JP6901715B2 - How to treat pineapple residue - Google Patents

Description

The present invention relates to a method for treating pineapple residue.

In Patent Document 1 (Japanese Unexamined Patent Publication No. 2016-044129), as a method for treating a pineapple fruit core, a cellulase enzyme treatment agent is used for cutting or crushed pineapple fruit core under the conditions of pH 4 to 8, preferably pH 5 to 7. A method of enzymatic treatment is shown. By this method, a new moisturizer that can be used as a raw material for cosmetics can be obtained, and the pineapple fruit core that has been treated as waste can be effectively used.

By the way, the cellulase enzyme agent provided as an industrial enzyme agent is an enzyme obtained from a microorganism and contains one or more types of cellulase, and in addition to cellobiohydrolase and endoglucanase, β-glucosidase and xylanase. , Includes hydrolases that decompose polysaccharides such as xylanase from reducing or non-reducing ends into monosaccharides. Therefore, it is expected that the product obtained by the method described in Patent Document 1 contains a large amount of glucose, and the actual measurement shows that the glucose content is high. Therefore, when the product obtained by this method is added to foods or the like, the sweetness is increased and the taste quality is affected, and the calorie intake is increased, which causes restrictions on use and inappropriate cases. Therefore, in order to use the processed product of pineapple fruit core in foods and the like, a treatment method in which the amount of glucose produced is suppressed as much as possible is desired. For that purpose, it was necessary to separate and purify only hemicellulase containing no cellulase, or to remove glucose after decomposition with an enzymatic agent.

By the way, there is known a method of obtaining a processed product having a high oligosaccharide content from a cellulosic raw material containing lignin, cellulose, hemicellulose and the like by using various cellulase enzyme agents. For example, in Patent Document 2 (Republished Patent WO2017 / 170919), β-xylosidase and β-glucosidase are prepared by incubating a cellulase enzyme preparation containing xylanase adjusted to pH 5.5 to 8 at 35 to 60 ° C. A method has been shown in which an inactivated cellulosic enzyme agent is used and hydrolyzed preferably at pH 6.0-8.0 to obtain a xylan-oligosaccharide. Patent Document 3 (Japanese Unexamined Patent Publication No. 2009-189293) describes a cellulase enzyme agent in which β-glucosidase activity is suppressed by suspending or dissolving the cellulase enzyme agent in an aqueous medium and heating the cellulase enzyme agent to 30 ° C. or higher at pH 7 or higher. It has been shown how to obtain cellooligosaccharides. Further, in Patent Document 4 (Japanese Unexamined Patent Publication No. 2006-204294), cellulase is preferably added to a cellulose slurry adjusted to pH 5.5 to 9.0, and cellulase other than β-glucosidase is added at a temperature of 5 to 60 ° C. A method of adsorbing an enzyme required for production on cellulose to further carry out an enzymatic reaction, and Patent Document 5 (Japanese Unexamined Patent Publication No. 63-226294) describes a substance containing insoluble cellulose and cellulase in an aqueous medium. The β-glucosidase is removed by separating the solid fraction after keeping it warm in an aqueous medium having a pH of 3 to 7, and further preferably washing the obtained solid fraction with a water-soluble solution having a pH of 3 to 7. After that, a method is shown in which the solid fraction is hydrolyzed preferably at pH 3 to 7 to obtain a product containing a high content of cellobiose.

However, in the method shown in Patent Document 2, although the β-glucosidase activity and the β-xylosidase activity are reduced, the cellulase activity remains, so that when the pineapple fruit core is treated with them, the pineapple fruit core is treated. It could not be said that oligosaccharides having a suppressed glucose content were produced. Patent Document 3 shows that the cellulase activity is not lost by heating on the alkaline side, but when the enzyme obtained by this method is used and treated in the same manner as in Patent Document 1. Cannot suppress the production of glucose. Further, in Patent Document 4, pulp from which most of lignin and hemicellulose have been removed is used, and in Patent Document 5, synthesized cellulose is used. Therefore, it was not considered that oligosaccharides other than cellobiose were produced even when the pineapple fruit core to which lignin or hemicellulose was attached was treated by the same method. Not only that, in Patent Documents 4 and 5, since cellulose is finally decomposed by cellulase, it is easily predicted that glucose will be produced from cellulose in the pineapple fruit core even with this treatment method.

