WO2015199521A1 - Enzymatic method for extracting dry fruit kernels from a lignocellulosic shell - Google Patents

Enzymatic method for extracting dry fruit kernels from a lignocellulosic shell Download PDF

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Publication number
WO2015199521A1
WO2015199521A1 PCT/MA2015/000006 MA2015000006W WO2015199521A1 WO 2015199521 A1 WO2015199521 A1 WO 2015199521A1 MA 2015000006 W MA2015000006 W MA 2015000006W WO 2015199521 A1 WO2015199521 A1 WO 2015199521A1
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Prior art keywords
nuts
laccase
shell
enzymatic
layers
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PCT/MA2015/000006
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French (fr)
Inventor
Imane WAHBY
Redouane BENHIMA
Kadmiri Issam MEFTAH
Badreddine SIJILMASSI
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Moroccan Foundation For Advanced Science, Innovation & Research (Mascir)
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Publication of WO2015199521A1 publication Critical patent/WO2015199521A1/en

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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Y—ENZYMES
    • C12Y110/00—Oxidoreductases acting on diphenols and related substances as donors (1.10)
    • C12Y110/03—Oxidoreductases acting on diphenols and related substances as donors (1.10) with an oxygen as acceptor (1.10.3)
    • C12Y110/03002—Laccase (1.10.3.2)
    • 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
    • A23L25/00—Food consisting mainly of nutmeat or seeds; Preparation or treatment thereof
    • A23L25/20—Food consisting mainly of nutmeat or seeds; Preparation or treatment thereof consisting of whole seeds or seed fragments
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23N—MACHINES OR APPARATUS FOR TREATING HARVESTED FRUIT, VEGETABLES OR FLOWER BULBS IN BULK, NOT OTHERWISE PROVIDED FOR; PEELING VEGETABLES OR FRUIT IN BULK; APPARATUS FOR PREPARING ANIMAL FEEDING- STUFFS
    • A23N5/00—Machines for hulling, husking or cracking nuts
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P19/00—Preparation of compounds containing saccharide radicals
    • C12P19/14—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
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Y—ENZYMES
    • C12Y111/00—Oxidoreductases acting on a peroxide as acceptor (1.11)
    • C12Y111/01—Peroxidases (1.11.1)
    • C12Y111/01013—Manganese peroxidase (1.11.1.13)
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Y—ENZYMES
    • C12Y111/00—Oxidoreductases acting on a peroxide as acceptor (1.11)
    • C12Y111/01—Peroxidases (1.11.1)
    • C12Y111/01014—Lignin peroxidase (1.11.1.14)

