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WO1989012648A1 - Procede d'amelioration des proprietes de gelification de pectine hautement esterifiee - Google Patents

Procede d'amelioration des proprietes de gelification de pectine hautement esterifiee Download PDF

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Publication number
WO1989012648A1
WO1989012648A1 PCT/DK1989/000159 DK8900159W WO8912648A1 WO 1989012648 A1 WO1989012648 A1 WO 1989012648A1 DK 8900159 W DK8900159 W DK 8900159W WO 8912648 A1 WO8912648 A1 WO 8912648A1
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WIPO (PCT)
Prior art keywords
pectin
enzyme
molecular weight
esterified
reaction
Prior art date
Application number
PCT/DK1989/000159
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English (en)
Inventor
Robin Rolin
Jørgen Robert SØDERBERG
Original Assignee
A/S Københavns Pektinfabrik
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 A/S Københavns Pektinfabrik filed Critical A/S Københavns Pektinfabrik
Publication of WO1989012648A1 publication Critical patent/WO1989012648A1/fr

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Classifications

    • C—CHEMISTRY; METALLURGY
    • C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
    • C08B—POLYSACCHARIDES; DERIVATIVES THEREOF
    • C08B37/00—Preparation of polysaccharides not provided for in groups C08B1/00 - C08B35/00; Derivatives thereof
    • C08B37/0006—Homoglycans, i.e. polysaccharides having a main chain consisting of one single sugar, e.g. colominic acid
    • C08B37/0045—Homoglycans, i.e. polysaccharides having a main chain consisting of one single sugar, e.g. colominic acid alpha-D-Galacturonans, e.g. methyl ester of (alpha-1,4)-linked D-galacturonic acid units, i.e. pectin, or hydrolysis product of methyl ester of alpha-1,4-linked D-galacturonic acid units, i.e. pectinic acid; Derivatives thereof
    • 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
    • C12P19/00—Preparation of compounds containing saccharide radicals
    • C12P19/04—Polysaccharides, i.e. compounds containing more than five saccharide radicals attached to each other by glycosidic bonds

