refining.

Abstract

Der Gehalt an phosphorhaltigen Bestandteilen, inbesondere Phosphatiden, wie Lecithin, sowie der Eisengehalt in pflanzlichen und tierischen, vorzugsweise vorentschleimten Ölen, z.B. Sojaöl, werden erfindungsgemäß durch enzymatischen Abbau mittels Phospholipase A1, A2 oder B vermindert.The content of phosphorus-containing components, in particular phosphatides such as lecithin, and the iron content in vegetable and animal, preferably pre-degummed, oils, e.g. Soybean oil is reduced according to the invention by enzymatic degradation using phospholipase A1, A2 or B.

Description

Die Erfindung betrifft ein Verfahren zur Verminderung des Gehaltes an phosphorhaltigen Bestandteilen in Speiseölen durch enzymatischen Abbau und Abtrennung der Abbauprodukte. Der Begriff Speiseöle umfaßt pflanzliche und tierische, vorzugsweise vorentschleimte Öle.The invention relates to a process for reducing the content of phosphorus-containing components in edible oils by enzymatic degradation and separation of the degradation products. The term edible oils includes vegetable and animal oils, preferably pre-degummed oils.

Stand der TechnikState of the art

Rohes Sojaöl und andere pflanzliche Rohöle werden einer Vorentschleimung unterworfen, bei der Phosphatide, wie Lecithin, und andere hydrophile Nebenbestandteile entfernt werden. Geschieht dies durch Extrahieren mit Wasser, spricht man auch von Naßentschleimung. Bei dieser Behandlung verbleibt ein Teil der Phosphatide im Öl, der unter dem Begriff "Nicht-hydratisierbare Phosphatide" (NHP) zusammengefaßt wird. Für die Herstellung von Speiseölen ist es unerläßlich, diesen Anteil zu entfernen; nach herrschender Meinung soll der Phosphorgehalt 5 ppm nicht überschreiten. (Vgl. Hermann Pardun, "Die Pflanzenlecithine", Verlag für chemische Industrie H. Ziolkowsky KG, Augsburg, 1988, Seiten 181-194)Crude soybean oil and other vegetable crude oils are subjected to preliminary degumming, in which phosphatides, such as lecithin, and other hydrophilic secondary components are removed. If this is done by extracting with water, one also speaks of wet degumming. In this treatment, part of the phosphatides remains in the oil, which is summarized under the term "non-hydratable phosphatides" (NHP). For the production of edible oils, it is essential to remove this portion; According to the prevailing opinion, the phosphorus content should not exceed 5 ppm. (See Hermann Pardun, "Die Pflanzenlecithine", publishing house for chemical industry H. Ziolkowsky KG, Augsburg, 1988, pages 181-194)

Die NHP entstehen durch die Wirkung pflanzeneigener Enzyme. Diese werden beim "Alcon-Verfahren" durch Dampfbehandlung der Sojaflocken inaktiviert, so daß die Bildung der NHP unterbunden wird und bei der Naßentschleimung des rohen Öles der Phosphatidanteil nahezu vollständig entfernt werden kann.The NHP arise from the action of the plant's own enzymes. In the "Alcon process", these are inactivated by steam treatment of the soy flakes, so that the formation of the NHP is prevented and the phosphatide content can be almost completely removed when the crude oil is degummed.

Aus dem vorentschleimten Öl kann mittels wäßriger Tensidlösungen ein wesentlicher Teil der NHP extrahiert werden, jedoch kommt man in der Regel nicht unter 30 ppm Phosphor. Erfolgreicher ist die Behandlung mit Säuren oder Alkalien, erfordert jedoch viele Arbeitsschritte.A substantial part of the NHP can be extracted from the pre-degummed oil using aqueous surfactant solutions are, but usually you do not get below 30 ppm phosphorus. Treatment with acids or alkalis is more successful, but requires many steps.

Bekannt ist die Behandlung pflanzlicher und tierischer Öle mit Enzymen, wodurch enzymatisch spaltbare Bestandteile zu wasserlöslichen, leicht extrahierbaren Stoffen abgebaut werden sollen. So werden nach DE-A 16 17 001 Fette für die Seifenherstellung mit proteolytischen Enzymen desodoriert. Zur Klärung von Pflanzenölen werden gemäß GB 1 440 462 amylolytische und pektolytische Enzyme eingesetzt. Nach EP-A 70 269 werden tierische oder pflanzliche Fette oder Öle im rohen, halbaufbereiteten oder raffinierten Zustand mit einem oder mehreren Enzymen behandelt, um alle Bestandteile, die keine Glyceride sind, zu spalten und abzutrennen. Als geeignete Enzyme werden Phosphatasen, Pektinasen, Zellulasen, Amylasen und Proteasen erwähnt. Als Beispiel einer Phosphatase wird Phospholipase C genannt. Die Anwendung von Enzymen zur Vollentlecithinierung oder Totalentschleimung, wie man die NHP-Entfernung aus vorentschleimten Ölen auch nennt, ist nicht bekannt.The treatment of vegetable and animal oils with enzymes is known, as a result of which enzymatically cleavable constituents are to be broken down into water-soluble, easily extractable substances. According to DE-A 16 17 001, fats for soap production are deodorized with proteolytic enzymes. According to GB 1,440,462, amylolytic and pectolytic enzymes are used to clarify vegetable oils. According to EP-A 70 269, animal or vegetable fats or oils in the raw, semi-processed or refined state are treated with one or more enzymes in order to cleave and separate off all non-glyceride components. Phosphatases, pectinases, cellulases, amylases and proteases are mentioned as suitable enzymes. Phospholipase C is mentioned as an example of a phosphatase. It is not known to use enzymes for complete deglecination or total degumming, as the NHP removal from pre-degummed oils is also called.

