FI127240B - Process for converting berry and fruit materials into fractions containing bioactive compounds - Google Patents

Process for converting berry and fruit materials into fractions containing bioactive compounds Download PDF

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Publication number
FI127240B
FI127240B FI20165506A FI20165506A FI127240B FI 127240 B FI127240 B FI 127240B FI 20165506 A FI20165506 A FI 20165506A FI 20165506 A FI20165506 A FI 20165506A FI 127240 B FI127240 B FI 127240B
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berry
fruit
seed
fraction
process according
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FI20165506A
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Finnish (fi)
Swedish (sv)
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FI20165506A (en
Inventor
Pimiae Riitta Puupponen
Liisa Nohynek
Veera Virtanen
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Teknologian Tutkimuskeskus Vtt Oy
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Priority to FI20165506A priority Critical patent/FI127240B/en
Priority to PCT/FI2017/050450 priority patent/WO2017216426A1/en
Priority to KR1020197001708A priority patent/KR20190019181A/en
Priority to EP17812815.3A priority patent/EP3474819A4/en
Publication of FI20165506A publication Critical patent/FI20165506A/en
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Publication of FI127240B publication Critical patent/FI127240B/en

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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P17/00—Preparation of heterocyclic carbon compounds with only O, N, S, Se or Te as ring hetero atoms
    • C12P17/18—Preparation of heterocyclic carbon compounds with only O, N, S, Se or Te as ring hetero atoms containing at least two hetero rings condensed among themselves or condensed with a common carbocyclic ring system, e.g. rifamycin
    • C12P17/181—Heterocyclic compounds containing oxygen atoms as the only ring heteroatoms in the condensed system, e.g. Salinomycin, Septamycin
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L19/00—Products from fruits or vegetables; Preparation or treatment thereof
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23K—FODDER
    • A23K10/00—Animal feeding-stuffs
    • A23K10/10—Animal feeding-stuffs obtained by microbiological or biochemical processes
    • A23K10/12—Animal feeding-stuffs obtained by microbiological or biochemical processes by fermentation of natural products, e.g. of vegetable material, animal waste material or biomass
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L3/00—Preservation of foods or foodstuffs, in general, e.g. pasteurising, sterilising, specially adapted for foods or foodstuffs
    • A23L3/34—Preservation of foods or foodstuffs, in general, e.g. pasteurising, sterilising, specially adapted for foods or foodstuffs by treatment with chemicals
    • A23L3/3454—Preservation of foods or foodstuffs, in general, e.g. pasteurising, sterilising, specially adapted for foods or foodstuffs by treatment with chemicals in the form of liquids or solids
    • A23L3/3463—Organic compounds; Microorganisms; Enzymes
    • A23L3/3472—Compounds of undetermined constitution obtained from animals or plants
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
    • A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
    • A23L33/105—Plant extracts, their artificial duplicates or their derivatives
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
    • A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
    • A23L33/105—Plant extracts, their artificial duplicates or their derivatives
    • A23L33/11—Plant sterols or derivatives thereof, e.g. phytosterols
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A—HUMAN NECESSITIES
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    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
    • A61K36/18—Magnoliophyta (angiosperms)
    • A61K36/185—Magnoliopsida (dicotyledons)
    • A61K36/45—Ericaceae or Vacciniaceae (Heath or Blueberry family), e.g. blueberry, cranberry or bilberry
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
    • A61K36/18—Magnoliophyta (angiosperms)
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    • A61K36/63—Oleaceae (Olive family), e.g. jasmine, lilac or ash tree
    • A—HUMAN NECESSITIES
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    • A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
    • A61K36/18—Magnoliophyta (angiosperms)
    • A61K36/185—Magnoliopsida (dicotyledons)
    • A61K36/73—Rosaceae (Rose family), e.g. strawberry, chokeberry, blackberry, pear or firethorn
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    • A61K36/18—Magnoliophyta (angiosperms)
    • A61K36/185—Magnoliopsida (dicotyledons)
    • A61K36/73—Rosaceae (Rose family), e.g. strawberry, chokeberry, blackberry, pear or firethorn
    • A61K36/736—Prunus, e.g. plum, cherry, peach, apricot or almond
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    • A61K36/18—Magnoliophyta (angiosperms)
    • A61K36/185—Magnoliopsida (dicotyledons)
    • A61K36/73—Rosaceae (Rose family), e.g. strawberry, chokeberry, blackberry, pear or firethorn
    • A61K36/738—Rosa (rose)
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    • A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
    • A61K36/18—Magnoliophyta (angiosperms)
    • A61K36/185—Magnoliopsida (dicotyledons)
    • A61K36/87—Vitaceae or Ampelidaceae (Vine or Grape family), e.g. wine grapes, muscadine or peppervine
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K8/00—Cosmetics or similar toiletry preparations
    • A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
    • A61K8/96—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution
    • A61K8/97—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution from algae, fungi, lichens or plants; from derivatives thereof
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K8/00—Cosmetics or similar toiletry preparations
    • A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
    • A61K8/96—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution
    • A61K8/97—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution from algae, fungi, lichens or plants; from derivatives thereof
    • A61K8/9783—Angiosperms [Magnoliophyta]
    • A61K8/9789—Magnoliopsida [dicotyledons]
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    • C12P1/00—Preparation of compounds or compositions, not provided for in groups C12P3/00 - C12P39/00, by using microorganisms or enzymes
    • C12P1/04—Preparation of compounds or compositions, not provided for in groups C12P3/00 - C12P39/00, by using microorganisms or enzymes by using bacteria
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    • C12P17/00—Preparation of heterocyclic carbon compounds with only O, N, S, Se or Te as ring hetero atoms
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    • C12P17/06—Oxygen as only ring hetero atoms containing a six-membered hetero ring, e.g. fluorescein
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    • C12P19/00—Preparation of compounds containing saccharide radicals
    • C12P19/14—Preparation of compounds containing saccharide radicals produced by the action of a carbohydrase (EC 3.2.x), e.g. by alpha-amylase, e.g. by cellulase, hemicellulase
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    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
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Abstract

The invention relates to process for converting berry and/or fruit materials to fractions comprising bioactive compounds, said process comprising the steps, where at least one berry material or fruit material selected from berries, fruits, by-products, side streams and waste materials originating from berries or fruits, and any combinations thereof is sieved whereby a seed fraction is separated from a skin fraction, the seed fraction is subjected to sanding, followed by separation of a seed coat fraction comprising surface layer of the seeds, which is subjected to hydrothermal extraction.

Description

PROCESS FOR CONVERTING BERRY AND FRUIT MATERIALS TO FRACTIONS
COMPRISING BIOACTIVE COMPOUNDS
20165506 prh 25 -09- 2017
FIELD OF THE INVENTION
The present invention relates to a hydrothermal process for converting berry and fruit materials to fractions comprising bioactive compounds. The invention also relates to fractions comprising bioactive compounds, originating from berries and fruit, obtainable by the process. The invention further relates to the use of said fractions comprising bioactive compounds in cosmetics, hygiene products, nutraceuticals, food products and food supplements, feeds, packages and in pharmaceutical products.