Definitions

  • This invention relates to a process of improving the gelling properties of high-esterified pectin.
  • pectin such as pectin produced of citrus fruits or apples, essentially consists of polymeric chains formed by polymeri ⁇ sation of galacturonic acid moieties the carbon atom in the 4-position in one moiety being bound to the carbon atom in the 1-po- sition in the neighbouring moiety.
  • the carbon atoms in 6-position are esterified with methanol and if over 50% of said carbon atoms are esterified with methanol the pectin is designated high-esterified.
  • the pectin is called low-esterified if less than 50% of the carbon atoms are esterified with methanol.
  • Pectin normally forms a gel if either sugar is added to obtain a concentration of more than about 60 weight percent and the pH-value is adjusted to less than about 3.1 or if a calcium solution or calcium containing fruit is added, the calcium ions being capable of functioning as a bridge between the pectin chains.
  • the first method is primarily used for gelling high- esterified pectins while the second method primarily is used for gelling low-esterified pectins.
  • pectin may react with calcium when a pectin solution and fruit juices containing calcium ions are mixed prior to the initiation of the gelling process thereby increasing the viscosity of the mixture. Consequently, it is difficult to obtain a uniform distribution of pectin, and the consequence is that the subsequent gelling process does not result in a gel having an optimum strength.
  • pectin essentially consists of chains of galacturonic acid moeities and methyl-esterified galacturonic acid moeities.
  • the sequence in which esterified and non-esterified moieties are present in the chains may vary. Segments may occur in which there is a local predominance of non-esterified moieties or in which esterified moieties may optionally be absent. It is assumed that the presence of such segments of a certain size in high-esterified pectins accounts for the calcium sensitivity of these pectins.
  • pectins having a high degree of polymerisation are preferred because a better gel quality is obtained with high molecular " pectins than with pectins having a lower degree of polymerisation.
  • Pectin produced by extraction of apple pulp has a relatively low calcium sensitivity.
  • apple pulp which i.a. is formed by production of apple juice and cider is present in relatively small amounts, and the pectin content of apple pulp is considerably lower than in the peel and residues obtained after pressing of citrus fruits in which pectin is also present and which are becoming increasingly common to use for the production of pectin by extraction.
  • High-esterified pectin produced from citrus fruits is, however, fairly calcium sensitive.
  • the object of the invention is to produce high-esterified pectin having a low calcium sensitivity and a relatively high degree of polymerization from raw materials which are readily available such as peels and press residues of citrus fruits.
  • This object is obtained by the process according to the invention which process is characterized in that the high-esterified pectin is reacted with polygalacturonase at a pH-value of 1-7 and at a temperature of 5-65 ⁇ C, and that the reaction is stopped before the average molecular weight is reduced to a value of less than 50 of the average molecular weight of the starting material.
  • an enzyme preparation containing a relatively large amount of polygalacturonase is capable of splitting low- esterified pectin and undissolved, high-molecular protopectin and thereby is suitable for extracting fruit juice from juicy fruits. It is also known that this enzyme preparation only to a limited degree splits high-esterified, soluble pectin.
  • the invention is based on the discovery that polygalacturonase even though it is capable of splitting high-esterified pectin to a low degree as mentioned above does have an effect on said pectin, and that the result is a considerable reduction in the calcium sensitivity of the pectin without a corresponding alteration of the degree of polymerization.
  • the reason for the change in the calcium sensitivity of the high-esterified pectin is that the enzyme can easily destroy the molecular regions which in particular account for the calcium sensitivity. Surprisingly, a depolymerization of these regions does not result in a drastic reduction in the molecular weight of the pectin. This unexpected phenomenon may be accounted for if it is assumed that the regions in question are situated mainly at the ends of the molecular chains. Another possible explanation could be that a relatively small portion of the pectin molecules, which differ considerably, is responsible for the calcium sensitivity and that particularly these molecules are decomposed by the enzyme treatment.
  • calcium ions are present in varying amounts in the fruit products which are used for manufacturing gels and jams by using pectin as a gelling agent.
  • the solidity of the gels can be measured by various methods which can be divided into two fundamentally different main groups:
  • the gel strength can be determined by the break stress method described in further detail in example 1.
  • Gels presenting a relatively close relationship between gel strength by destructive measurement and gel strength by non-destructive measurement have an elastic texture whereas gels showing the opposite trend have a crisp texture.
  • An elastic texture is desirable in connection with clear gels of a type particularly known in Denmark as currant gel and rowanberry gel; in the US a corresponding grape gel is widely used.
  • the pectin produced according to the method of the invention offers more elastic gels when employing calcium containing systems than normally obtained by a similar pectin which has not been treated with polygalacturonase.
  • the method according to the invention preferably uses high-esterified pectin extracted from citrus fruits.
  • suitable citrus fruits are lemons, oranges, lime fruits and grape fruits.
