WO2014178245A1 - Method for collecting protein contained in rice and protein obtained by said method - Google Patents

Method for collecting protein contained in rice and protein obtained by said method Download PDF

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WO2014178245A1
WO2014178245A1 PCT/JP2014/058913 JP2014058913W WO2014178245A1 WO 2014178245 A1 WO2014178245 A1 WO 2014178245A1 JP 2014058913 W JP2014058913 W JP 2014058913W WO 2014178245 A1 WO2014178245 A1 WO 2014178245A1
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protein
prolamin
rice
precipitate
added
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剛志郎 梶山
宏憲 藤竹
保坂　幸男
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株式会社サタケ
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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
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    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P3/00—Drugs for disorders of the metabolism
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
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    • A61P35/00—Antineoplastic agents
    • A61P35/02—Antineoplastic agents specific for leukemia

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  • the present invention relates to a method for recovering a protein contained in rice and a protein obtained by the method, and in particular, a protein recovery method that makes it possible to recover prolamin contained in rice with high purity and high recovery rate, and , Relating to the protein obtained by the method.
  • Patent Document 1 after dissolving rice in an alkaline solution, a solid-liquid separation operation is performed to separate the supernatant into a precipitate and a precipitate obtained by neutralizing the supernatant with an acid. A method of recovering proteins contained in rice by washing and drying is described.
  • Patent Document 2 ddH 2 O is added to rice to separate an albumin extract, NaCl is added to the remaining precipitate to separate the globulin extract, and then NaOH is added to the remaining precipitate to add gluten.
  • a method is described in which four proteins are sequentially recovered from rice by separating the extract and adding ethanol to the remaining precipitate to separate the prolamin extract.
  • Patent Document 1 According to the method described in Patent Document 1, it is expected that proteins contained in rice can be recovered at a high recovery rate. However, the method described in Patent Document 1 cannot selectively recover a protein, and when focusing on prolamin, there is a problem that the purity of the prolamin recovered is low.
  • an object of the present invention is to provide a protein recovery method capable of recovering prolamin contained in rice with high purity and high recovery rate, and a protein obtained by the method.
  • the protein recovery method of the present invention comprises: A protein dissolving step in which an alkaline solution and an alcohol solution are added to rice to dissolve the protein; A protein separation step of neutralizing the supernatant obtained in the protein dissolution step with an acid and precipitating unnecessary proteins in an alcohol solution other than prolamin; It is characterized by providing.
  • the protein recovery method preferably includes a prolamin recovery step of recovering prolamin from the supernatant obtained in the protein separation step.
  • the prolamin recovery step it is preferable to precipitate the prolamin dissolved in the alcohol solution by distilling off the alcohol solution from the supernatant obtained in the protein separation step.
  • the alcohol solution after adjusting the hydrogen ion concentration index, that is, pH by adding the alkaline solution.
  • the prolamin is preferably 13 kDa prolamin.
  • the protein recovery method preferably includes a protein recovery step of recovering a protein other than prolamin from the precipitate obtained in the protein separation step.
  • the protein of the present invention is prolamin obtained by any of the above methods.
  • the protein of the present invention is a protein obtained by any one of the above methods.
  • the purity of prolamin is the mass ratio (%) of prolamin contained in the recovered material recovered in the prolamin recovery step.
  • the recovery rate of prolamin is the mass ratio (%) of prolamin contained in the recovered product relative to prolamin contained in rice as a raw material.
  • the protein recovery method of the present invention includes a protein dissolution step in which an alkaline solution and an alcohol solution are added to rice to dissolve the protein, and the supernatant obtained in the protein dissolution step is neutralized with an acid, and is not required for alcohol solutions other than prolamin And a protein separation step of separating the protein into prolamin and a protein other than prolamin, so that prolamin contained in the rice can be recovered with high purity and high yield.
  • prolamin contained in rice can be recovered with high purity and high yield from the supernatant obtained in the protein separation step.
  • proteins other than prolamin contained in rice can be recovered from the precipitate obtained in the protein separation step.
  • Prolamin obtained by the protein recovery method can be widely used as a functional protein with high purity.
  • FIG. 1 is a flow diagram of a method for recovering protein contained in rice in an embodiment of the present invention.
  • FIG. 2 is a flowchart of a method for recovering proteins contained in rice in Comparative Example 1.
  • FIG. 3 is a flowchart of a method for recovering proteins contained in rice in Comparative Example 2.
  • 4 is a flowchart of a method for recovering proteins contained in rice in Comparative Example 3.
  • FIG. 5 is a flowchart of a method for recovering proteins contained in rice in Comparative Example 4.
  • FIG. 6 is an explanatory diagram of analysis results of raw materials and extracts in Examples and Comparative Examples 1 to 4.
