WO2001053511A1 - Process for preparing esters - Google Patents

Process for preparing esters Download PDF

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Publication number
WO2001053511A1
WO2001053511A1 PCT/DK2001/000039 DK0100039W WO0153511A1 WO 2001053511 A1 WO2001053511 A1 WO 2001053511A1 DK 0100039 W DK0100039 W DK 0100039W WO 0153511 A1 WO0153511 A1 WO 0153511A1
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WO
WIPO (PCT)
Prior art keywords
lipase
reaction
ester
carboxylic acid
mixture
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Application number
PCT/DK2001/000039
Other languages
French (fr)
Inventor
Kim Borch
Morten Würtz CHRISTENSEN
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Novozymes A/S
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Publication date
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Priority to AU2001226654A priority Critical patent/AU2001226654A1/en
Publication of WO2001053511A1 publication Critical patent/WO2001053511A1/en

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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P33/00—Preparation of steroids
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P7/00—Preparation of oxygen-containing organic compounds
    • C12P7/62—Carboxylic acid esters

Definitions

  • the present invention relates to a process for preparing a carboxylic acid ester of a sterol or a stanol.
  • Sterol and stanol esters are known to be useful, e.g. as ingredients in fat blends such as margarine. It is known to prepare such esters from a fatty acid together with a sterol or a stanol by a chemical process.
  • US 5,219,733 discloses a process for reacting a sterol and a fatty acid or fatty acid ester in contact with a lipase or cholesterol esterase, e.g. a lipase derived from Candida cylindracea.
  • a cholesterol esterase (EC 3.1.1.13) is an enzyme that can hydrolyze a steryl ester
  • a lipase (EC 3.1.1.3) is an enzyme that can hydrolyze thacylglycerol. It is known that some upases also have cholesterol esterase activity, but most do not.
  • WO 88/02775 discloses a lipase preparation from Candida antarctica and its separation into two distinct lipases (denoted lipase A and lipase B). It states that C. antarctica lipase is effective for ester synthesis, either as unseparated lipase preparation or as purified lipase A or B.
  • the invention provides a process for preparing a carboxylic acid ester comprising reacting a sterol or stanol with a carboxylic acid or a carboxylic acid ester in the presence of Candida antarctica lipase A or a variant thereof.
  • the process of the invention may be used to esterify any sterol or stanol of interest, e.g. of animal or plant origin.
  • sterol or stanol of interest e.g. of animal or plant origin.
  • examples are cholesterol, coprastanol, ergosterol, lanosterol, zymosterol, ⁇ -sitosterol, campesterol, campestanol, stigmasterol, ⁇ -sitostanol, and brassicasterol.
  • the process of the invention may be used to esterify the sterol or stanol with any carboxylic acid or ester thereof, including saturated straight-chain carboxylic acids, saturated branched carboxylic acids, mono- and poly-unsaturated fatty acids, hydroxy carboxylic acids and polycarboxylic acids.
  • the carboxylic acid may have 4-32 carbon atoms, e.g. 12-22.
  • the carboxylic acids may be a fatty acid found in natural fats and oils. Specific examples are lauric acid, palmitic acid, stearic acid, oleic acid, linolic acid, linolenic acid, conjugated linolenic acid (CLA), eicosapentaenoic acid (EPA), docosahexaenoic acid (DHA).
  • the carboxylic acid reactant may be a free carboxylic acid or an ester thereof, e.g. a methyl ester, an ethyl ester, a vinyl ester or a mono-, di- or th-glyceride. A mixture of such reactants may be used, e.g. a mixture of a triglyceride and free fatty acids.
  • Candida antarctica lipase A may be prepared as described in WO 88/02775.
  • variants thereof may be prepared by conventional methods introducing one or more alterations to the amino acid sequence; the alterations may be substitution, deletion or insertion of one or more amino acid residues. Examples of such variants and their preparation are described in WO 94/01541.
  • the lipase may optionally be used in immobilized form.
  • the immobilization may be done, e.g., by adsorption on a carrier such as a polymer-based carrier (e.g. polypropylene), an ion exchanger (e.g. a weakly basic anion exchanger) or silica, e.g. as described in EP 140542, WO 88/02775, WO 89/02916, WO 90/05778, WO
  • a carrier such as a polymer-based carrier (e.g. polypropylene), an ion exchanger (e.g. a weakly basic anion exchanger) or silica, e.g. as described in EP 140542, WO 88/02775, WO 89/02916, WO 90/05778, WO
  • the amount of lipase is typically in the range 10-1000 KLU/g sterol or stanol, e.g. 50-500 KLU/g.
  • the process will typically be performed so as to convert the stanol or sterol into the corresponding ester at a conversion of at least 50 %, e.g. at least 80 %, particularly at least 90 % conversion.
