METHOD OF PREPARING A CROP-BASED COMPOSITION

Abstract

A method of preparing a liquid crop-based composition for use as a food includes the steps of: i) rinsing a crop having at least one antinutritive compound with an aqueous liquid for a first time period, wherein the aqueous liquid has a temperature of 50 C.-90 C.; ii) disintegrating the rinsed crop, and mixing it with an aqueous liquid to obtain a liquid crop-based composition; iii) subjecting the liquid crop-based composition to an amylase so as to at least partially degrade starch in the liquid crop-based composition; and iv) subjecting the liquid crop-based composition to a betaglucosidase under first dosage, time and temperature conditions so as to reduce the content of the at least one antinutritive compound.

Claims

1-15: (canceled)

16. A method of preparing a liquid crop-based composition suitable for use in a food product or a feed product, the method comprising the steps of: i. rinsing a crop that includes at least one antinutritive compound with an aqueous liquid for a first time period, wherein the aqueous liquid has a temperature of 50 C.-90 C.; ii. disintegrating the rinsed crop, and mixing the disintegrated and rinsed crop with an aqueous liquid to obtain a liquid crop-based composition; iii. subjecting the liquid crop-based composition to an amylase so as to at least partially degrade starch in the liquid crop-based composition by an enzyme activity of the amylase; and iv. subjecting the liquid crop-based composition to a betaglucosidase under first dosage, time, and temperature conditions so as to reduce the content of the at least one antinutritive compound, thereby resulting in the liquid crop-based composition being suitable for use in a food product or a feed product.

17. The method according to claim 16, wherein the crop provided in step i comprises dehulled fava beans.

18. The method according to claim 16, wherein step ii comprises mixing the disintegrated crop with an amount of aqueous liquid sufficient to provide a liquid crop-based composition with a dry matter content of from 5% to 25%.

19. The method according to claim 16, further comprising the step of inactivating the enzyme activity of the amylase prior to performing step iv.

20. The method according to claim 16, wherein the first time period, and the first dosage, time and temperature conditions are selected so as to be sufficient to obtain a sum content of vicine and convicine, calculated as 1.5*content of convicine+content of vicine, in the liquid crop-based composition that is no more than 360 mg per kg of the liquid crop-based composition.

21. The method according to claim 16, wherein the first time period, and the first dosage, time and temperature conditions are selected so as to be sufficient to obtain a ratio of content to dry matter, calculated as (1.5*content of convicine+content of vicine)/dry matter content %, that is no more than 4.06 mg per percent dry matter.

22. The method according to claim 16, wherein the amylase comprises at least one amylase selected from the group consisting of alpha-amylase, beta-amylase, and gamma-amylase.

23. The method according to claim 16, wherein liquid crop-based composition is subjected to the amylase under second dosage, time, and temperature conditions such that the starch content, compared to the starch content of liquid crop-based composition obtained in step ii, is reduced by at least 50%.

24. The method according to claim 16, wherein the second dosage condition is at least 0.001 wt %, the second time condition is at least 30 minutes, and the second temperature condition is at least 50 C.

25. The method according to claim 16, wherein the first dosage condition is at least 0.1 wt % betaglucosidase per kg liquid crop-based composition, the first time condition is at least 2 hours, and the first temperature conditions is at least 60 C.

26. The method according to claim 16, wherein at least one of the crop and the aqueous liquid is agitated during the rinsing step i.

27. The method according to claim 26, wherein at least one of the crop and the aqueous liquid is agitated by a mechanical stirrer

28. The method according to claim 26, wherein at least one of the crop and the aqueous liquid is agitated by a flow of the aqueous liquid.

29. The method according to claim 16, further comprising the step v of manufacturing a food product or a feed product using the liquid crop-based composition.

