DE10119972A1 - Fatty acid steryl ester preparation in high yield, for use e.g. in pharmaceutical or cosmetic applications, by enzymatic conversion of untreated deodorizer distillates or tall oil using lipase - Google Patents

Abstract

Enzymatic synthesis of fatty acid steryl esters (I) comprises: (i) reacting fatty acids, fatty acid esters and sterols, present in untreated deodorizer distillates from fat refining, under stirring in presence of lipase; (ii) removing water and volatile by-products by distillation; and (iii) enriching and purifying (I) by deacidification. Enzymatic synthesis of fatty acid steryl esters (I) comprises: (i) reacting fatty acids, fatty acid esters and sterols, present in untreated deodorizer distillates from fat refining, under stirring in presence of lipase; (ii) removing the water of reaction and volatile by-products by distillation; and (iii) enriching and purifying the obtained (I) by deacidification with aqueous solution of (in)organic base and/or solvent fractionation with acetone, methanol, ethanol or other polar aprotic solvents (or their aqueous solutions). Independent claims are included for variants on the process, in which the starting mixture is untreated tall oil or a fraction obtained from untreated deodorizer distillates or tall oil.

Description

The invention is characterized by the solvent-free implementation of fatty acids and Fatty acid esters with sterols, used as mixtures in the steam refining distillates (Vapor oils) or in papermaking residues (tall oils), where the Reaction partner without solvent in the presence of lipases in vacuo to fatty acid react steryl esters. The resulting fatty acid sterylesters are removed by deacidification, Solvent fractionation and chromatographic processes enriched and purified.

When refining fats and oils, damper distillates (vapor oils) are used as a secondary factor products of the damping process (deodorization). Steam distillates from vegetable oils mainly contain free fatty acids, mono-, di- and triglycerides, phytosterols and their Fatty acid esters and tocopherols. Especially phytosterols, their fatty acid esters (fatty acid sterylester) and tocopherols are of economic value and are derived from the Damper distillates separated, in particular physical (different Distillation process) and chemical processes (extraction with solvents, Derivatization) can be used (cf. Kim, S.K. and Rhee, J.S., 1982. Isolation and purification of tocopherols and sterols from soya oil deodorization. Korean Journal of Food Science and Technology 14, 174-178; Sheabar, F.Z. and Neeman, L, 1988. Separation and concentration of natural antioxidants from the rape of olives. Journal of the American Oil Chemists' Society 65, 990-993; Lee, H., Chung, B.H. and Park, Y.H., 1991. Concentration of tocopherols from soybean sludge by supercritical carbon dioxide. Journal of the American Oil Chemists' Society 68, 571-573). More recently, there are also various biochemicals Process, e.g. B. lipase-catalyzed hydrolysis or transesterification, to obtain Phytosterols from steam distillates have been described (Ghosh, S. and Bhattacharyya, D.K., 1996. Isolation of tocopherol and sterol concentrate from sunflower oil deodorizer distillate. Journal of the American Oil Chemists' Society 73, 1271-1274; Ramamurthi, S. and McCurdy, A.R., 1993. Enzymatic pretreatment of deodorizer distillate for concentration of sterols and tocopherols. Journal of the American Oil Chemists' Society 70, 287-295; Ramamurthi, S., McCurdy, A.R. and Tyler, R.T., 1998. Deodorizer distillate: A valuable by-product In: Proceedings of the world conference on oilseed and edible oils processsing: Emerging technologies, current practices, quality control, technology transfer, and environmental issues, Vol. I [Koseoglu, S. S., Rhee, K. C. and Wilson, R. F., Eds.] S. 130-134, AOCS Press, Champaign; Shimada, Y., Nakai, S., Suenaga, M., T., Sugihara, A., Kitano, M. and Tominaga, Y., 2000. Facile purification of tocopherols from soybean oil deodorizer distillate in high yield  using lipase. Journal of the American Oil Chemists' Society 77, 1009-1013; Daguet, D. and Coic, J.-P., 1999. Phytosterol extraction: State of the art. Oleagineux Corps Gras Lipides 6, 25-28). In This process transforms the phytosterylesters of the damper distillates into free phytosterols or the free fatty acids converted to fatty acid methyl esters to make it easier and cheaper Separation of the commercially usable components of the steamer distillates, d. s. phytosterols and tocopherols.