  • the molar ratio of the carboxylic acid reactant to the sterol or stanol reactant will typically be greater than 1 (i.e. a molar excess of the fatty acid reactant), e.g. a ratio of 2-20, 5-10, 2-10, 5-20, 1 -4 or 1 -5.
  • 1 i.e. a molar excess of the fatty acid reactant
  • a ratio of 2-20, 5-10, 2-10, 5-20, 1 -4 or 1 -5 For the purpose of this calculation, only one fatty acid of a di- or triglyceride is assumed to react.
  • the reaction may conveniently be carried out at a temperature of 20-90°C, e.g. 50-90°C, particularly 60-80°C. This is advantageous because most reactants of interest are liquid at such temperatures.
  • the reaction time will typically be in the range of 20 minutes to 12 hours, e.g. 1 -4 hours.
  • the reaction may be carried out with or without an organic solvent, e.g. a liquid hydrocarbon such as n-heptane.
  • an organic solvent e.g. a liquid hydrocarbon such as n-heptane.
  • the solvent is generally not necessary provided the reactants are liquid at the process temperature, thus avoiding the need for subsequent separation of the solvent. If a solvent is used, one or both reactants may optionally be dissolved in the solvent.
  • water is generally not required, but water may optionally be added to the lipase or to the reactant mixture in any desired amount.
  • the reaction may be performed batch-wise or continuously.
  • a batch reaction may conducted be by mixing the reactants and the lipase (immobilized or not) with stirring, and holding for the required time, preferably with stirring.
  • a continuous process may be conducted by loading immobilized lipase into a column and letting a mixture of the reactants flow through the column.
  • reaction may be performed in analogy with WO 88/02775 or US 5,219,733.
  • ester formed by the reaction may be recovered and purified by separation from the reaction mixture and purification by conventional methods.
  • the reaction may be performed with a triglyceride reactant, and the resulting reaction mixture may be used directly without separation as an ingredient in a fat blend for cooking, such as margarine, shortening or salad dressing.
  • the fat blend may be prepared as described in WO 92/19640, WO 98/19556, WO 99/56558, WO 99/15546 (Raisio), WO 97/42830, WO 98/01126, EP 897971 (Unilever), WO 99/30569 (Henkel)
  • a substrate for lipase is prepared by emulsifying tributyrin (glycerin tributyrate) using gum Arabic as emulsifier.
  • tributyrin glycol tributyrate
  • gum Arabic gum Arabic
  • the products was analysed using a 400 MHz Varian Spectrometer.
  • the sterol ester was analysed using a YMC PVA-Sil (100 * 3,0 id, 5 ⁇ m) and a light scattering detector (Sedex 55), drift tube temperature 60°C, nitrogen pressure 2.3 bar.
  • Palmitic acid solution in n-heptane (0.04 M; 750 ⁇ l) was mixed with a cholesterol solution in n-heptane (0.04 M; 750 ⁇ l).
  • the mixture was added to 100 mg of an immobilised preparation of Candida antarctica lipase A.
  • the reaction mixture was heated to 50°C and stirred with a magnet stirrer. Sample from the mixture (50 ⁇ l) was withdrawn after 120 minutes reaction time and analysed by HPLC.
  • Palmitic acid solution in n-heptane (0.04 M; 750 ⁇ l) was mixed with a ⁇ - sitosterol solution in n-heptane (0.04 M; 750 ⁇ l).
  • the mixture was added to 100 mg of an immobilised preparation of Candida antarctica lipase A.
  • the reaction mixture was heated to 70°C and stirred with a magnet stirrer. Sample from the mixture (50 ⁇ l) was withdrawn after 120 minutes reaction time and analysed by 1 H-NMR.
  • Oleic acid solution in n-heptane (0.04 M; 1.50 ml) was mixed with a ⁇ - sitosterol solution in n-heptane (0.04 M; 1.50 ml).
  • the mixture was added to 250 mg of an immobilised preparation of Candida antarctica lipase A.
  • the reaction mixture was heated to 70°C and stirred with a magnet stirrer. Sample from the mixture (50 ⁇ l) was withdrawn after 120 minutes reaction time and analysed by 1 H-NMR.
  • Triolein solution in n-heptane (0.04 M; 1.50 ml) was mixed with a ⁇ -sitosterol solution in n-heptane (0.04 M; 1.50 ml).
  • the mixture was added to 250 mg of an immobilised preparation of Candida antarctica lipase A.
  • the reaction mixture was heated to 70°C and stirred with a magnet stirrer. Sample from the mixture (50 ⁇ l) was withdrawn after 120 minutes reaction time and analysed by H-NMR.
  • Palmitic acid solution in n-heptane (0.04 M; 1.50 ml) was mixed with a ⁇ - sitosterol solution in n-heptane (0.04 M; 750 ⁇ l).
  • the mixture was added to 100 mg of an immobilised preparation of Rhizomucor miehei lipase.
  • the reaction mixture was heated to 70°C. Sample from the mixture (50 ⁇ l) was withdrawn after 120 minutes reaction time and analysed by 1 H-NMR.
  • Triolein (3.00 g, 3.38 mmol) was added to cholesterol (0.300 g, 0.776 mmol).
  • 250 mg of an immobilised preparation of Candida antarctica lipase A was added and the mixture was heated to 70°C under stirring using a magnet stirrer. Sample was withdrawn after 120 minutes reaction time and analysed by 1 H-NMR. The conversion into cholesterol oleate was found to be 100 %.

