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DE10332065A1 - Enzymatic production of acids with intermediate formation of aldehydes in a single stage reaction from alcohols, involves addition of fresh arylalcohol- arylaldehyde-peroxidases and an oxidant - Google Patents

Enzymatic production of acids with intermediate formation of aldehydes in a single stage reaction from alcohols, involves addition of fresh arylalcohol- arylaldehyde-peroxidases and an oxidant Download PDF

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DE10332065A1
DE10332065A1 DE10332065A DE10332065A DE10332065A1 DE 10332065 A1 DE10332065 A1 DE 10332065A1 DE 10332065 A DE10332065 A DE 10332065A DE 10332065 A DE10332065 A DE 10332065A DE 10332065 A1 DE10332065 A1 DE 10332065A1
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Martin Prof. Dr. Hofrichter
Katrin Dr. Scheibner
Jörg Dr. Nüske
René Dipl.-Biol. Ullrich
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Friedrich Schiller Universtaet Jena FSU
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Abstract

Enzymatic production of acids from alcohols with intermediate formation of aldehydes in a single stage reaction involves the introduction by adding (or optionally dosing) into the reaction, fresh arylalcohol-arylaldehyde-peroxidases (AAP) and an oxidant (such as hydrogen peroxide) in an aqueous medium so as to oxidize the alcohol or aldehyde.

Description

Die Erfindung betrifft ein Verfahren zur enzymatischen Darstellung von Aldehyden, insbesondere aromatische Aldehyde (beispielsweise Benzaldehyd, Vanillin und Anisaldehyd), aus entsprechenden Alkoholen (wie Benzylakohol, Vanillylalkohol, Anisalkohol, Zimtalkohol) sowie zur enzymatischen Darstellung von Säuren, insbesondere aromatische Säuren (beispielsweise Benzoesäure, 4-Hydroxybenzoesäure und Zimtsäure), aus entsprechenden Aldehyden.The The invention relates to a process for the enzymatic preparation of Aldehydes, in particular aromatic aldehydes (for example benzaldehyde, vanillin and anisaldehyde), from corresponding alcohols (such as benzyl alcohol, Vanillyl alcohol, aniseed alcohol, cinnamyl alcohol) and to the enzymatic Representation of acids, in particular aromatic acids (for example benzoic acid, 4-hydroxybenzoic acid and cinnamic acid), from corresponding aldehydes.

Das Verfahren kann in verschiedensten Bereichen der Synthesechemie eingesetzt werden, u. a. zur Herstellung von Pharmazeutika oder Aromastoffen.The Process can be used in various fields of synthetic chemistry be, u. a. for the production of pharmaceuticals or flavorings.

Es ist allgemein bekannt (z. B. DE 197 23 961 A1 ; CH 71 329 A1 ), dass aromatische Aldehyde, wie Benzaldehyd, Anisaldehyd oder Vanillin, aus den entsprechenden alkoholischen Vorstufen durch chemische Oxidation (Friedel-Crafts-Alkanoylierung) hergestellt werden können. Dieser Prozess ist allerdings wenig umweltverträglich, da er chlorierte Reaktanden sowie Kohlenstoffdisulfid (CS2) erfordert.It is well known (eg DE 197 23 961 A1 ; CH 71 329 A1 ) that aromatic aldehydes, such as benzaldehyde, anisaldehyde or vanillin, can be prepared from the corresponding alcoholic precursors by chemical oxidation (Friedel-Crafts alkanoylation). However, this process is not environmentally friendly as it requires chlorinated reactants as well as carbon disulfide (CS 2 ).

Es ist weiterhin möglich, diese Verbindungen enzymatisch mit Hilfe pilzlicher Arylalkohol-Oxidasen (AAO) [EC 1.1.3.7] oder mikrobieller Arylalkohol-Dehydrogenasen [EC 1.1.1.90/91] aus geeigneten Alkoholen zu gewinnen (vgl. Enzyme Nomenclature (www.chem.qmul.ac.uk/iubmb/enzyme zu EC 1.1.3.7, EC 1.1.1.90 und EC 1.1.1.91).It is still possible these compounds enzymatically with the help of fungal aryl alcohol oxidases (AAO) [EC 1.1.3.7] or microbial aryl alcohol dehydrogenases [EC 1.1.1.90/91] from suitable alcohols (see Enzyme Nomenclature (www.chem.qmul.ac.uk/iubmb/enzyme to EC 1.1.3.7, EC 1.1.1.90 and EC 1.1.1.91).