Japanese Unexamined Patent Publication No. 2016-044129 Republished Patent WO2017 / 170919 Japanese Unexamined Patent Publication No. 2009-189293 Japanese Unexamined Patent Publication No. 2006-204294 Japanese Unexamined Patent Publication No. 63-226294

The invention according to the present application was made based on the above background technology, and the problem to be solved by the invention according to the present application is to reuse the pineapple residue treated as waste for foods and drinks, cosmetics, and the like. The purpose is to provide a processing method that can be performed.

Therefore, as a result of various trials and errors, the inventors of the present application first contacted the cellulase enzyme agent with the pineapple residue under neutral to alkaline conditions, and then added the solid portion obtained by solid-liquid separation in an acidic liquid. Upon warming, hemicellulose decomposition products were found to be produced in a state where the amount of glucose produced was suppressed, and the present invention was completed.

That is, the treatment method according to the present invention includes a first step of contacting the pineapple residue with a cellulase enzyme agent in an aqueous medium under neutral to alkaline conditions having a pH of 7 or higher, and a second step of solid-liquid separation after the contact. This is a method for treating a pineapple residue, which comprises a third step of leaving the solid portion obtained by separation in an aqueous medium.

According to the present invention, a treated product containing a hemicellulose decomposition product in which glucose production is suppressed can be obtained from the pineapple residue.

FIG. 1 is a flow chart showing an example of a processing method according to the present invention. FIG. 2 is an image of a thin layer chromatogram of the product obtained when the pineapple residues with different pretreatments were treated. Lanes 1 to 6 are treated according to the present invention, and lanes 1 and 4 are heat-treated at 100 ° C. for 10 minutes when unheated untreated pineapple fruit core residue is used. When the pineapple fruit core residue was used, lanes 3 and 6 showed the case of using the pineapple fruit core residue heat-treated by an autoclave at 121 ° C. for 15 minutes, and lanes 7 and 8 showed the pineapple fruit immediately under acidic conditions. In a comparative example in which the core residue was enzymatically treated, lane 7 used an unheated untreated pineapple fruit core residue, and lane 8 used a pineapple fruit core residue heat-treated by an autoclave at 121 ° C. for 15 minutes. Indicates the case where there was. FIG. 3 is an image of a thin layer chromatogram of the product obtained when the pineapple core residue was treated at different pH. FIG. 4 is an image of a thin layer chromatogram of the product obtained when the pineapple fruit core residue was treated with different types of cellulase enzyme agents.

The treatment method according to the present invention includes a first step of contacting the pineapple residue with a cellulase enzyme agent in an aqueous medium under neutral to alkaline conditions of pH 7 or higher, and a second step of solid-liquid separation after the contact. This is a method for treating a pineapple residue, which comprises a third step of leaving the solid portion obtained by separation in an aqueous medium.

The pineapple residue used in the method of the present application is a residue containing a so-called pineapple fruit core excluding the edible part and the peel from the fruit of pineapple, which is a plant belonging to the genus Bromeliad of the family Bromeliad, and is usually discarded after being processed into food. This is the part. Although the present invention utilizes this discarded portion, it may include an edible portion. For example, a fruit core portion obtained by removing the peel and edible portion from the pineapple and a residue (including the fruit core portion) after squeezing the juice from the pineapple fruit are used.

The pineapple residue is preferably cleaved and / or crushed in advance prior to contact with the cellulase enzyme preparation. Although it is possible to use a pineapple residue that has been heat-treated by autoclaving or boiling, it is preferable to use a pineapple residue that has not been heat-treated in the present invention. This is because the heat treatment tends to increase glucose in the treated product. On the other hand, a pineapple residue that has been subjected to a delignin treatment can also be used. The method of delignin treatment is not particularly limited, and for example, the pineapple residue is prepared in an alkaline aqueous solution of about 1 to 10% hypochlorous acid at room temperature or at a temperature of about 30 ° C. to 50 ° C. for 30 minutes to several hours. It is done by immersing.