Die Natur der NHP ist nicht genau bekannt. Nach Pardun (loc.cit) handelt es sich um Lysophosphatide und Phosphatidsäuren bzw. daraus gebildete Calcium- und Magnesium-Salze, die durch den Abbau von Phosphatiden unter der Einwirkung pflanzeneigener Phospholipasen entstehen.The nature of the NHP is not exactly known. According to Pardun (loc.cit), these are lysophosphatides and phosphatidic acids or calcium and magnesium salts formed therefrom, which result from the degradation of phosphatides under the influence of plant-specific phospholipases.

Aufgabe und LösungTask and solution

Ziel der Erfindung ist ein enzymatisches Verfahren zur Verminderung des Gehaltes an phosphor- und eisenhaltigen Bestandteilen in vorentschleimten Ölen.The aim of the invention is an enzymatic process for reducing the content of phosphorus and iron-containing components in pre-degummed oils.

Es wurde gefunden, daß sich die Behandlung der vorentschleimten Öle mit einer Phospholipase A₁, A₂ oder B für diesen Zweck eignet. Es wurden Phosphorgehalte unter 5 ppm und Eisengehalte unter 1 ppm erreicht. Der niedrige Eisengehalt ist für die Stabilität des Öles von Vorteil. Die Verminderung des Phosphorgehalts ist insofern überraschend, als Enzyme vom Typ der Phospholipasen für die Bildung der NHP verantwortlich gemacht werden. Mit Phospholipase C oder D läßt sich das Verfahrensziel nicht erreichen.It has been found that the treatment of the degummed oils with a phospholipase A₁, A₂ or B is suitable for this purpose. Phosphorus levels below 5 ppm and iron levels below 1 ppm were reached. The low iron content is beneficial for the stability of the oil. The reduction in the phosphorus content is surprising in that enzymes of the phospholipase type are held responsible for the formation of the NHP. The process goal cannot be achieved with phospholipase C or D.

Gegenstand der Erfindung ist daher ein Verfahren zur Verminderung des Gehaltes an phosphorhaltigen Bestandteilen in Speiseölen durch enzymatischen Abbau, bei dem als Enzym eine wäßrige Lösung einer Phospholipase A₁, A₂ oder B eingesetzt und die wäßrige Phase von dem behandelten Öl abgetrennt wird.The invention therefore relates to a process for reducing the content of phosphorus-containing constituents in edible oils by enzymatic degradation, in which an aqueous solution of a phospholipase A 1, A 2 or B is used as the enzyme and the aqueous phase is separated from the treated oil.

Ausführung der ErfindungImplementation of the invention

Da die Phospholipasen A₁, A₂ oder B Lecithin angreifen würden, ist es nicht sinnvoll, hoch lecithinhaltige Öle, wie rohes Sojaöl, in dem erfindungsgemäßen Verfahren einzusetzen. Ausgangsstoff sind daher vorentschleimte Öle, die sich in der Regel durch einen Phosphorgehalt zwischen 50 und 250 ppm auszeichnen. Öle schwankender Qualität können auf der gleichen Anlage verarbeitet werden. Vorzugsweise werden vorentschleimte Öle, vor allem Sonnenblumenöl, Rapsöl und insbesondere Sojaöl eingesetzt. Ein vorherige Trocknung des Öls ist entbehrlich.Since the phospholipases A₁, A₂ or B would attack lecithin, it does not make sense to use high-lecithin oils, such as crude soybean oil, in the process according to the invention. The source material is therefore pre-degummed oils, which are usually characterized by a phosphorus content between Mark 50 and 250 ppm. Oils of varying quality can be processed on the same system. Pre-degummed oils, especially sunflower oil, rapeseed oil and especially soybean oil, are preferably used. It is not necessary to dry the oil beforehand.

