DD295189A5 - PROCESS FOR PREPARING OXYGEN DERIVATIVES OF PHENYL ETHANE - Google Patents

Abstract

The invention relates to a process for the preparation of oxygen derivatives of phenylethane containing one to three oxygen atoms in the side chain by biotransformation. The aim is to carry out the oxidation of phenylethane, styrene or lower oxidation products of phenylethane with such microorganisms that allows the synthesis of almost all conceivable oxygen derivatives of the aromatic used. The inventive solution provides for the use of methanol-utilizing, but no methane monooxigenase contained, therefore, no methane-utilizing yeasts of the species Candida boidinii, of bacteria of the genus Aerobacter or Pseudomonas in the presence of 0.2 to 12 g methanol / 1 before. {Oxygen derivatives; Oxidation; Synthesis; Yeast; phenylethane; styrene; aromatics; Bacteria; biotransformation; microorganisms; methanol}

Description

The object is achieved in that the transformation of alkylbenzene particular styrene

or of phenylethane by means of methylotrophic bacteria or yeasts wM, which contain no Methanmonooxigenase, so have not previously been adapted to methane or can not use methane at all. This preferably occurs in the presence of methanol and in comparison to other processes while saving precultivation on methane.

As methanol utilizing, but not using methane microorganisms z. B. yeasts of the species Candida boidinii,

acidophilic bacteria of the genus Acetobacter methanolicus or neutrophilic bacteria of the genus Pseudomonas suitable.

The alkylbenzene transformation is carried out in the presence of methanol, a pH of 2.5-5.5 in acidophilic strains or

of 5.5-7.5 in less acidophilic or neutrophilic strains and a temperature of 30-50 ⁰ C carried out.

Under these conditions, the specified microorganisms are able to styrene and phenylethane in the most diverse To convert oxygenated and less oxidized compounds into more highly oxidized compounds. As oxidation products

arise here z. B. 1- and 2-phenylethanol, phenylacetaldehyde, acetophenone, phenylacetic acid, phenylglyoxal,

Mandelic acid, d. H. Compounds that z. T. in the pharmaceutical, especially cosmetic industry and in the Production of pesticides play an important role. Styrenes also form styrene oxide and styrene ethylene glycol. The percentage of said products is determined by both the aromatic to be oxidized Hydrocarbons as well as the set transformation conditions such as the presence or absence of individual Nutrients and in this or other known manner activated or deactivated fnzymes determined. The blocking of Enzymes by known enzyme inhibitors can lead to maximizing the yield of individual major products. in the

The individual procedure is such that in a methanol consuming fermentation process with one of said

Microorganisms and an extracellular methanol concentration of 0.5 to 10g methenol / l styrene or ethylbenzene in Portions of 0.5 ml / l fermenter content is registered. The microorganism concentration can be between 1 and 50 g BTS / I

and the styrene or ethylbenzene concentration should advantageously be below 0.5 g / l.

The invention will be explained in more detail with reference to the following examples: example 1 In a stirred tank reactor with a working volume of 21 is a discontinuous Verhefung of methanol

carried out:

Temperature around 32 ⁰ C, pH = 4.5, p O ₂ = 0-5ppm The methanol used as the yeast is Candida boidinii IMET M 131. Their starting concentration is 3 g HTS / kg, the specification of methanol about 10 g / l. The culture medium contained up to the nitrogen all vital nutrients and vitamins in the form of a small Proportion of yeast extract sufficient for an increase of 15g HTS / I. The N supply of the yeast took place autoregulativ over the

pH control with NH ₄ OH. The fermentation was carried out under protected conditions.

After the leaching of the methanol submit again, in total twice each 10g methanol / l accordingly

given the consumption growth substrate in the fermentor. As a result of the utilization of this amount of methanol by

Candida boidinii was given a biomass concentration of about 11 g HTS / I. Subsequently, another one Methanol amount of 5 g methanol / l presented and over a period of 11 h a styrene amount of 15 ml / l in portions of

0.5ml dosed. The amount of styrene was dependent on the d6r pOj display. After the last Styrengabe another half an hour was transformed and then stopped the fermentation process. The fermentor content was added and separated with 1 to 2% of the active surface active substance forms. The liquid phase was extracted with ethyl acetate, which

Extract dried using sodium sulfate and the solvent distilled off. In the concentrated extract amount of 2.2g

the following O-derivatives of ethylbenzene were determined by GC and GC / MS:

Styrene oxide, styrene ethylene glycol, 1- and 2-phenylethanol, acetophenone, phenylacetaldehyde, phenylacetic acid, mandelic acid. Example 2

Nearly the same product composition except for the first two was achieved in the transformation of ethylbenzene using the acidophilic bacterial culture Acetobacter methanolicus spec, under similar conditions of cultivation and transformation. 15 ml of ethylbenzene were reacted over a period of 14 hours and an extract amount of 1.9 g was obtained.