Similarly, tall oil is used as a by-product in the manufacture of pulp Digestion of resin-rich wood species after the sulfate process is obtained as a starting material used for the production of phytosterols. Tall oil mainly consists of one Mixture of fatty and resin acids, unsaponifiable fractions and fatty acid sterylesters (Drew, J. and Propst, M., ed., 1981. Tall oil. Pulp Chemical Association, New York; by Hellens, S., 1999. Benecol margarine enriched with stanol esters. Lipid Technology 11, 29-31).

Fatty acid steryl esters are used for a variety of purposes as pharmaceuticals, as components of Cosmetics and as liquid-crystalline compounds in technology (Koshiro, A., 1987. Cholesterol ester and its manufacture using microbial lipase. Japan. Kokai Tokkyo Koho 62 296 894 (Chemical Abstracts 110: 210969p, 1989]). Recently, fatty acid esters have become the Phytosterols are also used as dietary supplements in dietary margarines because they are the Can lower blood cholesterol levels (Miettinen, T., Vanhagen, H. and Wester, L, 1996. Use of stanol fatty acid ester for reducing serum cholesterol level. U.S. Patent 5,502,045; of Hellens, S., 1999. Benecol margarine enriched with stanol esters. Lipid Technology 11, 29-31; Wester, L, 2000. Cholesterol lowering effect of plant sterols. European Journal of Lipid Science and Technology, 37-44).

In previous chemical processes, phytosterols are obtained from steam distillates, by usually subjecting them to alkaline saponification to fatty acid converting steryl esters into free sterols (Daguet, D. and CoYc, J.-P., 1999. Phytosterol extraction: State of the art. Oleagineux Corps Gras Lipides 6, 25-28). The reaction mixtures were, among other things, by technical processes such as molecular distillation, Solvent fractionation and recrystallization from organic solvents cleaned. The mixture of free phytosterols obtained in this way served in esterification or Transesterification process for the production of the corresponding fatty acid sterylester. Dry Processes are primarily based on the transesterification of fatty acid esters with sterols or Reactions of carboxylic acid halides or anhydrides with hydroxyl groups of sterols (Mahadevan, V. and Lundberg, W., 1962. Preparation of cholesterol esters of long-chain fatty acids and characterization of cholesteryl arachidonate. Journal of Lipid Research 3, 106-110;  

Pinter, K.G., Hamilton, J.G. and Muldrey, J.E., 1964. A method for rapid microsynthesis of radioactive cholesterol esters. Journal of Lipid Research 5, 273-274; Piretti, M.V. and Pagliuca, G., 1996. Synthesis of highly pure dolichyl and cholesteryl esters. Italian Journal of Biochemistry 45, 67-69). Phytosterols are made from tall oil soaps or distillation residues of the tall oil obtained by extraction, catalytically hydrogenated and the resulting phytostanols converted into fatty acid stanyl ester by transesterification with fatty acid methyl esters (von Hellens, S., 1999. Benecol margarine enriched with stanol esters. Lipid Technology 11, 29-31).