Die Phospholipase wird zweckmäßig in wäßriger Lösung eingesetzt, die in dem Öl so fein wie möglich emulgiert wird. Die enzymatische Reaktion dürfte an der Grenzfläche zwischen der Ölphase und der Wasserphase stattfinden. Sie wird durch intensive Mischung, z.B. durch turbulentes Rühren und zusätzlich durch den Zusatz von Tensiden gefördert. Die Abbauprodukte der NHP haben eine höhere Hydrophilie und gehen daher in die Wasserphase über. Sie werden daher ebenso wie die Metallionen gleichzeitig mit der Wasserphase aus dem Öl entfernt.The phospholipase is advantageously used in an aqueous solution which is emulsified in the oil as finely as possible. The enzymatic reaction should take place at the interface between the oil phase and the water phase. It is mixed thoroughly, e.g. promoted by turbulent stirring and additionally by the addition of surfactants. The degradation products of the NHP have a higher hydrophilicity and therefore go into the water phase. Therefore, like the metal ions, they are removed from the oil at the same time as the water phase.

Phospholipase A₁, A₂ und B sind bekannte Enzyme; (Vgl. Pardun, loc.cit. Seiten 135-141). Phospholipase A₁ spaltet die Fettsäureestergruppe am C₁-Atom eines Phospholipidmoleküls. Sie findet sich z.B. in der Rattenleber und im Schweinepankreas. Aus Schimmelpilzkulturen von Rhizopus arrhizus konnte ein Enzym mit Phospholipase-A₁-Aktivität isoliert werden.Phospholipase A₁, A₂ and B are known enzymes; (See Pardun, loc.cit. Pages 135-141). Phospholipase A 1 cleaves the fatty acid ester group on the C 1 atom of a phospholipid molecule. It can be found e.g. in rat liver and pork pancreas. An enzyme with phospholipase A 1 activity could be isolated from mold cultures of Rhizopus arrhizus.

Phospholipase A₂, die früher auch als Lecithinase A bezeichnet wurde, spaltet die Fettsäureestergruppe am C₂-Atom eines Phospholipidmoleküls. Sie tritt, meist vergesellschaftet mit anderen Phospholipasen, in fast allen Tier- und Pflanzenzellen auf. Reichlich findet sie sich im Schlangengift der Klapperschlange und der Kobra, sowie im Bienen- und Skorpiongift. Technisch kann sie aus Pankreasdrüsen gewonnen werden, nachdem aktivitätsinhibierende Begleitproteine mit Trypsin abgebaut worden sind.Phospholipase A₂, which was formerly referred to as lecithinase A, cleaves the fatty acid ester group on the C₂ atom of a phospholipid molecule. It occurs, mostly in association with other phospholipases, in almost all animal and plant cells. She finds plenty in the snake venom of the rattlesnake and the cobra, as well as in the bee and scorpion venom. Technically, it can be obtained from pancreatic glands after activity-inhibiting accompanying proteins have been broken down with trypsin.

Phospholipase B kommt in der Natur weitverbreitet vor. Sie wirkt auf das durch Phospholipase A₁-Einwirkung entstandene Lysolecithin durch Abspaltung des zweiten Fettsäureesterrestes ein. Zum Teil wird sie auch als Gemisch der Phospholipasen A₁ und A₂ angesehen. Sie kommt in der Rattenleber vor und wird auch von manchen Schimmelpilzen, wie Penicillium notatum, erzeugt.Phospholipase B is widespread in nature. It acts on the lysolecithin caused by phospholipase A 1 action by splitting off the second fatty acid ester residue. In some cases it is also regarded as a mixture of the phospholipases A₁ and A₂. It occurs in rat liver and is also produced by some molds, such as Penicillium notatum.

Phospholipase A₂ und B sind als Handelsprodukte erhältlich. Für die technische Anwendung ist der Einsatz gereinigter Enzyme in der Regel nicht erforderlich. Für das Verfahren der Erfindung eignet sich ein Phospholipase-Präparat, das aus gemahlenen Pankreasdrüsenbrei gewonnen wird und vor allem Phospholipase A₂ enthält. Das Enzym wird - je nach Aktivität - in Mengen von 0,001 bis 1 Gew.-%, bezogen auf Öl, eingesetzt. Eine gute Verteilung des Enzyms im Öl wird gewährleistet, wenn es in einer Wassermenge von 0,5 bis 5 Gew.-%, bezogen auf Öl, gelöst und in dieser Form in dem Öl zu Tröpfchen von weniger als 10 Mikrometer Durchmesser (Gewichtsmittelwert) emulgiert wird. Turbulentes Rühren mit Radialgeschwindigkeiten über 100 cm/s hat sich bewährt. Stattdessen kann das Öl mit Hilfe einer externen Kreiselpumpe im Reaktor umgewälzt werden. Auch mittels Ultraschalleinwirkung läßt sich die enzymatische Reaktion fördern.Phospholipase A₂ and B are available as commercial products. The use of purified enzymes is generally not necessary for technical applications. A phospholipase preparation, which is obtained from ground pancreatic pulp and contains above all phospholipase A₂, is suitable for the process of the invention. Depending on the activity, the enzyme is used in amounts of 0.001 to 1% by weight, based on oil. A good distribution of the enzyme in the oil is ensured if it is dissolved in an amount of water of 0.5 to 5% by weight, based on oil, and emulsified in this form in the oil into droplets of less than 10 micrometers in diameter (weight average) becomes. Turbulent stirring at radial speeds above 100 cm / s has proven itself. Instead, the oil can be circulated in the reactor using an external centrifugal pump. The enzymatic reaction can also be promoted by means of ultrasound.

