AU730898B2

AU730898B2 - A method for fruit processing

Info

Publication number: AU730898B2
Application number: AU83348/98A
Authority: AU
Inventors: Yves Andre Gerard Coutel; Rodney Hugh Sharpe Dale
Original assignee: DSM NV
Current assignee: DSM IP Assets BV
Priority date: 1997-05-23
Filing date: 1998-05-25
Publication date: 2001-03-15
Anticipated expiration: 2018-05-25
Also published as: WO1998052423A2; NZ501237A; EP0987953A2; WO1998052423A3; AU8334898A

Description

WO 98/52423 PCT/EP98/03227 A method for fruit processing s Technical field The present invention provides a process to improve fruit or vegetable texture, firmness and integrity in fruit or vegetable preparations.

Background of the invention The texture of plant foods is attributed to the structural integrity of the primary cell wall and the middle lamella. The primary cell wall model consists of cellulose micro-fibrils aligned in parallel. Cellulosic microfibrils are cross linked with hemicellulosic xyloglucan polymers.

This domain is embedded in pectin matrix which form junctions with arabinogalacturonan and arabinogalactan side chains. Vincken, Thesis Wageningen Agricultural University, Ref ISBN 90-5485-510-X, 1996). The middle lamella is mainly composed of pectic substances partially esterified and cross linked with Ca 2 divalent cation. The middle lamella acts like adhesives between adjacent cells.

Ripened fruit tissue contains a higher degree of esterified pectin making it less cross-linked by Ca 2 and consequently more heat labile(D.T. Huber, Food Sci., 49: 1310 1315, 1988), Jackman and D.W. Standley, Trends Food Sci. and Technol. 6: 187-194, 1995).

The manufacturing of food compositions comprising entire fruit or fruit pieces is conventionally performed by mixing fruit, sucrose and other ingredients and heating the mix to prevent the possibility of microbiological 3s contamination (sterilisation).

Unfortunately, the heating needed for sterilisation has a negative effect on fruit texture and firmness and typically results in fruit with a fluffy appearance and mouthfeel.

-2- EP 0 624 062 describes a method for making food from fruit and vegetables with improved mouthfeel by in situ demethoxylation of the pectin backbone and allowing the thus treated fruit and vegetable pulp to form a gel.

The above discussion of documents, acts, materials, devices, articles and the like is included in this specification solely for the purpose of providing a context for the present invention. It is not suggested or represented that any or all of these matters formed part of the prior art base or were common general knowledge in the field relevant to the present invention as it existed in Australia before the priority date of each claim of this application.

Throughout the description and claims of the specification the word "comprise" and variations of the word, such as "comprising" and "comprises", is not intended to exclude other additives, components, integers or steps.

Summary of the Invention In one aspect the present invention provides a method for treating an entire fruit or vegetable or fruit or vegetable pieces, wherein the fruit or vegetable or fruit or vegetable pieces are subjected to a heat-treatment in the presence of pectin esterase and calcium ions.

In a further aspect the present invention provides a method for treating an 2o: 0 entire fruit or vegetable or fruit or vegetable pieces, wherein the fruit or vegetable or fruit or vegetable pieces is subjected to a heat-treatment in the presence of calcium ions.

e S"In an even further aspect the present invention provides a method for sterilising or preserving an entire fruit or vegetable or fruit or vegetable pieces, 25 wherein the fruit or vegetable or fruit or vegetable pieces are subjected to a heattreatment in the presence of pectin esterase and calcium ions.

The. Description of the Invention The present invention relates to a method for preserving firmness and good mouthfeel of an entire fruit or vegetable or of fruit or vegetable pieces which are subjected to a heat-treatment. More specifically, the present invention relates to a method in which an entire fruit or vegetable or fruit or vegetable pieces are subjected to a heat-treatment in the presence of calcium ions, more preferably also in the presence of pectin esterase.