Definitions

  • Enzymatic method for extracting almonds from dried fruits from a lignocellulosic shell Enzymatic method for extracting almonds from dried fruits from a lignocellulosic shell.
  • the present invention relates to a composition and a method for the recovery of edible almonds from nuts. More particularly, the invention relates to the treatment of hard outer layers, without prior treatment, with an enzyme-based composition. The recovery of almonds is then facilitated by the enzymatic treatment which is followed by a mechanical dehulling process of the treated layers, drying and recovery of the kernel.
  • Nuts have a strong and waterproof shell most of the time.
  • the shell of the cashew nut for example, is composed of two shells, one on the outside of green and thin color and the other inner of brown and hard color, separated by a cavity structure which contains a phenolic resin caustic called cashew shell oil or "Cashew Nut Liquid Shell CNSL".
  • the kernel or edible seed of cashew is covered by a thin skin called testa (Azam-Ali and Judge, 2001. Small scale cashew nut processing, FAO documents).
  • testa Azam-Ali and Judge, 2001. Small scale cashew nut processing, FAO documents.
  • the recovery of the kernel from the solid shells of walnuts is carried out by mechanical and / or physical or chemical methods.
  • the basic method of recovering almonds from the shells involves a separation of the edible almond from the outer shell and testa surrounding the almond without damaging it. This is a laborious process and must be done with care to ensure the quality of the kernel.
  • the patent WO2010092432 describes a hull depellicating device based on a mechanical breakage process by a chisel mounted on the device that allows to align the nuts. This device is confronted with the problems of the heterogeneous sizes of the nuts as well as the degree of the mechanical breakage by the chisel with a significant rate of damage of the almonds.
  • the heat treatment of the hard shells nuts which is another method of recovery, allows to weaken the outer shell whatever the caliber of the nuts (WO20010621 10 and EP1955603).
  • these heat treatments require a considerable energy which has repercussions on the cost of skinning.
  • the steam treatment ensures a softening of the shell of the nuts which allows easy recovery by hand or by longitudinal shear to obtain the kernel.
  • the prior drying of the nuts or their imbibition in water are adopted in order to increase the yield of intact kernels after the various treatments mentioned above.
  • Enzymes which hydrolyze lignocellulosic compounds for the production of fermentable sugars are described in several publications (CA 2786290, EP 2559768). These enzymes are not described in the treatment of dried fruit shells.
  • the present invention thus has several advantages in contrast to mechanical, physical or chemical methods through the use of an enzymatic treatment method.
  • the invention relates to a composition and method for the treatment of hard shells of dried fruits in general and cashew nuts in particular that have not undergone any pretreatment beforehand to easily recover edible almonds.
  • the invention applies to hulls that have a lignocellulosic nature with varying proportions.
  • the invention also describes the steps of the enzymatic post-treatment for the recovery of the intact kernel.
  • the enzymes used are those which are capable of oxidizing lignin or cellulose constituting the hard layer of nut shells. These enzymes are cellulase and its accessory enzymes, lignin peroxidase (LiP), manganese peroxidase (MnP) and laccase.
  • LiP lignin peroxidase
  • MnP manganese peroxidase
  • the treatment solution may contain the buffer, the mediator, the water and one or more of the laccase enzymes.
  • the enzyme used is laccase added simultaneously with a mediator.
  • the latter is defined as a chemical compound that is oxidized by laccase and whose oxidation product acts on the lignin of the shell to increase its hydrolysis.
  • the laccase mediator used in the invention and added to the enzyme solution is 2,2'-bis-bis (3-ethylbenzothiazoline-6-sulfonic acid (ABTS).
  • the enzymatic solution object of the invention is prepared in a sodium citrate or sodium acetate buffer or any other buffer with a concentration of between 50mM and 500mM.
  • the volume of the enzymatic solution is between 4 and 9 times the weight of the nuts to be treated.
  • the concentration of the enzyme of the laccase is 0.4 mg and 1 mg per gram of nuts this same concentration is used also for the mediator of laccase.
  • the enzymatic embrittlement of the hard layers of cashews can last between 4 and 8 days with continuous stirring at a temperature between 25 and 50 ° C.
  • the weakened shells are rinsed with water and the almond is collected directly by hand by applying pressure by the fingers to the center of the cashew nuts.
  • the enzymatic solution can be reused for 5 successive cycles with sustained weakening of the hard layers of cashews.
  • the hard layers weakened by the enzymatic treatment are rubbed against a rough wall or other means for removing the weakened layers.
  • the nuts are dried. Drying is done by evaporation of water by applying heat to the nuts. In the preferred embodiment of the invention the drying is carried out by a jet of hot air from 40 ° C to 65 ° C.
  • the last layer and the testa surrounding the kernel are easily removed by friction between the fingers, between two abrasive papers or any other mechanical application that mimics these movements.
  • the recovered almond is white in color and its edible quality is saved.
  • the invention offers several advantages, since the enzymatic treatment described is independent of the sizes of the nuts, the damage of the almonds is avoided and the recycling of the enzyme solution reduces the cost of this treatment.
  • FIG.1. Shows the steps of the process of enzymatic treatment of nut shells with the possibility of recycling the enzymatic solution.
  • FIG.2. Shows the spectra obtained by FTIR to characterize the chemical composition of the hard layers of cashews
  • FIG.3 Shows the appearance of the hull of cashews after enzymatic treatment with laccase by comparing it with a control hull
  • FIG.4 Shows the appearance of a nut dehulled its hard layers after enzymatic treatment and following a rub against a rough wall
  • FIG. 5 shows the monitoring of the absorbance of the oxidation of ABTS by the enzymatic solution of laccase recycled after 1 cycle of treatment of cashew nuts
  • Step 1 Determination of the composition of cashew shells: In order to characterize the chemical composition of cashew shells, the chemical composition of the shell of cashew nuts is determined by infrared spectroscopy Fourier transform (FTIR). The layers that constitute the shell and surrounding the kernel are four in number with diameters of between 2 and 3 mm. Each layer was isolated and analyzed. The ABB Bomem spectrometer in ATR mode with 16 scans and a pitch of 4 cm -1 was used and the spectra were recorded in the range 260-4000 cm -1 .
  • FTIR infrared spectroscopy Fourier transform
  • Fig.2 gives the spectra obtained for the 4 layers.
  • Table 1 shows characteristic peaks of lignin and cellulose for the upper layers of the shell. Lignin is therefore predominant in layer 1 and is responsible for the hardness of cashews. This result confirms the lignocellulosic nature of the hulls and explains the effect of laccase on the weakening of the hull of cashew nuts.
  • Step 2 Effect of treating cashew shells with the enzymatic solution:
  • the enzymatic reaction for the embrittlement of the cashew shell was conducted on untreated nuts having a weight of 15 g.
  • the treatment solution contains one enzyme among a laccase, a buffer from which sodium citrate with pH between 4.5 and 6 has been chosen.
  • the minimum concentration of the enzyme is 0.4 mg and the maximum concentration is lmg per gram of substrate. These same concentrations are used for the ABTS mediator.
  • the treatment cycle lasts between 4 and 10 days under constant stirring and at a temperature of 25-50 ° C.
  • the almonds are recovered by hand or other device for separating the nut from the rest of the layers.
  • the outer layers can also be rubbed off against a rough wall.
  • the deepest layer and testa that surrounds the kernel remain intact and thus protect the kernel from the residues of the enzyme.
  • the nuts After removal of the surface layers, the nuts are dried at a temperature between 40 and 100 ° C for 1 to 10 hours.
  • the layer 3 and the testa are removed mechanically.
  • Step 3 Recycling the enzymatic solution:
  • the enzyme solution was recycled for 5 cycles with a constant embrittlement of cashews.
  • the oils contained in the nuts are released into the solution which becomes more viscous and embrittlement of the shells is less.
  • This example shows the determination of the enzymatic activity after 1 cycle of reuse.
  • 0.1 ml of the enzymatic solution resulting from the treatment cycle is added to 1 ml of ABTS (Sigma, SA3219) and 1 ml of the pH 4.5 - 6 buffer.
  • the control samples contain 0.1 ml of distilled water at room temperature. the place of the enzymatic solution.
  • One unit of laccase activity is defined as the change in absorbance at 420nm (extinction coefficient in 10min which is equivalent to the ⁇ of ABTS oxidized in 10min.
  • Fig.5. The results of Fig.5. indicate a sustained laccase activity after 1 cashew cycle for 4 - 8 days. This activity resulted in oxidation of ABTS by laccase, the product of which is monitored at the absorbance of 420 nm. Based on these results, laccase activity after 1 cashew cycle is 2.2mmol ABTS oxidized per minute.