BACKGROUND OF THE INVENTION
In the industry dealing with processing of berries and fruits significant amounts of waste materials, side streams and by-products are formed. Food industry uses wild berries, cultured berries and fruits in the manufacture of wide range of products, such as pastes, beverages, alcohol products, jams, conserves, milk based products, sweets and the like. The use of berry and fruit fractions has also become very popular in cosmetic products and for example cloudberry seed oil, rich in polyunsaturated fatty acids, is regarded as a valuable component in cosmetic preparations.
Large volumes of waste are produced in the processing of berries and fruit, particularly in the food industry, which waste material is utilized to a very small extent. Most of the waste material is currently discarded or transported to landfill or dumping area, thus increasing the environmental burden. Some of this waste material is subjected to drying followed by extraction of seed oils, however only a very small portion of the material is used.
Typically, in the processing, berries and fruit are mechanically cleaned, followed by removing of the juice, pulp or paste by suitable methods, such as pressing. The remaining wastematerial, such as pomace, berry cake, fruit cake or press cake comprises berry or fruit skins, peels, seeds and pith, which contain bioactive phenolic compounds, fibers and other bioactive compounds.
FI 122664 B discloses a method for fractionating berries and separating nutrients from the fractions formed, in which method berry raw material is dried and ground lightly, so that the seeds of the berry separate from the fruit flesh and skin portion without
20165506 prh 25 -09- 2017 breaking, followed by a second light grinding, which is carried out on the formed seedless fruit flesh and skin fraction, whereby a fine powder is formed, which is screened or classified. Seeds are not fractionated by this method. Seed are discarded and peels are further fractionated.
US 2013/0040005 Al relates to an antihypertensive agent comprising boysenberry seed extract as active ingredient and to a method for obtaining said agent. In said method boysenberry pomace is dried, crushed and sieved to separate the seed, followed by grinding the seed to fine powder, which is extracted with water or organic solvent, followed by contacting the extraction solution with a polyphenol absorbent and eluting with alcohol based solvent to obtain the target extract.
Based on the above it can be seen that there still exists a need to provide improved methods for utilizing berry and fruit materials, particularly waste materials, side streams and by-products for providing fractions comprising valuable bioactive compounds contained in said materials.
SUMMARY
In the present invention it was found that fractions comprising bioactive compounds, particularly ellagic acid and/or derivatives thereof, originating from berry and/or fruit seeds, can be enriched with a simple and environmentally safe method.
The present invention is particularly based on studies on hydrothermal processing and converting of pomace, berry cake, fruit cake and press cake obtained from berries and fruit, and the use of the obtained products. The invention provides convenient and effective means particularly for utilizing waste materials, side streams and by-products from berry and fruit industry, such as pomace, press cake, berry cake and fruit cake, in the manufacture of fractions comprising bioactive compounds useful in cosmetics, hygiene products, food supplements, food products, feeds, packages and in pharmaceutical products, particularly as natural antimicrobials or natural preservatives.
Thus an object of the invention is to provide a process for converting berry and fruit materials to fractions comprising bioactive compounds, particularly ellagic acid and/or derivatives thereof.
20165506 prh 25 -09- 2017
A further object of the invention is to utilize waste materials, side streams and byproducts from berry industry and/or fruit industry without the need to discard said materials.
A further object of the invention is to provide fractions comprising bioactive compounds originating from berries and fruits.
A still further object of the invention is the use of said fractions comprising bioactive compounds in cosmetics, hygiene products, nutraceuticals, food products, food supplements, feeds, packages and in pharmaceutical products.
The invention is directed to a process for converting berry and fruit materials to fractions comprising bioactive compounds, which process comprises the steps, where in the first step at least one berry material or fruit material selected from berries, fruits, by-products, side streams and waste materials originating from berries or fruits selected from the genus Rubus, Sorbus, Rosa, Empetrum, Aronia and Hippophae and from combinations thereof, and the fruit are selected from the genus Vitis, Punica, Pyrus, Citrullus, Benincasa, Cucumis, Momordica, Olea, Malus and from species Argania spinosa, and combinations thereof, said berry material or fruit material having water content not more than 15 wt%, is sieved, whereby a seed fraction is separated from a skin fraction, in the second step the seed fraction is subjected to sanding where 2-40 wt% of the seed is removed and a seed coat fraction comprising surface layer of the seeds and a sanded seed fraction comprising the sanded seeds are obtained, in the third step, the seed coat fraction is mixed with an aqueous medium in a ratio from 1:3 to 1:80 by weight, at a temperature from 15 to 95°C for 20 min to 10 hours to obtain a mixture comprising bioactive compounds, and the mixture is subjected to separation whereby a hydrothermal extract is separated from a hydrothermal residue.
The bioacative compounds particularly comprise ellagic acid and/or derivatives thereof.
The invention is further directed to the use of the fractions comprising bioactive compounds, in cosmetics, hygiene products, nutraceuticals, food products, food supplements, feeds, packages and in pharmaceutical products.
Accordingly, the present invention provides simple and economic means for utilizing waste materials, side streams and by-products originating from berry industry and
20165506 prh 25 -09- 2017 fruit industry in the manufacture of fractions comprising bioactive compounds, whereby dumping of berry or fruit waste can be avoided or at least substantially decreased.
The characteristic features of the invention are presented in the appended claims.
BRIEF DESCRIPTION OF THE DRAWINGS
Figure 1 illustrates one embodiment of the process of the invention.
Figure 2 illustrates another embodiment of the process of the invention, where berry or fruit material is pretreated prior to sieving.
Figure 3 (3A-3E) shows antimicrobial activity of the fractions comprising bioactive compounds, obtained with the process.
DEFINITIONS
Unless otherwise specified, the terms, which are used in the specification and claims, have the meanings commonly used in the field of food industry. Specifically, the following terms have the meanings indicated below.
The term berry is understood here to mean all wild and cultivated berries comprising internal seeds, which berries belong to the genus Rubus, Sorbus, Empetrum, Rosa, Aronia or Hippophae, including all hybrid berries of these genera. Sea buckthorn is an example of the genus Hippophae. Raspberries, blackberries, arctic bramble, dewberries and cloudberries are examples of the Rubus species. As the Rubus species readily interbreed and are apomicts, the parentage of the hybrid plants is often highly complex, but it is generally agreed to include in the definition cultivars of blackberries and raspberries. Examples of said hybrid berries include loganberry, boysenberry, veitchberry, marionberry, silvanberry, tayberry, tummelberry and hildaberry.
The term fruit is understood here to mean all wild and cultivated fruit belonging to the genus Vitis, Punica, Pyrus, Malus, Citrullus, Benincasa, Cucumis, Momordica, Olea and species Argania spinosa, including all hybrid fruit of these genera or species.
Fruit of the genus Vitis mean grapes comprising internal seeds, including all hybrid grapes, which are primarily crosses between V. vinifera and another grapevine. Grapes are used for making wine, jam, beverages, jelly, seed extract, raisins, vinegar, and grape seed oil. Vitis is a genus of about 60 vining plants in the family Vitaceae.
20165506 prh 25 -09- 2017
Fruit of the genus Citrullus, Benincasa, Cucumis, Momordica mean melons comprising internal seeds, including all hybrid melons.
Fruit of the genus Olea means olives, particularly Olea europaea.
Fruits refers also to fruits of fruit trees, such as pomegranate (Punica granatum), pear tree (Pyrus communis) and apple tree (Malus family) and all hybrids thereof.
The terms berry industry and fruit industry refer to industry dealing with the development, refining and manufacture of products and processes relating to wild and cultivated berries and fruit.