Claims (4)

1. A process for the preparation of oxygen derivatives of phenylethane containing one to three oxygen atoms in the side chain by biotransformation, characterized in that the aromatic oxidation using methanol utilization but no methane monooxigenase contained, therefore no methane-utilizing, microorganisms is carried out in the presence of methanol , 2. The method according to claim 1, characterized in that as methylotrophic, no methane-utilizing microorganisms yeasts, for. As the Ait Candida boidini, or bacteria of the genera Aerobacter or Pseudomonas be used. 3. The method according to claim 1, characterized in that the biotransformation to styrene, phenyl ethane or to lower oxidation products of phenylethane such. B. is carried out on phenylethanols. 4. Process according to claims 1 to 3, characterized in that the aromatics oxidation in the presence of 0.2 to 12 g methanol / l is performed. Field of application of the invention The invention relates to a process for the preparation of oxygen derivatives of phenylethane having a proportion of one to three oxygen atoms in the side chain by microbial oxidation. The invention can be realized in continuous or semi-continuous fermentation processes. Characteristic of the known solutions Processes for the microbial oxidation of a wide variety of benzene derivatives are known. These are predominantly the production of styrene epoxies (Idemitsu Kazan Co., Ltd., 1.6.78, JP-PS 8000033, ICI, 297.81, DE-OS 3129935), of hydroxylated products such as phenols and phenylethanols (Phizer Co.). 1983, EP-PS 73134, ICI, 13.4.83, EP-PS 76606), ketones (Idemitsu Kazan Co. Ltd., 5.1.80, JP-PS 8000 012, DP Cox, CD Goldsmith, 1979, Appl. Environ Microbiol. 38 [3], 514-526) as well as the recovery of various carboxylic acids (Calanese Co. 18.1.84, EP-PS 98750, Hsieh, Jih Han, 10.1.82, US-PS 4608338, Horiguchi et al. Jigami, Yoshifumi et al., Agric., Biol. Chem., 1974, 38, [2], 401-8), 18.8.82, JP-PS 61185192; As a rule, strains as well as defective mutants of bacteria yeasts or fungi are used, which have special services with respect to individual target products or product mixtures and which must be protected from wild strains or foreign organisms and therefore cultivated under sterile conditions. Above all, microbial specialists usually require a great deal of effort and special procedures for cultivation, which are often time-consuming. Thus, by a process, methylotrophic bacteria are used for the oxidation of aromatic and aliphatic hydrocarbons (IJ Higgins, 22.4.82, DE-OS 3129935), which is prepared over several stages, first cultured on methane, then adapted to methanol vapor, further washed and then by oxidation process can be used. The microorganisms used in this method, according to the activity of methane monooxigenase on a relatively low growth rate and oxidation rate. Thus, according to a process variant, the adaptation time to methane is 4-5 days, the duration of cultivation on this carbon source alone 7 days. The methanol-adapted, a ϊ lethanmonooxigenase contained and therefore methane-utilizing bacteria of the species Methylosinus, Trichosporium and Methylococcus capsulatus are preferably used for the hydroxylation. However, they should also be able to be used for the epoxidation of styrene as well as for the synthesis of oxygen derivatives of phenylethane to a limited extent, such as for the production of 2-phenylethanol, p-hydroxyethylbenzene, benzoic acid and phenylacetic acid from ethylbenzene. For the synthesis of other target products other specialists must be used, whereby the cultivation regulations and thus the technical and material expenses can differ considerably, which is a financial disadvantage in a production conversion. Object of the invention The aim of the invention is a mlkroblelles method with which it is possible, inexpensive and in particular time-saving nahozu to synthesize all theoretically derivable oxygen derivatives of phenylethane. Explanation of the essence of the invention The object of the invention is a process which, by means of suitable microbial biomass and process control, makes it possible to obtain a broad spectrum of phenyiethane oxygen derivatives by oxidation of alkylbenzenes in comparison to known microbial processes.