Die Enzymwirkung wird durch den Zusatz einer Säure, vorzugsweise einer organischen Carbonsäure, gesteigert, die vor oder nach der Enzymbehandlung, am besten jedoch gleichzeitig zugegeben werden kann. Citronensäure ist bevorzugt. Sie kann in Form der freien Säure oder als Puffersystem in Kombination mit ihrem Salz, wie einem Alkalisalz (z.B. Natriumcitrat), einem Erdalkalisalz (z.B. Calciumcitrat) oder einem Ammoniumsalz, eingesetzt werden. Geeignete Mengen sind 0,01 - 1 Gew.-%, bez. auf Öl, optimal 0,1 Gew.-%. Mit der Säure wird der pH-Wert auf einen Wert von 3 bis 7, vorzugsweise von 4 bis 6, eingestellt. Das Optimum liegt etwa bei pH 5. Überraschenderweise ist dieser pH-Wert auch dann optimal, wenn die Phospholipase in Form eines Enzymkomplexes aus Pankreas eingesetzt wird. Der Pankreas-Enzymkomplex hat sonst ein pH-Optimum von 8 und ist bei pH 5 kaum noch wirksam ist. Anscheinend stellt sich an der Phasengrenzfläche, wo die Enzymwirkung eintritt, ein höherer pH-Wert als innerhalb der wäßrigen Phase ein.The enzyme effect is increased by the addition of an acid, preferably an organic carboxylic acid, which can be added before or after the enzyme treatment, but best at the same time. Citric acid is preferred. It can be used in the form of the free acid or as a buffer system in combination with its salt, such as an alkali salt (e.g. sodium citrate), an alkaline earth salt (e.g. calcium citrate) or an ammonium salt. Suitable amounts are 0.01-1% by weight, based on on oil, optimally 0.1% by weight. With the acid, the pH is adjusted to a value from 3 to 7, preferably from 4 to 6. The optimum is around pH 5. Surprisingly, this pH value is also optimal when the phospholipase is used in the form of an enzyme complex from the pancreas. The pancreas-enzyme complex otherwise has a pH optimum of 8 and is hardly effective at pH 5. Apparently, a higher pH value occurs at the phase interface where the enzyme action occurs than within the aqueous phase.

Um die Phospholipasen A₁, A₂ und B aus fetthaltigem Pankreatin oder Pankreasprodukten in Lösung zu bringen, sind emulgierende Zusätze hilfreich. Geeignet sind wasserlösliche Emulgatoren, insbesondere solche mit einem HLB-Wert über 9, wie Na-Dodecylsulfat. Es ist in einer Menge von z.B. 0,001 Gew.-%, bezogen auf Öl, wirksam, wenn es der Enzymlösung vor dem Emulgieren im Öl zugesetzt wird.To bring the phospholipases A₁, A₂ and B from fat-containing pancreatin or pancreatic products into solution, emulsifying additives are helpful. Water-soluble emulsifiers are suitable, in particular those with an HLB value above 9, such as sodium dodecyl sulfate. It is effective in an amount of, for example, 0.001% by weight based on oil if it is added to the enzyme solution prior to emulsification in the oil.

Der Zusatz weiterer Enzyme, vor allem Proteinasen und Amylasen, wirkt sich oft vorteilhaft aus. Auch Proteinzusätze können durch eine gewisse Tensidwirkung vorteilhaft sein.The addition of other enzymes, especially proteinases and amylases, often has an advantageous effect. Protein additives can also be advantageous due to a certain surfactant effect.

Die Temperatur bei der Enzymbehandlung ist nicht kritisch. Temperaturen zwischen 20 und 80oC sind geeignet. Optimal ist eine Temperatur von 50oC, jedoch kann kurzzeitig auch bis 70oC erwärmt werden. Die Behandlungsdauer hängt von der Temperatur ab und kann mit zunehmender Temperatur kürzer gehalten werden. Zeiten von 0,1 bis 10, vorzugsweise 1 bis 5 Stunden sind in der Regel ausreichend. Besonders bewährt hat sich ein Stufenprogramm, bei dem die erste Stufe bei einer Temperatur von 40 bis 60oC und die zweite Stufe bei einer höheren Temperatur im Bereich von 50 bis 80oC ausgeführt wird. Beispielsweise wird zuerst 3 Stunden bei 50oC, dann eine Stunde bei 75oC gerührt.The temperature in the enzyme treatment is not critical. Temperatures between 20 and 80 o C are suitable. A temperature of 50 o C is optimal, but it can also be heated up to 70 o C for a short time. The treatment time depends on the temperature and can be kept shorter with increasing temperature. Times of 0.1 to 10, preferably 1 to 5 hours are usually sufficient. A step program in which the first step is carried out at a temperature of 40 to 60 ° C. and the second step at a higher temperature in the range of 50 to 80 ° C. has proven particularly useful. For example, the mixture is first stirred at 50 ° C. for 3 hours, then at 75 ° C. for one hour.

