+

WO1991000918A1 - PROCEDE DE PREPARATION D'UN PHOSPHOLIPIDE A L'AIDE D'UN RESIDU D'ACIDE CARBOXYLIQUE EN POSITION 2, ET D'UN PHOSPHOLIPIDE A L'AIDE D'UN RESIDU D'ACIDE GRAS φ-3 EN POSITION 2 - Google Patents

PROCEDE DE PREPARATION D'UN PHOSPHOLIPIDE A L'AIDE D'UN RESIDU D'ACIDE CARBOXYLIQUE EN POSITION 2, ET D'UN PHOSPHOLIPIDE A L'AIDE D'UN RESIDU D'ACIDE GRAS φ-3 EN POSITION 2 Download PDF

Info

Publication number
WO1991000918A1
WO1991000918A1 PCT/SE1990/000481 SE9000481W WO9100918A1 WO 1991000918 A1 WO1991000918 A1 WO 1991000918A1 SE 9000481 W SE9000481 W SE 9000481W WO 9100918 A1 WO9100918 A1 WO 9100918A1
Authority
WO
WIPO (PCT)
Prior art keywords
phospholipid
fatty acid
weight
lysophospholipid
carboxylic acid
Prior art date
Application number
PCT/SE1990/000481
Other languages
English (en)
Inventor
Bo Ekstrand
Caj Eriksson
Krister Holmberg
Eva Österberg
Original Assignee
Berol Nobel Ab
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Berol Nobel Ab filed Critical Berol Nobel Ab
Publication of WO1991000918A1 publication Critical patent/WO1991000918A1/fr

Links

Classifications

    • 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
    • C12P7/64—Fats; Fatty oils; Ester-type waxes; Higher fatty acids, i.e. having at least seven carbon atoms in an unbroken chain bound to a carboxyl group; Oxidised oils or fats
    • C12P7/6436—Fatty acid esters
    • C12P7/6445—Glycerides
    • C12P7/6481—Phosphoglycerides
    • 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
    • C12P9/00—Preparation of organic compounds containing a metal or atom other than H, N, C, O, S or halogen