30. A liquid crop-based composition suitable for use in a food product or a feed product, wherein the liquid crop-based composition is obtained by a method comprising the steps of: i. rinsing a crop that includes at least one antinutritive compound with an aqueous liquid for a first time period, wherein the aqueous liquid has a temperature of 50 C.-90 C.; ii. disintegrating the rinsed crop, and mixing the disintegrated and rinsed crop with an aqueous liquid to obtain a liquid crop-based composition; iii. subjecting the liquid crop-based composition to an amylase so as to at least partially degrade starch in the liquid crop-based composition by an enzyme activity of the amylase; and iv. subjecting the liquid crop-based composition to a betaglucosidase under first dosage, time, and temperature conditions so as to reduce the content of the at least one antinutritive compound, thereby resulting in the liquid crop-based composition being suitable for use in a food product or a feed product.

31. A food product or feed product comprising a liquid crop-based composition obtained by a method comprising the steps of: i. rinsing a crop that includes at least one antinutritive compound with an aqueous liquid for a first time period, wherein the aqueous liquid has a temperature of 50 C.-90 C.; ii. disintegrating the rinsed crop, and mixing the disintegrated and rinsed crop with an aqueous liquid to obtain a liquid crop-based composition; iii. subjecting the liquid crop-based composition to an amylase so as to at least partially degrade starch in the liquid crop-based composition by an enzyme activity of the amylase; iv. subjecting the liquid crop-based composition to a betaglucosidase under first dosage, time, and temperature conditions so as to reduce the content of the at least one antinutritive compound, thereby resulting in the liquid crop-based composition being suitable for use in a food product or a feed product; and v. making a food product or a feed product containing the liquid crop-based composition resulting from the performance of the preceding step iv.

Description

BRIEF DESCRIPTION OF THE FIGURES AND DETAILED DESCRIPTION

[0081] A more complete understanding of the abovementioned and other features and advantages of the technology proposed herein will be apparent from the following detailed description of preferred embodiments in conjunction with the appended drawings, wherein:

[0082] FIG. 1A shows a first embodiment of the method of preparing a liquid crop-based composition according to the first aspect of the technology proposed herein,

[0083] FIG. 1B shows a second embodiment of the method of preparing a liquid crop-based composition according to the first aspect of the technology proposed herein.

[0084] In the below description of the figures the same reference numerals are used to designate the same features throughout the figures. Further, where present a added to a reference numeral indicates that the feature is a variant of the feature designated with the corresponding reference numeral not carrying the-sign.

[0085] FIG. 1A shows a first embodiment of the method of obtaining a liquid crop-based composition according to the first aspect of the technology proposed herein. The crops 2 are rinsed with water 4, which has a temperature of 50-90 C. The rinsing 3 may last for 2.5 hours. The duration of the rinsing may also be prolonged to last 3-12 hours (such as for 3 hours or 6 hours). As shown in the figure, the rinsed crops 2 and if necessary further water 4, is admitted to a mill 6 in which the rinsed crops are milled and finely divided together with the water. The crops are disintegrated to particles less than 5 mm in diameter. The milling in the mill 6 may take place at any suitable temperature. Typically, the temperature is at least 20 C. The temperature may be higher, such as at least 70 C. or 90 C., for example 95 C. temperature. The milling continues until of the crops 2 have been reduced in size to the size provided by the mill 6, i.e., less than 5 mm diameter. The mixture of milled crops, i.e., the crop material, and water is hereinafter referred to as a liquid crop-based composition. The liquid crop-based composition here has a dry matter content of about 9 wt %. The liquid crop-based composition may optionally, after the mill 6, be homogenized in a homogenizer 8 at elevated pressure and temperature, for example at more than 70 C. or more than 80 C. and at more than 100 bar or any other pressure suitable to further disintegrate the crop material in the crop-based composition and to increase the degree of extraction of the nutrients (carbohydrates, proteins, minerals, vitamins, etc.) from the crops material. The liquid crop-based composition is then optionally decanted in decanter 10 for separating out a solid fraction 12 of the liquid crop-based composition. This solid fraction 12 may be used as described further below.