Enzymatic processes for the production of fatty acid sterylesters are also known; they start from fatty acids and sterols (esterification) or fatty acid esters and sterols (transesterification), which are converted as biocatalysts in the presence of lipases. Previous methods (Koshiro, A., 1987. Cholesterol ester and its manufacture using microbial lipase. Japan. Kokai Tokkyo Koho 62 296 894 [Chemical Abstracts 110, 210969p, 1989]; Kosugi, Y., Tanaka, H., Tomizuka, N ., Akeboshi, K., Matsufune, Y. and Yoshikawa, S., 1989. Enzymic manufacture of sterol fatty acid esters. Japan. Kokai Tokkyo Koho 1 218 593 [Chemical Abstracts 113, 4707k, 1990]) describe the lipase-catalyzed esterification of organic carboxylic acids with hydroxyl groups of sterols, e.g. B. cholesterol, in organic solvents such as heptane, methyl tert-butyl ether, acetone and others, partly using molecular sieves in the reaction mixture to remove the water or alcohol formed (Shimada, Y., Hirota, Y., Baba , T., Sugihara, A., Moriyama, S., Tominaga, Y. and Terai, T., 1999. Enzymatic synthesis of steryl esters of polyunsaturated fatty acids. Journal of the American Oil Chemists' Society 76, 713-716; Haraldsson, GG, 1992. The application of lipases in organic synthesis. In: Supplement B: The chemistry of acid derivatives, Vol. 2 [Patai, S., Ed.] Pp. 1395-1473, John Wiley, Chichester; Kazlauskas, RJ and Bomscheuer, UT, 1998. Biotransformations with lipases In: Biotechnology [Rehm, H.-J. and Reed, G., Eds.] 2 ^nd edition, Vol. 8a: Biotransformations, pp. 37-191, Wiley- VCH, Weinheim; Riva, S. and Klibanov, AM, 1988. Enzymochemical regioselective oxidation of steroids without oxidoreductases, Journal of the American Chemical Society 110, 3291; Riva, S., Bovara, R., Ottolina, G., Secundo, F. and Carrea, G., 1989. Regioselective acylation of bile acid derivatives with Candida cylindracea lipase in anhydrous benzene. Journal of Organic Chemistry 54, 3161-3164; Faber, K. and Riva, S., 1992. Enzyme-catalyzed irreversible acyl transfer. Synthesis, 895-910).

Lipase catalyzed esterification and transesterification reactions in vacuo to remove the volatile reaction products have recently been used for the production of fatty acid sterylesters (Weber, N. and Mukherjee, K.D., Enzymatic Process for the Production of  Carboxylic acid steryl. Patent application DPMA-Az. 100 18 787.0 (4/15/00); Weber, N., Weitkamp, P. and Mukherjee, K.D., 2001. Fatty acid steryl, stanyl and steroid esters by esterification and transesterification in vacuo using Candida rugosa lipase as catalyst. Journal of Agricultural and Food Chemistry 49, 67-71).

In contrast to the known processes mentioned above, the enzymatic syntheses of the fatty acid steryl esters described in detail below are based on untreated by-products of fat processing (damper distillates) and paper production (tall oils) in the absence of solvents and desiccants such as e.g. B. molecular sieve or sodium sulfate. However, it is also possible to apply the process to treated and / or fractional steamer distillate and tall oil, which are usually produced for industrial applications, as starting materials (starting materials) for the enzyme-catalytic esterification and transesterification of fatty acid sterylesters. The mixtures of the reactants, ie fatty acids or fatty acid esters and phytosterols, such as those present in untreated or treated steam distillates from vegetable oils or in tall oils from paper processing and their fractions, are enzyme-catalytically converted in the presence of lipases and the water or other volatile reaction products formed in vacuo removed from the reaction mixture. This lipase catalyzes esterification and transesterification reactions in vacuo to produce fatty acid sterylesters ( Fig. 1) based on mixtures of fatty acids, fatty acid esters (mono-, di-, triglycerides) and phytosterols in damper distillates and tall oils with subsequent enrichment and purification of the fatty acid Sterile esters by deacidification, solvent fractionation and / or chromatographic processes have hitherto been unknown.