  • FIG. 1 shows a flow of a protein recovery method in an embodiment of the present invention.
  • prolamin which is an indigestible protein
  • brown rice of LGC soft variety name
  • prolamin was extracted and collected through the following steps.
  • Protein dissolution step 2000 mL of ion-exchanged water was added to 500 g of the rice flour, and a hydrogen ion concentration index was adjusted to pH 12.5 using a NaOH aqueous solution as a strong alkaline aqueous solution. Subsequently, 3000 mL of ethanol was added as an alcohol, and the mixture was stirred for 2 hours in a constant temperature chamber at 40 ° C. Thereafter, centrifugation was performed at a centrifugal force of 3080 ⁇ g for 5 minutes, and the supernatant was recovered.
  • FIG. 2 shows the flow of the protein recovery method in Comparative Example 1.
  • Comparative Example 1 is based on the method described in Patent Document 1 above.
  • the same rice flour as in the examples was used as a raw material, n-hexane was added to the rice flour for degreasing, water and NaOH were added to adjust the pH to 12.5, and the mixture was stirred for 4 hours. Thereafter, centrifugation was performed, and sulfuric acid was added dropwise to the supernatant to neutralize and adjust to pH 3, and further centrifugation was performed to collect a precipitate. And the extract was obtained by drying the said precipitate like an Example.
  • FIG. 3 shows the flow of the protein recovery method in Comparative Example 2.
  • Comparative Example 2 is based on the method described in Patent Document 2 above.
  • the same rice flour as in Example was used as a raw material, n-hexane was added to the rice flour for degreasing, deionized distilled water, that is, ddH 2 O was added and shaken for 4 hours, and then centrifuged. The precipitate was collected.
  • ddH 2 O was added to the precipitate, NaCl was added to the precipitate after repeating the above operation, and the mixture was shaken for 4 hours, and then centrifuged to collect the precipitate.
  • NaCl was added to the precipitate, NaOH was added to the precipitate after the above operation was repeated, and the mixture was shaken for 30 minutes, and then centrifuged to collect the precipitate.
  • FIG. 4 shows the flow of the protein recovery method in Comparative Example 3.
  • Comparative Example 3 ethanol was added to the extract obtained in Comparative Example 1, followed by centrifugation, and an extract was obtained from the collected supernatant through the prolamin recovery step shown in FIG.
  • FIG. 5 shows the flow of the protein recovery method in Comparative Example 4.
  • ethanol was directly added to the raw rice flour after defatted in Comparative Example 2, and the precipitate was collected by centrifugation.
  • work was repeated by adding ethanol to the said deposit, and the extract was obtained through the prolamin collection process similar to an Example from the extract which mixed the supernatant liquid of the 2 times of this operation
  • FIG. 6 shows analysis results of raw materials and extracts in Examples and Comparative Examples 1 to 4.
  • Each component amount of the raw material is a mass (g) contained in 100 g of the raw material
  • each component amount of the extract is a mass (g) contained in the extract per 100 g of the raw material.
  • the purity of prolamin is the mass ratio (%) of prolamin in the extract
  • the recovery rate of prolamin is the mass ratio (%) of prolamin contained in the extract with respect to prolamin contained in the raw material.
  • the prolamin obtained in Examples and Comparative Examples 1 to 4 is 13 kDa prolamin.
  • the prolamin obtained in Comparative Example 1 has a high recovery rate but a low purity. Moreover, it turns out that the prolamin obtained by the comparative example 2 is remarkably low, and the purity is not so high as expected.
  • Comparative Example 3 has a low recovery rate and Comparative Example 4 has a very low recovery rate.
  • the prolamin obtained in the examples has the highest purity of 91.0% and the recovery rate is as high as 46.8%. Therefore, it turns out that a present Example is excellent as a method of collect
  • the above examples utilize the property that prolamin dissolves in 60-90% ethanol as an alcohol.
  • ethanol is distilled off from the supernatant containing the prolamin to remove the ethanol.
  • the prolamin precipitate can also be recovered.
  • a prolamin precipitate may be recovered from the prolamin-containing supernatant by salting out.
  • prolamin was extracted and collected from the supernatant collected in the protein separation step, but proteins other than prolamin can also be collected from the precipitate collected in the protein separation step.
  • prolamin contained in rice can be recovered with high purity and high yield, and is extremely useful.

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Abstract

A protein collection method provided with: a protein solubilization process for solubilizing protein by adding an alkaline solution and an alcohol solution to rice; and a protein separation process for neutralizing the supernatant obtained in the protein solubilization process with acid and precipitating non-prolamine, alcohol solution-insoluble proteins.