  • the used pectin preferably has a degree of esterification of 50-85% and the pectin is preferably used in a concentration of 0.5-8%.
  • polygalacturonase examples include Rohament ® P (Rohm GmbH, BRD) and Pectinex MC (N0V0 Schweitzerische Ferment AG).
  • the enzymes are preferably used in a concentration of from 6 to 6250 PGU per gram of pectin.
  • the reaction takes place at a temperature of between 5 and 65 ⁇ C.
  • a certain destruction of the enzyme takes place concurrently with the interaction of the enzyme on the pectin thereby increasing the consumption of enzymes.
  • a particularly preferred temperature interval is 40 to 55°C.
  • the reaction time normally ranges from 5 minutes to 48 hours.
  • the reaction course is partly controlled by measuring the relative reduction in molecular weight, the reaction being terminated as mentioned above before the molecular weight of the pectin has been reduced to a value of 50% of the average molecular weight of the starting material. It is preferred to stop the reaction before the molecular weight has been reuced to 80%.
  • the reaction may be terminated by quickly heating the reaction mixture to about 70 ⁇ C thereby causing an irreversible destruction of the enzyme.
  • the process may also be stopped by precipitation of the pectin in a suitable solvent.
  • the enzyme is not or only partly destroyed by this treatment. If it is desired to reduce the reaction time prior to an actual termination of the reaction this may be effected by altering the pH-value of the reaction mixture away from the optimum value for enzyme efficiency.
  • reaction of pectin with polygalacturonase is not terminated until
  • U is the enzyme activity measured in PGU
  • T is the reaction time in minutes
  • P is the amount of pectin used in g.
  • PGU polygalacturonase activity
  • the molecular weight of the pectins thus treated is determined by measuring the relative viscosity of diluted solutions of pectin in a hexa etaphosphate buffer.
  • the amount of enzymes causing a 20% reduction in the initial molecular weight is determined. This amount of enzymes defines the activity of the preparation.
  • a pectin test sample system containing 1.05% (weight/volume) of pectin and 20 mM of CaCK and having a pH-value of 3.7 is prepared.
  • Pectin is used having a DE-value of from 70 to 72%, an AGA-value (anhydrogalacturonic acid content) of from 70 to 72%, and a molecular weight of from 125.000 to 135.000.
  • a series of enzyme solutions are prepared having various contents of enzyme preparation in a 20 mM CaC -solution.
  • Interpolation determines the enzyme concentration, C, which reduces the apparent molecular weight by 20% in relation to the molecular weight of the O-sample.
  • the specific activity of the enzyme preparations in PGU/mg is defined by dividing 200.000 with the specific value C.
  • the relative viscosity is determined by the formula
  • t and t are discharge times for the pectin solution and hexametaphosphate solution, respectively, and K is a constant depending on the used viscosimeter.
  • the molecular weight of the pectin is defined on the basis of the formula:
  • the value obtained is rounded up to an integer multiplum of 1000.
  • Pectin is extracted from orange-mash.
  • the extract is filtered through kieselguhr and the greater part of the calcium present is removed by ion exchange.
  • the resulting extract which has a pectin content of 9 g/1 is heated to 40 ⁇ C and divided into two fractions which are adjusted to a pH-value of 3.2 and 4.8, respectively.
  • pectins are produced in each series having an exponentially increasing degree of treatments i.e. the totals of products of enzyme concentration and treatment time in the relative ratio of 0, 1, 4, 11, 26, 57 and 120.
  • the pectins were analyzed for mol weight, USA-SAG, stress at break and SAG of gels at a pH-value of 3.1, partly in the presence and partly in the absence of calcium. The results obtained appear from table I and table 2.
  • the calcium sensitivity was also determined by determining the viscosity of pectin solutions of 1.5 weight percent with varying calcium dosages.
  • Example 2 Relationship between loss in molecular weight and gelling properties.
  • pectin solutions are subjected to treatment until a considerable but not complete reduction in the calcium sensitivity is considered 35. to have taken place. Once this stage has been reached the solution is poured into isopropanol in order to terminate the reaction and the pectin is isolated. The purpose of the test is to obtain pectins with a comparable molecular weight and calcium sensitivity.
  • This example uses a pectin extract in which the pectin to be precipitated has a lower degree of esterification than in the above examples.
  • a pectin extract was produced of lemon-mash.
  • the precipitation yield of the extract was 8.2 g/1.
  • Test data for the pectin which was precipitated from the non-enzyme treated extract appear from the table below. Prior to the enzyme treatment the pH-value of the extract was adjusted to 2.5.
  • the extract was continuously mixed with a solution of "Rohament® P" in the ratio 7 parts of extract to 1 part of enzyme solution and the mixture was passed through a tube reactor consisting of a 200 meter armed plastics tube with an inner diameter of 19 mm and with a flow velocity of 480 liter/hour.
  • the temperature of the tube reactor was 53 ⁇ C.
  • the enzyme solution contained 2 g of enzyme per 100 liter.
  • the extract was heated to 70 ⁇ C to terminate the reaction and to inactive the enzyme completely.
  • pectin which can be precipitated from the enzyme treated extract has higher SAG values and stresses at break at a pH value of 3.1 in the presence of calcium than the corresponding pectin of non-treated extract. In the absence of calcium the treated pectin has somewhat lower values than the non-treated.
  • This example tested the influence of temperature,the same pectin extract being treated in a series of different temperatures.
  • the extract was sampled from a conventional manufactory production. It was produced of lemon-mash.