Abstract

The invention relates to a composition and a method for weakening the layers of lignocellulosic nature which constitute the shell of nuts for the purpose of easy and non-destructive recovery of the edible kernel. The process is characterized in that the lignocellulosic layers are treated with an enzyme-based composition and the kernel is subsequently recovered manually or by mechanically stripping off the treated layers, followed by drying of the stripped nuts.

Description

Méthode enzymatique pour extraire les amandes des fruits secs à partir d'une coque lignocellulosique.  Enzymatic method for extracting almonds from dried fruits from a lignocellulosic shell.
Domaine de l'invention : Field of the invention
[0001] La présente invention concerne une composition et une méthode pour la récupération des amandes comestibles à partir des fruits à coque. Plus particulièrement, l'invention concerne le traitement des couches extérieures dures, sans traitement préalable, par une composition à base d'enzymes. La récupération des amandes est alors facilitée par le traitement enzymatique qui est suivi par un procédé de dépelliculage mécanique des couches traitées, séchage et récupération de l'amande. The present invention relates to a composition and a method for the recovery of edible almonds from nuts. More particularly, the invention relates to the treatment of hard outer layers, without prior treatment, with an enzyme-based composition. The recovery of almonds is then facilitated by the enzymatic treatment which is followed by a mechanical dehulling process of the treated layers, drying and recovery of the kernel.
Etat de l'invention : State of the invention
[0002] Les fruits à coques disposent d'une coque solide et imperméable la plupart du temps. La coque de la noix de cajou par exemple, est composée de deux coquilles l'une à l'extérieure de couleur verte et fine et l'autre interne de couleur brune et dure, séparées par une structure à cavités qui contient une résine phénolique caustique appelée huile de la coque de cajou ou « Cashew Nut Shell liquide CNSL ». L'amande ou la graine comestible de la noix de cajou est couverte par une fine peau appelée testa (Azam-Ali et Judge, 2001. Small scale cashew nut processing. FAO documents). Au jour d'aujourd'hui, la récupération de l'amande à partir des coques solides des noix est réalisée par des méthodes mécaniques et/ou physiques ou chimiques. [0002] Nuts have a strong and waterproof shell most of the time. The shell of the cashew nut, for example, is composed of two shells, one on the outside of green and thin color and the other inner of brown and hard color, separated by a cavity structure which contains a phenolic resin caustic called cashew shell oil or "Cashew Nut Liquid Shell CNSL". The kernel or edible seed of cashew is covered by a thin skin called testa (Azam-Ali and Judge, 2001. Small scale cashew nut processing, FAO documents). As of today, the recovery of the kernel from the solid shells of walnuts is carried out by mechanical and / or physical or chemical methods.
[0003] Le procédé basique de la récupération des amandes à partir des coques implique une séparation de l'amande comestible à partir de la coque externe et de la testa qui entoure l'amande sans l'endommager. Il s'agit d'un processus laborieux et qui doit être fait avec précaution pour garantir la qualité de l'amande. [0004] Parmi les méthodes suivies, le brevet WO2010092432 décrit un dispositif de dépelliculage des coques basé sur un procédé de casse mécanique par un ciseau monté sur le dispositif qui permet d'aligner les noix. Ce dispositif est confronté aux problèmes des calibres hétérogènes des noix ainsi que le degré de la casse mécanique par le ciseau avec un taux important d'endommagement des amandes. The basic method of recovering almonds from the shells involves a separation of the edible almond from the outer shell and testa surrounding the almond without damaging it. This is a laborious process and must be done with care to ensure the quality of the kernel. Among the methods followed, the patent WO2010092432 describes a hull depellicating device based on a mechanical breakage process by a chisel mounted on the device that allows to align the nuts. This device is confronted with the problems of the heterogeneous sizes of the nuts as well as the degree of the mechanical breakage by the chisel with a significant rate of damage of the almonds.
[0005] Le traitement thermique des coques dures des noix qui est une autre méthode de récupération, permet de fragiliser la coque externe quelque soit le calibre des noix (WO20010621 10 et EP1955603). Cependant ces traitements thermiques demandent une énergie considérable qui se répercute sur le coût de dépelliculage. Le traitement par la vapeur assure un adoucissement de la coque des noix qui permet une récupération aisée par la main ou par cisaillement longitudinale afin d'obtenir l'amande. Avant l'enlèvement des coques, le séchage préalable des noix ou leur imbibition dans l'eau sont adoptés afin d'augmenter le rendement en amandes intactes après les différents traitements cités précédemment. The heat treatment of the hard shells nuts which is another method of recovery, allows to weaken the outer shell whatever the caliber of the nuts (WO20010621 10 and EP1955603). However, these heat treatments require a considerable energy which has repercussions on the cost of skinning. The steam treatment ensures a softening of the shell of the nuts which allows easy recovery by hand or by longitudinal shear to obtain the kernel. Before removal of the hulls, the prior drying of the nuts or their imbibition in water are adopted in order to increase the yield of intact kernels after the various treatments mentioned above.
[0006] Ces différentes méthodes présentent par conséquent des inconvénients liés à l'hétérogénéité de la taille des noix, une consommation importante en énergie, un taux d'endommagement des amandes comestibles importants. These different methods therefore have disadvantages related to the heterogeneity of the size of the nuts, a high energy consumption, a damage rate of important edible almonds.
