DE2221534A1 - Process for the oxidation of hydrocarbons, alcohols and aldehydes - Google Patents

Description

DR.-ING. BY KRtISLER DR.-ING. SCHÖNWALD DR.-ING. TH. MEYER DR. FUES DIPL-CHEM. ALEK VON KREISLER DIPL.-CHEM. CAROLA KELLER DR.-ING. KLÖPSCH DIPL.-ING. SELTING

COLOGNE 1, DEICHMANNHAUS

Cologne, 2.5.1972. · ■■ ' Kl / Ax

The British Petroleum Company Limited, Britannic House, Moor Lane, London, E.C. 2 (England).

Process for the oxidation of hydrocarbons, alcohols and aldehydes

The invention relates to an improved process for the oxidation of hydrocarbons, alcohols and aldehydes, which are substantially immiscible or insoluble in water using a enzyme or enzyme complex extracted from the cells of microorganisms.

Process for the oxidation of hydrocarbons, alcohols and aldehydes using enzymes derived from the cells of microorganisms have been extracted have already been proposed. Method of extraction of enzymes or enzyme complexes suitable for use in these oxidation processes are also has been described in the literature.

The invention relates to an enzymatic oxidation process with increased reaction speed.

The method according to the invention for the enzymatic oxidation of hydrocarbons, alcohols and aldehydes, which are essentially immiscible with water or are essentially insoluble in water and hereinafter as "Substrate" is characterized by

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that a solution of the "substrate" is formed in an amide which is miscible with both water and the "substrate" is and the formula

has, in which R ^, Rp or R, stand for'a hydrogen atom or an aliphatic C.-Cp - radical »the solution of the" substrate "in the amide in an aqueous buffer, which has a Pjj value in the range of 6, 5 to 9.5, and dispersed in the presence of an enzyme extracted from a microorganism which oxidizes the "substrate" and a coenzyme, thereby forming a reaction mixture which, based on the solution, contains a trace amount of up to 50% by volume of the "substrate "Contains in the amide, and the reaction mixture for the oxidation of the" substrate in the presence of oxygen at a temperature in the range from 20 to 38 ⁰ C holds.

Aliphatic hydrocarbons, alcohols and aldehydes are particularly suitable as “substrate”, preferably with at least 6 to 20 carbon atoms. The carbon atoms are preferably arranged in a straight chain. The hydrocarbon is preferably saturated. Suitable for the treatment according to the invention are, for example, n-decane, n-hexadecane, 1-decanol, 1-tetradecanol, 1,10-decanediol, 1,10-hexadecanediol, 1-decanol and tetradeoanol.

The most suitable concentration of the substrate in the reaction mixture is in the range from 0.1 to 20 χ TO ^ mol.

The enzymes or enzyme complexes which are suitable for the method according to the invention can be obtained by culturing Microorganisms that consume hydrocarbons on a hydrocarbon as a source of assimilable Carbon and extraction of the enzyme or enzyme complex can be obtained in a known manner (see for example

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The Archives of Microbiology J ₀ Mo Lebeault et.al. 72.140-153, 1970). Suitable enzymes or enzyme complexes can be obtained from the cells of any bacteria, yeast or fungus which can oxidize hydrocarbons or which have been adapted for the oxidation of hydrocarbons.

If a particular oxidation is to be performed, a single enzyme can be used. For example can be an alcohol dehydrogenase for the oxidation of Alcohols can be used to form aldehydes. On the other hand, an enzyme complex can be used if more than one Oxidation stage is to be carried out. For example, an enzyme complex is used to convert alkanes to the corresponding To oxidize acids. The chemical and physical structure of the enzymes or enzyme complexes is not known in every detail. In the intact microorganism, however, these enzymes are mainly found in the mitochondria and in the cytoplasm.

The amount of enzyme or enzyme complex in the reaction mixture can be 0.1 to 10 g / l.

As a coenzyme it is suitable for the oxidation of decanes for example nicotinamide adenine dinucleotide (NAD or NADH), which is replaced by nicotinamide adenine dinucleotide phosphate (NADP or NADPH) can be replaced. The molarity of the

—2

Coenzyme in the reaction mixture can range from 10 to

10 ~ ⁵ lie.

The amides may contain the enzyme or the enzyme complex at least in the amounts and under the conditions under which they are present in the reaction mixture, do not deactivate. The amount of amidine used is usually 10 vol .- $, based on the volume of the reaction mixture “Preferably the amide is sourced in an amount of about $ 3 volume to the volume of the reaction mixture. Suitable amides are, for example, Ν, Ν-dimethylforma-

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mid, formamide, Ν, ΪΓ-diethylformamide, N ~ ethylformamide and N-methylformamide.

Any aqueous buffer commonly used for enzymatic reactions, e.g. a tris hydrochloric acid or phosphate buffers can be used.