The term antimicrobially active compound refers here to compounds being able to kill microorganisms or inhibit their growth. Examples of these compounds are phenolic compounds, such as ellagic acid and ellagitannins. In addition to antimicrobial and preservative activity, these compounds often have other biological activities, particularly antioxidant activity.
The term sanding of seeds refers here to mechanical, abrasive treatment where the surface layer of the seeds is removed as powder. Sanding is understood to also include polishing and pearling.
The term hydrothermal processing means here processing in the presence of water at elevated temperatures.
Hygiene products include here particularly diapers, female hygiene pads, tampons, etc, incontinence pads and products, and the like.
DETAILED DESCRIPTION OF THE INVENTION
The invention provides a convenient process for converting berry materials and fruit materials to fractions comprising valuable bioactive compounds, particularly to fractions comprising significant amounts of ellagic acid and/or derivatives of ellagic acid, particularly ellagitannins. Ellagitannins are esters of glucose with ellagic acid which, when hydrolysed, yield ellagic acid.
The invention is based on studies on the processing and converting of waste materials, side streams and by-products originating from processing of berries and fruit, such as
20165506 prh 25 -09- 2017 pomace and press cake, for providing means for utilizing these waste materials and by-products in the manufacture of fractions comprising bioactive compounds, useful for example in cosmetics, hygiene products, nutraceuticals, food products, food supplements, feeds, packages and in pharmaceutical products, as natural preservatives.
It was found that particularly the surface layer (seed coat fraction or seed peel layer) of the seeds of berries of the genus Rubus, Sorbus, Empetrum, Rosa, Aronia and Hippophae and seeds of the fruits of the genus Vitis, Punica, Pyrus, Malus, Citrullus,
Benincasa, Cucumis, Momordica, Olea, and species Argania spinosa, contains high concentrations of el leg ic acid and/or derivatives thereof, particularly ellagitannins. The fractions comprising ellagitannins show strong antimicrobial activity for example against the human pathogen Staphylococcus aureus. The seed coat fractions contain high concentrations of bioactive phenolic compounds. Particularly ellagic acid, ellagitannins and derivatives thereof are comprised in said fractions of Rubus berries.
Recent studies have shown that ellagic acid and ellagitannins possess many interesting biological activities and thus they may play preventive role in disease prevention.
Ellagic acid appears to have anti-cancer properties, especially against prostate cancer. In addition, pre-clinical work suggests a possible anti-inflammatory effect in obesityrelated diseases. Ellagitannins and ellagic acid may also have positive effects on glucose metabolism.
Remarkable amounts of biologically active phenolic compounds, particularly ellagic acid and/or ellagitannins and their derivatives remain in the waste material, which is currently not utilized. Ellagic acid and ellagitannins are natural antioxidants and preservatives having strong antimicrobial effect, thus useful as natural preservatives and antioxidants in various applications in the field of cosmetics, hygiene products, food industry and feed industry, as well as in packaging industry and pharmaceutical industry.
In skin care and hygiene products the invention may be utilized to convert berry and fruit materials to fractions that stabilize, balance and protect healthy skin microbiome, as these fractions are known to inhibit the growth of many pathogenic microbes without effecting the growth of beneficial microbes.
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With the process of the invention berry skins or fruit skins can be separated from the seeds and the seed surface layer can then be removed and subjected to hydrothermal extraction to obtain fractions having high antimicrobial activity. Further, optionally enzymes may be used in the hydrothermal extraction.
The hydrothermal residue separated after the hydrothermal extraction from the hydrothermal extract may also be used particularly in nutraceuticals, food products, food supplements and feeds because of high contents of ellagic acid, C18 fatty acids, lignin and fibers. Ellagic acid is more easily absorbed in the GI tract than ellagitannins typically present in the natural berries and fruit.
Berries
In the present invention, all wild berries, cultivated berries and all hybrid berries of the genus Rubus, Sorbus, Empetrum, Rosa, Aronia and Hippophae and any combinations thereof may be used. Raspberries, blackberries, arctic bramble (synonym arctic raspberries), dewberries and cloudberries, and hybrid berries including loganberry and boysenberry are examples of the Rubus species suitable for the invention. Rowanberry is an example of Sorbus species, crowberry of Empetrum species, rose hip and dog rose of Rosa species, chokeberry of Aronia species and sea buckthorn berry of Hippophae species suitable for the invention.
Cloudberries are valuable wild berries having high aroma content and they contain also valuable seed oil. Cloudberries are used in food, liqueur and cosmetic industry, however only the seed oil is presently utilized from the waste material remaining after pressing the berries. Ellagic acid content found in cloudberry is the following: berry fruit (fruit + seeds) 0.6 mg/g dry weight, skins 20.3 mg/g dry weight, sanded seed coat power 19.6 mg/g dry weight and polished seeds 12.6 mg/g dry weight. Ellagitannin content found in cloudberry is the following: berry fruit 24,6 mg/g dry weight, skins 11,0 mg/g dry weight, sanded seed coat powder 18,6 mg/g dry weight and polished seeds 10,9 mg/g dry weight. In the seeds ellagitannins are concentrated in the sanded seed coat fraction.
Arctic brambles contain very high ellagic acid and ellagitannin contents and thus they are also particularly suitable as raw material for the process of the invention. From the ecological point of view, wild berries, such as cloudberries and arctic bramble which have grown without any manmade fertilizers or pesticides, are particularly suitable.
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Fruits
All fruits of the genus Vitis, Punica, Pyrus, Mai us, Citrullus, Benincasa, Cucumis, Momordica, Ole, and of species Argania spinosa are suitable for the process of the invention. All grapes and all hybrid grapes, containing seeds and belonging to the genus Vitis may be used in the present invention. Typically huge amounts of waste material are obtained from processing of grapevines, such as from pressing grapevines and thus grapes offer also a particularly suitable raw material source for the present invention.
Fruits of fruit tree, such as pomegranate [Punica granatum), pear tree (Pyrus communis) and apple tree (Malus family), including all hybrids thereof are also suitable raw material source. Remarkable amounts of waste material are obtained during processing of these fruits.
Berry materials and fruit materials
The berry materials and fruit materials suitable for the process of the invention may be selected from whole berries, whole fruits, by-products, side streams and waste materials, originating from processing of berries or fruit. Examples of such byproducts, side streams and waste materials are press cakes, pomaces, berry cakes, fruit cakes and fractionating residues. Said by-products, side streams and waste material typically comprise berry or fruit skins or peels, seeds, some pulp, occasionally some leaves, arbors and conifer needles, depending also how well the berries or fruit are cleaned mechanically before processing.
Processing of berries or fruit may be carried out for example at a facility carrying out processing or refining or fractionating of berries or fruit, or at a facility in the food or feed processing industry, from the manufacture of beverages, pastes, purees, wines, jams, conserves, sweets and the like. Particularly preferably by-products, side streams and waste materials are used in the present invention.
Typically in a juicing line, berries or fruit are pressed and the remaining press cake is frozen and stored at approx. -20°C for further use, or alternatively it may be dried.
Berry and fruit pastes and purees are obtained for example by squeezing berries or fruit through a sieve and the remaining berry cake or fruit cake is frozen and stored at approx. -20°C, or alternatively it may be dried. The obtained frozen pomace, berry
20165506 prh 25 -09- 2017 cake or fruit cake may contain from 40 to 70 % by weight of water, typically from 50 to 60 % by weight of water.