Description

米に含まれるタンパク質の回収方法及び該方法により得られるタンパク質Method for recovering protein contained in rice and protein obtained by the method
　本発明は、米に含まれるタンパク質を回収する方法及び該方法により得られるタンパク質に関し、特に、米に含まれるプロラミンを高純度かつ高回収率で回収することを可能とする、タンパク質回収方法、及び、該方法により得られるタンパク質に関するものである。 The present invention relates to a method for recovering a protein contained in rice and a protein obtained by the method, and in particular, a protein recovery method that makes it possible to recover prolamin contained in rice with high purity and high recovery rate, and , Relating to the protein obtained by the method.
　従来、米に含まれるタンパク質が、コレステロールの低下や白血病の抑制等、人体に有用な機能を有することが知られている（特許文献１，２参照）。 Conventionally, it is known that proteins contained in rice have useful functions for the human body, such as lowering cholesterol and suppressing leukemia (see Patent Documents 1 and 2).
　特許文献１には、米をアルカリ溶液に溶解させた後、固液分離操作を行って上澄液と沈殿物に分離し、該上澄液を酸で中和して得られた沈殿物を洗浄・乾燥することによって、米に含まれるタンパク質を回収する方法が記載されている。 In Patent Document 1, after dissolving rice in an alkaline solution, a solid-liquid separation operation is performed to separate the supernatant into a precipitate and a precipitate obtained by neutralizing the supernatant with an acid. A method of recovering proteins contained in rice by washing and drying is described.
　また、特許文献２には、米にｄｄＨ２Ｏを加えてアルブミン抽出液を分離し、残留した沈殿物にＮａＣｌを加えてグロブリン抽出液を分離した後、残留した沈殿物にＮａＯＨを加えてグルテン抽出液を分離し、残留した沈殿物にエタノールを加えてプロラミン抽出液を分離することによって、４種のタンパク質を米から順次回収する方法が記載されている。 Further, in Patent Document 2, ddH 2 O is added to rice to separate an albumin extract, NaCl is added to the remaining precipitate to separate the globulin extract, and then NaOH is added to the remaining precipitate to add gluten. A method is described in which four proteins are sequentially recovered from rice by separating the extract and adding ethanol to the remaining precipitate to separate the prolamin extract.
　上記特許文献１に記載された方法によれば、米に含まれるタンパク質を高回収率で回収できることが期待される。
　しかしながら、上記特許文献１に記載された方法は、タンパク質を選択的に回収できるものでなく、プロラミンに着目した場合、回収されるプロラミンの純度が低くなる問題がある。 According to the method described in Patent Document 1, it is expected that proteins contained in rice can be recovered at a high recovery rate.
However, the method described in Patent Document 1 cannot selectively recover a protein, and when focusing on prolamin, there is a problem that the purity of the prolamin recovered is low.
　他方、上記特許文献２に記載された方法によれば、プロラミンを高純度で回収できることが期待される。
　しかしながら、上記特許文献２に記載された方法では、工程の数だけ回収ロスが発生し、前記プロラミンの回収率が低下する懸念がある。 On the other hand, according to the method described in Patent Document 2, it is expected that prolamin can be recovered with high purity.
However, in the method described in the above-mentioned Patent Document 2, there is a concern that a recovery loss occurs as many as the number of steps, and the recovery rate of the prolamin is reduced.
特開２００６－２７３８４０号公報JP 2006-273840 A 特開２０１１－１５７３４１号公報JP 2011-157341 A
　そこで、本発明は、米に含まれるプロラミンを高純度かつ高回収率で回収することを可能とするタンパク質回収方法及び該方法により得られるタンパク質を提供することを目的とする。 Therefore, an object of the present invention is to provide a protein recovery method capable of recovering prolamin contained in rice with high purity and high recovery rate, and a protein obtained by the method.
　本発明者は、鋭意検討した結果、米にアルカリ溶液及びアルコール溶液を加えてタンパク質を溶解させることが、米に含まれるプロラミンを高純度かつ高回収率で回収することを可能とすることを見出し、以下の発明を完成するに至った。 As a result of intensive studies, the present inventor has found that the addition of an alkaline solution and an alcohol solution to rice to dissolve the protein makes it possible to recover prolamin contained in the rice with high purity and high recovery rate. The following inventions have been completed.
　即ち、本発明のタンパク質回収方法は、
　米にアルカリ溶液及びアルコール溶液を加えてタンパク質を溶解させるタンパク質溶解工程と、
　前記タンパク質溶解工程で得られる上清を酸で中和し、プロラミン以外のアルコール溶液に不要なタンパク質を沈殿させるタンパク質分離工程と、
を備えることを特徴とする。 That is, the protein recovery method of the present invention comprises:
A protein dissolving step in which an alkaline solution and an alcohol solution are added to rice to dissolve the protein;
A protein separation step of neutralizing the supernatant obtained in the protein dissolution step with an acid and precipitating unnecessary proteins in an alcohol solution other than prolamin;
It is characterized by providing.
