DD215091A1 - METHOD OF OBTAINING UBICHINONES - Google Patents

Abstract

The invention relates to a process for obtaining the ubiquinones from lipid extracts which are obtained in the extraction of microbially produced biomasses and have a hydrocarbon content of <1%. The aim of the invention is the cost-effective production of ubiquinones from lipid extracts of microbial origin with minimal consumption of excipients. The task d. Erfindg. consists of enriching the ubiquinone before extraction. According to the invention, this object is achieved in that the lipid extract or the acetone-soluble fraction of the lipid extract of a discontinuous or continuous multiplicative distribution in a solvent system consisting of a hydrocarbon or a hydrocarbon mixture, preferably in the boiling range 50 to 160 degrees C, an alcohol (chain length C deep 1 to C deep 4) u. a water addition of 0 to 5%, is subjected. From the ubiquinone-containing phases, the ubiquinones are separated and optionally purified.

Description

250535 2

Title of the invention Process for the production of ubiquinones Field of application of the invention

The invention relates to a process for the production of ubiquinones from lipid extracts which are obtained by extraction of microbially produced biomasses with isolar, polar or "combinations of polar and / nonpolar solvents and which have a hydrocarbon content of '4L 1 % "

It is classified in the IPK C 12 P * Characteristic of the known technical solutions

Of the lipophilic constituents of biomasses, the ubiquinones which have coenzyme activity in the electron transport system of the respiratory chain are of particular interest. Numerous pharmacological effects have been reported for ubiquinone. For example, remarkable effects have been demonstrated on congestive heart failure, coronary insufficiency, malnutrition-induced muscular dystrophy, and anemia.

25 05 3 5 2

For the production of ubiquinones, microbial biomasses are particularly suitable, since they have higher ubiquinone contents compared to other biological starting materials (FJ, * Crane, Progr * Chem «Fats Lipids 7 (1964), 257), methods for obtaining ubiquinones from micro ~ For example, OP-PS 37-9290 describes a process "which aims at isolating ubiquinones from microbial biomass"

The lipid extract isolated by extraction with a polar solvent, for example methanol, is saponified in the presence of pyrogallol with alcoholic alkali hydroxide solution and the ubiquinones are separated from the unsaponifiable fraction by column chromatography using silica gel. Further purification of the ubiquinones is carried out by recrystallization.

DD-PS 115 899 reports on a process for the simultaneous recovery of ergosterol and ubiquinone-9 from lipid extracts of microbial origin

After alkaline-ethanolic saponification in the presence of a reducing agent, the unsaponifiable constituents are isolated from the lipid extract and separated therefrom by cooling ergosterol and also by chromatography on Al_O, ubiquinone-9. A disadvantage is found in the inOP-PS 37-9290 and DD-PS 115 899 that most of the lipids contained in the lipid extracts, such as phosphatides and fats, undergo a metabolic process. "Saponification, extraction of the soap solution, and isolation of the ubiquinones from the unsaponifiable materials require a high amount of chemicals, labor, and energy In DE-OS 28 34 952 described method, a material conversion is avoided. From moist or spray-dried bacterial biomass, especially of the species Xanthomones _{β W rc} | _m it i ~ _e nem mixture of methanol and butanol lipid extract ge ~ gained. From the alcoholic solution, the Ubi ~ become

25 05 3 5 2

extracted with hexane and then separated by acetone precipitation the phosphatides »from the acetone-soluble fraction are obtained by multiple recrystallization the ubiquinones« The method described also has a number of disadvantages * The method is only applicable to bacterial lipid extracts, the ubiquinone-8, ubiquinone Or which contain no further constituents which likewise accumulate in the ubiquinone-containing fraction, such as waxes, alcoholeu,

For lipid extracts containing the difficult to crystallize ubiquinones of lower chain lengths and / or other ingredients »additional chromatographic processes for separation and purification are required * For the recovery of ubiquinones from lipid extracts, which are obtained from yeast, that is described in DE-OS 28 34 952 described procedures due to the high proportion of fat and ergosterol principally not applicable «

Object of the invention

The aim of the invention is a process which allows the recovery of ubiquinones from lipid extracts which have been obtained by extraction of microbially produced biomasses with non-polar, polar or combinations of polar and non-polar solvents and have a hydrocarbon content <1%, cost-effective with minimal consumption of excipients and labor "

Presentation of the essence of the invention

The object of the invention is to _enrich ubiquinone from _lipid extracts by utilizing the different solubilities of the individual components of the lipid extract in lipophilic and hydrophilic solvents. _E

250535 2

According to the invention, this object is achieved in that from the lipid extract or acetone-isolated by acetone treatment acetone-soluble fraction, the ubiquinones are enriched by multiplicative distribution in a solvent system to the extent that a subsequent simple isolation is possible.