[0007] Devant ces méthodes mécaniques, physiques et chimiques le traitement enzymatique adéquat peut présenter plusieurs avantages. Les enzymes qui hydrolysent les composés ligno- cellulosiques pour la production de sucres fermentescibles sont décrites dans plusieurs publications (CA 2786290 ; EP 2559768). Ces enzymes ne sont pas décrites dans le traitement des coques de fruits secs. Faced with these mechanical, physical and chemical methods the appropriate enzymatic treatment can have several advantages. Enzymes which hydrolyze lignocellulosic compounds for the production of fermentable sugars are described in several publications (CA 2786290, EP 2559768). These enzymes are not described in the treatment of dried fruit shells.
[0008] la présente invention présente ainsi plusieurs avantages contrairement aux méthodes mécaniques, physiques ou chimiques grâce au recours à une méthode de traitement enzymatique. The present invention thus has several advantages in contrast to mechanical, physical or chemical methods through the use of an enzymatic treatment method.
Description de l'invention : Description of the invention
[009] L'invention concerne une composition et méthode pour le traitement des coques dures des fruits secs en général et des noix de cajou en particulier qui n'ont subit aucun prétraitement préalable afin de récupérer aisément les amandes comestibles. L'invention s'applique aux coques qui ont une nature ligno-cellulosique avec des proportions variables. L'invention décrit également les étapes du post-traitement enzymatique pour la récupération de l'amande intacte. [009] The invention relates to a composition and method for the treatment of hard shells of dried fruits in general and cashew nuts in particular that have not undergone any pretreatment beforehand to easily recover edible almonds. The invention applies to hulls that have a lignocellulosic nature with varying proportions. The invention also describes the steps of the enzymatic post-treatment for the recovery of the intact kernel.
[0010] Dans certains modes de réalisation, les enzymes utilisées sont celles qui sont capables d'oxyder la lignine ou la cellulose constituant la couche dure des coques de noix. Ces enzymes sont la cellulase et ses enzymes accessoires, la lignine peroxydase (LiP), la manganèse peroxydase (MnP) et la laccase. In some embodiments, the enzymes used are those which are capable of oxidizing lignin or cellulose constituting the hard layer of nut shells. These enzymes are cellulase and its accessory enzymes, lignin peroxidase (LiP), manganese peroxidase (MnP) and laccase.
[0011] Dans un mode de réalisation préféré, la solution de traitement peut contenir le tampon, le médiateur, l'eau et une ou plusieurs enzymes parmi les laccases. In a preferred embodiment, the treatment solution may contain the buffer, the mediator, the water and one or more of the laccase enzymes.
[0012] Dans un mode de réalisation préféré, l'enzyme utilisée est la laccase ajoutée simultanément avec un médiateur. Ce dernier est définit comme un composé chimique qui est oxydé par la laccase et dont le produit d'oxydation agit sur la lignine de la coque pour augmenter son hydrolyse. Le médiateur de la laccase utilisé dans l'invention et ajouté à la solution enzymatique est le 2,2'azine-bis-(3-ethylbenzothiazoline-6-sulfonique acide (ABTS). In a preferred embodiment, the enzyme used is laccase added simultaneously with a mediator. The latter is defined as a chemical compound that is oxidized by laccase and whose oxidation product acts on the lignin of the shell to increase its hydrolysis. The laccase mediator used in the invention and added to the enzyme solution is 2,2'-bis-bis (3-ethylbenzothiazoline-6-sulfonic acid (ABTS).
[0013] La solution enzymatique objet de l'invention est préparée dans un tampon de citrate de sodium ou d'acétate de sodium ou tout autre tampon avec une concentration comprise entre 50mM et 500mM. Le volume de la solution enzymatique est compris entre 4 et 9 fois le poids des noix à traiter. La concentration de l'enzyme de la laccase est 0,4 mg et lmg par gramme de noix cette même concentration est utilisée également pour le médiateur de la laccase. The enzymatic solution object of the invention is prepared in a sodium citrate or sodium acetate buffer or any other buffer with a concentration of between 50mM and 500mM. The volume of the enzymatic solution is between 4 and 9 times the weight of the nuts to be treated. The concentration of the enzyme of the laccase is 0.4 mg and 1 mg per gram of nuts this same concentration is used also for the mediator of laccase.
[0014] Dans certain mode de réalisation la fragilisation enzymatique des couches dures des noix de cajou peut durer entre 4 et 8 jours avec agitation permanente à une température comprise entre 25 et 50°C. Les coques fragilisées sont rincées par l'eau et l'amande est récupérée directement par la main en appliquant une pression par les doigts au centre des noix de cajou. Après un premier cycle de traitement, la solution enzymatique peut être réutilisée pendant 5 cycles successifs avec une fragilisation maintenue des couches dures des noix de cajou. In some embodiments, the enzymatic embrittlement of the hard layers of cashews can last between 4 and 8 days with continuous stirring at a temperature between 25 and 50 ° C. The weakened shells are rinsed with water and the almond is collected directly by hand by applying pressure by the fingers to the center of the cashew nuts. After a first treatment cycle, the enzymatic solution can be reused for 5 successive cycles with sustained weakening of the hard layers of cashews.