Abstract

Candida antarctica lipase A is effective for esterifying sterols with carboxylic acids, whereas C. antarctica lipase B and several other lipases are inactive. The invention provides a process for preparing a carboxylic acid ester comprising reacting a sterol or stanol with a carboxylic acid or a carboxylic acid ester in the presence of Candida antarctica lipase A or a variant thereof.

Description

PROCESS FOR PREPARING ESTERS
FIELD OF THE INVENTION
The present invention relates to a process for preparing a carboxylic acid ester of a sterol or a stanol.
BACKGROUND OF THE INVENTION
Sterol and stanol esters are known to be useful, e.g. as ingredients in fat blends such as margarine. It is known to prepare such esters from a fatty acid together with a sterol or a stanol by a chemical process.
US 5,219,733 (Yoshikawa Oil & Fat) discloses a process for reacting a sterol and a fatty acid or fatty acid ester in contact with a lipase or cholesterol esterase, e.g. a lipase derived from Candida cylindracea.
A cholesterol esterase (EC 3.1.1.13) is an enzyme that can hydrolyze a steryl ester, and a lipase (EC 3.1.1.3) is an enzyme that can hydrolyze thacylglycerol. It is known that some upases also have cholesterol esterase activity, but most do not. WO 88/02775 discloses a lipase preparation from Candida antarctica and its separation into two distinct lipases (denoted lipase A and lipase B). It states that C. antarctica lipase is effective for ester synthesis, either as unseparated lipase preparation or as purified lipase A or B.
K. Myojo and Y. Matsufune, Yukagaku 44(10), 883-96 (1995) states that a lipase from Candida antarctica is active for sterol fatty acid ester synthesis.
T. Sugai et al., Biosci. Biotechnol. Biochem., 60, 2059-2063 (1996) discloses the use of C. antarctica lipase B for the preparation of long-chain fatty acid esters of methyl cholate. E.M. Anderson et al., Biocatalysis and Biotransformation, vol. 16, pp.
181-204 (1998) gives a review of the use of C. antarctica lipase B in organic synthesis.
SUMMARY OF THE INVENTION
After testing a number of lipases, the inventors found that Candida antarctica lipase A is effective for esterifying sterols with carboxylic acids, whereas C. antarctica lipase B and several other lipases are inactive. Accordingly, the invention provides a process for preparing a carboxylic acid ester comprising reacting a sterol or stanol with a carboxylic acid or a carboxylic acid ester in the presence of Candida antarctica lipase A or a variant thereof.
DETAILED DESCRIPTION OF THE INVENTION
Sterol or stanol
The process of the invention may be used to esterify any sterol or stanol of interest, e.g. of animal or plant origin. Examples are cholesterol, coprastanol, ergosterol, lanosterol, zymosterol, β-sitosterol, campesterol, campestanol, stigmasterol, β-sitostanol, and brassicasterol.
Carboxylic acid reactant
The process of the invention may be used to esterify the sterol or stanol with any carboxylic acid or ester thereof, including saturated straight-chain carboxylic acids, saturated branched carboxylic acids, mono- and poly-unsaturated fatty acids, hydroxy carboxylic acids and polycarboxylic acids. The carboxylic acid may have 4-32 carbon atoms, e.g. 12-22.
The carboxylic acids may be a fatty acid found in natural fats and oils. Specific examples are lauric acid, palmitic acid, stearic acid, oleic acid, linolic acid, linolenic acid, conjugated linolenic acid (CLA), eicosapentaenoic acid (EPA), docosahexaenoic acid (DHA). The carboxylic acid reactant may be a free carboxylic acid or an ester thereof, e.g. a methyl ester, an ethyl ester, a vinyl ester or a mono-, di- or th-glyceride. A mixture of such reactants may be used, e.g. a mixture of a triglyceride and free fatty acids.