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  • Chemical & Material Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Organic Chemistry (AREA)
  • Engineering & Computer Science (AREA)
  • Health & Medical Sciences (AREA)
  • Wood Science & Technology (AREA)
  • Biochemistry (AREA)
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  • Biotechnology (AREA)
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  • Polymers & Plastics (AREA)
  • Medicinal Chemistry (AREA)
  • Materials Engineering (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • General Engineering & Computer Science (AREA)
  • Genetics & Genomics (AREA)
  • Polysaccharides And Polysaccharide Derivatives (AREA)
  • Jellies, Jams, And Syrups (AREA)
  • Preparation Of Compounds By Using Micro-Organisms (AREA)

Abstract

Un procédé d'amélioration des propriétés de gélification de pectine hautement estérifiée consiste à faire réagir la pectine avec de la polygalacturonase à une valeur de pH comprise entre 1 et 7 et à une température comprise entre 5 et 65°C. La réaction est terminée avant que la masse moléculaire moyenne de la pectine n'ait été réduite à une valeur égale à 50 % de la masse moléculaire moyenne de la matière de départ.
PCT/DK1989/000159 1988-06-24 1989-06-26 Procede d'amelioration des proprietes de gelification de pectine hautement esterifiee WO1989012648A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
DK3500/88 1988-06-24
DK350088A DK350088D0 (da) 1988-06-24 1988-06-24 Fremgangsmaade til forbedring af hoejforestret pektins geleringsegenskaber

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Cited By (13)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0574066A3 (en) * 1992-06-09 1994-09-28 Quest Int Method for the recovery of enzymes used in polysaccharide modification processes
EP0664300A1 (fr) * 1993-12-02 1995-07-26 Hercules Incorporated Pectine, procédé et composition
US5895784A (en) * 1994-07-07 1999-04-20 Michigan Cancer Foundation Method for treatment of cancer by oral administration of modified pectin
EP0868854A3 (fr) * 1992-01-20 1999-05-12 Japan Tobacco Inc. Pectine à bas poids moléculaire, et aliment et boisson contenant la pectine à bas poids moléculaire
WO1999037685A1 (fr) * 1998-01-20 1999-07-29 Hercules Incorporated Pectine utilisee dans des materiaux pateux, son procede de preparation, materiaux pateux la contenant ainsi que son utilisation
WO2000008952A1 (fr) * 1998-08-14 2000-02-24 Danisco A/S Utilisation d'une composition
WO2000027888A1 (fr) * 1998-11-05 2000-05-18 Cp Kelco Aps Pectine pour la stabilisation de proteines
WO2000058367A1 (fr) * 1999-03-31 2000-10-05 Hercules Incorporated Pectine peu sensible au calcium
US6159721A (en) * 1997-08-20 2000-12-12 Hercules Incorporated Amine modified polysaccharides
US6159503A (en) * 1993-12-02 2000-12-12 Hercules Incorporated Pectin process and composition
US6428837B1 (en) 2000-06-09 2002-08-06 Cp Kelco Aps Deesterified pectins, processes for producing such pectins, and stabilized acidic liquid systems comprising the same
US6699977B1 (en) 2000-06-09 2004-03-02 Cp Kelco Aps Low methoxyl pectins, processes thereof, and stabilized aqueous systems comprising the same
WO2005102262A3 (fr) * 2004-04-26 2005-12-08 Cp Kelco Aps Composition de protection cutanee a controle de l'alcalinite et son utilisation

Citations (1)

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Publication number Priority date Publication date Assignee Title
DE2843351A1 (de) * 1977-10-08 1979-04-12 Kikkoman Shoyu Co Ltd Neue pectinesterase, verfahren zu deren herstellung und verfahren zur herstellung von demethoxiliertem pectin unter verwendung der neuen pectinesterase

Patent Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
DE2843351A1 (de) * 1977-10-08 1979-04-12 Kikkoman Shoyu Co Ltd Neue pectinesterase, verfahren zu deren herstellung und verfahren zur herstellung von demethoxiliertem pectin unter verwendung der neuen pectinesterase