　前記タンパク質回収方法は、前記タンパク質分離工程で得られる上清からプロラミンを回収するプロラミン回収工程を備えることが好ましい。 The protein recovery method preferably includes a prolamin recovery step of recovering prolamin from the supernatant obtained in the protein separation step.
　前記プロラミン回収工程は、前記タンパク質分離工程で得られる上清からアルコール溶液を留去することで、アルコール溶液に溶解したプロラミンを沈殿させることが好ましい。 In the prolamin recovery step, it is preferable to precipitate the prolamin dissolved in the alcohol solution by distilling off the alcohol solution from the supernatant obtained in the protein separation step.
　前記タンパク質溶解工程は、前記アルカリ溶液を加えて水素イオン濃度指数、すなわちｐＨを調整した後に前記アルコール溶液を加えることが好ましい。 In the protein dissolving step, it is preferable to add the alcohol solution after adjusting the hydrogen ion concentration index, that is, pH by adding the alkaline solution.
　前記プロラミンは、１３ｋＤａプロラミンであることが好ましい。 The prolamin is preferably 13 kDa prolamin.
　前記タンパク質回収方法は、前記タンパク質分離工程で得られる沈殿物からプロラミン以外のタンパク質を回収するタンパク質回収工程を備えることが好ましい。 The protein recovery method preferably includes a protein recovery step of recovering a protein other than prolamin from the precipitate obtained in the protein separation step.
　本発明のタンパク質は、前記いずれかの方法により得られるプロラミンである。 The protein of the present invention is prolamin obtained by any of the above methods.
　また、本発明のタンパク質は、前記いずれかの方法により得られるタンパク質である。 The protein of the present invention is a protein obtained by any one of the above methods.
　ここで、本発明において、プロラミンの純度とは、プロラミン回収工程で回収される回収物中に含まれるプロラミンの質量割合（％）である。また、プロラミンの回収率とは、原料となる米に含まれるプロラミンに対する前記回収物に含まれるプロラミンの質量割合（％）である。 Here, in the present invention, the purity of prolamin is the mass ratio (%) of prolamin contained in the recovered material recovered in the prolamin recovery step. Moreover, the recovery rate of prolamin is the mass ratio (%) of prolamin contained in the recovered product relative to prolamin contained in rice as a raw material.
　本発明のタンパク質回収方法は、米にアルカリ溶液及びアルコール溶液を加えてタンパク質を溶解させるタンパク質溶解工程と、前記タンパク質溶解工程で得られる上清を酸で中和し、プロラミン以外のアルコール溶液に不要なタンパク質を沈殿させて、前記タンパク質をプロラミンとプロラミン以外のタンパク質に分離させるタンパク質分離工程と、を備えるので、米に含まれるプロラミンを高純度かつ高収率で回収することが可能となる。 The protein recovery method of the present invention includes a protein dissolution step in which an alkaline solution and an alcohol solution are added to rice to dissolve the protein, and the supernatant obtained in the protein dissolution step is neutralized with an acid, and is not required for alcohol solutions other than prolamin And a protein separation step of separating the protein into prolamin and a protein other than prolamin, so that prolamin contained in the rice can be recovered with high purity and high yield.
　そして、前記タンパク質回収方法によれば、前記タンパク質分離工程で得られる上清から、米に含まれるプロラミンを高純度かつ高収率で回収することができる。 And according to the protein recovery method, prolamin contained in rice can be recovered with high purity and high yield from the supernatant obtained in the protein separation step.
　また、前記タンパク質回収方法によれば、前記タンパク質分離工程で得られる沈殿物から、米に含まれるプロラミン以外のタンパク質を回収することができる。 Further, according to the protein recovery method, proteins other than prolamin contained in rice can be recovered from the precipitate obtained in the protein separation step.
　前記タンパク質回収方法により得られるプロラミンは、純度の高い機能性タンパク質として広く利用できる。
Prolamin obtained by the protein recovery method can be widely used as a functional protein with high purity.