Definitions

  • the present invention relates to a method for the preparation of a phospholipid with a selected carboxylic acid residue in the 2-position by esterification in a microemulsion with the aid of a specific enzyme, phospho- lipase A2.
  • the invention also concerns a phospholipid with an ⁇ -3-fatty acid residue in the 2-position.
  • hydrolytic enzymes have both high activity and high stability in microemulsions, i.e. thermodynamically stable solutions of a hydrophobic component, water, and a surface-active component.
  • thermodynamically stable solutions of a hydrophobic component, water, and a surface-active component In studies focussing on lipase, it has been found that a high water content in the system promotes hydrolysis, whereas a low water content, prefer ⁇ ably below 1% by weight, results in the opposite reaction, i.e condensation.
  • Lipase-catalysed esterification of gly- cerol and fatty acid in a microemulsion with low water content is described in literature (Fletcher et al., Bio- chim. Biophys. Acta 912(1987)278; Bello et al., Biochem. Biophys. Res. Comm., 146(1987)361).
  • triglycerides As to triglycerides, it has been found that a small addition of a triglyceride with a different fatty acid composition, e.g. containing an unusually short, an unusually long, a branched or an unusually unsaturated fatty acid, markedly affects such physico-chemical pro ⁇ perties as crystallisation and softening ranges, as well as lubricating and friction-reducing qualities. Further ⁇ more, it would seem that the nutritional properties are much altered already by minor changes in the fatty acid composition of the triglycerides.
  • a different fatty acid composition e.g. containing an unusually short, an unusually long, a branched or an unusually unsaturated fatty acid
  • ⁇ -3- fatty acids which is the generic term for polyunsaturated fatty acids which have 18-22 carbon atoms and whose last double bond, as counted from the carboxyl group, is between the third and the fourth carbon atom as counted from the methyl group end of the fatty acid molecule.
  • a connection has been shown between a high intake of ⁇ -3- fatty acids and a reduced frequency of heart and vascular diseases.
  • An augmented intake of ⁇ -3-fatty acids reduces the cholesterol content of the blood, and ⁇ -3-fatty acids are therefore often prescribed for people with blood counts indicating an increased risk of thrombosis and infarct of the heart.
  • the ⁇ -3-fatty acids are normally available not only in the form of triglycerides from e.g. cod-liver oil, but also in the form of free fatty acids usually extracted from fish oils.
  • the triglycerides are metabolised, and part of the fatty acids are incorporated in the cell membranes of the body, a main component of these membranes being phospholipids.
  • this incorporation is a slow process and only a minor amount of the added ⁇ -3-fatty acids is incorporated in the membranes, regardless of whether they originally had the form of triglycerides or free fatty acids. Therefore, there is a great need for products which contain ⁇ -3-fatty acids and can be taken up by the body in a more efficient manner.
  • phospholipase A2 With the aid of a specific enzyme, phospholipase A2, it has now proved to be feasible to esterify the 2-posi ⁇ tion of a lysophospholipid by adding a carboxylic acid. Normally, phospholipase A2 hydrolyses the ester bond of the phospholipid in the 2-position, but under the condi ⁇ tions prevalent during the inventive esterfication, the enzyme esterifies a lysophospholipid in the 2-position, surprisingly enough. This reaction takes place in a micro ⁇ emulsion. Since the phospholipid is surface-active in itself, it should theoretically be able to form by itself a microemulsion together with the hydrophobic component and the water.
  • the lysophospholipid is supplemented with at least one other surface-active compound, preferably an anionic ten- side.
  • the hydrophobic component consists of aliphatic hydrocarbons, but it may also be a supercritical solvent, e.g. carbon dioxide.
  • the water content is low, usually 0.1-2% by weight, preferably 0.5-2% by weight.
  • the added carboxylic acid which preferably is ali ⁇ phatic, may be straight or branched, saturated or unsatu- rated.
  • the number of carbon atoms of the fatty acid may vary within wide limits, but the range of 10-22 carbon atoms has attracted the greatest interest.
  • Suitable carboxylic acids include decanoic acid, dodecanoic acid, tetradecanoic acid, hexadecanoic acid, octadecanoic acid, oleic acid, ricinoleic acid, linoleic acid, linolenic acid, abietic acid, and dehydroabietic acid.
  • the ⁇ -3-fatty acids which above all are to be found in algae and fish oils, are especially preferred. The most common ⁇ -3-fatty acids, i.e.
  • EPA eicosapentaenoic acid
  • DHA docosahexaenoic acid
  • the phospholipids with ⁇ -3-fatty acid residues in the 2-position obtained according to the invention are extremely advantageous compared with conventional fatty acid and triglyceride products. Since phopholipids are a biologically active part of the cell membranes of the cellular body, no metabolism need take place in the body, which should increase the efficiency of the o-3-fatty acid. It is to b understood that the invention does not require pure products.