Arylalkohol-Dehydrogenasen sind instabile intrazelluläre Enzyme, die NAD bzw. NADP als Coenzyme benötigen, was eine industrielle Verwendung erschwert. Arylalkohol-Oxidasen können als extrazelluläre oder intrazelluläre Biokatalysatoren bei bestimmten Pilzen vorkommen (z. B. Pleurotus spp: Guillen et al: Eur. J. Biochem. 209, 1992, 603-611; Botrytis cinerea: Goetghebeur et al: Biosci. Biotech. Biochem. 56, 1992, 298-303). Diese Enzyme nutzen Luftsauerstoff als Oxidationsmittel; können allerdings nur eine begrenzte Zahl von Aldehyden umsetzen und sind nicht in der Lage, die Folgereaktion zu den entsprechenden aromatischen Säuren zu katalysieren.Aryl alcohol dehydrogenases are unstable intracellular Enzymes that require NAD or NADP as coenzyme, which is an industrial Use difficult. Aryl alcohol oxidases can be called extracellular or intracellular Biocatalysts occur in certain fungi (eg Pleurotus spp: Guillen et al: Eur. J. Biochem. 209, 1992, 603-611; Botrytis cinerea: Goetghebeur et al: Biosci. Biotech. Biochem. 56, 1992, 298-303). These enzymes use atmospheric oxygen as the oxidant; but only implement a limited number of aldehydes and are not in the Able to add the subsequent reaction to the corresponding aromatic acids catalyze.

Aromatische Säuren, wie insbesondere die in der Lebensmittelindustrie als Konservierungsmittel verwendete Benzoesäure, können auf chemischen Wege über die Oxidation methylierter Vorstufen (z. B. Toluol) mit Hilfe aggressiver Oxidationsmittel (beispielsweise Kaliumpermanganat) gewonnen werden. Dieses Verfahren ist wiederum umweltgefährdend und führt zu einer Reihe unerwünschter Nebenprodukte.aromatic acids, especially as used in the food industry as a preservative benzoic acid, can in chemical ways the oxidation of methylated precursors (eg toluene) with the help of aggressive Oxidizing agent (for example, potassium permanganate) are obtained. This Process is again environmentally hazardous and leads to a series of undesirable ones By-products.

Enzymatisch werden Benzoesäuren aus Benzaldehyden unter Vermittlung spezifischer intrazellulärer Dehydrogenasen [EC 1.2.1.28/38] gebildet (vgl. Enzyme Nomenclature (www.chem.qmul.ac.uk/iubmb/enzyme zu EC 1.2.1.28 und EC 4.1.2.38), welche ebenfalls kostenintensives NAD oder NADP als Coenzyme benötigen. Weiterhin ist die enzymatische Oxidation von Aldehyden durch intrazelluläre Aldehyd-Oxidasen (EC 1.2.3.1) möglich, die u.a. in Leberzellen gebildet werden.enzymatic become benzoic acids from benzaldehydes mediating specific intracellular dehydrogenases [EC 1.2.1.28 / 38] (see Enzyme Nomenclature (www.chem.qmul.ac.uk/iubmb/enzyme to EC 1.2.1.28 and EC 4.1.2.38), which are also costly Need NAD or NADP as coenzyme. Furthermore, the enzymatic oxidation of aldehydes by intracellular aldehyde oxidases (EC 1.2.3.1) possible, the u.a. be formed in liver cells.

Auch diese Enzyme sind instabil und können nur unter hohem apparativen Aufwand (Zellaufschluss) isoliert werden.Also These enzymes are unstable and can only be isolated under high expenditure on equipment (cell disruption).