Nach Abschluß der Behandlung wird die Enzymlösung mitsamt der darin aufgenommenen Abbauprodukte der NHP von der Ölphase abgetrennt, vorzugsweise durch Zentrifugieren. Da sich die Enzyme durch eine hohe Stabilität auszeichnen und die Menge der aufgenommenen Abbauprodukte gering ist, kann die gleiche Enzymlösung mehrmals wiederverwendet werden.After completion of the treatment, the enzyme solution together with the degradation products of the NHP contained therein is separated from the oil phase, preferably by centrifugation. Since the enzymes are characterized by high stability and the amount of degradation products absorbed is small, the same enzyme solution can be reused several times.

Vorzugsweise wird das Verfahren kontinuierlich durchgeführt. Bei einer zweckmäßigen kontinuierlicher Arbeitsweise wird das Öl in einem ersten Mischgefäß mit der Enzymlösung emulgiert, in einem oder mehreren nachfolgenden Reaktionsgefäßen, gegebenenfalls bei steigender Temperatur, unter turbulenter Bewegung reagieren gelassen, und anschließend in einer Zentrifuge die wäßrige Enzymlösung abgetrennt. Um eine Anreicherung der Abbauprodukte in der Enzymlösung zu vermeiden, kann ein Teil davon laufend durch frische Enzymlösung ersetzt und der Rest in den Prozeß zurückgeführt werden.The method is preferably carried out continuously. In an expedient, continuous mode of operation, the oil is mixed in a first mixing vessel emulsified the enzyme solution, allowed to react in turbulent agitation in one or more subsequent reaction vessels, optionally at increasing temperature, and then the aqueous enzyme solution was separated off in a centrifuge. In order to avoid an accumulation of the degradation products in the enzyme solution, part of it can be continuously replaced by fresh enzyme solution and the rest can be returned to the process.

Das gewonnene Öl hat einen Phosphor-Gehalt unter 5 ppm und ist damit zur physikalischen Raffination zu Speiseöl geeignet. Dank des erreichten niedrigen Eisengehaltes hat es gute Voraussetzungen, um beim Raffinieren eine hohe Oxydationsbeständigkeit zu erreichen.The oil obtained has a phosphorus content of less than 5 ppm and is therefore suitable for physical refining to edible oil. Thanks to the low iron content achieved, it has good prerequisites for achieving high oxidation resistance when refining.

BEISPIELEEXAMPLES Beispiel 1example 1

1 l naßentschleimtes Sojaöl mit einem Rest-Phosphorgehalt von 130 ppm wird in einem Rundkolben auf 50oC erwärmt. 0,1 g einer reinen Phospholipase A₂ mit einer Aktivität von 10 000 Einheiten/g (1 Phospholipase-A₂-Einheit setzt bei 40oC, pH 8 aus Eigelb 1 Mikro-Mol Fettsäure pro Minute frei), 1 g Na-Citrat, und 20 mg Na-Dodecylsulfat werden in 33,3 g Wasser gelöst, mit Citronensäure auf pH 5,2 eingestellt und die Lösung in dem Öl zu Tröpfchen mit einem Durchmesser von 0,1 Mikrometer emulgiert. Zu diesem Zweck wird das Öl mittels einer externen Kreiselpumpe etwa 3 mal pro Minute umgewälzt. Nach einer Behandlungsdauer von 3 Stunden wird in einer abzentrifugierten Probe ein NHP-Gehalt von 34 ppm P gefunden. Nach Steigerung der Temperatur auf 75oC und einstündiger Weiterbehandlung ist der NHP-Gehalt auf 3 ppm P gesunken. Das so behandelte Öl kann nunmehr zur physikalischen Raffination eingesetzt werden.1 liter of wet-degummed soybean oil with a residual phosphorus content of 130 ppm is heated to 50 ° C. in a round bottom flask. 0.1 g of a pure phospholipase A₂ with an activity of 10,000 units / g (1 phospholipase A₂ unit releases 1 micro-mole of fatty acid per minute from egg yolk at 40 ° C., pH 8), 1 g of sodium citrate, and 20 mg Na dodecyl sulfate are dissolved in 33.3 g water, adjusted to pH 5.2 with citric acid and the solution in the oil emulsified into droplets with a diameter of 0.1 micrometer. For this purpose, the oil is circulated about 3 times per minute using an external centrifugal pump. After a treatment period of 3 hours, an NHP content of 34 ppm P is found in a centrifuged sample. After increasing the temperature to 75 o C and further treatment for one hour, the NHP content dropped to 3 ppm P. The oil treated in this way can now be used for physical refining.