The first step is a step of bringing the pineapple residue and the cellulase enzyme preparation into contact with each other in an aqueous medium under neutral to alkaline conditions having a pH of 7 or higher. The cellulase enzyme agent that can be used in the present invention may be an enzyme agent containing cellulase and hemicellulase. Cellulase is an enzyme having a cellulose-degrading activity, hemicellulase is an enzyme having a hemicellulose-degrading activity, and examples of the enzyme having a hemicellulose-degrading activity include xylanase, xylosidase, glucanase, pectinase, alabanase, mannanase, and mannosidase. The cellulase enzyme preparation may also include an enzyme that hydrolyzes a polysaccharide such as xylosidase, galactosidase, or glucosidase from a reducing end or a non-reducing end to produce a monosaccharide. Further, the cellulase enzyme agent that can be used in the present invention may be an enzyme agent called cellulase or hemicellulase, as well as an enzyme agent referred to by a name other than cellulase or hemicellulase such as pectinase or ferulate esterase. This is because enzyme agents called pectinase, ferulic acid esterase, and the like are labeled with enzyme names having strong enzymatic activity, and these enzyme agents derived from microorganisms may contain cellulase as a by-product. The origin of the cellulase enzyme preparation is not limited, and as exemplified in Patent Documents 2 and 3, various cellulase enzyme preparations derived from the genus Aspergillus, for example, Aspergillus niger, and various types derived from Trichoderma reesei. Enzymatic agents can be used.

The aqueous medium used in the first step is preferably water. Organic solvents such as alcohols may also be included, as long as the hemicellulase activity in the cellulase enzyme preparation is not significantly inactivated. The pH of the aqueous medium is 7 or higher. On the other hand, when the alkalinity is high, the production of glucose tends to be suppressed, but the amount of hemicellulose decomposition products produced tends to be small. Therefore, the pH is preferably adjusted to 10 or less, more preferably 9 or less, and preferably 7 or more and 8 or less. To. If the pH is less than 7, a large amount of glucose can be produced by contact between the cellulase enzyme preparation and the pineapple residue. Further, when the pH exceeds 10, the production of glucose tends to be suppressed, but the amount of hemicellulose decomposition products produced also tends to decrease.

The contact between the cellulase enzyme agent and the pineapple residue in the aqueous medium may be carried out under a temperature condition of 1 ° C. or higher, preferably about 20 ° C. to 40 ° C. Warming improves the contact between the cellulase enzyme agent and the pineapple residue, and tends to reduce the amount of glucose produced in the final product. However, if it is heated too much, not only glucose is produced by the activity of cellulase, but also the cellulase enzyme agent may be inactivated and the amount of hemicellulose decomposition product produced in the processed product may be reduced.

The contact time is also a matter that can be appropriately determined by those skilled in the art, and varies depending on the contact temperature, pH, and the amount of treatment, but is generally about 5 minutes to 1 hour. In this first step, it is considered that hemicellulase in the cellulase enzyme preparation is mainly adsorbed, so that sufficient time is required for the adsorption. Therefore, it is preferable to contact them for at least 5 minutes, preferably 10 minutes or more. On the other hand, there is little significance in contacting for a long time, and the amount of glucose produced tends to increase as the contact time becomes longer, so a maximum of 2 to 3 hours is sufficient.

The second step is a step of contacting the cellulase enzyme agent with the pineapple residue and then separating the solid portion and the liquid portion. By this step, the pineapple residue, the enzyme adsorbed on the residue, and the enzyme not adsorbed are separated. The method of solid-liquid separation is not particularly limited, and a method of filtration or centrifugation is exemplified. Moreover, you may take out the pineapple residue visually. After that, the solid portion separated into solid and liquid is subjected to a third step. At this time, it is preferable to wash the obtained solid portion with an aqueous medium such as water, if necessary.

The third step is a step of leaving the solid portion obtained by solid-liquid separation in an aqueous medium. In this step, the pineapple residue, which is a solid component, is decomposed by an enzymatic reaction, and the decomposition of the pineapple residue proceeds. Since it is considered that hemicellulase remains in the solid portion without being separated by solid-liquid separation, hydrolysis of the pineapple residue occurs and a hemicellulose decomposition product is produced. Therefore, in this third step, a hemicellulose decomposition product can be produced without newly adding a cellulase enzyme agent from the outside. The aqueous medium used in the third step is also preferably water. The aqueous medium may also contain an organic solvent such as an alcohol, as long as the hemicellulase activity is not significantly inactivated.