Fig. 1

Production of fatty acid steryl esters, e.g. B. fatty acid sitosterylesters, lipase-catalyzed esterification and / or transesterification of the mixtures of fatty acids, fatty acid esters (e.g. triglycerides) and phytosterols (R = alkyl residues of fatty acids or fatty acid esters) contained in steamer distillates and tall oils

Phytosterols are found in steam distillates and tall oils as natural mixtures, for example evidence of sitosterol, stigmasterol and campesterol. The composition of phytosterols in steam distillates or tall oils is typical for every product; for rapeseed oil, for example Characteristic of brassicasterin occurrence.

The free fatty acids are mixtures of fatty acids as described in Damper distillates of vegetable oils and naturally in tall oils are available with one for the each product typical composition.

For the lipase-catalyzed synthesis of fatty acid sterylesters by transesterification stand as Fatty acid esters, especially those in the steam distillates of vegetable oils, e.g. B. rapeseed oil, Sunflower oil and soybean oil, contain mono-, di- and triglycerides. tall oils usually contain only small amounts of triglycerides. The transesterification (alcoholysis) of In contrast, mono-, di- and triglycerides with hydroxy groups of phytosterols are required basically no vacuum for the esterification of fatty acids described above, since none Water of reaction is formed. Nevertheless, here too the yield increases when the reaction is carried out in a vacuum.

All known lipases can be used as enzymes for the production of the fatty acid steryl esters catalysts are used, especially lipases from microorganisms, such as. B. from Rhizopus arrhizus, Candida antarctica, Candida nigosa (= C. cylindracea), Rhizomucor miehei and Geotrichum candidum, pancreatic lipases from various animal species as well as lipases and Acyl hydrolases from plants, such as. B. papaya, rapeseed, castor, potato, rice, Vernonia and Pineapple.

The lipase-catalyzed syntheses of fatty acid sterylesters can be used in different Reaction conditions are carried out, in particular by the selected lipases depend. In general, temperatures between 10 and 100 ° C are used, preferred those between 30 and 80 ° C. The molar ratios between the reactants, i.e. s. Fatty acids or fatty acid esters and sterols are in the raw products (steam distillates, tall oils) specified. But you can also by adding other components (free sterols, Fatty acids, fatty acid esters) are specifically changed and are subject to the proportions of added lipases no restrictions. The response time is also not limited. Stirring the reaction mixture and increasing the amount of lipase in the test batch increase the Reaction speed and thus accelerate the formation of the fatty acid sterylester. Repeated use of the lipase used in one reaction batch in a new batch is possible without significant loss of performance of the biocatalyst. Suppress between  900 mbar and 0.1 mbar can be used for the reaction; usually become Pressures between 200 mbar and 10 mbar maintained.

The fatty acid steryl esters can be cleaned in various ways. Usually the enzyme catalyst is after the reaction by centrifugation or Filtration separated from the reaction mixture or the reaction mixture with an organic, preferably extracted apolar solvent. The enrichment and purification of the fatty acid Steryl ester is first carried out by removing the free fatty acids from the reaction mixture by extraction with the aqueous solution of a strong inorganic or organic base, z. B. with 5% aqueous sodium carbonate solution (deacidification). Alternatively, the free ones Fatty acids can also be separated by distillation.

Then solvent fractionation in polar aprotic solvents and made their aqueous solutions, in which preferably the fraction of the neutral polar components - such as triglycerides - separated from the fatty acid sterylesters becomes. For fractionation, for example, water-miscible polar organic Solvents such as acetone, methanol or ethanol are used in which the raw products or intermediates are solved. Then phase separation by adding Water and / or cooling initiated. Alternatively, the enrichment of fatty acid sterylesters by distributing the components of the reaction mixture between apolar organic Solvents such as As petroleum ether, hexane or iso-hexane, and polar aprotic Solvents or their aqueous solutions possible. In these fractionation processes the apolar fatty acid sterylester accumulate in the less polar organic phases that more polar constituents (e.g. triglycerides) in the polar, for example organic-aqueous Phases.