図１は本発明の実施例における米に含まれるタンパク質の回収方法のフロー図である。FIG. 1 is a flow diagram of a method for recovering protein contained in rice in an embodiment of the present invention. 図２は比較例１における米に含まれるタンパク質の回収方法のフロー図である。FIG. 2 is a flowchart of a method for recovering proteins contained in rice in Comparative Example 1. 図３は比較例２における米に含まれるタンパク質の回収方法のフロー図である。FIG. 3 is a flowchart of a method for recovering proteins contained in rice in Comparative Example 2. 図４は比較例３における米に含まれるタンパク質の回収方法のフロー図である。4 is a flowchart of a method for recovering proteins contained in rice in Comparative Example 3. FIG. 図５は比較例４における米に含まれるタンパク質の回収方法のフロー図である。FIG. 5 is a flowchart of a method for recovering proteins contained in rice in Comparative Example 4. 図６は実施例及び比較例１～４における原料及び抽出物の分析結果の説明図である。FIG. 6 is an explanatory diagram of analysis results of raw materials and extracts in Examples and Comparative Examples 1 to 4.
　以下、実施例に基づいて本発明を説明するが、本発明はこれに限定されるものではない。 Hereinafter, the present invention will be described based on examples, but the present invention is not limited thereto.
　図１は本発明の実施例におけるタンパク質回収方法のフローを示す。
　本実施例では、難消化性タンパク質であるプロラミンを効率良く得るため、高難消化性タンパク質米であるＬＧＣソフト（品種名）の玄米を使用することとし、前記玄米を９０％精米した後に製粉機で米粉にした。そして、前記米粉を原料とし、以下の各工程を経てプロラミンを抽出・回収した。 FIG. 1 shows a flow of a protein recovery method in an embodiment of the present invention.
In this example, in order to efficiently obtain prolamin, which is an indigestible protein, brown rice of LGC soft (variety name), which is a highly indigestible protein rice, is used. I made rice flour. Then, using the rice flour as a raw material, prolamin was extracted and collected through the following steps.
　（１）タンパク質溶解工程
　前記米粉５００ｇに対しイオン交換水２０００ｍＬを加え、強アルカリ水溶液としてＮａＯＨ水溶液を使い水素イオン濃度指数をｐＨ１２．５に調整した。次いで、アルコールとしてエタノール３０００ｍＬを加え、４０℃の恒温庫内で２時間攪拌した。その後、３０８０×ｇの遠心力で５分間遠心分離を行い、上清を回収した。 (1) Protein dissolution step 2000 mL of ion-exchanged water was added to 500 g of the rice flour, and a hydrogen ion concentration index was adjusted to pH 12.5 using a NaOH aqueous solution as a strong alkaline aqueous solution. Subsequently, 3000 mL of ethanol was added as an alcohol, and the mixture was stirred for 2 hours in a constant temperature chamber at 40 ° C. Thereafter, centrifugation was performed at a centrifugal force of 3080 × g for 5 minutes, and the supernatant was recovered.
　（２）タンパク質分離工程
　前記上清に、強酸として硫酸を滴下して中和しｐＨ３に調整した。その後、３０８０×ｇの遠心力で５分間遠心分離を行い、プロラミンを含む上清と、プロラミン以外のエタノールに不溶なタンパク質を含む沈殿物とを分離回収した。 (2) Protein separation step The supernatant was neutralized by adding sulfuric acid as a strong acid dropwise to adjust to pH 3. Thereafter, centrifugation was performed at a centrifugal force of 3080 × g for 5 minutes to separate and collect a supernatant containing prolamin and a precipitate containing a protein insoluble in ethanol other than prolamin.
　（３）プロラミン回収工程
　湯浴４０℃、圧力９０ｈＰａの減圧濃縮機を使い、前記プロラミンを含む上清からエタノールを留去した後、３０８０×ｇの遠心力で５分間遠心分離を行い、沈殿物を回収した。そして、前記沈殿物にエタノール５０ｍＬを加え、ボルテックスミキサーで混合した後、３０８０×ｇの遠心力で５分間遠心分離を行い、沈殿物を回収した（洗浄、脱脂の準備）。その後、前記沈殿物にｎ－ヘキサン５０ｍＬを加え、ボルテックスミキサーで混合した後、３０８０×ｇの遠心力で５分間遠心分離を行い、沈殿物を回収した（脱脂）。さらに、前記沈殿物にエタノール５０ｍＬを加え、ボルテックスミキサーで混合した後、３０８０×ｇの遠心力で５分間遠心分離を行い、沈殿物を回収した（洗浄、乾燥準備）。そして、前記沈殿物を室温で一晩、つまり約１６時間、予備乾燥し、前記予備乾燥した固形物を微粉末化して４０℃で２４時間乾燥することで抽出物を得た。 (3) Prolamin recovery step Using a vacuum concentrator with a hot water bath of 40 ° C and a pressure of 90 hPa, ethanol was distilled off from the supernatant containing prolamin, and then centrifuged for 5 minutes at a centrifugal force of 3080 xg to obtain a precipitate. Was recovered. And after adding ethanol 50mL to the said deposit and mixing with a vortex mixer, it centrifuged for 5 minutes with the centrifugal force of 3080 * g, and collect | recovered deposits (preparation of washing | cleaning and degreasing). Thereafter, 50 mL of n-hexane was added to the precipitate, mixed with a vortex mixer, and then centrifuged for 5 minutes at a centrifugal force of 3080 × g to collect the precipitate (degreasing). Furthermore, 50 mL of ethanol was added to the precipitate, mixed with a vortex mixer, and then centrifuged for 5 minutes with a centrifugal force of 3080 × g to collect the precipitate (washing and preparation for drying). Then, the precipitate was preliminarily dried at room temperature overnight, that is, about 16 hours, and the pre-dried solid was pulverized and dried at 40 ° C. for 24 hours to obtain an extract.