Non-Patent Citations (5)

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Title
CHEMICAL ABSTRACT, Vol. 95, (1981), abstract No. 40943c, & Sci. Aliments 1981, 1(1), 81-9 (Eng). *
CHEMICAL ABSTRACTS, Vol. 103, (1985), abstract No. 67444m; & DD,A,216955. *
CHEMICAL ABSTRACTS, Vol. 107, (1987), abstract No. 171224h, Prikl. Biokhim., & Mikrobiol. 1987, 23(4), 561-7 (Russ). *
CHEMICAL ABSTRACTS, Vol. 76, (1972), abstract No. 96126p, J. Gen Appl., & Microbiol. 1971, 17(5), 421-7 (Eng). *
CHEMICAL ABSTRACTS, Vol. 90, (1979), abstract No. 181916e, & J. Food Sci. 1979, 44(2), 611-14 (Eng). *

Cited By (22)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0868854A3 (fr) * 1992-01-20 1999-05-12 Japan Tobacco Inc. Pectine à bas poids moléculaire, et aliment et boisson contenant la pectine à bas poids moléculaire
EP0574066A3 (en) * 1992-06-09 1994-09-28 Quest Int Method for the recovery of enzymes used in polysaccharide modification processes
EP0664300A1 (fr) * 1993-12-02 1995-07-26 Hercules Incorporated Pectine, procédé et composition
US6207194B1 (en) 1993-12-02 2001-03-27 Hercules Incorporated Pectin process and composition
US6159503A (en) * 1993-12-02 2000-12-12 Hercules Incorporated Pectin process and composition
US6143346A (en) * 1993-12-02 2000-11-07 Hercules Incorporated Pectin process and composition
US5895784A (en) * 1994-07-07 1999-04-20 Michigan Cancer Foundation Method for treatment of cancer by oral administration of modified pectin
US6159721A (en) * 1997-08-20 2000-12-12 Hercules Incorporated Amine modified polysaccharides
WO1999037685A1 (fr) * 1998-01-20 1999-07-29 Hercules Incorporated Pectine utilisee dans des materiaux pateux, son procede de preparation, materiaux pateux la contenant ainsi que son utilisation
WO2000008952A1 (fr) * 1998-08-14 2000-02-24 Danisco A/S Utilisation d'une composition
US6528085B2 (en) 1998-08-14 2003-03-04 Danisco A/S Use of a composition
US7371402B2 (en) 1998-08-14 2008-05-13 Danisco A/S Use of a composition
WO2000027888A1 (fr) * 1998-11-05 2000-05-18 Cp Kelco Aps Pectine pour la stabilisation de proteines
US6221419B1 (en) 1998-11-05 2001-04-24 Hercules Incorporated Pectin for stabilizing proteins
WO2000058367A1 (fr) * 1999-03-31 2000-10-05 Hercules Incorporated Pectine peu sensible au calcium
JP2002540258A (ja) * 1999-03-31 2002-11-26 シーピー・ケルコ・エイピーエス 低いカルシウム感度を有するペクチン
KR100791157B1 (ko) 1999-03-31 2008-01-02 씨피 켈코 에이피에스 칼슘 감수성이 감소된 펙틴
US6428837B1 (en) 2000-06-09 2002-08-06 Cp Kelco Aps Deesterified pectins, processes for producing such pectins, and stabilized acidic liquid systems comprising the same
US6699977B1 (en) 2000-06-09 2004-03-02 Cp Kelco Aps Low methoxyl pectins, processes thereof, and stabilized aqueous systems comprising the same
WO2005102262A3 (fr) * 2004-04-26 2005-12-08 Cp Kelco Aps Composition de protection cutanee a controle de l'alcalinite et son utilisation
EP2196242A1 (fr) 2004-04-26 2010-06-16 CP Kelco ApS Composition de controle de l'alcalinite et de protection pour la peau et son utilisation
CN1976677B (zh) * 2004-04-26 2012-10-24 Cp科尔克公司 保护皮肤的碱度控制组合物及其应用

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