\F:\on Spcci\8334X.doc 2a The major advantage of the method of the invention is that damage and disintegration of the heat-treated fruit is minimised. This effect is markedly strong where a combination of pectin esterase and calcium ions is used. Neither the pectin esterase nor the calcium ions gives by itself the same positive effect which is experienced when a combination of both is used. The amount of pectin esterase needed exceeds the amounts disclosed in EP 0 624 062.

Another advantage is that the integrity and firmness and the texture of the fruit is preserved as much as possible.

WAF-i;i\Spcci W 349.doc WO 98/52423 PCT/EP98/03227 3- Many types of fruit may be used in the method of the present invention, including strawberries, apples, pears, raspberries, rhubarb and so on. The method of the invention may be used on fruit, whether it is in one piece or in pieces, such as sliced or diced fruit; whether it is fresh, frozen or thawed. However, if the fruit is in pieces, these pieces are preferably not smaller than 5 mm 3 Although the method of the invention may be used for subjecting fruit to any kind of heat treatment, such as for example blanching, cooking, baking, it is especially advantageous for the sterilisation or pasteurisation of fruit.

Any pectin esterase (EC 3.1.1.11) which is suitable for the de-esterification of pectin, may be used in the method of the invention. Such pectin esterases are obtainable from plants, bacteria or fungi. The pectin esterase is preferably from fungal origin. More preferably, the pectin esterase is obtainable from Aspergilli, especially preferred is the use of a pectin esterase obtainable from Aspergillus niger.

Although both randomly and blockwise de-esterifying pectin esterases may be used, blockwise de-esterifying pectin esterases are preferred.

The pectin esterase may be partially or completely purified from other enzyme activities, provided that pectin depolymerising activites, such as pectin/pectate .lyase or polygalacturonase activity are kept at levels which leave the pectin backbone intact. In a preferred embodiment, a pectin esterase preparation from Aspergillus niger is used.

The calcium ions which are present in the method of the invention, may be added in any form. In a preferred embodiment of the invention, CaCl 2 .2H 2 0 is used to supply the calcium ions.

The pectin esterase and the calcium ions should be present in sufficient amounts. Therefore, at least 560 pectin esterase units per kg fruit (PE U/kg) are present and preferably, at least 2000 PE u/kg fruit. One PE unit is as defined in EP 0 624 062. The minimum amount of Ca 2 which WO 98/52423 PCT/EP98/03227 4 should be present is 0.6 mmol Ca 2 per kg fruit. Preferably, at least 1.0 mmol Cd 2 per kg fruit is present.

The invention also relates to a method for preparing a food composition which comprises an entire fruit or vegetable or fruit or vegetable pieces that are heat-treated according to the method of the invention, and to food compositions thus obtained.

The method of the invention is particularly suitable for the preparation of dairy and bakery food compositions, which need a heat treatment (for sterilisation and baking, respectively) Examples of such food compositions according to the invention include but are not limited to yogurt fruit preparations, bakery fruit preparations, such as oven stable conserves and canned fruits in syrups.

The amounts of calcium ions and pectin esterase may be varied as long as at least 560 pectin esterase units per kg fruit (PE U/kg) are present and at least 0.6 mmol Ca 2 per kg fruit. The person skilled in the art will understand that the effect of the pectin esterase and the calcium ions will not only depend on the amount of these compounds present, but also on the time of incubation and the presence of other compounds. One such compound which will have a negative effect is citrate, or its derivatives. In one preferred embodiment it is shown how the effect of citrate/citric acid may be neutralised by the addition of extra calcium ions.

The person skilled in the art will understand that the method of the invention is also applicable to plants, especially to dicots, which have similar cell wall structure. Therefore, the method of the invention may also be used to minimise heat damage during sterilisation of vegetables.

The invention will be illustrated by the following examples.