　［比較例１］
　図２は比較例１におけるタンパク質回収方法のフローを示す。比較例１は、上記特許文献１に記載された方法に基づくものである。
　比較例１では、実施例と同じ米粉を原料とし、該米粉にｎ－ヘキサンを加えて脱脂を行い、水とＮａＯＨを加えてｐＨ１２．５に調整し、４時間撹拌した。その後、遠心分離を行い、上清に硫酸を滴下して中和しｐＨ３に調整し、さらに遠心分離を行い、沈殿物を回収した。そして、前記沈殿物を実施例と同様に乾燥することで抽出物を得た。 [Comparative Example 1]
FIG. 2 shows the flow of the protein recovery method in Comparative Example 1. Comparative Example 1 is based on the method described in Patent Document 1 above.
In Comparative Example 1, the same rice flour as in the examples was used as a raw material, n-hexane was added to the rice flour for degreasing, water and NaOH were added to adjust the pH to 12.5, and the mixture was stirred for 4 hours. Thereafter, centrifugation was performed, and sulfuric acid was added dropwise to the supernatant to neutralize and adjust to pH 3, and further centrifugation was performed to collect a precipitate. And the extract was obtained by drying the said precipitate like an Example.
　［比較例２］
　図３は比較例２におけるタンパク質回収方法のフローを示す。比較例２は、上記特許文献２に記載された方法に基づくものである。
　比較例２では、実施例と同じ米粉を原料とし、該米粉にｎ－ヘキサンを加えて脱脂した後、脱イオン蒸留水、すなわちｄｄＨ２Ｏを加えて４時間揺らし、その後、遠心分離を行い、沈殿物を回収した。
　次に、前記沈殿物にｄｄＨ２Ｏを加え、前記作業を繰り返した後の沈殿物にＮａＣｌを加え、４時間揺らし、その後、遠心分離を行い、沈殿物を回収した。
次に、前記沈殿物にＮａＣｌを加え、前記作業を繰り返した後の沈殿物にＮａＯＨを加え、３０分間揺らし、その後、遠心分離を行い、沈殿物を回収した。 [Comparative Example 2]
FIG. 3 shows the flow of the protein recovery method in Comparative Example 2. Comparative Example 2 is based on the method described in Patent Document 2 above.
In Comparative Example 2, the same rice flour as in Example was used as a raw material, n-hexane was added to the rice flour for degreasing, deionized distilled water, that is, ddH 2 O was added and shaken for 4 hours, and then centrifuged. The precipitate was collected.
Next, ddH 2 O was added to the precipitate, NaCl was added to the precipitate after repeating the above operation, and the mixture was shaken for 4 hours, and then centrifuged to collect the precipitate.
Next, NaCl was added to the precipitate, NaOH was added to the precipitate after the above operation was repeated, and the mixture was shaken for 30 minutes, and then centrifuged to collect the precipitate.
　次に、前記沈殿物にＮａＯＨを加え、前記作業を繰り返した後の沈殿物にエタノールを加え、４時間揺らし、その後、遠心分離を行い、沈殿物を回収した。
　そして、前記沈殿物にエタノールを加え、前記作業を繰り返し、該作業の２回の上清を混合した抽出液から、実施例と同様のプロラミン回収工程を経て抽出物を得た。 Next, NaOH was added to the precipitate, ethanol was added to the precipitate after repeating the above operation, and the mixture was shaken for 4 hours, and then centrifuged to collect the precipitate.
And the ethanol was added to the said precipitate, the said operation | work was repeated, and the extract was obtained through the prolamin collection process similar to an Example from the extract which mixed the supernatant liquid of 2 times of this operation | work.
　［比較例３］
　図４は比較例３におけるタンパク質回収方法のフローを示す。
比較例３では、比較例１において得られた抽出物にエタノールを加えた後、遠心分離を行い、回収した上清から、図４に示されるプロラミン回収工程を経て抽出物を得た。 [Comparative Example 3]
FIG. 4 shows the flow of the protein recovery method in Comparative Example 3.