[0015] Pour une industrialisation de l'invention, les couches dures fragilisées par le traitement enzymatique sont frottées contre une paroi rugueuse ou un autre moyen permettant d'enlever les couches fragilisées. [0016] Après enlèvement des couches supérieures dures les noix sont séchées. Le séchage est réalisé par évaporation d'eau en appliquant une chaleur sur les noix. Dans le mode préféré de l'invention le séchage est réalisé par un jet d'air chaud de 40°C à 65°C. For industrialization of the invention, the hard layers weakened by the enzymatic treatment are rubbed against a rough wall or other means for removing the weakened layers. After removal of the hard top layers, the nuts are dried. Drying is done by evaporation of water by applying heat to the nuts. In the preferred embodiment of the invention the drying is carried out by a jet of hot air from 40 ° C to 65 ° C.
[0017] La dernière couche et la testa qui entourent l'amande sont aisément enlevées par frottement entre les doigts, entre deux papiers abrasifs ou tout autre application mécanique qui mime ces mouvements. L'amande récupérée est de couleur blanche et sa qualité comestible est sauvegardée. The last layer and the testa surrounding the kernel are easily removed by friction between the fingers, between two abrasive papers or any other mechanical application that mimics these movements. The recovered almond is white in color and its edible quality is saved.
[0018] L'invention offre plusieurs avantages, vu que le traitement enzymatique décrit est indépendant aux calibres des noix, l'endommagement des amandes est évité et le recyclage de la solution enzymatique réduit le coût de ce traitement. The invention offers several advantages, since the enzymatic treatment described is independent of the sizes of the nuts, the damage of the almonds is avoided and the recycling of the enzyme solution reduces the cost of this treatment.
Brève description des figures Brief description of the figures
[0019] FIG.1. Montre les étapes du procédé de traitement enzymatique des coques des noix avec la possibilité de recycler la solution enzymatique. FIG.1. Shows the steps of the process of enzymatic treatment of nut shells with the possibility of recycling the enzymatic solution.
[0020] FIG.2. Montre les spectres obtenus par FTIR pour caractériser la composition chimique des couches dures des noix de cajou FIG.2. Shows the spectra obtained by FTIR to characterize the chemical composition of the hard layers of cashews
[0021] FIG.3 Montre l'aspect de la coque des noix de cajou après traitement enzymatique par la laccase en le comparant avec une coque témoin FIG.3 Shows the appearance of the hull of cashews after enzymatic treatment with laccase by comparing it with a control hull
[0022] FIG.4 Montre l'aspect d'une noix dépelliculée de ses couches dures après le traitement enzymatique et suite à un frottement contre une paroi rugueuse FIG.4 Shows the appearance of a nut dehulled its hard layers after enzymatic treatment and following a rub against a rough wall
[0023] FIG.5 Montre le suivi de l'absorbance de l'oxydation de l'ABTS par la solution enzymatique de laccase recyclée après 1 cycle de traitement de noix de cajou FIG. 5 shows the monitoring of the absorbance of the oxidation of ABTS by the enzymatic solution of laccase recycled after 1 cycle of treatment of cashew nuts
Mode de réalisation préféré de l'invention Preferred embodiment of the invention
Etape 1: Détermination de la composition des coques de noix de cajou: [0024] Afin de caractériser la composition chimique des coques des noix de cajou la composition chimique de la coque des noix de cajou est déterminée par spectroscopie infrarouge à transformée de fourrier (FTIR). Les couches qui constituent la coque et qui entourent l'amande sont au nombre de 4 avec des diamètres compris entre 2 et 3 mm. Chacune des couches a été isolée et analysée. Le spectromètre ABB Bomem en mode ATR avec 16 scans et un pas de 4 cm"1 a été utilisé et les spectres ont été enregistrés dans la gamme 260-4000 cm-1. Step 1: Determination of the composition of cashew shells: In order to characterize the chemical composition of cashew shells, the chemical composition of the shell of cashew nuts is determined by infrared spectroscopy Fourier transform (FTIR). The layers that constitute the shell and surrounding the kernel are four in number with diameters of between 2 and 3 mm. Each layer was isolated and analyzed. The ABB Bomem spectrometer in ATR mode with 16 scans and a pitch of 4 cm -1 was used and the spectra were recorded in the range 260-4000 cm -1 .
[0025] La Fig.2 donne les spectres obtenus pour les 4 couches. L'analyse des spectres FTIR présentée dans le tableau 1 montre des pics caractéristiques de la lignine et de la cellulose pour les couches supérieures de la coque. La lignine est par conséquent majoritaire dans la couche 1 et qui est responsable de la dureté des noix de cajou. Ce résultat confirme la nature lignocellulosique des coques et explique l'effet de la laccase sur la fragilisation de la coque des noix de cajou. Fig.2 gives the spectra obtained for the 4 layers. The analysis of FTIR spectra presented in Table 1 shows characteristic peaks of lignin and cellulose for the upper layers of the shell. Lignin is therefore predominant in layer 1 and is responsible for the hardness of cashews. This result confirms the lignocellulosic nature of the hulls and explains the effect of laccase on the weakening of the hull of cashew nuts.
Tableau 1: Analyse et interprétation des spectres FTIR des couches constituant la coque des noix de cajouTable 1: Analysis and interpretation of the FTIR spectra of the shell layers of cashews
Figure imgf000007_0001
Etape 2: Effet du traitement des coques de noix de cajou par la solution enzymatique:
Figure imgf000007_0001
Step 2: Effect of treating cashew shells with the enzymatic solution:
[0026] La réaction enzymatique pour la fragilisation de la coque de noix de cajou a été menée sur des noix non traitées ayant un poids de 15 g. La solution de traitement contient une enzyme parmi une laccase, un tampon parmi lesquels on a choisi le citrate de sodium avec pH compris entre 4,5 et 6. La concentration minimale de l'enzyme est de 0,4mg et la concentration maximale est de lmg par gramme de substrat. Ces mêmes concentrations sont utilisées pour le médiateur ABTS. The enzymatic reaction for the embrittlement of the cashew shell was conducted on untreated nuts having a weight of 15 g. The treatment solution contains one enzyme among a laccase, a buffer from which sodium citrate with pH between 4.5 and 6 has been chosen. The minimum concentration of the enzyme is 0.4 mg and the maximum concentration is lmg per gram of substrate. These same concentrations are used for the ABTS mediator.
[0027] Le cycle de traitement dure entre 4 et 10 jours sous une agitation constante et à la température de 25 - 50°C. The treatment cycle lasts between 4 and 10 days under constant stirring and at a temperature of 25-50 ° C.
[0028] Les résultats du traitement enzymatique de la coque de noix de cajou montrent un changement de coloration de la solution qui devient noirâtre. Ce changement de coloration indique l'effet de l'enzyme et du médiateur sur les couches superficielles de la coque avec libération de l'huile de coque de noix de cajou (CSNL) qui donne cette couleur noirâtre à la solution. The results of the enzymatic treatment of the cashew shell show a color change of the solution which becomes blackish. This change in color indicates the effect of the enzyme and mediator on the surface layers of the shell with release of cashew nut shell oil (CSNL) which gives this solution a blackish color.
[0029] L'aspect des couches externes traitées est illustré par la Fig.3. The appearance of the outer layers treated is illustrated in Fig.3.
[0030] Après fragilisation, les amandes sont récupérées par la main ou un autre dispositif permettant la séparation de la noix du reste des couches. Les couches externes peuvent également être enlevées par frottement contre une paroi rugueuse. La couche la plus profonde et la testa qui entoure l'amande restent intactes et protègent ainsi l'amande contre les résidus de l'enzyme. After embrittlement, the almonds are recovered by hand or other device for separating the nut from the rest of the layers. The outer layers can also be rubbed off against a rough wall. The deepest layer and testa that surrounds the kernel remain intact and thus protect the kernel from the residues of the enzyme.
[0031] L'aspect d'une noix dépelliculée de ses couches dures par frottement contre une paroi rugueuse est illustré par la Fig.4. The appearance of a walnut skinned hard layers by friction against a rough wall is illustrated in Fig.4.
[0032] Après enlèvement des couches superficielles, les noix sont séchées à une température comprise entre 40 et 100°C pendant 1 à 10 heures. After removal of the surface layers, the nuts are dried at a temperature between 40 and 100 ° C for 1 to 10 hours.
[0033] La couche 3 et la testa sont enlevées mécaniquement. The layer 3 and the testa are removed mechanically.
[0034] Le traitement enzymatique décrit dans l'exemple 2 a été appliqué sur 1 kg de noix de cajou, le rendement de récupération est supérieur à 95%. Etape 3: Recyclage de la solution enzymatique : The enzymatic treatment described in Example 2 was applied to 1 kg of cashew nuts, the recovery yield is greater than 95%. Step 3: Recycling the enzymatic solution:
[0035] Dans le but d'étudier la possibilité de réutiliser l'enzyme plusieurs fois, la solution enzymatique a été recyclée pendant 5 cycles avec une fragilisation constante des noix de cajou. Au bout du sixième cycle, les huiles contenues dans les noix sont libérées dans la solution qui devient plus visqueuse et la fragilisation des coques est moindre. In order to study the possibility of reusing the enzyme several times, the enzyme solution was recycled for 5 cycles with a constant embrittlement of cashews. At the end of the sixth cycle, the oils contained in the nuts are released into the solution which becomes more viscous and embrittlement of the shells is less.
[0036] Cet exemple montre la détermination de l'activité enzymatique après 1 cycle de réutilisation. 0,1 mL de la solution enzymatique qui résulte du cycle de traitement est est additionné à 1ml d'ABTS (Sigma, SA3219) et 1ml du tampon pH 4,5 - 6. Les échantillons témoins contiennent 0,1ml d'eau distillée à la place de la solution enzymatique. Une unité de l'activité de laccase est définie comme le changement d'absorbance à 420nm (coefficient d'extinction
Figure imgf000009_0001
dans 10min qui est équivalent au μΜ d'ABTS oxydé en 10min. This example shows the determination of the enzymatic activity after 1 cycle of reuse. 0.1 ml of the enzymatic solution resulting from the treatment cycle is added to 1 ml of ABTS (Sigma, SA3219) and 1 ml of the pH 4.5 - 6 buffer. The control samples contain 0.1 ml of distilled water at room temperature. the place of the enzymatic solution. One unit of laccase activity is defined as the change in absorbance at 420nm (extinction coefficient
Figure imgf000009_0001
in 10min which is equivalent to the μΜ of ABTS oxidized in 10min.
[0037] Les résultats de suivi de l'absorbance sont présentés sur Fig.5.  The results of monitoring the absorbance are shown in Fig.5.
[0038] Les résultats de la Fig.5. indiquent une activité maintenue de la laccase après 1 cycle de traitement des noix de cajou pendant 4 - 8 jours. Cette activité a résulté en une oxydation de l'ABTS par la laccase dont le produit est suivi à l'absorbance de 420nm. En se basant sur ces résultats l'activité de la laccase après 1 cycle de traitement des noix de cajou est de 2,2mmol d'ABTS oxydé par minute.  The results of Fig.5. indicate a sustained laccase activity after 1 cashew cycle for 4 - 8 days. This activity resulted in oxidation of ABTS by laccase, the product of which is monitored at the absorbance of 420 nm. Based on these results, laccase activity after 1 cashew cycle is 2.2mmol ABTS oxidized per minute.