Aufgabe der vorliegenden Erfindung ist es, die Prozesse zur enzymatischen Darstellung von Aldehyden aus entsprechenden Alkoholen sowie zur enzymatischen Darstellung von Säuren aus entsprechenden Aldehyden mit möglichst geringem Aufwand verfahrenstechnischer und apparativer Art sowie unter Vermeidung umweltbelastender Chemikalien durchzuführen.task The present invention is the processes for enzymatic Preparation of aldehydes from corresponding alcohols as well as enzymatic Representation of acids from appropriate aldehydes with the least possible effort procedural and apparatus-type and to avoid polluting chemicals.

Die Ausgangsverbindungen sollen insbesondere mit geringem Energie- und Chemikalieneinsatz, schadstoffarm, ohne erhöhte Anforderungen an sterile bzw. semisterile Reaktionsführungen und mit möglichst kurzen Inkubationszeiten in wässrigen Medien umgesetzt werden.The Starting compounds are intended in particular with low energy and Chemical use, low in pollutants, without increased demands on sterile or semisterile reaction guides and with as possible short incubation times in aqueous Media are implemented.

Erfindungsgemäß wird ein Verfahren zur enzymatischen Darstellung von Säuren aus Alkoholen über die Zwischenbildung von Aldehyden in einem Einstufen-Reaktionsverfahren vorgeschlagen, bei dem die Substanzen bzw. Substanzgemische zumindest durch Zugabe und ggf. Zudosierung von neu gefundenen Arylalkohol-Arylaldehyd-Peroxidasen (AAP) und zumindest eines Oxidationsmittels, wie beispielsweise Wasserstoffperoxid, in einem wässrigen Milieu zur Reaktion gebracht werden, wobei die Alkohol- bzw. Aldehydfunktion jeweils oxidiert wird.According to the invention is a Process for the enzymatic preparation of acids from alcohols via the Intermediate formation of aldehydes in a one-step reaction process proposed in which the substances or mixtures of substances, at least by adding and if necessary, addition of newly found aryl alcohol-aryl aldehyde peroxidases (AAP) and at least an oxidizing agent, such as hydrogen peroxide, in an aqueous Milieu reacted, the alcohol or aldehyde function each oxidized.

Das zellfreie, enzymatische Verfahren beruht dabei auf einem neu gefundenen extrazellulären Pilzenzym, wobei diese Peroxidase (EC 1.11.1.) in Gegenwart katalytischer Mengen des Oxidationsmittels in vorzugsweise gepufferten wässrigen Lösungen Alkohole (wie Benzylakohol, Vanillylalkohol, Anisalkohol, Zimtalkohol) zu entsprechenden Aldehyden und weiter zu den daraus entstehenden Säuren unmittelbar, d. h. in dem besagten Einstufen-Reaktionsverfahren, umsetzt. Prinzipiell werden auf die gleiche Weise Aldehyde (beispielsweise Benzaldehyd, Vanillin und Anisaldehyd) direkt zu den entsprechenden Säuren oxidiert.The Cell-free, enzymatic method is based on a newly found extracellular Fungal enzyme, said peroxidase (EC 1.11.1.) In the presence of catalytic Amounts of the oxidizing agent in preferably buffered aqueous solutions Alcohols (such as benzyl alcohol, vanillyl alcohol, aniseed alcohol, cinnamyl alcohol) to corresponding aldehydes and further to the resulting acids directly, d. H. in said one-step reaction process, implements. In principle, in the same way aldehydes (for example Benzaldehyde, vanillin and anisaldehyde) directly to the corresponding acids.

Das neu gefundene und als Arylalkohol-Arylaldehyd-Peroxidase (AAP) bezeichnete Enzym wird vorzugsweise von Basidiomyceten aus der Familie Bolbitiaceae (z. B. Agrocybe sp.) gebildet und zeichnet sich durch besondere katalytische Eigenschaften aus, die keine der bisher bekannten Peroxidasen besitzt.The newly discovered and designated as aryl alcohol-aryl aldehyde peroxidase (AAP) Enzyme is preferably derived from Basidiomycetes of the family Bolbitiaceae (eg Agrocybe sp.) and is characterized by special catalytic properties, none of the previously known peroxidases has.