In Comparative Example 3, ethanol was added to the extract obtained in Comparative Example 1, followed by centrifugation, and an extract was obtained from the collected supernatant through the prolamin recovery step shown in FIG.
　［比較例４］
　図５は比較例４におけるタンパク質回収方法のフローを示す。
　比較例４では、比較例２において脱脂した後の原料米粉に、直接エタノールを加え、遠心分離を行い、沈殿物を回収した。
　そして、前記沈殿物にエタノールを加えることで、前記作業を繰り返し、該作業の２回の上清を混合した抽出液から、実施例と同様のプロラミン回収工程を経て抽出物を得た。 [Comparative Example 4]
FIG. 5 shows the flow of the protein recovery method in Comparative Example 4.
In Comparative Example 4, ethanol was directly added to the raw rice flour after defatted in Comparative Example 2, and the precipitate was collected by centrifugation.
And the said operation | work was repeated by adding ethanol to the said deposit, and the extract was obtained through the prolamin collection process similar to an Example from the extract which mixed the supernatant liquid of the 2 times of this operation | work.
　図６は、実施例及び比較例１～４における原料及び抽出物の分析結果を示す。
　原料の各成分量は、原料１００ｇ中に含まれる質量（ｇ）であり、抽出物の各成分量は、原料１００ｇ当たりの抽出物中に含まれる質量（ｇ）である。
　また、プロラミンの純度は、抽出物におけるプロラミンの質量割合（％）であり、プロラミンの回収率は、原料に含まれるプロラミンに対する抽出物に含まれるプロラミンの質量割合（％）である。
　ここで、実施例及び比較例１～４により得られるプロラミンは１３ｋＤａプロラミンである。 FIG. 6 shows analysis results of raw materials and extracts in Examples and Comparative Examples 1 to 4.
Each component amount of the raw material is a mass (g) contained in 100 g of the raw material, and each component amount of the extract is a mass (g) contained in the extract per 100 g of the raw material.
The purity of prolamin is the mass ratio (%) of prolamin in the extract, and the recovery rate of prolamin is the mass ratio (%) of prolamin contained in the extract with respect to prolamin contained in the raw material.
Here, the prolamin obtained in Examples and Comparative Examples 1 to 4 is 13 kDa prolamin.
　図６に示されるように、比較例１により得られたプロラミンは、回収率は高いものの純度が低いことが分かる。また、比較例２により得られたプロラミンは、回収率が著しく低く、純度も期待したほど高くないことが分かる。 As can be seen from FIG. 6, the prolamin obtained in Comparative Example 1 has a high recovery rate but a low purity. Moreover, it turns out that the prolamin obtained by the comparative example 2 is remarkably low, and the purity is not so high as expected.
　他方、比較例３及び４により得られたプロラミンは、純度はある程度高いものの、比較例３では回収率が低く、比較例４では回収率が著しく低いことが分かる。 On the other hand, the prolamins obtained in Comparative Examples 3 and 4 have high purity to some extent, but it can be seen that Comparative Example 3 has a low recovery rate and Comparative Example 4 has a very low recovery rate.
　これに対し、実施例により得られたプロラミンは、純度が９１．０％と最も高く、回収率も４６．８％と高い。
　したがって、本実施例は、高純度のプロラミンを効率よく回収する方法として優れていることが分かる。 On the other hand, the prolamin obtained in the examples has the highest purity of 91.0% and the recovery rate is as high as 46.8%.
Therefore, it turns out that a present Example is excellent as a method of collect | recovering highly pure prolamins efficiently.
　そして、図６に示される実施例と比較例１～４の分析結果の比較から、原料米粉にＮａＯＨとエタノールを加えてタンパク質を溶解させることが、米に含まれるプロラミンを高純度かつ高収率で回収することを可能とすることを見出すことができる。 From the comparison of the analysis results of the example shown in FIG. 6 and Comparative Examples 1 to 4, the addition of NaOH and ethanol to the raw rice flour to dissolve the protein results in high purity and high yield of prolamin contained in the rice. It can be found that it can be recovered with.
　なお、上記実施例では、米粉にＮａＯＨを加えた後にエタノールを加えたが、ＮａＯＨとエタノールを同時に加えた場合、エタノールを加えた後にＮａＯＨを加えた場合、及び硫酸で中和する前の上清にエタノールを加えた場合でも、米に含まれるプロラミンを高純度かつ高収率で回収することができる。 In addition, in the said Example, ethanol was added after adding NaOH to rice flour, but when adding NaOH and ethanol simultaneously, when adding NaOH after adding ethanol, and the supernatant before neutralizing with a sulfuric acid Even when ethanol is added to, prolamin contained in rice can be recovered with high purity and high yield.