Examples Fruits texture analysis WO 98/52423 PCT/EP98/03227 5 Texture analysis were made on a TA.HD 50 kg F Texture Analyser (Stable Micro Systems, Vienna Court. Lammas Road, Godalming, Surrey, England) using the OTTAWA Cell system. We have used two tests: The first, Maximum Extrusion Force measurement is realised to evaluate fruit firmness. For that 240 g of fruits are extruded with a constant velocity displacement through an extrusion plate located in the base of the cell.

The maximum extrusion force is calculated by average of io forces registered at the constant part of the curve.

The second, springiness measurement is realised in a range of forces situated in the viscoelastic deformation phenomenous domain. A constant force is applied on fruit sample to obtain a planned regular surface. On a second time a stress at constant displacement is applied on the sample.

The forces ratio between stress maximum force and the lowest force value registered at the end of the relaxation time gives us an estimated value of fruits springiness properties.

The value of Max Extrusion Force (Firmness) is calculated by average of 5 measurements. Springiness value is calculated by average of 3 measurements.

Example 1 Pectin esterase and calcium treatment effect on strawberries firmness Washed frozen graded Turkish strawberries were used (size between 18-22 mm) and the process is presented in Scheme 1.

Treatments were carried out in distilled water (30 w/w and in agitated vessel at 20 rpm. 600 g of strawberries were treated with pectin esterase activity from 0 up to 8600 PE U/kg treated fruits. The pH of the reaction mixture was not regulated but is estimated at 3.47 0.05 pH unit after average calculation. The added calcium level was fixed at 0.05% of CaCI2-2H20 except for the control.

The enzyme is inactivated by heat treatment at 85 0 C for mn. All the trials were duplicated.

WO 98/52423 PCT/EP98/03227 6 The results summarised in Table 1 show the effect of enzyme dosage combined with calcium at constant concentration on strawberries firmness.

PE in combination with calcium increase the firmness s of strawberries and significantly preserve the firmness of fruits during cooking.

Table 1: The effect of pectin esterase on the firmness of heat treated strawberries No. PE CaCl 2 2H2O Calcium FIRMNESS activity (g/kg fruits) (mmole/k (F-Max kg (PE g) unit/kg) 1 0 0 0 8.7 2 0 0.75 5.1 14 3 560 0.75 5.1 13 4 750 0.75 5.1 2240 0.75 5.1 21 6 4480 0.75 5.1 7 8950 0.75 5.1 18 It is possible to determine an optimum at a dosage of 2240 PE Unit/kg of fruit.

enzyme effect Example 2 Calcium dosage effect on PE treated strawberries.

Using the same material and process as in Example (1) we have studied calcium dosage effect from 680 up to 6800 meq kg of treated fruits at a constant PE activity.

The Pectin esterase activity is fixed at 2240 PE U/ kg of fruit, except for the control. The firmness measurements of such treated strawberries is summarized in Table II. They indicate an optimum calcium level located around 0.05% w/w CaC1 2 *2H 2 0 per kg reaction mixture. This optimum corresponds to 136 mg of CaCl 2 or to 3400 pmole Ca2+ per kg treated strawberries.

WO 98/52423 PCT/EP98/03227 7- Table 2 The effect of calcium on the firmness of PE treated strawberries N° PE CaCl 2 2H20 Calcium FIRMNESS SPRINGINESS activity (g/kg (mmole/ (F-Max ratio) (PE fruits) kg) kg) unit/kg) s 1 0 0 0 9.6 5.8 (ref)_ 2 2240 0.15 1.025 13 3 2240 0.30 2.03 15 11 4 2240 0.75 5.07 22 11 5 2240 1.5 10.15 21 13 Example 3 Citrate buffering agent effect on fruit firmness treatment is Strawberries are treated in the same way as in Examples 1 and 2, except higher PE activities up to 2250 and 2700 U/kg of treated fruit were used to reduce the holding time during fruit cooking. Firmness values measured on such treated fruits according to conditions summarised in Table 3 show us a negative effect of Citrate buffering agent on texture improvement (Trials 1 to 3).