Die Reaktionen mit dieser Peroxidase sind umweltfreundlich (d. h. sie erfordern keine aggressiven und umweltbelastenden Chemikalien). So sind Oxidationsmittel nur in katalytischen Mengen zur Gewährleistung des Peroxidasezyklus, nicht aber zur direkten Umsetzung der Ausgangsverbindungen erforderlich. Die chemische Oxidation von Alkoholen und Aldehyden erfordert hingegen äquimolare Mengen an umweltgefährdenden Oxidationsmitteln (Peroxide, Ozon, Permanganate, Chromate).The Reactions with this peroxidase are environmentally friendly (i.e. do not require aggressive and polluting chemicals). Thus, oxidizing agents are only in catalytic amounts to ensure the peroxidase cycle, but not for direct reaction of the starting compounds required. The chemical oxidation of alcohols and aldehydes however, requires equimolar Amounts of environmentally hazardous Oxidizing agents (peroxides, ozone, permanganate, chromates).

Des weiteren laufen die rein chemischen Oxidationen nur in Gegenwart geeigneter Lösungsmittel (Methanol, Dimethylsulfoxid, Aceton) ab und erfolgen in wässrigen, lediglich gepufferten Reaktionslösungen nicht mit befriedigender Ausbeute. Während chemische Umsetzungen in der Regel eine Prozessführung bei höheren Temperaturen (Heizquelle) und/oder höheren Drücken benötigen, zeigt sich, dass die erfindungsgemäße AAP-katalysierte Umsetzung jeweils bei Raumtemperatur realisiert werden kann und keine speziellen Apparaturen (wie Druckreaktoren o. ä.) oder apparative Aufwände erfordert.Of Further, the purely chemical oxidations run only in the presence suitable solvents (Methanol, dimethyl sulfoxide, acetone) and take place in aqueous, only buffered reaction solutions not with satisfactory yield. During chemical reactions usually a litigation at higher Temperatures (heat source) and / or higher pressures, it turns out that the AAP-catalyzed reaction according to the invention can be implemented at room temperature and no special Equipment (such as pressure reactors o. Ä.) Or equipment required.

Die Vorteile der zellfreien, enzymatischen AAP-Umsetzungen gegenüber einer ebenfalls möglichen Oxidation von Alkoholen und Aldehyden durch ganze Zellen (Pilze, Bakterien) bestehen in den relativ kurzen Reaktionszeiten, die keine sterile oder semisterile Reaktionsführung erforderlich machen.The Advantages of cell-free, enzymatic AAP reactions over one also possible Oxidation of alcohols and aldehydes by whole cells (fungi, Bacteria) exist in the relatively short reaction times, the no require sterile or semisterile reaction.

Mit den AAP-katalysierten Reaktionen ist erstmals möglich, Alkohole mit Hilfe eines einzelnen, extrazellulären Biokatalysators in einem einstufigen Prozess bis zu den entsprechenden Säuren zu oxidieren; gleichzeitig ermöglicht die Prozeßführung aber auch, die als Zwischenprodukte vorübergehend gebildeten Aldehyde gezielt zu gewinnen. Die stufenweise enzymatische Synthese von Säuren aus Alkoholen ist ebenfalls durch andere Biokatalysatoren auf umweltschonende Weise möglich; allerdings müssen hierfür mindestens zwei verschiedene Enzyme (z. B. Alkohol-Oxidase und Aldehyd-Oxidase oder Alkohol-Dehydrogenase und Aldehyd-Dehydrogenase) eingesetzt werden, da sie nur jeweils einen der beiden Oxidationschritte katalysieren können.With The AAP-catalyzed reactions is possible for the first time, alcohols with the help of a single, extracellular Biocatalyst in a one-step process up to the corresponding acids to oxidize; simultaneously possible but the process control also, the intermediately formed aldehydes to gain targeted. The stepwise enzymatic synthesis of acids Alcohol is also environmentally friendly through other biocatalysts Way possible; however, this must be at least two different enzymes (eg, alcohol oxidase and aldehyde oxidase or alcohol dehydrogenase and aldehyde dehydrogenase) because they catalyze only one of the two oxidation steps can.