  • the lysophospholipids are mixtures of several different components, and most of them have a structure which resembles that of the diglycerides but in which a fatty acid group has been replaced by an organic group derived from phosphoric acid and a nitrogen base.
  • Phosphatidyl choline is usually a main component.
  • the added ⁇ -3-fatty acid is not pure, but consists of a mixture of different fatty acids, such as EPA and DHA, and further contains a fairly significant amount of fatty acids other than the ⁇ -3-type. Even if pure ⁇ -3- fatty acid were to be used in the inventive reaction, the incorporation in the phospholipid would not be complete, since the esterification is an equilibrium reaction. All in all, the ⁇ -3-fatty acid-containing phospholipid referred to in this context may consist of a large number of different substances. However, a distinctive feature is that a fairly significant proportion of the phospholipid, i.e. at least 10% and usually 15% or more, contains ⁇ -3- fatty acid in the 2-position.
  • the inventive reaction may be illustrated by the reaction formula of lysophosphatidyl choline.
  • R ⁇ is an acyclic hydrocarbon residue which contains 11-21 carbon atoms and is not being of ⁇ -3-type
  • R 2 is a polyunsaturated ⁇ -3-fatty alkyl group with 17-21 carbon atoms.
  • the esterification according to the present invention is normally carried out at a temperature of between 20°C and 60 C C.
  • the aliphatic hydrocarbons, such as isooctane and nonane, forming the hydrophobic component constitute 65-98% by weight of the composition.
  • Supercritical sol ⁇ vents, such as carbon dioxide, can be used as hydrophobic component instead of the aliphatic hydrocarbons, in which case the reaction takes place at an elevated pressure.
  • the water is buffered to pH 6-10.
  • both ionic and nonionic tensides may serve as surface-active component.
  • Suitable ionic tensides include sodium dioctyl sulphosuccinate and fatty acid soaps
  • suitable nonionic tensides include monoglyce- rides, sucrose fatty acid esters, sorbitan esters, and ethoxylated sorbitan esters. Frequently, mixtures of dif ⁇ ferent tensides are to be preferred when formulating microemulsions.
  • the amount of lysophospholipid and ⁇ -3-fatty acid added makes up 1-20% of the total com ⁇ position.
  • the surface-active components used should meet food standard requirements.
  • a suitable con ⁇ tent is 0.1-10% by weight of the total composition.
  • a conventional phospholipid first undergoes enzymatic hydrolysis to become a lysophos ⁇ pholipid, whereupon esterification as above is carried out.-
  • the net result of the two reaction steps is a trans- esterification, i.e. a replacement of an ordinary fatty acid residue in the 2-position by another carboxylic acid residue, e.g. an ⁇ -3-fatty acid residue.
  • the same enzyme, phospholipase A2 serves as catalyst in both the hydro- lysis step and the condensation step, and microemulsions are a suitable medium for both processes.
  • the first step the hydrolysis reaction
  • the hydrolysis reaction is suitably carried out in a microemulsion with a higher water content, e..g. 2-5%, than the second step, the esterification reaction, which may take place in a medium with a water content of 0.5-2%.
  • the two reaction steps may be combined to form a single process in which phospholipid and car ⁇ boxylic acid together with an enzyme are added to a micro ⁇ emulsion with the higher water content and in which the water content is gradually reduced by stripping under vacuum, or by adding a hydrophilic substance, e.g. zeo ⁇ lite.
  • the reaction is made to take place in a proctective atmosphere and in the presence of an anti- oxidant in order to avoid autoxidation of the polyunsa ⁇ turated fatty acids.
  • Suitable antioxidants include toco- pherol, butyl hydroxyanisole, butyl hydroxytoluene, and ascorbic acid. Combinations of at least one lipophilic and at least one hydrophilic antioxidant have at times proved advantageous.
  • Example 1 The invention will be illustrated in more detail by the following Examples.
  • Example 1 The invention will be illustrated in more detail by the following Examples.
  • Aqueous buffer pH 8.2 1.3
  • Example 2 The same composition as in Example 1 was used, except that the tenside was made up of a mixture of sorbitan mono stearate and polyoxyethylene (20) sorbitan monostearate, the molar ratio being 1:2.
  • Example 3 The same composition as in Example 1 was used, except that isooctane was replaced by n-heptane, and a phosphate buffer of pH 7.0 was substituted for the phosphate buffer of pH 8.5.
  • Example 4 The following composition was used:
  • Aqueous buffer pH 8.2 1.3
  • Example 5 The same composition as in Example 5 was used, except that dodecanoic acid was replaced by y-linolenic acid, and the contents of fatty acid and lysophosphatidyl choline were altered from 3.0% and 5.0%, respectively, to 4.0% for both.
  • Example 7 After a 16-hour reaction during which the same amount of enzyme was added and the same conditions prevailed as in Example 5, a phospholipid fraction, of which more than 90% contained 2C-linolenic acid residues, was obtained in a yield of 11%.
  • Example 7 After a 16-hour reaction during which the same amount of enzyme was added and the same conditions prevailed as in Example 5, a phospholipid fraction, of which more than 90% contained 2C-linolenic acid residues, was obtained in a yield of 11%.
  • Example 5 The same composition as in Example 5 was used, except that 11-methyl dodecanoic acid was substituted for dode- canoic acid.