Lipase Candida antarctica lipase A may be prepared as described in WO 88/02775.
Its amino acid sequence is described in WO 94/01541. Variants thereof may be prepared by conventional methods introducing one or more alterations to the amino acid sequence; the alterations may be substitution, deletion or insertion of one or more amino acid residues. Examples of such variants and their preparation are described in WO 94/01541.
The lipase may optionally be used in immobilized form. The immobilization may be done, e.g., by adsorption on a carrier such as a polymer-based carrier (e.g. polypropylene), an ion exchanger (e.g. a weakly basic anion exchanger) or silica, e.g. as described in EP 140542, WO 88/02775, WO 89/02916, WO 90/05778, WO
90/15868, WO 95/22606, or WO 99/33964.
The amount of lipase is typically in the range 10-1000 KLU/g sterol or stanol, e.g. 50-500 KLU/g.
Process conditions
The process will typically be performed so as to convert the stanol or sterol into the corresponding ester at a conversion of at least 50 %, e.g. at least 80 %, particularly at least 90 % conversion.
The molar ratio of the carboxylic acid reactant to the sterol or stanol reactant will typically be greater than 1 (i.e. a molar excess of the fatty acid reactant), e.g. a ratio of 2-20, 5-10, 2-10, 5-20, 1 -4 or 1 -5. For the purpose of this calculation, only one fatty acid of a di- or triglyceride is assumed to react.
The reaction may conveniently be carried out at a temperature of 20-90°C, e.g. 50-90°C, particularly 60-80°C. This is advantageous because most reactants of interest are liquid at such temperatures. The reaction time will typically be in the range of 20 minutes to 12 hours, e.g. 1 -4 hours.
The reaction may be carried out with or without an organic solvent, e.g. a liquid hydrocarbon such as n-heptane. Advantageously, the solvent is generally not necessary provided the reactants are liquid at the process temperature, thus avoiding the need for subsequent separation of the solvent. If a solvent is used, one or both reactants may optionally be dissolved in the solvent.
The addition of water is generally not required, but water may optionally be added to the lipase or to the reactant mixture in any desired amount.
The reaction may be performed batch-wise or continuously. A batch reaction may conducted be by mixing the reactants and the lipase (immobilized or not) with stirring, and holding for the required time, preferably with stirring. A continuous process may be conducted by loading immobilized lipase into a column and letting a mixture of the reactants flow through the column.
The reaction may be performed in analogy with WO 88/02775 or US 5,219,733.
Use of reaction product
If desired, the ester formed by the reaction may be recovered and purified by separation from the reaction mixture and purification by conventional methods.
Alternatively, the reaction may be performed with a triglyceride reactant, and the resulting reaction mixture may be used directly without separation as an ingredient in a fat blend for cooking, such as margarine, shortening or salad dressing. The fat blend may be prepared as described in WO 92/19640, WO 98/19556, WO 99/56558, WO 99/15546 (Raisio), WO 97/42830, WO 98/01126, EP 897971 (Unilever), WO 99/30569 (Henkel)
METHODS
Lipase activity (LU)
A substrate for lipase is prepared by emulsifying tributyrin (glycerin tributyrate) using gum Arabic as emulsifier. The hydrolysis of tributyrin at 30 °C at pH
7 is followed in a pH-stat titration experiment. One unit of lipase activity (1 LU) equals the amount of enzyme capable of releasing 1 μmol butyric acid/min at the standard conditions. 1 KLU = 1000 LU.
1H-NMR:
The products was analysed using a 400 MHz Varian Spectrometer.
HPLC analysis:
The sterol ester was analysed using a YMC PVA-Sil (100 * 3,0 id, 5 μm) and a light scattering detector (Sedex 55), drift tube temperature 60°C, nitrogen pressure 2.3 bar.
20 μl of diluted sample was injected via autosampler. A calibration curve of commercially available cholesterol palmitate was used for the quantification of the product.
A tertiary gradient was used with the following mobile phases:
Solvent A:
Isooctane 980 ml
Methyltertbutylether 20 ml
Solvent B
2-propanol 700 ml
Acetonit le 200 ml
2-butanone 100 ml
Solvent C
2-propanol 560 ml
Acetonithle 140 ml
2-butanone 72 ml
Methanol 140 ml
Water 84 ml
N-ethylmorpholine 4,2 ml
Acetic acid 1 ,5 ml
Gradient:
Figure imgf000006_0001
EXAMPLES
Example 1.
Palmitic acid solution in n-heptane (0.04 M; 750 μl) was mixed with a cholesterol solution in n-heptane (0.04 M; 750 μl). The mixture was added to 100 mg of an immobilised preparation of Candida antarctica lipase A. The reaction mixture was heated to 50°C and stirred with a magnet stirrer. Sample from the mixture (50 μl) was withdrawn after 120 minutes reaction time and analysed by HPLC.
Two other lipases in immobilized form were tested for comparison. All lipases were immobilised onto a porous polypropylene carrier (Accurel EP 100, from Akzo) by simple adsorption. The amount of lipase adsorbed to the carrier is given in KLU (kilo lipase units). The conversion into cholesterol palmitate is also given in the table below. The enzyme load indicates the amount of lipase removed from the liquid during immobilization (in KLU defined above) divided by the weight of the carrier.
Figure imgf000007_0001
Example 2.
Palmitic acid solution in n-heptane (0.04 M; 750 μl) was mixed with a β- sitosterol solution in n-heptane (0.04 M; 750 μl). The mixture was added to 100 mg of an immobilised preparation of Candida antarctica lipase A. The reaction mixture was heated to 70°C and stirred with a magnet stirrer. Sample from the mixture (50 μl) was withdrawn after 120 minutes reaction time and analysed by 1H-NMR.
The conversion into β-sitosterol palmitate was found to be 45 %. Example 3.
Oleic acid solution in n-heptane (0.04 M; 1.50 ml) was mixed with a β- sitosterol solution in n-heptane (0.04 M; 1.50 ml). The mixture was added to 250 mg of an immobilised preparation of Candida antarctica lipase A. The reaction mixture was heated to 70°C and stirred with a magnet stirrer. Sample from the mixture (50 μl) was withdrawn after 120 minutes reaction time and analysed by 1H-NMR.
The conversion into β-sitosterol oleate was found to be 26 %.
Example 4.
Triolein solution in n-heptane (0.04 M; 1.50 ml) was mixed with a β-sitosterol solution in n-heptane (0.04 M; 1.50 ml). The mixture was added to 250 mg of an immobilised preparation of Candida antarctica lipase A. The reaction mixture was heated to 70°C and stirred with a magnet stirrer. Sample from the mixture (50 μl) was withdrawn after 120 minutes reaction time and analysed by H-NMR.
The conversion into β-sitosterol oleate was found to be 57 %.
Example 5 (reference)
Palmitic acid solution in n-heptane (0.04 M; 1.50 ml) was mixed with a β- sitosterol solution in n-heptane (0.04 M; 750 μl). The mixture was added to 100 mg of an immobilised preparation of Rhizomucor miehei lipase. The reaction mixture was heated to 70°C. Sample from the mixture (50 μl) was withdrawn after 120 minutes reaction time and analysed by 1H-NMR.
The conversion into β-sitosterol palmitate was found to be 0 %.
Example 6.
Triolein (3.00 g, 3.38 mmol) was added to cholesterol (0.300 g, 0.776 mmol). 250 mg of an immobilised preparation of Candida antarctica lipase A was added and the mixture was heated to 70°C under stirring using a magnet stirrer. Sample was withdrawn after 120 minutes reaction time and analysed by 1H-NMR. The conversion into cholesterol oleate was found to be 100 %.