Die Erfindung soll nachstehend anhand von in der Zeichnung dargestellten Ausführungsbeispielen näher erläutert werden, wobei die Erfindung nicht auf die behandelten Aromaten und Aromate an sich beschränkt sein soll.The Invention will be described below with reference to the drawing embodiments be explained in more detail, the invention is not limited to the aromatics and aromatics treated limited in itself should be.

Es zeigen:It demonstrate:

1: Umsetzung von Benzylalkohol (⦁) zu Benzaldehyd

Figure 00040001
und Benzoesäure
Figure 00040002
mittels Arylalkohol-Arylaldehyd-Peroxidase (AAP) von Agrocybe sp.. 1 : Reaction of benzyl alcohol (⦁) to benzaldehyde
Figure 00040001
and benzoic acid
Figure 00040002
using aryl alcohol-aryl aldehyde peroxidase (AAP) from Agrocybe sp.

2: Umsetzung von Veratrylalkohol (⦁) zu Veratrylaldehyd

Figure 00040003
und Veratrylsäure
Figure 00040004
2 : Conversion of veratryl alcohol (⦁) to veratrylaldehyde
Figure 00040003
and veratrylic acid
Figure 00040004

3: Umsetzung von Anisalkohol (⦁) zu Anisaldehyd

Figure 00040005
3 : Reaction of Anisalcohol (⦁) to Anisaldehyde
Figure 00040005

4: Formelschema zu den in 1-3 dargestellten Arylalkohol-Arylaldehyd-Peroxidase katalysierten Oxidationen aromatischer Alkohole (PO = Peroxidase) 4 : Formula Scheme to the in 1 - 3 Aryl alcohol-aryl aldehyde peroxidase catalyzed oxidations of aromatic alcohols (PO = peroxidase)

Ausführunsbeispiel 1:Embodiment 1:

200 nmol Benzylalkohol werden in Natriumphosphat-Citrat-Puffer (150 mM, pH 6,8) zusammen mit Glukose (15 mM), Glukose-Oxidase (0,2 Units) und Arylalkohol-Arylaldehyd-Peroxidase (Agrocybe sp., 1 Unit) in einem Gesamtvolumen von 1 ml bei 24 °C in einem offenem Glasgefäß gerührt (Magnetrührwerk). In Abständen von zwei bis acht Minuten werden 20 μl des Versuchsansatzes entnommen und mittels High Performance Liquid Chromatography (HPLC) vermessen [Säule: LiChrospher® RP18 5 μm 125/4 (Firma Merck Darmstadt), Trennbedingungen: 30 % Acetonitril, 70 % Essigsäure in Reinstwasser (0,05 %), Flußrate 1 ml/min] (vgl. 1).200 nmol of benzyl alcohol are dissolved in sodium phosphate citrate buffer (150 mM, pH 6.8) together with glucose (15 mM), glucose oxidase (0.2 units) and aryl alcohol-aryl aldehyde peroxidase (Agrocybe sp., 1 unit). stirred in a total volume of 1 ml at 24 ° C in an open glass vessel (magnetic stirrer). At intervals of two to eight minutes of the test batch are 20 ul removed and analyzed by High Performance Liquid Chromatography (HPLC) measured [column: LiChrospher ® RP18 5 micron 125/4 (Merck Darmstadt), separation conditions: 30% acetonitrile, 70% acetic acid in Ultrapure water (0.05%), flow rate 1 ml / min] (cf. 1 ).

Ausführungsbeispiel 2:Embodiment 2:

200 nmol Veratrylalkohol (3,4-Dimethoxybenzylalkohol) werden in Natriumphosphat-Citrat-Puffer (150 mM, pH 6,8) zusammen mit Glukose (15 mM), Glukose-Oxidase (0,2 Units) und Arylalkohol-Arylaldehyd-Peroxidase (Agrocybe sp., 1 Unit) in einem Gesamtvolumen von 1 ml bei 24 °C in einem offenem Glasgefäß gerührt (Magnetrührwerk). In Abständen von zwei bis acht Minuten werden 20 μl des Versuchsansatzes entnommen und mittels High Performance Liquid Chromatography (HPLC) vermessen [Säule: LiChrospher® RP18 5 μm 125/4 (Firma Merck Darmstadt), Trennbedingungen: 30 % Acetonitril, 70 % Essigsäure in Reinstwasser (0,05 %), Flußrate 1 ml/min] (2).200 nmol of veratryl alcohol (3,4-dimethoxybenzyl alcohol) are dissolved in sodium phosphate citrate buffer (150 mM, pH 6.8) together with glucose (15 mM), glucose oxidase (0.2 units) and aryl alcohol-aryl aldehyde peroxidase ( Agrocybe sp., 1 unit) in a total volume of 1 ml at 24 ° C in an open glass vessel stirred (magnetic stirrer). At intervals of two to eight minutes of the test batch are 20 ul removed and analyzed by High Performance Liquid Chromatography (HPLC) measured [column: LiChrospher ® RP18 5 micron 125/4 (Merck Darmstadt), separation conditions: 30% acetonitrile, 70% acetic acid in Ultrapure water (0.05%), flow rate 1 ml / min] ( 2 ).

Ausführungsbeispiel 3Embodiment 3

200 nmol Anisalkohol (4-Methoxybenzylalkohol) werden in Natriumphosphat-Citrat-Puffer (150 mM, pH 6,8) zusammen mit Glukose (15 mM), Glukose-Oxidase (0,2 Units) und Arylalkohol-Arylaldehyd-Peroxidase (Agrocybe sp., 1 Unit) in einem Gesamtvolumen von 1 ml bei 24 °C in einem offenem Glasgefäß gerührt (Magnetrührwerk). In Abständen von zwei bis acht Minuten werden 20 μl des Versuchsansatzes entnommen und mittels High Performance Liquid Chromatography (HPLC) vermessen [Säule: LiChrospher® RP18 5 μm 125/4 (Firma Merck Darmstadt), Trennbedingungen: 30 % Acetonitril, 70 % Essigsäure in Reinstwasser (0,05 %), Flußrate 1 ml/min] (3).200 nmol of anisalcohol (4-methoxybenzyl alcohol) are dissolved in sodium phosphate citrate buffer (150 mM, pH 6.8) together with glucose (15 mM), gluco Se oxidase (0.2 units) and aryl alcohol-aryl aldehyde peroxidase (Agrocybe sp., 1 unit) in a total volume of 1 ml at 24 ° C in an open glass vessel stirred (magnetic stirrer). At intervals of two to eight minutes of the test batch are 20 ul removed and analyzed by High Performance Liquid Chromatography (HPLC) measured [column: LiChrospher ® RP18 5 micron 125/4 (Merck Darmstadt), separation conditions: 30% acetonitrile, 70% acetic acid in Ultrapure water (0.05%), flow rate 1 ml / min] ( 3 ).

Claims (10)