　また、上記実施例は、プロラミンが、アルコールである６０～９０％エタノールに溶解する性質を利用するものであり、プロラミン回収工程において、前記プロラミンを含む上清からエタノールを留去して該エタノールの濃度を下げることでプロラミンの沈殿物を回収することとしたが、前記上清に大量の水を加えてエタノールの濃度を低下させたり、エタノールを加えて濃度を９０％以上に上昇させたりすることでプロラミンの沈殿物を回収することもできる。 The above examples utilize the property that prolamin dissolves in 60-90% ethanol as an alcohol. In the prolamin recovery step, ethanol is distilled off from the supernatant containing the prolamin to remove the ethanol. We decided to collect the prolamin precipitate by lowering the concentration, but adding a large amount of water to the supernatant to lower the ethanol concentration, or adding ethanol to increase the concentration to 90% or more. The prolamin precipitate can also be recovered.
　さらに、上記実施例におけるプロラミン回収工程では、エタノール濃度を調整することに代えて、塩析により前記プロラミンを含む上清からプロラミンの沈殿物を回収することとしてもよい。 Furthermore, in the prolamin recovery step in the above example, instead of adjusting the ethanol concentration, a prolamin precipitate may be recovered from the prolamin-containing supernatant by salting out.
　上記実施例では、前記タンパク質分離工程で回収される上清からプロラミンを抽出・回収したが、前記タンパク質分離工程で回収される沈殿物からプロラミン以外のタンパク質を回収することもできる。 In the above examples, prolamin was extracted and collected from the supernatant collected in the protein separation step, but proteins other than prolamin can also be collected from the precipitate collected in the protein separation step.
　本発明のタンパク質回収方法では、米に含まれるプロラミンを高純度かつ高収率で回収することが可能であり、極めて利用価値が高い。 In the protein recovery method of the present invention, prolamin contained in rice can be recovered with high purity and high yield, and is extremely useful.

Claims (8)

  1. 　米にアルカリ溶液及びアルコール溶液を加えてタンパク質を溶解させるタンパク質溶解工程と、
    　前記タンパク質溶解工程で得られる上清を酸で中和し、プロラミン以外のタンパク質を沈殿させるタンパク質分離工程と、
    を備えることを特徴とするタンパク質回収方法。     A protein dissolving step in which an alkaline solution and an alcohol solution are added to rice to dissolve the protein;
    A protein separation step of neutralizing the supernatant obtained in the protein dissolution step with an acid and precipitating proteins other than prolamin;
    A protein recovery method comprising:
  2. 　前記タンパク質分離工程で得られる上清からプロラミンを回収するプロラミン回収工程を備える請求項１記載のタンパク質回収方法。 The protein recovery method according to claim 1, further comprising a prolamin recovery step of recovering prolamin from the supernatant obtained in the protein separation step.
  3. 　前記プロラミン回収工程は、前記タンパク質分離工程で得られる上清からアルコール溶液を留去し、前記プロラミンを沈殿させる請求項２記載のタンパク質回収方法。 The protein recovery method according to claim 2, wherein in the prolamin recovery step, an alcohol solution is distilled off from the supernatant obtained in the protein separation step to precipitate the prolamin.
  4. 　前記タンパク質溶解工程は、前記アルカリ溶液を加えて水素イオン濃度指数を調整した後に前記アルコール溶液を加える請求項１乃至３のいずれか一項記載のタンパク質回収方法。 The protein recovery method according to any one of claims 1 to 3, wherein in the protein dissolution step, the alcohol solution is added after adjusting the hydrogen ion concentration index by adding the alkaline solution.
  5. 　前記プロラミンは、１３ｋＤａプロラミンである請求項１乃至４のいずれか一項記載のタンパク質回収方法。 The protein recovery method according to any one of claims 1 to 4, wherein the prolamin is 13 kDa prolamin.
  6. 　前記タンパク質分離工程で得られる沈殿物からプロラミン以外のタンパク質を回収するタンパク質回収工程を備える請求項１乃至５のいずれか一項記載のタンパク質回収方法。 The protein recovery method according to any one of claims 1 to 5, further comprising a protein recovery step of recovering a protein other than prolamin from the precipitate obtained in the protein separation step.
  7. 　前記請求項１乃至６のいずれか一項記載の方法により得られるプロラミン。 A prolamin obtained by the method according to any one of claims 1 to 6.
  8. 　前記請求項１乃至６のいずれか一項記載の方法により得られるタンパク質。 A protein obtained by the method according to any one of claims 1 to 6.
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