Table 3 The effect of citrate levels on the firmness of PE treated strawberries N' PE PE Citrate CaC1,2H20 Calcium FIRM- SPRINGI amount acti- (mmole/ (g/kg (mmole/ NESS NESS (g/kg vity kg) fruits kg) (F-Max fruits) (PE kg) ratio) unit/ kg) 1 Control 0 0 0 0 8.7 6.3

I

2 2.2 3360 0 0.75 5.1 15.5 7.1 3 2.2 3360 24.9 0.75 5.1 9.8 7.6 4 Control 0 0 0 0 9.9 nd

II

2.7 4029 24.9 0.75 5.1 17 7.7 6 2.7 4029 24.9 2.24 15.2 29 10.5 7 2.7 4029 24.9 4.48 30.4 26.5 9.1 WO 98/52423 PCT/EP98/03227 8 This effect may be eliminated by addition of a sufficient amount of calcium chloride for complete citrate anibn saturation (Trials 5 to 7) Example 4 Use of the invention for processing strawberries conserves for use with yogurts and dairy desserts Many trials have been made at pilot and industrial scale. We describe in the following examples a few of them to illustrate each point which constitute key points for the present invention.

Enzyme preparation is We have used to perform the following trials a pectin esterase preparation from Aspergillus niger at 950 PEUnit/g.

Texture analysis The texture analysis was performed in the same manner as described previously.

For fruit piece texture measurement we used a retroextrusion cell (50 mm Internal diameter with a 45 mm compression disc. With this kind of probe it is not possible to measure the fruit pieces springiness.

Pilot scale process descriptions The two processes we used at pilot scale for strawberries yogurt preparation with entire fruits are described briefly in Schemes 2 and 3.

The enzyme level needed was calculated according to our previous work Examples 1, 2, 3) as was other biochemical data in regard of the amount of fruit, the pH, the temperature and the incubation time used in the trials.

The calcium chloride level was also calculated according to our previous work.

We used two kinds of frozen entire strawberries. The first one was the same as in previous example (Turkish origin, graded between 18 and 22 mm) the second one consisted of Polish strawberries (graded 15 mm 22 mm). The WO 98/52423 PCT/EP98/03227 9- Turkish strawberry batch was used to produce the diced fruit pieces.

s First experiment: Trials on entire Turkish and Polish strawberries using process I (scheme 2) and II (scheme 3).

Trial details and texture analysis measurements are summarised in Table 4.

Table 4 The effect of PE and calcium on the firmness of whole strawberries No Process Straw- PE Citrate Calcium FIRMNESS SPRING- No. berries activity (mmole/ (mmole/ (F-Max INESS Type (PE kg) kg) kg unit/kg) ratio) 1 I turkish 0 Control 24.9 0 3.6 5.8

I

2 I turkish 0 24.9 30.3 9.2 13 Control

II

3 I turkish 5700 24.9 30.3 15 12 4 I turkish 11400 24.9 30.3 14 12 5 I polish 5700 24.9 30.3 22 15.5 6 II turkish 0 24.9 0 4.4 nd control

III

7 II turkish 4880 24.9 30.3 17 17 Trials 1 and 2 demonstrate the improved firmness obtained by 2s the use of calcium addition alone. However, trials 3, 4 and show the synergistic effect of PE and calcium. Trials 3 and 5 show that the effect is reproducible for different strawberry varieties sourced from different countries.

WO 98/52423 Scheme 1 PCT/EP98/03227 10 Laboratory combined pectin esterase and calcium treatment on crude entire calibrated strawberries.

Frozed Strawberries, 600 g weighted into the reactor Thawed at 5°C, one night Water (with CaC 2 2HO according the experiment) Heated at Enzyme solution (50 ml) Incubation at 50'C 60 mn at 20 tr/mn.