Verfahren zur enzymatischen Darstellung von Säuren aus Alkoholen über die intermediäre Bildung von Aldehyden in einem Einstufen-Reaktionsverfahren, bei dem die Substanzen bzw. Substanzgemische zumindest durch Zugabe und ggf. Zudosierung von neu gefundenen Arylalkohol-Arylaldehyd-Peroxidasen (AAP) und zumindest eines Oxidationsmittels, wie beispielsweise Wasserstoffperoxid, in einem wässrigen Milieu zur Reaktion gebracht werden, wobei die Alkohol- bzw. Aldehydfunktion jeweils oxidiert wird.Process for the enzymatic preparation of acids Alcohols over the intermediaries Formation of aldehydes in a one-step reaction process, at the substances or substance mixtures at least by adding and, if appropriate, addition of newly found aryl alcohol-aryl aldehyde peroxidases (AAP) and at least one oxidizing agent, such as Hydrogen peroxide, in an aqueous Milieu reacted, the alcohol or aldehyde function each oxidized. Verfahren nach Anspruch 1, dadurch gekennzeichnet, dass als Arylalkohol-Arylaldehyd-Peroxidasen (AAP) die Enzyme von Agrocybe aegerita (Syn. Pholiota cylindracea, Südlicher Ackerling) verwendet werden.Method according to claim 1, characterized in that that as aryl alcohol-aryl aldehyde peroxidases (AAP) the enzymes of Agrocybe aegerita (Syn. Pholiota cylindracea, southern Ackerling) can be used. Verfahren nach Anspruch 1, dadurch gekennzeichnet, dass als Arylalkohol-Arylaldehyd-Peroxidasen (AAP) die Enzyme von anderen Vertretern der Gattung Agrocybe verwendet werden.Method according to claim 1, characterized in that that as aryl alcohol-aryl aldehyde peroxidases (AAP) uses the enzymes of other members of the genus Agrocybe become. Verfahren nach Anspruch 1, dadurch gekennzeichnet, dass als Arylalkohol-Arylaldehyd-Peroxidasen (AAP) die Enzyme der Familie Bolbitiaceae verwendet werden.Method according to claim 1, characterized in that that as aryl alcohol-aryl aldehyde peroxidases (AAP) the enzymes of the family Bolbitiaceae be used. Verfahren nach einem oder mehreren der vorgenannten Ansprüche, dadurch gekennzeichnet, dass dem Reaktionsgemisch zur weiteren Beschleunigung und Regulation der Umsetzung H2O2-generierende Enzyme, vorzugsweise Oxidasen (z. B. Glucose-Oxidase) zugesetzt werden.Method according to one or more of the preceding claims, characterized in that H 2 O 2 -generating enzymes, preferably oxidases (eg glucose oxidase) are added to the reaction mixture for further acceleration and regulation of the reaction. Verfahren nach einem oder mehreren der vorgenannten Ansprüche, dadurch gekennzeichnet, dass zur Stabilisierung der Reaktion Puffer auf Basis organischer Säuren, vorzugsweise Zitronensäure, und Phosphaten, vorzugsweise Natriumhydrogenphosphate, zugesetzt werden.Method according to one or more of the aforementioned Claims, characterized in that to stabilize the reaction buffer based on organic acids, preferably citric acid, and Phosphates, preferably sodium hydrogenphosphates, are added. Verfahren nach einem oder mehreren der vorgenannten Ansprüche, dadurch gekennzeichnet, dass zur Verbesserung der Produktbildung organische Lösungsmittel (z. B. Ethanol) zugesetzt werden.Method according to one or more of the aforementioned Claims, characterized in that to improve product formation organic solvents (For example, ethanol) are added. Verfahren nach einem oder mehreren der vorgenannten Ansprüche, dadurch gekennzeichnet, dass die beteiligten Enzyme zur Stabilisierung der Reaktionen in immobilisierter Form eingesetzt werden.Method according to one or more of the aforementioned Claims, characterized in that the enzymes involved for stabilization the reactions are used in immobilized form. Verfahren nach Anspruch 1, dadurch gekennzeichnet, dass zwecks Herstellung eines Aldehyds nur der Teilprozess der Darstellung des Aldehyds aus dem Alkohol genutzt wird, indem der Aldehyd vor einer weiteren enzymatischen Umsetzung zur Säure vollständig oder teilweise aus der Reaktionslösung entnommen wird.Method according to claim 1, characterized in that that for the production of an aldehyde only the sub-process of the representation of the aldehyde from the alcohol is used by the aldehyde before a further enzymatic conversion to the acid completely or partially from the Removed reaction solution becomes. Verfahren nach Anspruch 1, dadurch gekennzeichnet, dass zwecks Herstellung der Säure aus einem Aldehyd nur der Teilprozess der Darstellung der Säure aus dem Aldehyd genutzt wird, indem der Aldehyd als Ausgangsstoff zur Reaktion mit der Arylalkohol-Arylaldehyd-Peroxidase (AAP) eingebracht wird.Method according to claim 1, characterized in that that in order to manufacture the acid from an aldehyde only the sub-process of representation of the acid the aldehyde is used by the aldehyde as the starting material for Reaction with the aryl alcohol-aryl aldehyde peroxidase (AAP) is introduced.
DE10332065A 2003-07-11 2003-07-11 Enzymatic production of acids with intermediate formation of aldehydes in a single stage reaction from alcohols, involves addition of fresh arylalcohol- arylaldehyde-peroxidases and an oxidant Withdrawn DE10332065A1 (en)

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