Claims

1. A process for preparing an ester comprising reacting: a) a sterol or stanol with b) a carboxylic acid or an ester thereof in the presence of Candida antarctica lipase A or a variant thereof.
2. The process of the preceding claim wherein reactant b) is present in a greater molar amount than reactant a), e.g. such that the molar ratio of b) to a) is 2-20, particularly 5-10.
3. The process of either preceding claim wherein the lipase is in immobilized form.
4. The process of any preceding claim wherein the reaction is carried out in the absence of an organic solvent.
5. The process of any preceding claim wherein the reaction is carried out without addition of water.
6. The process of any preceding claim wherein the reaction is carried out at a temperature of 60-80°C.
7. The process of any preceding claim wherein the carboxylic acid ester is a triglyceride.
8. The process of the preceding claim wherein the mixture resulting from the reaction is used without separation as a component for making a fat blend for cooking.
PCT/DK2001/000039 2000-01-21 2001-01-19 Process for preparing esters WO2001053511A1 (en)

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Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP1285968A2 (en) * 2001-08-22 2003-02-26 Härting, Thomas Francis Selective transesterification of stanols in mixtures comprising sterols and stanols
EP1285969A2 (en) * 2001-08-22 2003-02-26 Härting, Thomas Francis Process for producing sterol or stanol esters by enzymatic transesterification in solvent and water-free media
WO2008032007A1 (en) 2006-09-14 2008-03-20 Tmo Renewables Ltd Lipase
EP2886657A1 (en) 2013-12-19 2015-06-24 ETH Zurich Enzymatic synthesis of phenolic acid esters and steryl phenolates

Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1988002775A1 (en) * 1986-10-17 1988-04-21 Novo Industri A/S Positionally non-specific lipase from candida sp, a method for producing it, its use and a recombinant dna process for producing it
WO1990015868A1 (en) * 1989-06-21 1990-12-27 Novo Nordisk A/S Immobilized lipase preparation and use thereof for ester synthesis
US5658769A (en) * 1993-07-02 1997-08-19 Unichem Chemie Bv Process for the esterification of carboxylic acids with tertiary alcohols using a lipase from Candida antarctica

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1988002775A1 (en) * 1986-10-17 1988-04-21 Novo Industri A/S Positionally non-specific lipase from candida sp, a method for producing it, its use and a recombinant dna process for producing it
WO1990015868A1 (en) * 1989-06-21 1990-12-27 Novo Nordisk A/S Immobilized lipase preparation and use thereof for ester synthesis
US5658769A (en) * 1993-07-02 1997-08-19 Unichem Chemie Bv Process for the esterification of carboxylic acids with tertiary alcohols using a lipase from Candida antarctica

Cited By (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP1285968A2 (en) * 2001-08-22 2003-02-26 Härting, Thomas Francis Selective transesterification of stanols in mixtures comprising sterols and stanols
EP1285969A2 (en) * 2001-08-22 2003-02-26 Härting, Thomas Francis Process for producing sterol or stanol esters by enzymatic transesterification in solvent and water-free media
EP1285969A3 (en) * 2001-08-22 2003-07-23 Härting, Thomas Francis Process for producing sterol or stanol esters by enzymatic transesterification in solvent and water-free media
EP1285968A3 (en) * 2001-08-22 2003-07-23 Härting, Thomas Francis Selective transesterification of stanols in mixtures comprising sterols and stanols
US7078544B2 (en) * 2001-08-22 2006-07-18 Harting, S.A. Process for producing sterol or stanol esters by enzymatic transesterification in solvent and water free media
WO2008032007A1 (en) 2006-09-14 2008-03-20 Tmo Renewables Ltd Lipase
EP2886657A1 (en) 2013-12-19 2015-06-24 ETH Zurich Enzymatic synthesis of phenolic acid esters and steryl phenolates

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