[0086] The remainder of the liquid crop-based composition 14, also called the liquid fraction, is transferred into a mixing tank 16. Here an amylase 18 at a dosage of at least 0.001 wt % is added to at least partially degrade starch in the liquid crop-based composition for a time of for example 45 minutes at a temperature of for example 60 C. Following an optional inactivation of the amylase, and in order to reduce the content of the antinutritive compounds, such as vicine, divicine, convicine and isouramil in the liquid crop-based composition 14, a betaglucosidase enzyme 20 is added to the mixing tank 16. The addition of the betaglucosidase enzyme 20 is typically 0.1 to 1 wt % of the crop-based composition and the enzyme is allowed to process the liquid crop-based composition 14 for about at least 1 hour, such as for example 2-4 hours at a temperature of 40-70 C., for example from 55 to 65 C., at which the enzyme is active. The enzyme may, as here, be provided as a solution containing 20 wt % enzyme, whereby the dosage of 0.1 to 1 wt % refers to the enzyme solution and corresponds to 0.02 to 0.2 wt % of isolated enzyme being added.

[0087] The so treated liquid crop-based composition comprises a reduced content of at least one of the compounds vicine and convicine.

[0088] Optionally, to reduce the microbial content, the liquid crop-based composition 14 may for example be heat treated, for example as illustrated here using a UHT device 22 wherein it is heat treated at temperatures above 100 C. for sufficient time.

[0089] If the betaglucosidase and the amylase is not inactivated in the UHT device 20, then the enzyme activity may be deactivated in the mixing tank 16, after the enzymation process has concluded by cooling the mixing tank 16 to a temperature below room temperature, or alternatively be heating the mixing tank 16 to a temperature sufficient to cause heat denaturing of the enzymes. Finally, the liquid crop-based composition may be homogenized in a second homogenizer 24 before being stored, for example in a sterile tank 26, or used further. The liquid crop-based composition 14 so obtained may be used as a substitute to cream and whipping cream, as a base for soups etc. The solid fraction 12 may be used as a meat substitute. Both the solid fraction 12 and the liquid crop-based composition 14 contain reduced contents of antinutritive compounds such as vicine and divicine compared to the crops 2 in the beginning of the process.

[0090] Although in FIG. 1A only the liquid crop-based composition 14 enters the mixing tank 16 to be subjected to the amylase and the betaglucosidase 18 and 20, it is also possible to position the decanter 10 after the mixing tank 16 such that the enzymation using the enzymes is performed on the liquid crop-based composition prior to the optional separation of the solid fraction 12 from the liquid crop-based composition 14.

[0091] Additionally, the amylase 18 may be added earlier in the process, such as in the mill 6. This is advantageous as the retention time in the process provides the amylase 18 with the time needed to at least partially degrade the starch in the liquid crop-based composition 14 prior to the betaglucosidase 20 being added, and thereby decreases the risk for interaction between the amylase 18 and the betaglucosidase 20 without having to inactivate the enzymatic activity of the amylase 18 in the mixing tank 16.

[0092] Further, whereas FIGS. 1A (and 1B below) show and describe the use of a number of devices and steps, including the mill 6, the homogenizer 8, decanter 10, UHT device 22, homogenizer 24, sterile tank 26, which are advantageous, these devices and steps are not essential for achieving the objects of the technology described herein.

[0093] FIG. 1B shows a second embodiment of the method of obtaining a liquid crop-based composition according to the first aspect of the technology proposed herein. This embodiment differs in addition of different further nutrients to the liquid crop-based composition. Accordingly, in certain cases, it may be desired to further adjust or increase the nutritious properties of the crop-based composition. Edible oil, such as rape seed oil, may in particular be added to the mixing tank upon conclusion of the enzymation. This option is illustrated in FIG. 1B by the addition of edible oil 30 either in the beginning of the process, or the addition of edible oil 32 to the mixing tank 16. Edible oil may additionally be added at other steps of the method.