The by-products, side streams or waste material obtained from the berries of the genus 5 Rubus or grapes from the genus Vitis, such as press cake or pomace contains predominantly seeds, skins or peels and some pulp.
Process
The present invention is directed a process for converting berry and fruit materials to fractions comprising bioactive compounds, which process comprises the steps, where in the first step at least one berry material or fruit material selected from berries, fruits, by-products, side streams and waste materials originating from berries or fruits selected from the genus Rubus, Sorbus, Rosa, Empetrum, Aronia and Hippophae and from combinations thereof, and the fruit are selected from the genus Vitis, Punica,
Pyrus, Malus, Citrullus, Benincasa, Cucumis, Momordica, Olea and from species Argania spinosa, and combinations thereof, said berry material or fruit material having water content not more than 15 wt%, is sieved, whereby a seed fraction is separated from a skin fraction, in the second step the seed fraction is subjected to sanding where 2-40 wt% of the seed is removed and a seed coat fraction comprising surface layer of the seeds and a sanded seed fraction comprising the sanded seeds are obtained, in the third step, the seed coat fraction is mixed with an aqueous medium, in a ratio from 1:3 to 1:80 by weight, at a temperature from 15 to 95°C for 20 min to 10 hours to obtain a mixture comprising bioactive compounds, and the mixture is subjected to separation whereby a hydrothermal extract is separated from hydrothermal residue.
The bioactive compounds comprise ellegic acid and/or derivatives thereof, particularly ellagitannins. Preferably the bioactive compounds comprise ellagic acid and/or ellagitannins.
The berry material or fruit material having water content not more than 15 wt% means here dry berry material or fruit material.
In an embodiment in the third step the ratio of the seed coat fraction to the aqueous medium is from 1:5 to 1:50, respectively.
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In an embodiment in the third step the ratio of the seed coat to the aqueous medium is from 1:5 to 1:40, respectively.
In an embodiment the seed coat fraction is mixed with the aqueous medium using 5 mixing speed 100-1500rpm. Any suitable mixing devices may be used.
In an embodiment in the aqueous medium is selected from water or a mixture of water with an acid selected from ascorbic acid oxalic acid, citric acid, acetic acid, malic acid, benzoic acid and HCI, preferably ascorbic acid. The concentration of the acid in the aqueous medium is from 0.01 to 0.2 % by weight, preferably from 0.05 to 0.1 % by weight. The mild acidic conditions have stabilizing effect on anthocyanins.
In an embodiment in the third step the mixing temperature may range from 20 to 95°C, preferably from 25 to 90°C.
In an embodiment in the third step at least one enzyme may be added to the aqueous medium. The enzyme is suitably selected from carbohydrate hydrolyzing enzymes, suitably from cellulose, pectinase, xylanase and combinations thereof. The enzymes are dosed based on their main activity (e.g. 100-200 nkat/g or 0.01-1 % by weight).
The enzyme is diluted in water before mixing with the aqueous medium. Depending on the enzyme the treatments are carried out either at a pH optimal for each enzyme or at the intrinsic pH of the used material. When an enzyme is used, the third step is carried at a temperature of 40-47°C for 1-4 hours, and then from 30 min to 7 hours at 50-90°C.
In the third step the separation of the hydrothermal extract from the hydrothermal residue (solid materials), is carried out using suitable methods, such as centrifuging, filtration, decanting and the like.
In an embodiment the hydrothermal extract, obtained in the third step is dried. The drying may be carried out by freeze-drying, spray-drying, drying at elevated temperature, in drying cabinets, and the like. Suitably the drying is carried out at a temperature from 40 to 150oC, spray-drying at 80- 95°C.
The process of the invention is illustrated in Figure 1, where berry and/or fruit material 10 having water content not more than 15 wt%, is subjected to sieving 100, whereby a seed fraction 30 is separated from a skin fraction 20, the seed fraction 30 is subjected
20165506 prh 25 -09- 2017 to sanding 200 where 2-40 wt% of the seed is removed and a seed coat fraction 50 comprising surface layer of the seeds and a sanded seed fraction 40 comprising the sanded seeds are obtained, the seed coat fraction 50 and an aqueous medium 60 are subjected to mixing 300 to obtain a mixture 70 comprising bioactive compounds with antimicrobial activity. The mixture 70 is subjected to separation 400 to obtain hydrothermal extract 80 and hydrothermal residue 82. The hydrothermal extract 80 is subjected to drying 800 to obtain dry fraction 81 comprising bioactive compounds.
The seed fraction comprises seeds. Occasionally it may comprise small amounts of any 10 of skin, peel and pulp.
The skin fraction comprises skin, peel and pulp. Occasionally it may comprise small amounts of any of leaves, arbors and conifer needles.
In a preferable embodiment, the berry and/or fruit material has water content of 0.115 10 wt%, particularly preferably 0.1-8 wt%.
In the process sieving (separation of seeds) is carried out, where the berry material or fruit material having water content not more than 15 wt% is sieved, whereby the seeds remain on the sieve and the skin, peels, pulp etc. pass through the sieve.
Suitably a sieving device, air classification device, air jet sieve device, screening device or rotary screen is used, preferably a vibratory or a shaking sieving/screening device is used, where the sieve size is selected according to the seed size of the berry or fruit.
For example, when sieving berry material obtained from the genus Rubus, suitably a sieve is used where the sieve has a mesh opening of 0.5-2.0 mm, preferably 0.6-1.6 mm.
The sanding is carried out using a sanding or polishing apparatus selected from grain polishing machines (e.g. barley), rice etc. polishing machines, pearling machines, dehullers and polishers/hullers.
In the sanding from 2 to 40 wt%, preferably 3-35 wt%, particularly preferably from 3 to 30 wt% of the seed surface layer is removed as finely divided powder fraction (seed coat fraction or seed peel fraction) from the seeds and a sanded seed fraction is also obtained.
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Pretreatment of berry material or fruit material
In an embodiment of the invention the berry material or fruit material is pretreated prior to sieving. The pretreatment may be carried by subjecting the berry material or fruit material to methods selected from heat treatment, fermentation, enzymatic treatment, pressing, squeezing, drying, crushing and combinations thereof.
According to one embodiment of the invention the berry material or fruit material is pressed, separated, decanted or centrifuged to separate the juice from solid matter (peels and seeds).
According to one embodiment of the invention the berry material or fruit material is heat treated for the removal of harmful microbes. Suitably the heat treatment is carried out at 80°C for 5 min.
According to one embodiment of the invention the berry material or fruit material is fermented using lactic acid bacteria to modify the phenolic compounds and the carbohydrate components of the seed. Preferably the starter culture is selected from the genera Lactococcus, Lactobacillus, Pediococcus and Oenococcus.