Beispiel 2Example 2

Das Verfahren gemäß Beispiel 1 wird mit dem Unterschied wiederholt, daß anstelle von Phospholipase A₂ 1 g eines Phospholipase B-Präparats aus Corticium species (Firma Amano, Versuchsprodukt ohne Aktivitätsangabe) eingesetzt wird. Der Phosphorgehalt im Sojaöl wird auf unter 1 ppm reduziert.The process according to Example 1 is repeated with the difference that, instead of phospholipase A₂, 1 g of a phospholipase B preparation from Corticium species (Amano company, test product without activity specification) is used. The phosphorus content in soybean oil is reduced to below 1 ppm.

VergleichsversucheComparative tests

Das Verfahren gemäß Beispiel 1 wird mit dem Unterschied wiederholt, daß anstelle von Phospholipase A₂ 1 g eines Phospholipase C-Präparats (Firma Amano Pharmaceutical Co, Ltd., Nagoya, Tokyo, Japan, Versuchsprodukt ohne Aktivitätsangabe) eingesetzt wird. Der Phosphorgehalt im Sojaöl vermindert sich nur auf 45 ppm.The process according to Example 1 is repeated with the difference that instead of phospholipase A₂ 1 g of a phospholipase C preparation (Amano Pharmaceutical Co, Ltd., Nagoya, Tokyo, Japan, trial product without activity information) is used. The phosphorus content in soybean oil only drops to 45 ppm.

Bei Einsatz von 1 g eines Phospholipase D-Präparats mit einer Aktivität von 1250 Phospholipase-Einheiten pro Gramm (Firma Sigma Chemie GmbH, Deisenhofen) wurde ein Phosphorgehalt von 48 ppm erreicht. Der Einsatz von 1 g einer sauren Phosphatase (Firma Sigma Chemie GmbH, Deisenhofen) ergab einen Phosphorgehalt von 47 ppm.When 1 g of a phospholipase D preparation with an activity of 1250 phospholipase units per gram (Sigma Chemie GmbH, Deisenhofen) was used, a phosphorus content of 48 ppm was achieved. The use of 1 g of an acidic phosphatase (Sigma Chemie GmbH, Deisenhofen) resulted in a phosphorus content of 47 ppm.

Etwa der gleiche Phosphorgehalt wird gefunden, wenn das Verfahren ohne Enzymzusatz durchgeführt wird.Approximately the same phosphorus content is found if the process is carried out without the addition of enzymes.

Beispiel 3Example 3

1 l naßentschleimtes Sojaöl mit einem Rest-Phosphorgehalt von 110 ppm wird in einem Rundkolben auf 75oC erwärmt. Unter starkem Rühren mit einem Flügelrührer von 5 cm Durchmesser bei 700 UpM werden 10 ml Wasser, enthaltend 1 g Citronensäure, zugesetzt und 1 Stunde weitergerührt. Dann wird auf 40oC abgekühlt und eine Lösung von 0,1 g Phospholipase A₂ der in Beispiel 1 angegebenen Qualität sowie 50 mg Calciumchlorid in 20 ml 0,1 molarem Acetatpuffer pH 5,5 zugegeben. Nach weiteren 5 Stunden intensiver Rührung wird die Wasserphase abzentrifugiert. Das erhaltene Öl enthält 2 ppm P und ist zur physikalischen Raffination geeignet. Die Veränderung der anderen Kennzahlen geht aus folgender Tabelle hervor: Ausgangsöl behandeltes Öl Phosphor 110 ppm 2 ppm Eisen 3,3 ppm <0,1 ppm Calcium 65,4 ppm 5,3 ppm Magnesium 38,4 ppm <0,1 ppm Peroxidzahl 18,3 18,50 Säurezahl 0,91 1,10 Verseifungszahl 191,2 190,4 1 l wet-degummed soybean oil with a residual phosphorus content of 110 ppm is heated to 75 o C in a round bottom flask. With vigorous stirring using a 5 cm diameter paddle stirrer at 700 rpm, 10 ml of water containing 1 g of citric acid are added and stirring is continued for 1 hour. Then it is cooled to 40 ° C. and a solution of 0.1 g of phospholipase A₂ of the quality given in Example 1 and 50 mg of calcium chloride in 20 ml of 0.1 molar acetate buffer pH 5.5 are added. After a further 5 hours of intensive stirring, the water phase is centrifuged off. The oil obtained contains 2 ppm P and is suitable for physical refining. The change in the other key figures is shown in the following table: Starting oil treated oil phosphorus 110 ppm 2 ppm iron 3.3 ppm <0.1 ppm Calcium 65.4 ppm 5.3 ppm magnesium 38.4 ppm <0.1 ppm Peroxide number 18.3 18.50 Acid number 0.91 1.10 Saponification number 191.2 190.4