In the third step, since the solid portion gradually proceeds by the enzymatic reaction, the solid portion may be suspended in an aqueous medium and left to stand. At this time, it may be left under the condition that the enzymatic reaction occurs, and it may be only kept at room temperature of, for example, about 1 to 30 ° C. However, it is preferable to promote the enzymatic reaction for the purpose of producing and obtaining a hemicellulose decomposition product. From the viewpoint of promoting the enzyme reaction, it is preferably left under suitable reaction conditions for the remaining enzyme. Since hemicellulase generally has many types of enzymes having an optimum pH under pH 7 or less, particularly pH 4 to 6, the aqueous medium is under acidic pH below pH 7, preferably pH 4 or higher, for the production of hemicellulose decomposition products. Adjusted to 6 or less. Further, it is preferable to heat the temperature to 30 ° C. or higher and 60 ° C. or lower. From the viewpoint of further promoting decomposition, stirring is desired. The leaving time can be set as appropriate, but if it is heated for a long time, a large amount of xylose, glucose and other monosaccharides may be produced.

By this third step, a processed product composed of a solid portion derived from the pineapple residue and a liquid portion other than that is obtained. The solid portion contains fiber, which is a decomposition product of pineapple residue, and the liquid portion contains a hemicellulose decomposition product in which hemicellulose is decomposed by hemicellulose. This hemicellulose decomposition product includes oligosaccharides composed of 2 to 10 monosaccharides and polysaccharides having a sugar chain length of up to about 30 or more. The processed material is separated into a solid part and a liquid part as needed. The method of separation is also carried out by an appropriate method, and a method of filtration or centrifugation is exemplified. Further, regardless of whether or not the solid portion and the liquid portion are separated, the processed product may be subjected to a treatment such as heating to inactivate the enzyme, if necessary.

Most of the hemicellulose decomposition products contained in the liquid portion contain arabinose, xylose, and galactose as constituent sugars. Since it is considered that cellulase has been removed through the first and second steps, it is considered that they are mostly derived from hemicellulose. Further, the liquid portion may contain monosaccharides such as glucose in addition to xylose and galactose, but the amount of glucose produced can be suppressed as compared with the case where the treatment is performed without going through the first and second steps. Therefore, even when glucose is used without being separated from the liquid, a composition having less sweetness and low calories can be obtained. Further, since the liquid portion of the treated product obtained by the treatment method of the present invention contains various kinds of hemicellulose decomposition products containing arabinose, xylose, galactose, glucose and the like as constituent sugars, it can be used as a food composition aimed at health consciousness. Can be preferably used. Not only that, it can be suitably used as an additive for pharmaceutical compositions and as a material for cosmetic compositions, and further effective utilization of pineapple residues, particularly pineapple-derived wastes such as pineapple fruit cores, can be achieved.

The liquid portion obtained by solid-liquid separation of the processed product obtained by the above three steps can be used as it is as a raw material for each of the above compositions, or in the form of a dried product by concentration, freeze-drying or spray-drying. Needless to say, it can be used as a hemicellulose decomposition product having different constituent sugars by fractionation and purification by various methods. On the other hand, the solid portion in the processed product can be used as dietary fiber as it is or in the form of a dried product, or can be used as animal feed. Of course, the liquid portion and the solid portion may be used without being separated, and a method of obtaining a dried product containing a hemicellulose decomposition product after drying the obtained processed product can also be adopted.

Hereinafter, the invention according to the present application will be described based on examples, but it goes without saying that the present invention is not limited to the following examples.

Using the pineapple fruit core produced as waste, treatment with a cellulase enzyme agent was attempted. As the cellulase enzyme preparation, the commercially available trade name Multifect GC Extra (manufactured by DuPont: hereinafter referred to as "Multifect") was used. This cellulase enzyme agent comprises at least (hemi) cellulase, xylanase and glucanase from Trichoderma reesei.