Figure 2 illustrates an embodiment where the berry material or fruit material is pretreated prior to sieving. Berry material or fruit material 90 is subjected to heat treatment and fermentation 500 whereby fermented berry or fruit material 91 is obtained, which is then subjected to pressing 600, whereby juice fraction 92 is separated from press cake 93. The press cake 93 is subjected to drying 700 to obtain berry material or fruit material 10 having water content not more than 15 wt%, which is subjected to sieving 100, whereby a seed fraction 30 is separated from a skin fraction 20, the seed fraction 30 is subjected to sanding 200 where 2-40 wt% of the seed is removed and a seed coat fraction 50 comprising surface layer of the seeds and a sanded seed fraction 40 comprising the sanded seeds are obtained. The seed coat fraction 50 and an aqueous medium 60 are subjected to mixing 300 to obtain a mixture 70 comprising bioactive compounds with antimicrobial activity. The mixture 70 is subjected to separation 400 to obtain hydrothermal extract 80 and hydrothermal residue 82. The hydrothermal extract 80 is subjected to drying 800 to obtain dry fraction 81 comprising bioactive compounds with antimicrobial activity. The hydrothermal residue may be subjected to drying at mild conditions, such as freezedrying or drying in a fluid-bed drier at a temperature from 35 to 70°C preferably 3545°C. (Not shown in the figure)
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In the fermentation typically frozen berry or fruit material and water, suitably ultrapure water, are mixed together (1:1) and heated, suitably at 80°C for 5 min. The mixture is cooled, suitably in an ice bath and if needed berry or fruit material is crushed. The pH of the mixture is adjusted to approx. pH 5.0, suitably with 5 N sodium hydroxide. The microbes are pre-grown in food-grade media. The fermentation is carried out in a bioreactor (a vessel) etc., for example for 3 days at 30°C under constant mixing. Lactic acid bacteria fermentations are purged with sterile nitrogen gas to create anaerobic conditions.
According to one embodiment of the invention the berry material or fruit material is treated with carbohydrate hydrolyzing enzymes. The berry material or fruit material is preferably pressed or squeezed after the enzyme incubation. Suitably the enzyme is selected from cellulose, pectinase, xylanase and combinations thereof. The juice yield is increased and the press cake or berry cake contains decreased amounts of sugars and water.
In the enzyme treatment, the enzymes are dosed based on their main activity (e.g. 100-200 nkat/g or 0.01-1 % by weight). Thawed, mashed and heated (40-45 °C) berry or fruit material is incubated at 40-45 °C for 2-4 hours. Enzyme is diluted in water before mixing with the mashed and heated berry and fruit materials. The treatments are carried out at the intrinsic pH of the used material (about pH 3). After enzyme incubations, the berry or fruit juice is extracted by a juice pressing device. Enzyme treatment typically increases the yields of the hydrothermal extract and compounds therein.
According to one embodiment of the invention the berry material or fruit material is dried prior to introducing to the process for removing excess water, until it has water content of not more than 15 wt%, preferably 0.1-10 wt%. The drying may be carried out as convective drying, such as hot-air drying, vacuum drying or steam drying, microwave drying with or without vacuum drying, or freeze-drying. The drying may be carried out using a fluid-bed drier at a temperature from 35 to 70°C preferably 3545°C. Suitably the freeze drying is carried out at a temperature from -40 to 0°C, and convective drying at a temperature from 40 to 70°C, preferably from 40 to 50°C. Any conventional drying devices suitable for the drying can be used.
According to one embodiment the berry material or fruit material is crushed prior to sieving, suitably using compression crushing to break lumps of skin, peels, pulp etc
20165506 prh 25 -09- 2017 from the seeds and to cause minimum damage to the seeds. The crusher may be selected from roll crusher, a ball crusher, manual type crusher, a kneader grinder or a combination thereof. A kneader grinder may be used as the crusher, by which the seeds of the berry are detached from the dry pulp and skin portion without breaking the seeds. A suitable grinder is a falling number mill containing a rotating rotor and a stationary stator. This mill provides a kneading and slightly cutting and striking effect, wherein the grinding energy is, however, not sufficient to break the seeds, but they are detached from the matrix. A disc mill or an impact mill with a guided impact is preferably used, whereby a gentler grinding process is achieved.
The sanded seeds comprising the seed core and the seed oil may be used as such in food and animal feed applications as healthy nutritional components. Alternatively the sanded seeds may be treated with an enzyme.
The sanded seed fraction may be further treated with an enzyme selected from cellulase, pectinase, xylanase and combinations thereof to soften the still existing seed coat, preferably combinations are used. As the seed coat is thinner and further softer after the enzyme treatment, the healthy fatty acids can be delivered more easily from the seeds during consumption and the mouth-feel is also better as the seed coat is softer. These sanded and enzyme treated seeds are particularly useful in various food applications, for example in snacks, cereals, muesli, bakery products, etc, and is animal feeds.
The enzyme treated sanded seeds may also be subjected to extraction of phenolics or seed oil using suitable extraction technique, such as conventional super critical or solvent extraction methods or combinations thereof. The enzyme treatment improves the extraction of fatty acids from the sanded seeds, whereby valuable seed oil may be obtained.
Alternatively the sanded seeds may be milled, followed by optional dry fractionation.
The fiber rich berry skin or fruit skin fraction separated from the seeds may be used as such or it may be further ground finer by a suitable mill, which is preferably a pin crusher. The amount of the skin fraction is typically 2-10 wt% of the berry material or fruit material, calculated on dry basis. As an example the skin fraction of cloudberry contains typically ellagic acid 20 mg/g dry weight. It may be used as an ingredient in cosmetics, food products and animal feeds.
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The hydrothermal extracts (aqueous mixtures or dry fractions) comprising bioactive compounds, obtained with process of the invention are rich in phenolic compounds, such as ellagic acid, ellagitannins and their derivatives and other bioactive compounds, and also C18 fatty acids. Said hydrothermal extracts can be used as natural preservatives in cosmetics, hygiene products, nutraceuticals, food products, food supplements, animal feeds, packages and in pharmaceutical products. The hydrothermal extracts comprising bioactive compounds with antimicrobial activity, particularly ellagitannins, obtained with process of the invention have particularly good effect against Staphylococcus aureus, as can be seen from the examples, also effect against Pseudomonas aeruginosa and Escherichia coli was shown.
In skin care and hygiene products the invention could also be utilized to convert berry and fruit materials to fractions that stabilize, balance and protect healthy skin microbiome, as these fractions inhibit the growth of many pathogenic microbes without effecting the growth of beneficial microbes.
The hydrothermal extracts comprising bioactive compounds with antimicrobial activity, obtained with process of the invention are suitably incorporated in food products, in cosmetic products, in hygiene products, in pharmaceutical products, in animal feeds, in packaging materials, particularly in packaging materials of products, such as food which is easily spoiled, and in pharmaceuticals, such as topical products like creams, ointments, etc.
Examples of said easily spoiled food products are poultry products, such as marinades, milk based products, such as yoghurts, drinks, sour cream products, fermented milk based products; berry or fruit containing products, such as jams, beverages, berry soups, conserves, pastes, purees, babyfood; nutritional food products, particularly for special use, such as hospital use and hose administration; grain products, such as bread, cereals, snack products, muesli, precooked porridge, fermented grain based products and gluten-free products.
The hydrothermal residues contain high amounts of ellagic acid (approx. 80 g/kg), C18 fatty acids, fibers and lignin, and thus they provide an excellent additive particularly for nutraceuticals, food products, food supplements and feeds.
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The present invention provides several advantages. Particularly the by-products, side streams and waste materials originating from berry and fruit processing industry can be effectively utilized in the simple and economic process of the invention, for obtaining mixtures and dry fractions comprising bioactive compounds, particularly ellagic acid and/or derivatives thereof, obtained with process of the invention, as well as skin fractions and sanded seed fractions, which also find several valuable uses. With the process of the invention practically all the waste and by-product material can be utilized effectively.