[0094] Other foodstuffs may also be added to the liquid crop-based composition. One example is oats and other cereals. Foodstuffs may in particular be added prior to or together with the addition of the enzymes (amylase and/or betaglucosidase) for example as indicated by 34 in FIG. 1B.

Example 1Determination of Generally Safe Levels of Vicine and Convicine

[0095] According to the initially identified papers by Duc G et al and Gallo V et al, the fava bean cultivar Divine does not produce symptoms of favism when consumed at an amount of 500 g raw beans per 70 kg body weight. The Divine cultivar contains 0.16 g vicine and convicine per kg wet weight. The typical value for general fava bean cultivars is 4.75 g vicine and convicine per kg wet weight, and Gallo et al also found that this amount of vicine and convicine did produce symptoms of favism. The ratio between the divine cultivar and the general fava bean cultivar thus becomes (0.16/4.7)=0.034.

[0096] In order to account for differing dry matter contents, the present inventors disintegrated beans of the Divine cultivar in order to obtain a paste. Further analysis showed that the paste had a dry matter content of 88.6% and contained 360 mg/kg of vicine and undetectable amounts of convicine.

[0097] In order to account for differing toxicity between vicine and convicine, the present inventors then considered the monograph: Toxicants of Plant Origin, Vol 2 Glycosides by Petter R. Cheeke published 2020. This work shows that the LD50 value in rats and the non-toxic dosage of vicine is generally 1.21-1.5 times that of convicine, i.e. convicine is more toxic than vicine. From this work a factor of 1.5 can be determined, whereby the content of convicine is to be multiplied by 1,5 and summed with the content of vicine to determine a combined value of vicine and convicine.

[0098] Accordingly, a product ready for consumption should have a total amount of vicine and convicine, calculated as 1.5*content of convicine+content of vicine that is less than 360 mg/kg product.

[0099] The concentration of convicine and vicine in a liquid composition will depend on the dry matter content. A low dry matter content of fava beans will thus result in a liquid composition having a low content of vicine and convicine, however, such a composition would have a correspondingly low nutritional value. Accordingly, the present inventors defined a dry matter independent value for the combined content of vicine and convicine expressed as the ratio of (1.5*content of convicine+content of vicine)/% TS. Applying this formula to the results obtained for the Divine cultivar bean paste and noting that the paste did not contain any detectable amounts of convicine, yielded a value of (360)/88.6=4.06. Accordingly, a product ready for consumption should therefore preferably have a ratio of (1.5*content of convicine+content of vicine)/% TS that is equal to or lesser than 4.06 in order to be both safe and nutritious.

Example 2Preparing Liquid Crop-Based Compositions

[0100] Table 1 discloses the results obtained on reducing the content of vicine, divicine, convicine and isouramil in a liquid crop-based composition, in this case a liquid fava bean-based composition, dependent on the treatment method used.

[0101] The content of the compounds was determined as follows.

[0102] Vicine and convicine were determined as based on the method described by Gutierrez et al: CAPs markers to assist selection for low vicine and convicine contents in faba bean (Vicia faba L.). Theor. Appl. Genet. 2006. The samples were extracted with water in a hot water bath for 3.5 hours, after which concentrated HCl was added to the sample extracts. Samples were analyzed by an Agilent 1100 series high performance liquid chromatography device with a diode array detector (HPLC-DAD). A C18 column was used with a gradient of phosphate buffer and methanol. The breakdown products divicine and isouramil were identified by their UV-spectra and quantitated using a calibration curve of vicine and correcting the result by molar mass ratio.

[0103] For Method/samples no. 2-4, the respective methods were carried out as described in table 1, and samples for the analysis above were collected about 24 hours after conclusion of the enzymation. The collected samples were then frozen, stored for 1-2 weeks, and then thawed before being extracted with water and further prepared for analysis as described above.

[0104] The results listed in table 1 are typical values from several runs of each of the respective methods (sample no. 2-3), and from several samples collected from each run.