Heated to min)

\D

Transferred in boxes of 900 g

I

Stored 48 h in cold chamber 4 Texture analysis measurements WO 98/52423 Scheme 2 PCT/EP98/03227 11 PROCESS I (BATCH) WATER PE (5700/11400 PE Unit/kg Fruits) CaC1 2 -2H 2 0 4.46 g/kg Fruits)

*C)

Agitation 8 rpm FRUIT SUGAR BRING TO 50*C HOLD 2Omn THICKENING AGENT BRING TO

'I

ADD

CITRIC ACID SODIUM CITRATE FILL

COOL

WO 98/52423 Scheme 3 PCT/EP98/03227 12 PROCESS II (Continuous) WATER PE (4885 U/kg Fruits) CaC1 2 2H 2 0 4.46 g/kg Fruits)

*C)

Agitation 8rpm FRUIT SUGAR THICKENING AGENT BRING TO 25*C HOLD 6Omn BRING TO CITRIC ACID SODIUM CITRATE FILL

COOL

WO 98/52423 PCT/EP98/03227 13 Second experiment: using process Trials on diced Turkish strawberries Table 5 The effect of PE and calcium on the firmness of s fruit pieces No Process Strawber- PE Citrate Calcium FIRMNESS No ries activity (mmole/kg) (mmoie/kg) (F-Max Type (PE kg) unit/kg) 1 I turkish 0 24.9 0 4.8 Control

I

2 I turkish 5700 24.9 30.3 23 Results of those trials clearly demonstrate improvement to firmness and fruit piece integrity (Table Third experiment: Scaling up of the process Table 6 The effect of scale-up on the firmness of strawberries treated with PE and calcium No Process Size PE Citrate Calcium FIRMNESS SPRING- No. (kg) activity (mmole/ (mmole/ (F-Max INESS (PE kg) kg) kg) ratio) unit/kg) 1 I 50 5400 24.9 30.3 22 18 2 I 200 5400 24.9 30.3 22 22 3 I 600 5400 24.9 30.3 23 4 II 50 4880 24.9 30.3 17 17 II 500* Control 24.9 0 8.9 11 6 II 500* 4880 24.9 30.3 20 21 continuous process Scaling up does not reveal negative effect on fruit texture and identity improvement in the two processes studied for both batch and continuous production systems.

WO 98/52423 PCT/EP98/03227 14 Example 5 Treated fruit texture stability during prolonged storage of the fruit preparation Preparations made in Example 4 (first experiment) were stored in a dark room at 15°C for 14 weeks. Texture measurements were performed after 1, 8 and 14 weeks storage.

The texture measurement results are summarised in Table 7.

The positive effect on fruit texture between first and 8th weeks measurements on trials 3 and 4 may be caused by non achieved equilibrium between the processed fruits and their medium.

A longer storage time does not alter fruits texture properties. Consequently we can expect to apply the treatment described in the present invention to fruits and vegetables long shelf life products sterilised by heat treatment like canned fruits.

Table 7 Strawberry texture stability during storage Samples FIRMNESS (F-MAX kg) SPRINGINESS Example 4 1 week 8 wks 14 wks 1 week 8 wks 14 wks No.

1 3.6 5.1 3.4 5.8 4.1 6.3 2 9.2 11 11 13 11 3 15 19 18 12 13 18 4 14 20 20 12 18 19 22 26 26 15.5 nd 23 6 4.4 5.7 4.4 nd 11 9 7 17 28 23 17 28 29 nd: Not determined.

Example 6 Treated fruit texture stability mixed in yogurt PE treated strawberry preparation as described in Example 4, first experiment, was mixed with yogurt at an inclusion level of 17%. The mix and the original sample were stored under refridgerated condition. Texture analysis was WO 98/52423 PCT/EP98/03227 15 performed after one and three weeks storage time on washed fruits.

The texture analysis measurement summarised in the following table show good fruit stability in such conditions. The fruit firmness seems to increase slightly in yogurt which may be due to calcium equilibria within the system.