Alternatively, reaction mixtures or those enriched with fatty acid sterylesters Fractions from deacidification or solvent fractionation enriched by chromatography and be cleaned. For this purpose, the mixtures are packed on a short column Sorbent, preferably silica gel, adsorbed and then by the fatty acid sterylester Elution with mixtures of apolar and less polar, aprotic organic solvents, e.g. B. iso-hexane / diethyl ether mixtures, of residues of unreacted fatty acids or of Fatty acid esters and phytosterols separated.

Various exemplary embodiments are given below for the production of Fatty acid sterylesters, which is carried out in heated stirred vessels under vacuum:  

example 1 Characterization of steam distillates and crude tall oil

Damper distillates (vapor oils) and crude tall oils are thin-layer and gas chromatographic analyzed and the content of free phytosterols, phytosterylesters, free fatty acids and Triglycerides determined (Table 1). Implementation of thin-layer chromatography and gas chromatographic analyzes are described in Example 2.

Table 1

Composition of steam distillates and crude tall oil, which were used for the lipase-catalyzed esterification and transesterification of the fatty acids, fatty acid esters and phytosterols contained therein

Example 2 Analysis of the fatty acid sterylester Thin layer chromatography on silica gel layers

Proportions of steam distillates from vegetable oils and crude tall oil or proportions of Reaction mixtures, dissolved in iso-hexane / diethyl ether, are on thin-layer silica gel plates (0.3 mm layer thickness) applied in dots or lines. The plates are in one Mixture of iso-hexane / diethyl ether (95: 5, v / v) developed and the different Connection classes of the reaction mixture separated. By spraying the plates with a aqueous sulfuric acid solution and subsequent heating or by steaming with iodine the different connection classes are then made visible. The identification of the individual connection classes are made by comparison with known standards. The following  Rf values are given for the different classes of compounds in the reaction mixture Found use of the above eluent: fatty acid sterylester 0.6-0.7; fatty acid methyl ester 0.4-0.5; Triglycerides 0.3-0.35; Phytosterols 0.1-0.15; free fatty acids and others organic carboxylic acids <0.1.

Gas chromatography (GC)

Proportions of steam distillates from vegetable oils and crude tall oil or proportions of Reaction mixtures are suspended in diethyl ether and the insoluble constituents (e.g. Lipase catalysts) by centrifugation and subsequent filtration through a syringe filter (Pore size 0.35 µm) separated. Free fatty acids in the reaction mixture are Derivatization with diazomethane converted into the corresponding methyl ester. The resulting Mixture of fatty acid methyl esters, phytosterols and fatty acid steryl esters is gas chromato analyzed graphically. The separation on a 0.1 µm Quadrex 400-1HT high temperature Capillary column, 15 m × 0.25 mm i. D. (Hydrogen as a carrier gas with a linear flow speed of 25-35 cm / sec; Temperature program: 130 ° C [5 min isothermal] followed by a linear temperature increase from 130 to 180 ° C with 5 ° C / min. then at 20 ° C / min to 410 ° C [10 min isothermal]) provides for the different compounds of the reaction mixtures and standards, for example the following retention times (Rt): methyl palmitate (1.0 min); Methyl stearate (1.7 min); Methyl oleic acid (1.6 min), methyl linoleic acid (1.4 min); Cholesterol (13.4 min); Campesterin (14.1 min); Stigmaster (14.3 min); sitosterol (15.4 min); Sitostanol (15.6 min); Cholesteryl myristic acid ester (19.7 min); oleic acid campesteryl ester (20.74 min); Stigmasteryl oleic acid ester (20.77 min); Oleic acid sitosterylester (20.95 min); Oleic acid sitostanyl ester (21.0 min); Linoleic acid sitostanyl ester (20.95 min); triolein (22.3).