Claims

Revendications : Claims:
1. Méthode pour l'extraction des amandes comestibles à partir des noix d'un fruit à coque comprenant les étapes suivantes : 1. Method for the extraction of edible kernels from the nuts of a tree nut comprising the following steps:
a) Mélange des noix avec une solution contenant des enzymes pendant une durée de temps.  a) Blend the nuts with a solution containing enzymes for a period of time.
b) Frottement mécanique des couches fragilisées de la coque des noix  b) Mechanical friction of the weakened layers of the shell of the nuts
c) Séchage des noix dépelliculées par frottement  c) Drying of dehulled nuts by rubbing
d) Récupération mécanique des amandes comestibles  d) Mechanical recovery of edible almonds
2. Méthode selon la revendication 1 caractérisée en ce que l'enzyme utilisée peut être la laccase, la lignine peroxydase, la manganèse peroxydase, la cellulase ou un mélange de ces enzymes ou tout enzyme qui dégrade les substrats ligno-cellulosiques. 2. Method according to claim 1 characterized in that the enzyme used may be laccase, lignin peroxidase, manganese peroxidase, cellulase or a mixture of these enzymes or any enzyme that degrades the lignocellulosic substrates.
3. Méthode selon les revendications 1 et 2 caractérisée en ce que la solution contient un ou plusieurs composée parmi lesquels une laccase, un tampon et un médiateur de la laccase. 3. Method according to claims 1 and 2 characterized in that the solution contains one or more compound among which a laccase, a buffer and a mediator of laccase.
4. Méthode selon les revendications 1, 2 et 3 caractérisée en ce que le médiateur de la laccase peut être l'ABTS ou tout autre produit chimique qui est oxydé par la laccase plusieurs cycles de réaction et avec un potentiel redox élevé. 4. Method according to claims 1, 2 and 3 characterized in that the mediator of the laccase can be ABTS or any other chemical that is oxidized by laccase several reaction cycles and with a high redox potential.
5. Méthode selon les revendications 1, 2, 3 et 4 caractérisée en ce que le tampon peut être toute solution aqueuse avec pH stable compris entre 4,5 et 6. 5. Method according to claims 1, 2, 3 and 4, characterized in that the buffer can be any aqueous solution with a stable pH of between 4.5 and 6.
6. Méthode selon les revendications précédentes caractérisée en ce que la durée du traitement enzymatique nécessaire pour fragiliser la coque est comprise entre 4 et 10 jours. 6. Method according to the preceding claims characterized in that the duration of the enzymatic treatment necessary to weaken the shell is between 4 and 10 days.
7. Méthode selon les revendications 1 à 6 caractérisée en ce qu'une agitation constante est assurée pendant toute la durée du traitement et peut être réalisée par hélice, par agitation orbital, par compression d'air ou tout autre dispositif de mélange des solutions aqueuses. 7. Method according to claims 1 to 6 characterized in that constant agitation is provided throughout the duration of the treatment and can be carried out by helix, orbital shaking, air compression or any other mixing device aqueous solutions .
8. Méthode selon les revendications 1 à 7 caractérisée en ce que le séchage est réalisé par un jet d'air chaud ou en étuve ou tout autre mécanisme d'évaporation d'eau. 8. Method according to claims 1 to 7 characterized in that the drying is carried out by a jet of hot air or in an oven or any other mechanism of water evaporation.
9. Méthode selon les revendications 1 à 7 caractérisée en ce que la récupération des amandes comestibles se fait par frottement mécanique des couches fragilisées de la coque avec des parois rugueuses ou des brosses métalliques. 9. Method according to claims 1 to 7 characterized in that the recovery of edible almonds is by mechanical friction of the weakened layers of the shell with rough walls or metal brushes.
10. Méthode selon les revendications 1 à 9 caractérisée en ce que la solution enzymatique est réutilisable jusqu'à 5 fois. 10. Method according to claims 1 to 9 characterized in that the enzymatic solution is reusable up to 5 times.
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Citations (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
FR1561968A (en) * 1967-11-30 1969-04-04
WO2001062110A2 (en) 2000-02-25 2001-08-30 Berlein Process Technology (Pty) Limited Process for shelling fruits with shells
WO2004060072A1 (en) * 2002-12-27 2004-07-22 Chi's Research Corporation Vegetable tenderizer
EP1955603A1 (en) 2007-02-06 2008-08-13 Guy Hervé Jean Method of shelling the seeds of nuts, associated device and food products obtained
WO2010092432A1 (en) 2009-02-10 2010-08-19 Mahipala Balasooriya Kankanamage Apparatus or machine for de-shelling cashew nuts or any other nuts
CA2786290A1 (en) 2011-08-18 2013-02-18 Andritz Inc. Enzymatic hydrolysis pretreatment of lignocellulosic materials
EP2622967A1 (en) * 2012-02-02 2013-08-07 Bühler Barth AG Device for processing plant remnants
WO2013182827A1 (en) * 2012-06-08 2013-12-12 Institut National Polytechnique De Toulouse Method of enzymatic treatment of a solid lignocellulosic material