Said bioactive mixtures and fractions may be used as effective antioxidants, antimicrobial agents and preservatives, particularly in the field of cosmetics, hygiene products, food products and animal feeds, as well as in packages and in pharmaceutical products.
The process of the invention provides enrichment of phenolic compounds, such as of ellagic acid and ellagitannin and their derivatives, whereby said compounds are concentrated in specific fractions for further use. The specific fractions can be added as such to various products. Dumping of the waste materials to the landfills can be avoided or at least significantly reduced. This is also a clear environmental and ecological benefit. Nutritionally rich and valuable waste materials and by-products from the berry industry and fruit industry can be utilized in a simple and efficient way in food products and animal feeds, as well as in packages and in pharmaceutical products.
The antimicrobial effect of the hydrothermal extract is dose dependent.
The invention provides improved storage and microbiological safety to the products, as these fractions comprising the bioactive compounds act as preservatives. It is possible to decrease the amount of synthetic preservatives in the products and replace them by these natural compounds. In addition, in cosmetic products these natural compounds also balance skin microbiota, as they effectively inhibit the growth of skin pathogens, such as Staphylococcus.
The invention provides improved microbiological preservation, improved inhibition of oxidation reactions and increased antioxidant status to the products where they are incorporated. In general this means improved stability and microbiological safety. The hydrothermal extracts are readily soluble in water, which is an important technological benefit. The hydrothermal extracts contain no or only very limited amounts of free sugars, which improves stability of the products. Thus the hydrothermal extracts are not sticky and easily stored in frozen form. Whole procedure to prepare hydrothermal extracts is food grade, no toxic or harmful reagents or process steps are used. Thus the extracts are particularly suitable also for food purposes.
EXAMPLES
The following examples are illustrative of embodiments of the present invention, as described above, and they are not meant to limit the invention in any way.
Example 1: Sanding of cloudberry seeds separated from press cake
A dried pressed cake of cloudberry from a commercial juice pressing process was sieved. The seeds were separated from the skin fraction by using a vibratory sieve shaker with a 1.6 mm screen at settings of 10 minutes sieving time and 1.5 mm amplitude. The seeds were sanded by using an abrasive machine (barley pearling machine) for 15 minutes sanding time. Table 1 shows yields obtained by the different process steps and distribution of fractions obtained by sieving and sanding.
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Table 1
Process: Sample: Yields:
- Dried cloudberry press cake 100%
Separation of seeds from press cake by sieving Sieved seeds 93%
Skin fraction 7%
Sanding of sieved seeds (sanding time 15 min) Sanded seeds 69%
Seed coat powder 11%
Loss 13%
Example 2: Sanding of cloudberry seeds separated from press cake
A dried pressed cake of cloudberry from a commercial juice pressing process was 25 sieved. The seeds were separated from the skin fraction by using a vibratory sieve shaker with a 1.60 mm screen at settings of 5 minutes sieving time and 1.5 mm amplitude. The seeds were sanded by using an abrasive machine (barley pearling machine) for 15 minutes sanding time. The yields of the sieving and sanding fractions and distribution of fractions obtained by sieving and sanding are shown in Table 2.
Table 2
Process: Sample: Yields:
- Dried cloudberry press cake 100%
Separation of seeds from press cake by sieving Sieved seeds 96%
Skin fraction /Dry fraction A 4%
Sanding of sieved seeds (sanding time 15 min) Sanded seeds 74%
Seed coat powder/Dry fraction B 18%
Loss 4%
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Example 3: Sanding of raspberry seeds separated from press cake
Frozen raspberries were thawed and crushed with a pestle. The crushed raspberries were warmed up to 45°C and iopectinase Super 8X enzyme was added. The enzyme dosage was 100 nkat/g berries (i.e. 1.98 ml/1 kg berries, active 51000 nkat/ml). After the incubation time (4 hours), juice pressing was performed by a High Pressure Tincture Press H P5 presser. The amount of press cake was 16% by weight and the dry matter of the press cake was 48% by weight. The press cake was dried by a quick drying machine with an air flow at 45°C to a dry matter of 89%.
Seeds were separated from the dried press cake by using a vibratory sieve shaker with a 0.63 mm screen at settings of 5 minutes sieving time and 1.0 mm amplitude. The seeds were sanded by using an abrasive machine (barley pearling machine) for 15 minutes sanding time. The yields obtained by the different process steps and distribution of fractions obtained by the sieving and the sanding machines are shown in Table 3.
Table 3
Process: Sample: Yields:
- Dried raspberry press cake 100%
Separation of seeds from raspberry press cake by sieving Sieved seeds 96%
Skin fraction/Dry fraction A 4%
Sanding trial I:
Sanding of sieved seeds (sanding time 1 min) Sanded seeds 73%
Seed coat powder/Dry fraction B 4%
Loss 19%
Sanding trial II:
Sanding of sieved seeds (sanding time 2 min) Sanded seeds 72%
Seed coat powder/Dry fraction B 7%
Loss 17%
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Example 4: Sanding of seeds separated from cloudberries
Cloudberries were frozen and freeze-dried. The dried cloudberries (less than 15 wt% of water) were crushed by hand to separate the skin, fruit flesh and seed portion from the whole berries. The seeds were separated from the skin and fruit flesh portion by using a vibratory sieve shaker with a 1.6 mm screen. The cloudberry material was first sieved using the 5 minutes sieving time and 1.0 mm amplitude and after that sieved by using the same sieve settings with ten glass balls. The glass balls assisted to separate the fruits flesh and skin from the seeds. After that the seeds were sanded using an abrasive machine (barley pearling machine) for 15 and 30 minutes sanding time. The yields obtained by the different process steps and distribution of fractions obtained by sieving and sanding are shown in Table 4.
Table 4
Process: Sample: Yields:
- Freeze-dried cloudberries 100%
Seeds separating from berries by sieving twice (without and with 10 glass balls) Sieved seeds 48%
Skin and fruit flesh fraction 52%
Sanding trial I:
Sanding of sieved seeds (sanding time 15 min) Sanded seeds 38%
Seed coat powder/Dry fraction B 4%
Loss 6%
Sanding trial II:
Sanding of sieved seeds (sanding time 30 min) Sanded seeds 39%
Seed coat powder/Dry fraction B 6%
Loss 3%
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EXAMPLE 5: Fermentation of cloudberries, pressing and sieving
Fermentation
Frozen, ripe cloudberries (Rubus chamaemorus) were used as the berry material. The berry material was first heat treated and then inoculated with approximately 105 6 cfu g_1 of washed LAB cells. Pedicoccus pentosaceus VTT E-072742 from VTT Culture
Collection was used as a starter culture in the fermentation of cloudberries (http://culturecollection.vtt.fi/). Prior to fermentations, the strain was refreshed in de Man Rogosa Sharpe broth for 1 day in a 100% carbon dioxide atmosphere which was created using anaerobic jars and Anaerocult C strips. The cells were collected from refreshed cultures by centrifugation and washed once in Ringer's solution. The fermentations were performed in a 6 kg scale in a 15-1 capacity bioreactor for 3 days at 30 °C under constant mixing (130 rpm). The bioreactor was purged with sterilefiltered nitrogen gas in order to create anaerobic conditions. The viable counts of lactic acid bacteria and yeasts were determined before and after the fermentations using plate count technique. The results were expressed as colony-forming units (CFU) per gram of wet weight. The fermented berry mash was stored frozen.