Landscapes

  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Engineering & Computer Science (AREA)
  • Zoology (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Wood Science & Technology (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Microbiology (AREA)
  • General Chemical & Material Sciences (AREA)
  • Biotechnology (AREA)
  • Health & Medical Sciences (AREA)
  • Biochemistry (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • General Engineering & Computer Science (AREA)
  • General Health & Medical Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Oil, Petroleum & Natural Gas (AREA)
  • Preparation Of Compounds By Using Micro-Organisms (AREA)

Abstract

L'invention concerne des phospholipides comportant un résidu d'acide carboxylique voulu, tel qu'un résidu d'acide gras φ-3 en position 2. On produit ces composés par estérification d'un lysophospholipide classique à l'aide de l'acide carboxylique correspondant en présence de la phospholipase A2 de catalyse, l'estérification ayant lieu dans une microémulsion ayant une teneur en eau ne dépassant pas 0,1 à 2 % en poids.
PCT/SE1990/000481 1989-07-12 1990-07-04 PROCEDE DE PREPARATION D'UN PHOSPHOLIPIDE A L'AIDE D'UN RESIDU D'ACIDE CARBOXYLIQUE EN POSITION 2, ET D'UN PHOSPHOLIPIDE A L'AIDE D'UN RESIDU D'ACIDE GRAS φ-3 EN POSITION 2 WO1991000918A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
ATPCT/SE89/00409 1989-07-12
SE8900409 1989-07-12

Publications (1)

Publication Number Publication Date
WO1991000918A1 true WO1991000918A1 (fr) 1991-01-24

Family

ID=20374967

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/SE1990/000481 WO1991000918A1 (fr) 1989-07-12 1990-07-04 PROCEDE DE PREPARATION D'UN PHOSPHOLIPIDE A L'AIDE D'UN RESIDU D'ACIDE CARBOXYLIQUE EN POSITION 2, ET D'UN PHOSPHOLIPIDE A L'AIDE D'UN RESIDU D'ACIDE GRAS φ-3 EN POSITION 2

Country Status (1)

Country Link
WO (1) WO1991000918A1 (fr)

Cited By (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2002090560A3 (fr) * 2001-05-03 2004-02-19 Enzymotec Ltd Procede de production de phospholipides
WO2005017142A1 (fr) * 2003-08-14 2005-02-24 Cognis Ip Management Gmbh Utilisation d'emulsions pit dans des reactions enzymatiques
WO2005038037A3 (fr) * 2003-10-22 2005-12-01 Enzymotec Ltd Methodes de preparation de phospholipides contenant des fractions omega-3 et omega-6
EP2453020A4 (fr) * 2009-07-06 2013-06-05 Kaneka Corp Procédé de production d un phospholipide
CN104531790A (zh) * 2014-12-17 2015-04-22 中国科学院天津工业生物技术研究所 一种磷脂dha的制备方法

Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
SE452166B (sv) * 1986-03-10 1987-11-16 Berol Kemi Ab Forfarande for transesterifiering av triglycerider
FR2599382A1 (fr) * 1986-05-29 1987-12-04 Inst Corps Gras Iterg Procede de preparation de diglycerides contenant en position 2 un reste d'acide gras poly-insature, et produits obtenus
EP0265699A2 (fr) * 1986-10-01 1988-05-04 The Nisshin Oil Mills, Ltd. Graisses et huiles ayant une digestibilité et une absorptivité améliorées

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
SE452166B (sv) * 1986-03-10 1987-11-16 Berol Kemi Ab Forfarande for transesterifiering av triglycerider
FR2599382A1 (fr) * 1986-05-29 1987-12-04 Inst Corps Gras Iterg Procede de preparation de diglycerides contenant en position 2 un reste d'acide gras poly-insature, et produits obtenus
EP0265699A2 (fr) * 1986-10-01 1988-05-04 The Nisshin Oil Mills, Ltd. Graisses et huiles ayant une digestibilité et une absorptivité améliorées

Non-Patent Citations (4)