Beispiel 4Example 4

Das Verfahren gemäß Beispiel 3 wird mit dem Unterschied wiederholt, daß anstelle von Phospholipase A₂ 1 g eines Pankreaspräparats (Pankreatin, 800 Phospholipase-Einheiten/g) eingesetzt wird. Das Präparat enthält Phospholipase A₂, Proteinase, Amylase, Lipase. Der Phosphorgehalt sinkt unter 1 ppm. Durch den Einfluß der Lipase tritt nur ein geringfügiger Anstieg der Säurezahl von 0,91 auf 1,49 ein.The process according to Example 3 is repeated with the difference that instead of phospholipase A₂ 1 g of a pancreatic preparation (pancreatin, 800 phospholipase units / g) is used. The preparation contains phospholipase A₂, proteinase, amylase, lipase. The phosphorus content drops below 1 ppm. Due to the influence of lipase, there is only a slight increase in the acid number from 0.91 to 1.49.

Beispiel 5Example 5

9 l naßentschleimtes Rapsöl mit einem Phosphorgehalt von 72 ppm wird mit einer Lösung von 8.6 g Citronensäure in 250 ml Wasser vermischt und auf 60oC erhitzt. Die Mischung wird homogenisiert, indem man sie mittels einer externen Kreiselpumpe einmal pro Minute umwälzt. Dann wird der pH-Wert der wäßrigen Phase mit 30 g 10 %iger Natronlauge auf 5,0 eingestellt. Nun werden 9 g Phospholipase A₂ mit einer Aktivität von 400 U/g zusammen mit etwas Calciumchlorid zugesetzt und die Mischung 3 Stunden bei 60oC in der angegebenen Weise umgewälzt. Nach dem Abzentrifugieren wird ein Phosphorgehalt von 3 ppm gefunden.9 l wet-degummed rapeseed oil with a phosphorus content of 72 ppm is mixed with a solution of 8.6 g citric acid in 250 ml water and heated to 60 ° C. The mixture is homogenized by circulating it once per minute using an external centrifugal pump. Then the pH of the aqueous phase is adjusted to 5.0 with 30 g of 10% sodium hydroxide solution. Now 9 g of phospholipase A₂ with an activity of 400 U / g are added together with some calcium chloride and the mixture is circulated for 3 hours at 60 o C in the manner specified. After centrifugation, a phosphorus content of 3 ppm is found.

Beispiel 6Example 6

Die Arbeitsweise gemäß Beispiel 5 wird mit dem Unterschied wiederholt, daß rohes (nicht naßentschleimtes) Sonnenblumenöl mit einem Wachsgehalt von 1,64 Gew.-% eingesetzt wird. Der Phosphorgehalt sinkt durch die Enzym-Behandlung von 223 auf 3 ppm.The procedure according to Example 5 is repeated with the difference that crude (not wet-degummed) sunflower oil with a wax content of 1.64% by weight is used. The enzyme treatment reduces the phosphorus content from 223 to 3 ppm.

Claims (15)

Verfahren zur Verminderung des Gehaltes an phosphorhaltigen Bestandteilen in Speiseölen durch enzymatischen Abbau,

   dadurch gekennzeichnet,

daß als Enzym eine wäßrige Lösung einer Phospholipase A₁, A₂ oder B eingesetzt und die wäßrige Phase von dem behandelten Öl abgetrennt wird.Process for reducing the content of phosphorus-containing components in edible oils by enzymatic degradation,