Pressing and drying
After fermentation the berry mash was treated with a hydraulically operated highpressure tincture press using 5 litres filling material to separate juice and insoluble press cake.
The press cake from juice pressing was dried in a fluid bed dryer using +45°C air flow, until the water content of the berry press cake was below 15 % by weight. After that, the dried berry press cake was dry sieved using different sieve sizes or using a suction apparatus. A skin fraction having average particle less than 1250 pm was separated and seed fraction having average particle size of more than 750 pm was collected.
The mass yields of cloudberry press cake (fermented and non-fermented samples) were 8-10 %. About 5 % of the press cake consisted of peels and pulp, and remaining
95% were seeds.
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EXAMPLE 6: Hydrothermal extraction of cloudberry and sea buckthorn seed coat material
Cloudberry and sea buckthorn seed coat materials, obtained with the method as described in example 1, were processed, each as follows:
A.
g of the seed coat material was extracted with 1 I of water for 1 hour at 80°C (with 500 rpm mixing). The mixture was filtered using Miracloth® filter material with 22-25 pm pore size. The filtrate was freeze dried.
B.
50 g of the seed coat material was mixed with 1 I of water and 0.1 wt% of ascorbic acid agitated for 1 hour at 80°C (with 500rpm mixing). The mixture was filtered using Miracloth® filter material with 22-25 pm pore size. The filtrate was freeze dried. The extraction was carried out in acidic conditions in order to stabilize anthocyanins.
The antimicrobial activity of the cloudberry and sea buckthorn freeze dried materials was tested against selected microbes including Staphylococcus aureus and Pseudomonas aeruginosa.
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Antimicrobial activities were measured in liquid cultures. Freeze-dried materials of 1 7.5 mg ml’1 were suspended into microbial cultures. Microbial culture without berry material was used as positive control, and culture with antibiotic (chloramphenicol for bacteria or hygromycin B for C. albicans) was used for negative growth control. The microbial cultures were incubated in their optimal growth conditions, and growth by cell counts was followed by plate count during 24 h of cultivation.
The results show clear bactericidal activity of the cloudberry and sea buckthorn 10 fractions against Staphylococcus aureus compared to positive control culture with no antimicrobial agents. Cloudberry freeze dried material prepared by hydrothermal extraction (1 hour extraction in 80°C water) was the best inhibitor. Moderate growth inhibition was detected with Pseudomonas aeruginosa and Escherichia coli too.
Figure 3A shows the antimicrobial effect of hydrothermal extracts and water extracts without heating of cloudberry seed coat fractions against Staphylococcus aureus in liquid culture during cultivation for 24 hours. Culture with antibiotic chloramphenicol was used as negative control. Two different batches of sanded cloudberry seed coat powder were compared (A and B). Effect of extract was tested in two concentrations in the culture: 1 mg/ml and 5 mg/ml.
Figure 3B shows antimicrobial effect of hydrothermal extracts of cloudberry seed coat fraction (B) against Staphylococcus aureus. Concentration of extract tested was 5 mg/ml microbial culture. Abbreviations:
· cloudberry seed coat/ cold = extraction 1 h room temperature, freeze-drying • cloudberry seed coat = extraction lh, 80°C, freeze-drying • cloudberry seed coat/ +enzyme = 2 h 45°C + enzymes followed by extraction lh 80°C, freeze-drying • cloudberry seed coat/ no enzyme = 2 h 45°C, followed by extraction lh, 80°C, freeze-drying • cloudberry seed coat/ oven = extraction lh 80°C, oven drying (85°C)
Figure 3C shows antimicrobial effect of hydrothermal extracts of sea buckthorn seed coat fraction against Staphylococcus aureus. Concentrations of extracts tested were
2.5 mg/ml, 5 mg/ml and 7.5 mg/ml microbial culture.
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Figure 3D shows antimicrobial activity of hydrothermal extracts of cloudberry seed coat fraction (B) against Pseudomonas aeruginosa. Concentration of extracts tested were 5 mg/ml and 7.5 mg/ml microbial culture. Abbreviations:
• cloudberry seed coat/ cold = extraction 1 h room temperature, freeze-drying • cloudberry seed coat = extraction lh, 80°C, freeze-drying • cloudberry seed coat/ +enzyme = 2 h 45°C + enzymes followed by extraction lh 80°C, freeze-drying • cloudberry seed coat/ no enzyme = 2 h 45°C, followed by extraction lh, 80°C, freeze-drying • cloudberry seed coat/ oven = extraction lh 80°C, oven drying (85°C)
Figure 3E shows antimicrobial activity of hydrothermal extracts of cloudberry seed coat fraction B and buckthorn seed materials against Escherichia coli. Concentrations of extracts tested were 5.0 mg/ml microbial culture.
Abbreviations:
• cloudberry seed coat/ cold = extraction 1 h room temperature, freeze-drying • cloudberry seed coat = extraction lh, 80°C, freeze-drying • cloudberry seed coat/ +enzyme = 2 h 45°C + enzymes followed by extraction lh 80°C, freeze-drying • cloudberry seed coat/ no enzyme = 2 h 45°C, followed by extraction lh, 80°C, freeze-drying • cloudberry seed coat/ oven = extraction lh 80°C, oven drying (85°C)
While the invention has been described with respect to specific examples including presently preferred modes of carrying out the invention, those skilled in the art will appreciate that there are numerous variations and permutations of the above described embodiments that fall within the spirit and scope of the invention. It should be understood that the invention is not limited in its application to the details of construction and arrangements of the components set forth herein. Variations and modifications of the foregoing are within the scope of the present invention.