Table 8 The firmess of PE/calcium treated strawberries throughout the shelf-life of yogurt TRIALS Firmness F-Max (kg) PE treated Stawberries 18.5 (1 week) Yogurt's Strawberries (1 week) PE treated Stawberries 22 (3 weeks) Yogurt's Strawberries 24 (3 weeks) Example 7 Effect of pectin esterase treatment on fruit pectin Pectin esterase effect on processed pectin has been studied using the following methods on samples issued from trials 1, 2, 3, 6 and 7 example 4 (Use of the invention for processing strawberries conserves for use with yoghurts and dairy desserts).

Strawberries Insoluble Alcohol Material were extracted according to the method described by D.J. Huber et al., Huber, Food Sci., 49: 1310-1315, 1988).

Pectin breakdown according to pectin solubility properties were estimated using the method AFNOR NF 05-128 (Nov.1984).

Esterification Degree (ED) of extracted IAM were determined after saponification using High Performance Liquid Chromatography (HPLC) method according to WO 98/52423 PCT/EP98/03227 16 A.G.J. Voragen et al. (Food Hydrocolloids, Vol. 1, 1986) According to the results summarized in Table 9, pectin s esterase decreases the methylation degree of strawberry IAM, whereby fruit firmness is improved. Especially if we compare the fruits processed with the same physical parameters (Trials 1, 2, 3 and 4 process I and trials 4 and 6 for process II).

Table 9 Pectin esterase and pectin calcium effect on strawberry Trials IAM Galac Mety- Solu- Pec- Proto Fruit from Con- turo- lation ble tate pectin Firm- (Exam- tent nic Degree pectin ratio ratio ness pie 4) washed Acid% ratio kg F fruits IAM GalA GalA GalA GalA 1 Control 5.8 7.5 46 46.7 22.7 30.7 3.6

I

mn, 0

C

2 Control 5.1 9 43 33.3 28.9 37.8 9.2 Calcium 3 PE 5.6 10.4 25 20.2 48.1 31.7 14.8 0,35%, mn, 0

C

6 Control 7.4 6.6 54 40.9 28.8 30.3 4.4

II

mn, 250C 7 PE 0,3% 5.4 9.8 35 19.4 51 29.6 16.8 mn, 0

C__

Pectin esterase Methylation Degree (MD) firmness improvement.

treatment decreases fruit pectin with a concommittant effect on fruit The pectate ratio increases and water soluble pectin ratio decreases due to pectin esterase treatment and calcium WO 98/52423 PCT/EP98/03227 17 chloride addition. The use of calcium salt alone as additive did not affect IAM methylation degree. However, it has an effect on pectin breakdown and improves fruit textural properties, but not to such a significant level as when used s in combination with pectin esterase.

Firmness measurement values of those processed fruits revealed a correlation between pectate and soluble pectin ratios and fruit textural properties.

This shows that the ratio of soluble polyuronides on total polyuronides, which is known to be a good indicator of fruit softening during maturation, can be modified using pectin esterase and calcium treatment during processing.

Example 8 Pectin esterase and Calcium process adaptability First Experiment: Process regulation using physico chemical enzyme properties.

It is well known that enzymatic process can be regulated according to an enzyme's physico-chemical properties which influence enzyme kinetics. The following experiments illustrate this point.

Experiments were carried out according to an experimental design in order to determine the rl1e of each factor which has an effect on enzyme activity in regard to strawberries firmness improvement. The pectin esterase preparation is the same as previously. All trials and firmness measurements are presented in Table WO 98/52423 Table 10 PCT/EP98/03227 18 Experimental design and results.

No PE Holding Holding Firmness Springiness (PE U/kg time Temperature (kg) fruits) (min) 1 6650 5 40 10.9 13 2 6650 15 50 19.0 21 3 13300 5 50 21.4 21 4 13300 15 40 17.5 23 9975 10 45 17.6 19 6 9975 10 45 22.7 19 7 9975 10 45 19.1 18 From the results of trials 1 to 4 the effects of each factor on firmness may be determined.