characterized,

that an aqueous solution of a phospholipase A₁, A₂ or B is used as the enzyme and the aqueous phase is separated from the treated oil. Verfahren nach Anspruch 1, dadurch gekennzeichnet, daß vorentschleimtes, insbesondere naßentschleimtes Öl eingesetzt wird.A method according to claim 1, characterized in that pre-degummed, in particular wet degummed oil is used. Verfahren nach Anspruch 1 oder 2, dadurch gekennzeichnet, daß zusätzlich ein Zitrat, wie Alkali-, Calcium- oder Ammoniumzitrat, eingesetzt wird.A method according to claim 1 or 2, characterized in that a citrate such as alkali, calcium or ammonium citrate is additionally used. Verfahren nach einem oder mehreren der Ansprüche 1 bis 3, dadurch gekennzeichnet, daß zusätzlich ein Emulgator eingesetzt wird.Method according to one or more of claims 1 to 3, characterized in that an emulsifier is additionally used. Verfahren nach einem oder mehreren der Ansprüche 1 bis 4, dadurch gekennzeichnet, daß es bei Temperaturen von 20 bis 80oC ausgeführt wird.Method according to one or more of claims 1 to 4, characterized in that it is carried out at temperatures of 20 to 80 o C. Verfahren nach einem oder mehreren der Ansprüche 1 bis 5, dadurch gekennzeichnet, daß es bei einem pH-Wert von 3 bis 7, vorzugsweise 4 bis 6, ausgeführt wird.Method according to one or more of claims 1 to 5, characterized in that it is carried out at a pH of 3 to 7, preferably 4 to 6. Verfahren nach einem oder mehreren der Ansprüche 1 bis 6, dadurch gekennzeichnet, daß vorentschleimtes Sojaöl verarbeitet wird.Method according to one or more of claims 1 to 6, characterized in that pre-degummed soybean oil is processed. Verfahren nach einem oder mehreren der Ansprüche 1 bis 6, dadurch gekennzeichnet, daß vorentschleimtes Raps- oder Sonnenblumenöl verarbeitet wird.Method according to one or more of claims 1 to 6, characterized in that pre-degummed rapeseed or sunflower oil is processed. Verfahren nach einem oder mehreren der Ansprüche 1 bis 8, dadurch gekennzeichnet, daß die Phospholipase in Form einer wäßrigen Lösung eingesetzt wird und die Lösung in dem Öl zu Tröpfchen einer Größe unter 10 Mikrometer emulgiert wird.Method according to one or more of claims 1 to 8, characterized in that the phospholipase is used in the form of an aqueous solution and the solution in the oil is emulsified into droplets of a size below 10 micrometers. Verfahren nach Anspruch 9, dadurch gekennzeichnet, daß die wäßrige Lösung der Phospholipase nach der Einwirkung von dem behandelten Öl abgetrennt und erneut verwendet wird.Process according to claim 9, characterized in that the aqueous solution of the phospholipase is separated from the treated oil after exposure and is reused. Verfahren nach einem oder mehreren der Ansprüche 1 bis 10, dadurch gekennzeichnet, daß es chargenweise durchgeführt wird.Method according to one or more of claims 1 to 10, characterized in that it is carried out in batches. Verfahren nach einem oder mehreren der Ansprüche 1 bis 10, dadurch gekennzeichnet, daß es kontinuierlich durchgeführt wird.Method according to one or more of claims 1 to 10, characterized in that it is carried out continuously. Verfahren nach einem oder mehreren der Ansprüche 1 bis 12, dadurch gekennzeichnet, daß es in zweistufig durchgeführt wird und daß die erste Stufe bei einer Temperatur von 40 bis 60oC und die zweite Stufe bei einer höheren Temperatur im Bereich von 50 bis 80oC ausgeführt wird.Method according to one or more of claims 1 to 12, characterized in that it is carried out in two stages and in that the first stage at a temperature of 40 to 60 o C and the second stage at a higher temperature in the range of 50 to 80 o C. is performed. Verfahren nach einem oder mehreren der Ansprüche 1 bis 13, dadurch gekennzeichnet, daß zusätzlich Zitronensäure eingesetzt wird.Method according to one or more of claims 1 to 13, characterized in that citric acid is additionally used. Verfahren nach einem oder mehreren der Ansprüche 1 bis 14, dadurch gekennzeichnet, daß gleichzeitig mit der Verminderung des Gehalts an phosphorhaltigen Bestandteilen auch der Eisengehalt des Öles vermindert wird.Method according to one or more of claims 1 to 14, characterized in that the iron content of the oil is reduced at the same time as the content of phosphorus-containing constituents is reduced.
EP92107888A 1991-05-16 1992-05-11 Enzymatic method for reducing the amount of phosphorous-containing components in vegetable and animal oils Expired - Lifetime EP0513709B2 (en)

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ATE120482T1 (en) 1995-04-15
AR245193A1 (en) 1993-12-30
EP0513709B2 (en) 1999-10-06
GR3031804T3 (en) 2000-02-29
DK0513709T3 (en) 1995-07-24
CA2068933A1 (en) 1992-11-17
CN1034587C (en) 1997-04-16
PL170548B1 (en) 1996-12-31
HU9201630D0 (en) 1992-08-28
HU213754B (en) 1997-09-29
ES2072043T5 (en) 2000-02-01
DK0513709T4 (en) 1999-12-27
BR9201859A (en) 1993-01-05
DE4115938A1 (en) 1992-11-19
ES2072043T3 (en) 1995-07-01
GR3015920T3 (en) 1995-07-31
DE59201753D1 (en) 1995-05-04
PL294543A1 (en) 1993-01-25
HUT64578A (en) 1994-01-28
EP0513709B1 (en) 1995-03-29
RU2033422C1 (en) 1995-04-20
EP0513709A3 (en) 1992-12-30
TW203625B (en) 1993-04-11
US5264367A (en) 1993-11-23
CA2068933C (en) 1995-12-19
CN1066679A (en) 1992-12-02

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