FIG. 1 shows a method for treating pineapple residue. That is, after adding a phosphate buffer solution having a pH of 7 to 1 g of a fruit core residue obtained by crushing the fruit core of pineapple, the pH was adjusted to 7 with an aqueous sodium hydroxide solution. A cellulase enzyme preparation was added thereto, and the mixture was stirred and then left at 30 ° C. for 30 minutes. Then, the residue (R1) and the filtrate (S1) which are solid portions were obtained by filtration. After washing the residue (R1) with water, water was added to the residue to adjust the pH to 5 with hydrochloric acid. Then, it was heated at 50 degreeC for about 2 hours to obtain a processed product, and then it was filtered and separated into a filtrate (S2) which is a liquid part and a residue (R2) which is a solid part. Then, the filtrate (S2) was freeze-dried to obtain a dried product. Here, raw fruit core residue (untreated fruit core residue) that has not been heat-treated and heat-treated at 100 ° C. for 10 minutes or heat-treated at 121 ° C. for 15 minutes (autoclave treatment: AC treatment) was performed. The same treatment was performed on the heat treatment residue. As a comparative example, the untreated fruit core residue and the heat-treated residue that had been heat-treated at 121 ° C. for 15 minutes were immediately subjected to enzyme treatment under acidic conditions of pH 5.

The sugar component in the filtrate (S2) obtained from the pineapple fruit core was analyzed by thin layer chromatography (TLC). An image of the obtained thin layer chromatogram is shown in FIG. Moreover, when the sugar component of (S2) in the filtrate was analyzed, the results shown in Table 1 were obtained.

Next, the change in the constituent sugars was examined by acid hydrolysis of the filtrate (S2) in the untreated fruit core residue of Example. The results are shown in Table 2. The content of constituent sugars was measured using HPAEC-PAD (High Performance Anion Exchange Chromatography-Pulsed Amperometric Detection). Although not shown, when sugar in the filtrate (S2) was analyzed by high performance liquid chromatography, a sugar component (polysaccharide) having an average degree of polymerization of about 10 to 30 was detected, and this was acid hydrolyzed. Only the peak of monosaccharides was detected.

As shown in FIG. 2, in the filtrates (lanes 1, 2, and 3) left at pH 7, the amount of glucose produced in each of the fruit core residues was small, and oligosaccharide spots and other oligos were also produced. The formation of origin components, which are thought to be hemicellulose decomposition products containing sugars and polysaccharides, was also observed, but the amount was small. However, when the residue (R1) obtained in the second step was heated in an acidic liquid (third step), monosaccharides and oligosaccharides were found in any of the fruit core residues (lanes 4, 5, 6). Although an increase in the origin component was observed, the amount of increase in glucose was significantly higher in the untreated fruit core (lane 4), which was not heat-treated, than in the case of immediate enzymatic decomposition at pH 5 (lanes 7 and 8). It was suppressed. As shown in Table 1, when the first to third steps were performed using the autoclaved fruit core residue, the amount of glucose produced increased as compared with the case where the autoclave treatment was not performed. When the enzyme treatment was performed without going through the first and second steps, the amount of glucose produced was further increased. Further, as shown in Table 2, the filtrate (S2) contains arabinose, galactose, and xylose in addition to glucose, but glucose is hardly increased by acid hydrolysis, whereas arabinose, galactose, and xylose are contained. It increased significantly. From these facts, it was considered that the hemicellulose decomposition product was generated by the decomposition of hemicellulose by hemicellulose rather than the decomposition of cellulose by cellulase in the third step.

The same treatment as in Example 1 was carried out by changing the contact time in the third step. The results are shown in Table 3. This is the case when the untreated fruit core residue is used. The average degree of polymerization estimated from the amount of glucose, the total amount of sugar and the amount of reducing sugar reaches a peak at a contact time of 1 to 2 hours, and thereafter, the degree of polymerization tends to decrease and the amount of reducing sugar and the amount of glucose tend to increase. Was done. Therefore, the contact time is about 1 to 2 hours, and the contact time is about 4 hours at the longest, and a hemicellulose decomposition product with less glucose production can be obtained.