The gas chromatograms are made using a Hewlett-Packard GC ChemStation program evaluated and from the proportions of free sterols to fatty acid sterylesters the sales determined, the sum of the proportions of free sterols and fatty acid sterylesters as 100% Is accepted.

Example 3

Educt: 1 g steam distillate from conventional deodorization of rapeseed oil
Lipase: Candida rugosa lipase, immobilized, 50 or 100 mg
Temperature: 40 ° C
Pressure: 50 mbar
Duration: 24 hours
Analysis: The analysis of the product is carried out by GC as described in Example 2.
Conversion (GC): 87 or 93% fatty acid sterylester (based on the total free phyto sterols; see Table 2)
Purity (GC): ≧ 90%

Working up the reaction mixture

The workup of the reaction mixture is described below as an example in the case of lipase-catalyzed implementation of a conventional deodorization of rapeseed oil resulting steam distillate is obtained.

deacidification

Proportions of excess free fatty acids are removed from the reaction mixture removed by deacidification with dilute aqueous sodium carbonate solution. This will be Reaction mixture described above from the lipase-catalyzed synthesis of fatty acid extracted twice with 10 mL diethyl ether and the lipase catalyst in each case Centrifugation separated. Free fatty acids are extracted from the ether solution three times Extraction with 6 mL 5% aqueous sodium carbonate solution and subsequent washing removed with 10 mL water. The ether phase is dried over anhydrous sodium sulfate, evaporated and the residue (350 mg) dried at 50 ° C in a vacuum. An aliquot will analyzed by gas chromatography as described in Example 2.

Solvent fractionation

The deacidified reaction mixture (310 mg) is added Warm to 50 ° C dissolved in 2 mL anhydrous acetone and dropwise while stirring with 0.2 mL distilled water added. After cooling to + 7 ° C, the acetone-water The mixture was centrifuged and the supernatant removed. The lower pasty phase, which the contains enriched fatty acid sterylester, is dissolved again in 2 mL acetone and that Repeated procedure. After three repetitions, 90 mg of fatty acid steryl ester are obtained (Purity 89%) and after column chromatography (analogous to that described below) 85 mg Fatty acid sterylester (purity 93%), whose fatty acid composition is described in Table 3 is. The combined acetone / water mixtures are evaporated, and later Solvent fractionations reused.

column

The deacidified reaction mixture (350 mg) is iso- Hexane / diethyl ether (9: 1, v / v) dissolved and packed on a short column (20 × 1 cm ID) Silica gel (Kieselgel 60, Merck) adsorbed in iso-hexane. Elution with 30 mL iso- Hexane / diethyl ether (95: 5, v / v) gives 130 mg of fatty acid sterylester (purity 67%).  

Table 2

Proportions of phytosterols and fatty acid sterylesters before and after the esterification and / or transesterification of fatty acids, fatty acid esters and phytosterols in steam distillates of vegetable oils under the catalysis of Candida rugosa lipase in vacuo ^a

Table 3

Fatty acid pattern of fatty acid sterylesters produced by lipase-catalyzed esterification and / or transesterification of fatty acids or fatty acid esters with phytosterols in the steam distillate of conventionally deodorized rapeseed oil compared to the fatty acid pattern of the total lipids of the steam distillate

Example 4

Educt: 1 g steam distillate from the physical refining of rapeseed oil
Lipase: Candida rugosa lipase, immobilized, 50 mg
Temperature: 40 ° C
Pressure: 50 mbar
Duration: 18 hours

The product is worked up as in Example 3, the analysis by GC as described in Example 2.
Conversion (GC): 73% fatty acid sterylester (based on the total free phytosterols, see Table 2)

Example 5

Educt: 1 g steam distillate mixture from the physical refining of rapeseed oil and soybean oil
Lipase: Candida rugosa lipase, immobilized, 50 mg
Temperature: 40 ° C
Pressure: 30 mbar
Duration: 18 hours