Claims (13)

1. Prosessi marja- ja hedelmämateriaalien konvertoimiseksi bioaktiivisia yhdisteitä 5 käsittäviksi fraktioiksi, tunnettu siitä, että prosessi käsittää vaiheet, missä ensimmäisessä vaiheessa vähintä yksi marja- tai hedelmämateriaali, joka valitaan marjojen, hedelmien, marjoista tai hedelmistä peräisin olevien sivutuotteiden, sivuvirtojen ja jätemateriaalien joukosta, jotka marjat ja hedelmät on valittu sukujen Rubus, Sorbus, Rosa, Empetrum, Aronia ja Hippophae ja niiden yhdistelmien joukostaA process for converting berry and fruit materials into fractions containing bioactive compounds 5, characterized in that the process comprises the first step of at least one berry or fruit material selected from berries, fruits, berry or fruit by-products, by-products and waste materials, selected from the genera Rubus, Sorbus, Rosa, Empetrum, Aronia and Hippophae and combinations thereof 10 ja hedelmät on valittu sukujen Vitis, Punica, Pyrus, Malus, Citrullus, Benincasa, Cucumis, Momordica, Olea ja lajin Argania spinosa ja niiden yhdistelmien joukosta, jonka mainitun marja- tai hedelmämateriaan vesipitoisuus on enintään 15 p-%, seulotaan, jolloin siemenfraktio erotetaan kuorifraktiosta, toisessa vaiheessa siemenfraktiolle suoritetaan hionta, missä 2-40 p-% siemenestä10 and the fruit is selected from the genera Vitis, Punica, Pyrus, Malus, Citrullus, Benincasa, Cucumis, Momordica, Olea and Argania spinosa and combinations thereof having a water content of up to 15% by weight of said berry or fruit material, wherein the seed fraction in a second step, the seed fraction is subjected to grinding, wherein 2-40% by weight of the seed 15 poistetaan ja saadaan siemenenpintafraktio, joka käsittää siementen pintakerroksen, ja hiottu siemenfraktio, joka käsittää hiotut siemenet, kolmannessa vaiheessa siemenenpintafraktio sekoitetaan vesipitoisen väliaineen kanssa painosuhteessa 1:3 ja 1:80 välillä, lämpötilassa välillä 15 ja 95°C, aikana, joka on 20 min ja 10 h välillä, jolloin saadaan bioaktiivisia yhdisteitä käsittävä seos, jaRemoving the seed surface fraction comprising the seed surface layer and the ground seed fraction comprising the ground seed, the third step mixing the seed surface fraction with an aqueous medium in a weight ratio of 1: 3 to 1:80 at a temperature between 15 and 95 ° C. between min and 10 h to give a mixture of bioactive compounds, and 20 seokselle suoritetaan erotus, missä hydrotermaalinen uute erotetaan hydrotermaalisesta jäämästä.The mixture is subjected to separation, whereby the hydrothermal extract is separated from the hydrothermal residue. 2. Patenttivaatimuksen 1 mukainen prosessi, tunnettu siitä, että bioaktiiset yhdisteet käsittävät ellagihappoa ja/tai sen johdannaisia, edullisesti ellagihappoa ja/taiProcess according to claim 1, characterized in that the bioactive compounds comprise ellagic acid and / or derivatives thereof, preferably ellagic acid and / or 25 ellagitanniineja.25 ellagitannins. 3. Patenttivaatimuksen 1 tai 2 mukainen prosessi, tunnettu siitä, että hydrotermaalinen uute kuivataan.Process according to claim 1 or 2, characterized in that the hydrothermal extract is dried. 3030 4. Jonkin patenttivaatimuksen 1-3 mukainen prosessi, tunnettu siitä, että kolmannessa vaiheessa siemenenpintafraktion suhde vesipitoiseen väliaineeseen on 1:5 ja 1:50 välillä, vastaavasti.Process according to one of Claims 1 to 3, characterized in that in the third step the ratio of the seed surface fraction to the aqueous medium is between 1: 5 and 1:50, respectively. 5. Jonkin patenttivaatimuksen 1-4 mukainen prosessi, tunnettu siitä, ettäProcess according to one of Claims 1 to 4, characterized in that 35 vesipitoinen väliaine valitaan veden ja veden ja askorbiinihapon seosten joukosta.The aqueous medium is selected from water and mixtures of water and ascorbic acid. 20165506 prh 25 -09- 201720165506 prh 25 -09- 2017 6. Jonkin patenttivaatimuksen 1-5 mukainen prosessi, tunnettu siitä, että kolmannessa vaiheessa vähintään yhtä entsyymiä, joka valitaan hiilihydraatteja hydrolysoivien entsyymien joukosta, edullisesti selluloosan, pektinaasin, ksylanaasin ja niiden yhdistelmien joukosta, lisätään vesipitoiseen väliaineeseen.Process according to one of Claims 1 to 5, characterized in that in the third step at least one enzyme selected from carbohydrate hydrolyzing enzymes, preferably cellulose, pectinase, xylanase and combinations thereof, is added to the aqueous medium. 7. Jonkin patenttivaatimuksen 1-6 mukainen prosessi, tunnettu siitä, että marjatai hedelmämateriaali esikäsitellään ennen seulontaa, ja marja- tai hedelmämateriaalin esikäsittely suoritetaan menetelmällä, joka valitaan lämpökäsittelyn, fermentoinnin, entsymaattisen käsittelyn, puristuksen, puserruksen,Process according to any one of claims 1 to 6, characterized in that the berry or fruit material is pretreated before screening, and the berry or fruit material is pretreated by a process selected from heat treatment, fermentation, enzymatic treatment, compression, pressing, 10 kuivauksen, murskauksen ja niiden yhdistelmien joukosta.10 drying, crushing and combinations thereof. 8. Patenttivaatimuksen 1-7 mukainen prosessi, tunnettu siitä, että marja- tai hedelmämateriaali fermentoidaan maitohappobakteerin avulla, edullisesti mainittu maitohappobakteeri valitaan sukujen Lactococcus, Lactobacillus, Pediococcus jaProcess according to Claims 1 to 7, characterized in that the berry or fruit material is fermented by means of a lactic acid bacterium, preferably said lactic acid bacterium is selected from the genera Lactococcus, Lactobacillus, Pediococcus and 15 Oenococcus joukosta.15 of Oenococcus. 9. Patenttivaatimuksen 1-8 mukainen prosessi, tunnettu siitä, että marja- tai hedelmämateriaali käsitellään hiilihydraatteja hydrolysoivien entsyymien avulla, edullisesti mainitut entsyymit valitaan selluloosan, pektinaasin, ksylanaasin ja niidenProcess according to claims 1-8, characterized in that the berry or fruit material is treated with carbohydrate hydrolysing enzymes, preferably said enzymes are selected from cellulose, pectinase, xylanase and their 20 yhdistelmien joukosta.20 combinations. 10. Jonkin patenttivaatimuksen 1-9 mukainen prosessi, tunnettu siitä, että seulonta suoritetaan seulontalaitteella, ilmaluokituslaitteella, ilmasuihkuseulalaitteella, verkkoseulalaitteella, rotaatioseulalaitteella tai värinäseulalaitteella.Process according to one of Claims 1 to 9, characterized in that the screening is carried out with a screening device, an air grading device, an air jet screening device, a mesh screening device, a rotary screening device or a vibrating screening device. 11. Jonkin patenttivaatimuksen 1-10 mukainen prosessi, tunnettu siitä, että hionta suoritetaan hionta- tai kiillotuslaitteella, joka valitaan viljankiillotuslaitteiden, riisinkiillotuslaitteiden, helmimyllyjen, kuorienpoistimien ja kiillotuslaitteiden joukosta.Process according to one of Claims 1 to 10, characterized in that the grinding is carried out with a grinding or polishing device selected from cereal polishers, rice polishers, pearl mills, peelers and polishers. 3030 12. Jonkin patenttivaatimuksen 1-11 mukainen prosessi, tunnettu siitä, että marjatai hedelmämateriaali kuivataan, kunnes sen vesipitoisuus on 0.1-10 p-%, edullisesti 0.1-8 p-%.Process according to one of Claims 1 to 11, characterized in that the berry or fruit material is dried until it has a water content of 0.1 to 10% by weight, preferably 0.1 to 8% by weight. 13. Jonkin patenttivaatimuksen 1-12 mukainen prosessi, tunnettu siitä, että marja35 tai hedelmämateriaali murskataan ennen seulontaa, edullisesti murskaimella, joka valitaan rullamurskaimen, pallomurskaimen, manuaalisen murskaimen, vaivainjauhimen ja niiden yhdistelmien joukosta.A process according to any one of claims 1 to 12, characterized in that the berry or fruit material is crushed before screening, preferably with a crusher selected from roll crusher, ball crusher, manual crusher, kneading crusher and combinations thereof.
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