* Cited by examiner, † Cited by third party
Title
CHEMICAL ABSTRACTS, Vol. 69, (1968), Abstract No. 168k. H.P. FRANCK et al.; & Z. NATURFORSCH. B 1968, 23(4). 43948, (Ger.). *
CHEMICAL ABSTRACTS, Vol. 73, (1970), Abstract No. 84125n, H.P. FRANCK et al.; & Z. NATURFORSCH B 1970, 25(6), 581-6, (Ger.). *
CHEMICAL ABSTRACTS, Vol. 82 (1975), Abstract No. 53462b, RONALD L. MISIOROWSKI et al.; & BIOCHEMISTRY 1974, 13(24), 4921-7, (Eng.). *
CHEMICAL ABSTRACTS, Vol. 90, (1979), Abstract No. 50071c, R.W. EVANS et al.; & CHEM. PHYS. LIPIDS 1978, 22(3), 207-20, (Eng.). *

Cited By (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2002090560A3 (fr) * 2001-05-03 2004-02-19 Enzymotec Ltd Procede de production de phospholipides
US7034168B2 (en) 2001-05-03 2006-04-25 Enzymotec Ltd. Process for the production of phospholipids
WO2005017142A1 (fr) * 2003-08-14 2005-02-24 Cognis Ip Management Gmbh Utilisation d'emulsions pit dans des reactions enzymatiques
US8318468B2 (en) 2003-08-14 2012-11-27 Cognis Ip Management Gmbh Emulsions in enzymatic reactions
WO2005038037A3 (fr) * 2003-10-22 2005-12-01 Enzymotec Ltd Methodes de preparation de phospholipides contenant des fractions omega-3 et omega-6
EP2453020A4 (fr) * 2009-07-06 2013-06-05 Kaneka Corp Procédé de production d un phospholipide
CN104531790A (zh) * 2014-12-17 2015-04-22 中国科学院天津工业生物技术研究所 一种磷脂dha的制备方法

Similar Documents

Publication Publication Date Title
EP0749468B1 (fr) Raffinage de compositions d'huile
US5128251A (en) Immobilized lipolytic enzyme for esterification and interesterification
US5935828A (en) Enzymatic production of monoglycerides containing omega-3 unsaturated fatty acids
EP0307154B1 (fr) Préparation de diglycérides
JPH08291188A (ja) ホスフォリピドのエステル交換方法
US20030130533A1 (en) Conjugated fatty acid containing monoglycerides and process for producing them
US20020120159A1 (en) Process for the preparation of a monoglyceride
JP2749587B2 (ja) 固定化酵素の製造方法
WO1991000918A1 (fr) PROCEDE DE PREPARATION D'UN PHOSPHOLIPIDE A L'AIDE D'UN RESIDU D'ACIDE CARBOXYLIQUE EN POSITION 2, ET D'UN PHOSPHOLIPIDE A L'AIDE D'UN RESIDU D'ACIDE GRAS φ-3 EN POSITION 2
WO2003060139A1 (fr) Procede de production de diglycerides
JP4079516B2 (ja) トリグリセリドの製造方法
EP0349606B1 (fr) Procede d'obtention d'acide oleique de grande purete par hydrolyse d'huile de graines de tournesol
JP3813585B2 (ja) ジグリセリドの製造方法
KR100850646B1 (ko) 인지질 조성물 제조방법
JP3929890B2 (ja) ジグリセリドの製造方法
JP3853767B2 (ja) 共役脂肪酸メントールエステルおよびその製造方法
EP1285968B1 (fr) Transestérification sélective de stanoles dans des mélanges contenant des stérols et des stanols
JP2570774B2 (ja) 油脂の改質法
JP2004208546A (ja) 脂肪酸の製造方法
JP3764793B2 (ja) ジグリセリド類の製造方法
JPS63214184A (ja) 固定化酵素およびその製造方法
JP2886628B2 (ja) モノアシルグリセロリン脂質の製造方法
EP1978101A1 (fr) Procédé d'enrichissement en acides gras polyinsaturés
JP3527502B1 (ja) ステロール脂肪酸エステル含有組成物およびその製造方法
JP2004331607A (ja) 共役脂肪酸のポリグリセリンエステルおよびその製造方法

Legal Events

Date Code Title Description
AK Designated states

Kind code of ref document: A1

Designated state(s): CA JP US

AL Designated countries for regional patents

Kind code of ref document: A1

Designated state(s): AT BE CH DE DK ES FR GB IT LU NL SE

NENP Non-entry into the national phase

Ref country code: CA

点击 这是indexloc提供的php浏览器服务，不要输入任何密码和下载