Factors Effect on firmness improvement Holding temperature from 40 to 50 0 C 6 kg Pectin esterase dosage from 6650 to 13300 UPE/kg Hold time 5-15 min.

4.5 kg 2.2 kg The temperature increase has the strongest effect on firmness improvement which is shown by statistics. The enzyme amount cannot be considered as significant and holding time has also little effect in our process.

Second Experiment: Enzyme preparation from Genetically Modified Aspergillus niger origin To perfom this experiment, we compared strawberries from Example 8, first experiment trials 5,6 and 7 which were treated with pectin esterase from the classical organism Aspergillus niger, with strawberries treated with pectin esterase obtained from a genetically modified Aspergillus niger strain in which the homologous gene has been amplified. The results reported in Table 11 illustrate the WO 98/52423 PCT/EP98/03227 19 similarity between fruit identity texture improvement obtained with those two enzyme preparations.

Table 11 The effect of the source of pectin esterase on the firmness of strawberries No. PE origin Firmness Firmness Springiness (kg) std deviation (kg) 5,6,7 Classical 22.5 2.2 22.5 Strain 8 Gen Mod 19.8 1.3 22 Strain Third Experiment: Combined PE/Calcium treatment on other fruits We have optimised the process I (Scheme 2; Example 4) to apply it to different kinds of fruits for fruit preparation manufacturing. The first trial was performed on diced apple pieces. The results reported in Table 12 demonstrate the beneficial effects of the combined PE and calcium on the fruit texture.

The second was performed on entire thawed raspberries.

Raspberries texture measurements were performed on full fruit preparation using retro extrusion cell, 50 mm Internal diameter with a 45 mm compression disc. The value of Max extrusion Force is calculated by average of 15 measurements.

The results reported respectively in Tables 12 and 13 demonstrate the beneficial effects of combined PE and calcium on fruit texture.

WO 98/52423 PCT/EP98/03227 20 Table 12 The effect of PE and calcium on apple pieces No Trial PE activity Citrate Calcium FIRMNESS (F-Max kg) (PE unit/kg) (mmole/kg) Value Std Deviation 9 Control 0 24.9 No 6.0 0.9 Calcium 0 24.9 Yes 9.1 0.9 11 PE+Calcium 9975 24.9 Yes 16.9 0.85 Table 13. PE and calcium effect on entire thawed raspberries.

No Trial PE Citrate Calcium Firmness Std activity m mole/ mmole/ (gF) Deviatio (PE kg Fruit kg Fruit n (gF) Unit/kg Fruit) 12 Control 0 24.9 30 506 13 PE.

Calcium 4750 24.9 30 1584 172 chloride 14 PE.

Calcium 7125 24.9 30 1653 482 chloride

PE.

Calcium 9500 24.9 30 1973 210 chloride 16 PE.

Calcium 4750 24.9 30 1325 190 phosphate Example 9 Pectin esterase and calcium treatment effect on tomato pieces texture This example was performed with canned diced peeled tomatoes as raw material. Trials have been done using the process described in scheme Tomato pieces were washed with distilled water before treatment to remove tomato sauce. The citric acid content in washed tomato pieces was checked using the enzymatic method according to the procedure described by AFNOR, Contr8le de la qualit6 des WO 98/52423 PCT/EP98/03227 21 produits alimentaires, produits derives de fruits et lgumes, p300-303, (1990 AFNOR., Paris.

Calcium chloride in mono hydrated form was used as calcium salt. We added a sufficient amount of calcium to s improve tomato pieces firmness and to counterbalance the citrate complexation effect as described respectively in examples 8 and 3).

Pectin esterase activity was as high as 6000 U /kg of fruits, using the same enzymatic preparation as previously.

Enzymatic incubat-ion has been done between 20 and 48°C during 50 min. The pH of the reaction mixture was not regulated and was found to be 4.3.