TABLE-US-00001 TABLE 1 Sample no. Method/sample Contents and comment 1 Freshly harvested fava In total about 10740 mg/kg of convicine and bean vicine. Reference value for a non-processed fava bean. Divicine and isouramil are degradation products/metabolites of convicine and vicine, and therefore not present in any significant amount in the fava bean. From comparison with Example 1 it can be noted that the content of convicine and vicine is clearly above the generally safe level of 360 mg/kg. 2 Liquid fava bean-based 360 mg/kg of convicine composition as formed 791 mg/kg of vicine according to FIG. 1A but Less than 2 mg/kg of divicine and isouramil without the rinsing and The liquid fava bean-based composition enzymation step. comprises a lower content of convicine and vicine The temperature of the compared to the freshly harvested fava bean. liquid crop-based This is due to the dilution effected by the addition composition during of the water 4. In this case the ratio of fava bean: preparation was about 90 water was about 1:9, i.e. about 10% dry matter C. content. Although the content of convicine and vicine has been reduced compared to the freshly harvested fava bean, it however needs to be further reduced as it is above the generally safe level of 360 mg/kg established in Example 1. 3 The liquid fava bean-based 5 mg/kg of convicine composition obtained as 68 mg/kg of vicine shown in FIG. 1A, i.e., using divicine could not be detected (less than 2 mg/kg) only enzymation, but isouramil could not be detected (less than 2 without rinsing step mg/kg) The temperature of fava This process provides a significant reduction of bean composition in the convicine and vicine. No divicine or isouramil steps up to the enzymation could be detected in the fava bean composition. was about 90 C. The sum content of vicine and convicine for A first enzymation step comparison with the levels obtained for the Divine comprised simultaneous fava cultivar is (68 mg/kg + 5*1.5 mg/kg) = 75.5 addition of 0.0015 wt % mg/kg and thus within the generally safe level of alpha amylase (activity 480 360 mg/kg established in Example 1. KNU-B/g) and 0.01 wt % The dry matter content was 10%. beta amylase (Activity 5000 The ratio of content to dry matter could thus be BAMU/g) for 45 min at 60 determined to (1.5*5 mg/kg + 68 mg/kg)/10 = 7.55 C. mg per percentage dry matter The second enzymation The liquid fava bean-based composition is thus step comprises addition of safe to consume but is less preferable with 0.25 wt % betaglucosidase respect to the dry matter content percentage as (activity of 1500 AZO BBG the ratio of 7.55 is above the ratio for the Divine U/g or 6200 IU/g) for 2 hours fava bean cultivar. at 60 C.) Between the first and second enzymation steps the alpha and beta amylase was deactivated by heating the liquid fava bean-based composition to 95 C. 4 The liquid fava bean-based Dehulled fava beans: composition obtained as in 10 mg/kg of vicine FIG. 1A using both rinsing 11 mg/kg of convicine and enzymation. Non-dehulled fava beans: Rinsing for 6 hours at 40-60 11 mg/kg of vicine C. 20 mg/kg of convicine Enzymation as in No divicine or isouramil was detected. Method/sample 3 above. For dehulled fava beans, the sum content of vicine and convicine for comparison with the levels obtained for the Divine fava cultivar is 11*1.5 mg/kg + 10 mg/kg) = 26.5 mg/kg and thus within the generally safe level of 360 mg/kg established in Example 1. The dry matter content was 10%. The ratio of content to dry matter could thus be determined to (1.5*11 mg/kg + 10 mg/kg)/10 = 2.65 For non-dehulled fava beans the sum content of vicine and convicine was (1.5*20 mg/kg + 11 mg/kg) = 41 mg The ratio was (1.5*20 mg/kg + 11 mg/kg)/10 = 4.1 per percentage dry matter. These values show a liquid fava bean-based composition that is safe to consume, and that has a ratio of content to dry matter that is below (2.65) or similar (4.10) to the ratio of content to dry matter percentage obtained for the Divine fava bean cultivar.

[0105] As shown, the addition of the rinsing step further decreased the content of convicine and vicine. Dehulling of the fava beans decreased the content even further.