In the first step, the contacts were contacted at different pHs, and the effect of the pH at the time of contact was investigated. The pH was adjusted to 8, 9, and 10 with an aqueous sodium hydroxide solution, and the cellulase enzyme agent and the pineapple fruit core residue (untreated fruit core residue) were brought into contact with each other. The result is shown in FIG.

As a result, in the filtrate (S1), the amount of hemicellulose decomposition products produced decreased as the pH increased, and in the filtrate (S2), the amount of hemicellulose decomposition products produced did not change even if the pH at the time of contact increased, and glucose was produced. The amount has decreased. On the other hand, at around pH 10, the amount of glucose produced was remarkably small, but the amount of hemicellulose decomposition products produced was also small.

A treatment was attempted under the same conditions as in Example 1 using a cellulase enzyme agent different from the cellulase enzyme agent used in Example 1. Of the 24 types of enzyme preparations for the food industry obtained from Shin Nihon Kagaku Kogyo Co., Ltd., the decomposition activity for pineapple residue (the amount of reduction in the insoluble fraction when contacted with the untreated fruit core residue of pineapple under acidic conditions of pH 5). (Volume reduction ability)), three cellulase enzyme agents (Sumiteam C: cellulase derived from Trichoderma longibrachiatum, Sumiteam SPC: pectinase derived from Aspergillus niger, Sumiteam FE: ferulate esterase derived from Aspergillus niger) were used. The contact time in the third step was 1 hour and 2 hours. The result is shown in FIG. According to this, compared with the case where the treatment was carried out without going through the first and second steps, when the treatment according to the present invention was carried out, glucose production was slight or hardly observed, while on the other hand. It was confirmed that the amount of hemicellulose decomposition products produced increased. From this, it can be said that by contacting the pineapple residue with the cellulase enzyme agent under neutral to alkaline conditions having a pH of 7 or higher, decomposition by cellulase is suppressed and a treated product having a small amount of glucose produced can be obtained.

The present invention provides new uses for pineapple residues that have been treated as waste.

Claims (11)

The first step of contacting the pineapple residue containing the pineapple fruit core, which is obtained by removing the edible portion and the peel from the pineapple fruit , with the cellulase enzyme agent in an aqueous medium having a pH of 7 or higher.
The second step of solid-liquid separation after the contact and
A method for treating a pineapple residue, which comprises a third step of leaving the solid portion obtained by separation in an aqueous medium. The treatment method according to claim 1, wherein the contact is made in an aqueous medium having a pH of 7 or more and a pH of 10 or less in the first step. The treatment method according to claim 1 or 2, which is left at pH 4 or more and pH 6 or less in the third step. The first step of contacting the pineapple residue containing the pineapple fruit core, which is obtained by removing the edible portion and the peel from the pineapple fruit , with the cellulase enzyme agent in an aqueous medium having a pH of 7 or higher.
The second step of solid-liquid separation after the contact and
A method for producing a composition, which comprises a step of incorporating a processed product obtained by a treatment method having a third step of leaving the solid portion obtained by separation in an aqueous medium. The production method according to claim 4, wherein in the first step, contact is carried out in an aqueous medium having a pH of 7 or more and a pH of 10 or less. The production method according to claim 4 or 5, which is left at a pH of less than 7 in the third step. The treated product to be contained in the composition is a liquid portion and / or a solid portion obtained by solid-liquid separation of the standing mixture obtained in the third step, and / or a dried product obtained from them. The production method according to any one of claims 4 to 6. The production method according to any one of claims 4 to 6, wherein the processed product to be contained in the composition is a dried product of the neglected mixture obtained in the third step. The production method according to any one of claims 4 to 8, wherein the composition is any one of a food composition, a pharmaceutical composition, and a cosmetic composition. The first step of contacting the pineapple residue containing the pineapple fruit core, which is obtained by removing the edible portion and the peel from the pineapple fruit , with the cellulase enzyme agent in an aqueous medium having a pH of 7 or higher.
The second step of solid-liquid separation after the contact and
A method for producing a hemicellulose decomposition product, which comprises a third step of producing a hemicellulose decomposition product from the solid portion obtained by separation. The method for producing a hemicellulose decomposition product according to claim 10, wherein the third step is a step of bringing the solid portion obtained by separation into contact with an aqueous medium at a pH of less than 7.

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