In detail, the following procedure is followed: The lipid extract is separated in known manner into an aceto-insoluble fraction containing the phosphatides and an acetone-soluble fraction containing the ubiquinones. The acetone-soluble fraction is removed after distillative removal of the acetone a solvent mixture of a hydrocarbon or a hydrocarbon mixture, preferably in the boiling range 50 to 160 ⁰ C, an alcohol (chain length C ₁ to C ₄ ) and a water addition of O to 5 % of a multiplicative distribution subjected »It is also possible directly from the Lipid extract enrich the ubiquinones by multiplicative distribution »The multiplicative distribution can be carried out batchwise or continuously« From the ubiquinone-containing phases, which are obtained as a result of the multiplicative distribution »after removal of the solvent, the ubiquinones by recrystallization from alcohol However, it is also possible to optimize the multiplicative distribution so that ubiquinones with a purity of> 95 % are obtained without additional purification operations. From the values obtained in the Craig distribution, the continuous separation of the ubiquinones can be determined without difficulty Calculate O'Keeffe "The essential advantage of the method according to the invention is that the recovery of ubiquinones is possible to the exclusion of metabolic processes. Thus, the time and effort while reducing so "as the consumption of energy and chemicals in the isolation of ubiquinones ·

25 053 5 2

All of the raw materials used in the process can be fed into repeated use. "The method according to the invention can be used for the isolation of ubiquinones with different lengths of the isoprenoid side chain (ubiquinone-6 to ubiquinone-10) from lipid X-tracts of various origin" such as lipid extracts of bacteria, yeasts - or algae biomasses "used" if they have a hydrocarbon content 4L 1 % "Aue sterolhaltigen starting biomass can be isolated without additional effort, the sterols" because the multiplicative distribution of sterols and ubiquinones are completely separated. All fractions resulting from the work-up are economically usable, d. h, the method according to the invention works free from waste «

The method according to the invention is characterized by the following; At-, games explained:

example 1

A lipid extract was obtained from the methanol-utilizing bacterial strain Acetobacter spec. The lipid extract is characterized as follows:

Phosphatides 63.5 %

Lipids (MAINLY 32 _# 7% fatty acids quietly u., Glycerides)

Unsaponifiable 4.8 % ubiquinone-10 0.7% '

To prepare the upper and lower phase for the Craig distribution, 14 l of low-boiling point benzine are added to each other with 14 l of methanol. 50 g lipid extract are made up to 200 ml with upper phase and introduced into the first two distribution elements. Apparatus with an element volume of 200 ml »The following parameters were set for the distribution:

25 05 3 5 2

Elements for Saturation: 5 to Distribution: 100 settling time: 5 min Temperature: 15 ⁰ C

After concentration of the ubiquinone-containing elements, 480 mg of product are obtained with a ubiquinone content of 62 % »

Recrystallization from ethanol gives 253 mg of reinubichinone,

Example 2

To prepare the upper and lower phases for the Craig distribution, 14 l of low-boiling point benzine are added to each other with 14 l of methanol and 70 ml of water.

100 g of a lipid extract obtained from Acetobacter spec are mixed with 300 g of acetone at room temperature for 10 minutes. The lighter ubiquinone-containing phase is separated off, the acetone is removed by distillation and the residue is topped up to 200 ml and placed in the first at the distribution elements. For an element volume of 200 ml, the following parameters were set for the Craig distribution:

Elements for saturation: 5

for distribution: 100

Settling time: 4 min

Temperature: 22 to 25 ⁰ C

After concentrating the ubiquinone-containing elements, 1205 mg of product having a ubiquinone content of 58 % were obtained.

Recrystallization from ethanol gives 502 mg of pureubiquinone. "

25053 5 2 ⁷

Example 3

A lipid extract obtained from Saccharomyces cerevisiae is characterized as follows:

Phosphatides 55 %

Sterols g %. ,

Lipids (mainly 35.8 % fatty acids and esters)

Ubiquinone-6 O _s 15%

To prepare the upper and lower phase for the Vertexlung be 14 1 of low-boiling point benzene with 14 1 methanol and 140 ml Vlaseer added to the mutual saturation «IGO g lipid extract are mixed at room temperature for 10 minutes with 30Og of acetone. The lighter ubiquinone-containing phase is separated off, the acetone is removed by distillation and the residue is topped up to 200 ml and introduced into the first two distributing elements * With an element volume of 200 ml, the following parameters were set for the Crsic distribution:

Elements for saturation: 5

for distribution: 100

Settling time: 3 min

Temperature: 20 ^° C

After concentration of the ubiquinone-containing elements, 198 mg of product with a ubiquinone content of 74% were obtained.

Recrystallization from ethanol gives 133 mg of rhi- umubinone

Claims (2)

κ *. N ₁ / \ J \ j W sJ «wot Erf induncjsanepruch Process for the production of ubiquinones from lipid extracts obtained by extraction of microbially produced biomass with nonpolar, polar or combinations of polar and nonpolar solvents and having a hydrocarbon content of <1 % , characterized in that the lipid extract or one thereof isolated bcetonlösliche fraction of a multiplicative distribution in a solvent system consisting of a hydrocarbon or a hydrocarbon mixture, preferably in the boiling range 50 to 160 ⁰ C, an alcohol (chain length C ₁ to C ₄ ) and a water addition of O to 5 % for enrichment subjugates the ubiquinone, and from the enriched phases, it gains the supraquinone » 2, method according to item 1, characterized in that the multiplicative distribution is carried out discontinuously or continuously »