The oxidation is carried out under the relatively mild reaction conditions which are generally used for carrying out enzymatic reactions. For example, the temperature in the range of 20 to 38 may be ⁰ O. A temperature in the range of 22 to 25 ^0C, are, for the oxidation of n-alkanes to alcohols and for the oxidation of aldehydes to acids PJJ value in the range of 7 * 5 to 8.5 is most suitable is preferred. In contrast, a pjT value in the range from 8.5 to 9.5 is particularly suitable for the oxidation of alcohols to aldehydes.

The oxygen can be used as free oxygen, e.g. as atmospheric oxygen, are present. It is also possible to use bound oxygen, e.g. those present in hydroxyl groups or water Oxygen to use.

The method according to the invention is suitable for oxidation from diols to ω-hydroxy acids. For example, can Decanediol are oxidized to hydroxydecanoic acid. Hydrocarbons can either be in alcohols or in acids being transformed. The alcohols can either be in the aldehyde or in the acid and the aldehydes in the acid being transformed.

example 1

70 mg of decane were dissolved in 1 ml of dimethylformamide. 0.1 ml of this solution was mixed with 3 ml of an aqueous tris-hydrochloric acid buffer which contained a decane-oxidizing enzyme complex and a coenzyme. The buffer had a P value of 8.0 _H ~. The decane had a molarity of 18.0 χ 10 ~ ⁵ in the mixture.

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The decane oxidizing enzyme was obtained by cultivating the yeast Candida tropicalis on n-tetradecane as a carbon source been won. The fermentation medium was centrifuged, leaving a cream containing yeast cells was obtained. The cells were washed with an isotonic solution of sodium chloride and in a tris-hydrochloric acid buffer "suspended at Pu 7.8. The cell suspension was passed through a French pressure cell and the product centrifuged at 4000 g for 20 minutes. The protruding Liquid had a concentration of 10 to 20 mg protein / ml. 1 mg was added to the mixture described above added to the supernatant liquid (which contained the decane-oxidizing enzyme complex).

Nicotinamide adenine dinucleotide phosphate (NADPH) was added as a coenzyme to the mixture in such an amount that its molarity was 17 × 10. The mixture was incubated for 30 minutes at 25 ^° C. in the presence of atmospheric oxygen. Analysis of the mixture then indicated that 85% by weight of the decane had been oxidized to a mixture of decanol and decanoic acid. The proportion of the decanol was 20 wt _o - $ 6, and the proportion of the decanoic acid 65 wt .- ^.

Example 2

1,10-decanediol was converted to 10-hydroxydecanoic acid on the in Example 1 described oxidizes, but the Decane replaced by the same amount by weight of 1,10 decaudiol became. 10-hydroxydecanoic acid was obtained in a yield of 70 to 80 wt .- ^.

Example 3

Decanol was oxidized to decanoic acid in the manner described in Example 1, but the decane was replaced by decanol was replaced. The decanoic acid was obtained in a yield of 95 to 98% by weight.

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Comparative experiment

For comparison, decane was oxidized to decanol and decanoic acid in the manner described in Example 1 with the essential difference that the dimethylformamide was omitted and the mixture was stirred in an effort to accelerate the reaction. Analysis of the mixture showed that only 10 wt - ⁰ Jo of decane had been oxidized to a mixture of decanol and decanoic acid.. The proportion of decanol was 20% by weight and the proportion of decanoic acid was 65% by weight.

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Claims (5)

te η ta ns ρ r ti c h 1) Process on enzymatic oxidation ψ & ό hydrocarbons, "alcohols und.Aldehyden ₉ - which are essentially immiscible with water or are essentially insoluble in water and are hereinafter referred to as" Stfbstrai ⁵ ', characterized- that forming a solution of the "substrate" in an amide, which is miscible both with water and with the "** substrate and the formula O ₅ has to stand in the R ^, R ₂ or R * is a Y / asserstoffatom or an aliphatic Cj-Cc residue, the solution! of "substrate" in the amide in an aqueous buffering a _pH value in the range of 6, 5 to 9.5 has, VoI 50 .- ^ country in the presence of the "substrate" oxidizing dispersed from a microorganism extracted enzyme and a Goenayms and thereby a reaction mixture is the _s, based on the solution, of Spur'bis " Contains substrate "in the amide, and the reaction mixture for the oxidation of the" substrate "in the presence of oxygen at a temperature in the range of 2Q to 38 ° C. 2) Method according to claim 1, characterized in that the concentration of the "substrate" in the reaction mixture is in the range from 0.1 to 20 χ 10 ^J mol. 3) Method according to claims 1 and 2, characterized in that that the enzyme which oxidizes the "substrate" is a complex which carries out more than one oxidation stage capable of being used. 4) Process according to Claims 1 to 3, characterized in that the "substrates" are aliphatic hydrocarbons 209847/1247. substances with at least 6 to 20 carbon atoms are used. 5) Process according to claims 1 to 4, characterized thereby. draws that dimethylformamide is used as the amide will. 209847/1247