The product is worked up as in Example 3, the analysis by GC as described in Example 2.
Conversion (GC): 93% fatty acid sterylester (based on the total free phytosterols; see Table 2)

Example 6 Bleaching of the steam distillate

10 g of steam distillate are mixed intensively with 1 g of Tonsil Optimum NFF bleaching earth and heated to 100 ° C under a nitrogen atmosphere over a period of 30 minutes. After centrifugation and decanting of the still hot product, bleached steam distillate (8.5 g) is obtained, which is used for the lipase-catalyzed esterification and / or transesterification:
Educt: 1 g bleached steam distillate from conventional deodorization of rapeseed oil
Lipase: Candida rugosa lipase, immobilized, 50 mg
Temperature: 40 ° C
Pressure: 20 mbar
Duration: 24 hours
Analysis: The analysis of the product is carried out by GC as described in Example 2.
Sales (GC): 85% fatty acid sterylester (based on the total free phytosterols)

Example 7

Educt: 0.5 g of crude tall oil
Lipase: Candida rugosa lipase, immobilized, 100 mg
Temperature: 40 ° C
Pressure: 30 mbar
Duration: 24 hours

Table 4

Proportions of phytosterols and fatty acid sterylesters before and after the esterification and / or transesterification of fatty acids, fatty acid esters with phytosterols in crude tall oil with catalysis of various lipases in vacuo ^a

The product is worked up as in Example 3, the analysis by GC as described in Example 2.
Conversion (GC): 59% fatty acid sterylester (based on the total free phytosterols, see Table 4)

Example 8

Educt: 0.5 g of crude tall oil
Lipase: Candida antarctica Lipase B, immobilized (Novozym 435®) 100 mg
Temperature: 80 ° C
Pressure: 30 mbar
Duration: 24 hours

The product is worked up as in Example 3, the analysis by GC as described in Example 2.
Conversion (GC): 34% fatty acid sterylester (based on the total free phytosterols, see Table 4)

Example 9

Educt: 0.5 g of crude tall oil
Lipase: Rhizomucor miehei lipase, immobilized (Lipozyme IM®), 100 mg
Temperature: 80 ° C
Pressure: 20 mbar
Duration: 24 hours

The product is worked up as in Example 3, the analysis by GC as described in Example 2.
Conversion (GC): 12% fatty acid sterylester (based on the total free phytosterols, see Table 4)

Example 10

The bleaching of crude tall oil was carried out as described in Example 6 for the damper distillate.
Educt: 0.5 g bleached raw tall oil
Lipase: Candida rugosa lipase, immobilized, 100 mg
Temperature: 40 ° C
Pressure: 30 mbar
Duration: 24 hours

The product is worked up as in Example 3, the analysis by GC as described in Example 2.
Sales (GC): 55% fatty acid sterylester (based on the total free phytosterols)

Claims (5)

1. Enzymatic process for the synthesis of fatty acid sterylesters, being in untreated Damper distillates from fat refining (vapor oils) occurring fatty acids, fatty acid esters and sterols react with stirring in the presence of lipases and resulting Water of reaction and volatile by-products are removed by distillation in vacuo. The Fatty acid sterylesters are formed by deacidification with aqueous solutions inorganic or organic bases and / or solvent fractionation with acetone, Methanol, ethanol or other polar aprotic solvents and their aqueous ones Solutions enriched and cleaned. 2. The method according to claim 1, wherein the solvent fractionation with petroleum ether, Hexane, iso-hexane or other apolar solvents and polar aprotic Solvents and their aqueous solutions is carried out. 3. The method according to claim 1, which involves the deacidification process column chromatographic purification process connects. 4. The method according to claims 1 to 3, in which tall oils instead of steam distillates Fat refining can be used to obtain fatty acid sterylesters. 5. The method according to claims 1 to 4, in which fractions of steamer distillates Fat refining or tall oils instead of untreated raw products for the extraction of Fatty acid sterylesters are used.