Patent Citations (9)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
FR1561968A (en) * 1967-11-30 1969-04-04
WO2001062110A2 (en) 2000-02-25 2001-08-30 Berlein Process Technology (Pty) Limited Process for shelling fruits with shells
WO2004060072A1 (en) * 2002-12-27 2004-07-22 Chi's Research Corporation Vegetable tenderizer
EP1955603A1 (en) 2007-02-06 2008-08-13 Guy Hervé Jean Method of shelling the seeds of nuts, associated device and food products obtained
WO2010092432A1 (en) 2009-02-10 2010-08-19 Mahipala Balasooriya Kankanamage Apparatus or machine for de-shelling cashew nuts or any other nuts
CA2786290A1 (en) 2011-08-18 2013-02-18 Andritz Inc. Enzymatic hydrolysis pretreatment of lignocellulosic materials
EP2559768A1 (en) 2011-08-18 2013-02-20 Andritz, Inc. Enzymatic hydrolysis pretreatment of lignocellulosic materials
EP2622967A1 (en) * 2012-02-02 2013-08-07 Bühler Barth AG Device for processing plant remnants
WO2013182827A1 (en) * 2012-06-08 2013-12-12 Institut National Polytechnique De Toulouse Method of enzymatic treatment of a solid lignocellulosic material

Non-Patent Citations (6)

* Cited by examiner, † Cited by third party
Title
AZAM-ALI, SMALL SCALE CASHEW NUT PROCESSING, 2001
CHERIYAN S ET AL: "Enzymatic bioremediation of cashew nut shell liquid contamination", JOURNAL OF HAZARDOUS MATERIALS, ELSEVIER, AMSTERDAM, NL, vol. 176, no. 1-3, 15 April 2010 (2010-04-15), pages 1097 - 1100, XP027105096, ISSN: 0304-3894, [retrieved on 20091124] *
DONG YA-CHEN ET AL: "Biodegradation of chestnut shell and lignin-modifying enzymes production by the white-rot fungiDichomitus squalens,Phlebia radiata", BIOPROCESS AND BIOSYSTEMS ENGINEERING, SPRINGER, DE, vol. 37, no. 5, 8 September 2013 (2013-09-08), pages 755 - 764, XP035315935, ISSN: 1615-7591, [retrieved on 20130908], DOI: 10.1007/S00449-013-1045-9 *
ELEGIR G ET AL: "Laccase mediator system: Oxidation of recalcitrant lignin model structures present in residual kraft lignin", ENZYME AND MICROBIAL TECHNOLOGY, STONEHAM, MA, US, vol. 37, no. 3, 2 August 2005 (2005-08-02), pages 340 - 346, XP027765950, ISSN: 0141-0229, [retrieved on 20050802] *
HETTI PALONEN ET AL: "Role of oxidative enzymatic treatments on enzymatic hydrolysis of softwood", BIOTECHNOLOGY AND BIOENGINEERING, vol. 86, no. 5, 5 June 2004 (2004-06-05), pages 550 - 557, XP055074986, ISSN: 0006-3592, DOI: 10.1002/bit.20135 *
WUAN XIN TEH, MD. MOKARROM HOSSAIN, TRANG QUYNH TO, AND LEIGH ALDOUS: "Pretreatment of Macadamia Nut Shells with Ionic Liquids Facilitates Both Mechanical Cracking and Enzymatic Hydrolysis", ACS SUSTAINABLE CHEM. ENG, vol. 35, no. 1, 18 March 2015 (2015-03-18), pages 992 - 999, XP008177852, DOI: 10.1021/acssuschemeng.5b00126 *

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