The agitation mixture was agitated only every ten minutes during 20 sec at 20 rpm to reduce mechanical effect on tomato pieces integrity. The enzyme was inactivated at the end of the reaction time by heating the reaction mixture to 90 0 C during 20 0 C using a microwave oven.

Texture measurement were done for 20 h after processing with the same tests as previously. Texture measurement values summarised in Table 14 are in line with our previous results.

WO 98/52423 Scheme 4 PCT/EP98/03227 22 Laboratory pectin esterase and calcium treatment on canned tomato pieces.

Canned tomato pieces Washed with distilled water 600 g Water with CaC 2 1.2(H 2 0) according to the experiment 200 ml Enzyme solution or water (50 ml) Heated up to 48 0

C

during 50 min.

Heated to 90 0

C

min) 4 Transferred in boxes Stored one night in cold chamber Texture analysis measurements.

WO 98/52423 PCT/EP98/03227 23 Table 14 Effect of PE and calcium on tomato pieces texture.

Trial PE Citrate CaCl, Calcium Firm- Spring Disrup No. Activity mmole/ 2 (HO0) (mmole/ ness in ess tion PE U/ kg kg g k g kg) F.Max ratio Energy of kg kgFxm Fruit 1 Raw 27.3 0 0 16 6.5 0.064 material 2 Control 27.3 0 0 16 5.2 0.062

I

3 Control 27.3 3 20.4 28.5 8.9 0.147

II

4 6000 27.3 3 20.4 47 12.8 0.277 Pectin esterase reaction combined with calcium addition increase tomato pieces firmness and springiness significantly. Trials 3 improvement obtained by such significant as the and 4 demonstrate that texture calcium addition alone is not as combined treatment.

Claims

1. A method for treating an entire fruit or vegetable or fruit or vegetable pieces, wherein the fruit or vegetable or fruit or vegetable pieces are subjected to a heat-treatment in the presence of pectin esterase and calcium ions.

2. A method according to claim 1 wherein the pectin esterase is present in a concentration of at least 560 units per kg fruit or vegetable or fruit or vegetable pieces.

3. A method according to claim 1 or 2 wherein the calcium ions are present in a concentration of at least 0.6 mmol per kg fruit or vegetable or fruit or vegetable pieces.

4. A method according to claims 1-3 wherein 1,000-10,000, preferably 2,000-5,000 units pectin esterase and 1-30 mmol Ca++ are present per kg fruit or vegetable or fruit or vegetable pieces. 0000 0S •0 •l

5. A method for preparing a fruit or vegetable composition comprising an 20 entire fruit or vegetable or fruit or vegetable pieces, wherein the entire fruit or 00 vegetable or fruit or vegetable pieces are treated according to any one of the methods of claims 1-4. *too*:

6. A fruit or vegetable composition comprising an entire fruit or vegetable or 0004 q0• 25 fruit or vegetable pieces which is obtainable by a method according to claim

7. A fruit or vegetable composition according to claim 6 wherein the fruit is a strawberry, apple, raspberry or a tomato. 0000

8. The use of a fruit or vegetable composition according to claim 6 or 7 in a dairy, bakery or canned product. W:\fiona K1Spdie.\83348.doc

9. A method for treating an entire fruit or vegetable or fruit or vegetable pieces, wherein the fruit or vegetable or fruit or vegetable pieces is subjected to a heat-treatment in the presence of calcium ions.

10. A method for sterilising or preserving an entire fruit or vegetable or fruit or vegetable pieces, wherein the fruit or vegetable or fruit or vegetable pieces are subjected to a heat-treatment in the presence of pectin esterase and calcium ions.

11. Fruit or vegetable or pieces thereof which have been treated by a method o0 according to any one of claims 1-4, 9 or

12. A method according to claim 1, 9 or 10 substantially as hereinbefore described with reference to any of the examples. DATED: 4 January, 2001 PHILLIPS ORMONDE FITZPATRICK Attorneys for: 20 DSM N.V. W:ioll\NKI\Spccics\83348.do