JP7156627B2 - Method for producing krill component-containing composition - Google Patents
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Description
本発明は、医薬品素材や食品素材などとして用いることができるオキアミ成分含有組成物の製造方法に関する。 TECHNICAL FIELD The present invention relates to a method for producing a krill component-containing composition that can be used as a pharmaceutical material, food material, or the like.
オキアミ（Ｅｕｐｈａｕｓｉａｃｅａ）は、養殖魚のエサや釣りのエサなどとして利用されている海洋生物であることは周知の通りである。また、オキアミの一種であるツノナシオキアミ（Ｅｕｐｈａｕｓｉａ ｐａｃｉｆｉｃａ）は、三陸地方ではイサダと呼ばれ、食用として馴染み深い。 As is well known, krill (Euphausiacea) is a marine organism that is used as bait for farmed fish and bait for fishing. Also, Euphausia pacifica, which is a kind of krill, is called Isada in the Sanriku region and is familiar as food.
近年、オキアミには、エイコサペンタエン酸（ＥＰＡ）やドコサヘキサエン酸（ＤＨＡ）などのω－３系脂肪酸、１－アルキルエーテル型リン脂質をはじめとする様々な生理活性成分が含まれていることが知られるに至り、オキアミ由来の医薬品素材や食品素材の研究開発が精力的に行われている（例えば特許文献１や特許文献２）。しかしながら、オキアミに含まれている生理活性成分を医薬品素材や食品素材として提供するためのさらなる方法の探索は、今なお意義深い状況にある。 In recent years, krill has been found to contain various physiologically active components, including omega-3 fatty acids such as eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA), and 1-alkyl ether-type phospholipids. Research and development of krill-derived pharmaceutical materials and food materials have been vigorously conducted (for example, Patent Document 1 and Patent Document 2). However, the search for further methods for providing the physiologically active ingredients contained in krill as pharmaceutical materials and food materials is still in a significant situation.
そこで本発明は、医薬品素材や食品素材などとして用いることができるオキアミ成分含有組成物の製造方法を提供することを目的とする。 Accordingly, an object of the present invention is to provide a method for producing a krill component-containing composition that can be used as a pharmaceutical material, a food material, and the like.
本発明者らは上記の点に鑑みて鋭意検討を行った結果、水中に懸濁したオキアミの破砕物を、所定の酸性条件下で麹菌由来のプロテアーゼを用いて酵素処理することにより、医薬品素材や食品素材などとして用いることができるオキアミ成分含有組成物を製造することができることを見出した。 The present inventors conducted extensive studies in view of the above points, and found that a pharmaceutical material was obtained by enzymatically treating crushed krill suspended in water using a protease derived from Aspergillus oryzae under predetermined acidic conditions. It was found that a krill component-containing composition that can be used as a food material or the like can be produced.
上記の知見に基づいてなされた本発明のオキアミ成分含有組成物の製造方法は、請求項１記載の通り、水中に懸濁したオキアミの破砕物を、ｐＨ３～４の条件下で麹菌由来のプロテアーゼを用いて酵素処理することによって得られる処理液を、オイル画分を含む液分と固形分に分離した後、得られたオイル画分を含む液分をオイル画分と水画分に分離し、得られたオイル画分を乾燥することによる。
また、請求項２記載の製造方法は、請求項１記載の製造方法において、ｐＨの調整を有機酸および／または炭酸を用いて行うことによる。
また、請求項３記載の製造方法は、請求項２記載の製造方法において、有機酸がクエン酸である。
The method for producing a krill component-containing composition of the present invention, which has been made based on the above findings, is as described in claim 1. As described in claim 1, crushed krill suspended in water is treated with krill-derived protease under pH 3-4 conditions. After separating the treated liquid obtained by enzymatic treatment using , by drying the resulting oil fraction.
Moreover, the manufacturing method of claim 2 is based on the manufacturing method of claim 1, wherein the pH is adjusted using an organic acid and/or carbonic acid.
Moreover, the manufacturing method according to claim 3 is the manufacturing method according to claim 2, wherein the organic acid is citric acid.
本発明によれば、医薬品素材や食品素材などとして用いることができるオキアミ成分含有組成物の製造方法を提供することができる。 ADVANTAGE OF THE INVENTION According to this invention, the manufacturing method of the krill component containing composition which can be used as a pharmaceutical material, a food material, etc. can be provided.
本発明のオキアミ成分含有組成物の製造方法は、水中に懸濁したオキアミの破砕物を、ｐＨ３～４の条件下で麹菌由来のプロテアーゼを用いて酵素処理することによって得られるオイル画分を乾燥することによるものである。 In the method for producing a krill component-containing composition of the present invention, crushed krill suspended in water is enzymatically treated with krill-derived protease under pH 3-4 conditions, and the oil fraction obtained is dried. It is due to
本発明において原料として用いるオキアミは、特に限定されるものではなく、三陸地方でイサダと呼ばれているツノナシオキアミの他、ナンキョクオキアミ（Ｅｕｐｈａｕｓｉａ ｓｕｐｅｒｂａ）などであってもよい。オキアミは生のものを用いてもよいし、凍結したもの（例えば無加水や加水の冷凍ブロックであって－５０℃以下で保存したもの）を用いてもよい。 The krill used as a raw material in the present invention is not particularly limited, and may be Euphausia superba, in addition to the horned krill called Isada in the Sanriku region. The krill may be used fresh or frozen (for example, frozen block with no water or water and stored at −50° C. or below).
オキアミの破砕は、例えば、室温環境下において、－１０～－３℃に維持したオキアミを（凍結したオキアミはこの温度域まで解凍してから）、ミートチョッパーを用いて粗く行った後、マスコロイダーを用いてペースト状になるまで行うことが、優れた品質のオキアミ成分含有組成物を製造することができる点において望ましい。 The krill is crushed, for example, in a room temperature environment, the krill maintained at -10 to -3 ° C. (frozen krill is thawed to this temperature range), coarsely using a meat chopper, and then masscolloider. It is desirable in that it is possible to produce a krill component-containing composition of excellent quality.
こうして得られたオキアミの破砕物を、水中に懸濁し、ｐＨ３～４の条件下で麹菌（Ａｓｐｅｒｇｉｌｌｕｓ ｏｒｙｚａｅ）由来のプロテアーゼを用いて酵素処理する。オキアミの破砕物の水中への懸濁は、例えばオキアミの破砕物１重量部に０．３～０．７重量部の水を加えて撹拌することで行えばよい。麹菌由来のプロテアーゼは、麹菌由来の酸性プロテアーゼ、中性プロテアーゼ、アルカリ性プロテアーゼから選択される少なくとも１種であってよく、市販のものでは、これらの混合製剤であるコクラーゼＰ顆粒（三菱ケミカルフーズ社製）、中性プロテアーゼ製剤であるオリエンターゼＯＰ（エイチビィアイ社製）やスミチームＦＰ（新日本化学工業社製）などを用いることができる。麹菌由来のプロテアーゼは、例えばオキアミの破砕物１重量部に対して０．００１～０．０３重量部用い、その至適温度でオキアミの破砕物を酵素処理する。麹菌由来のプロテアーゼとしてコクラーゼＰ顆粒を用いる場合の至適温度は、５０～５５℃である。処理時間は、例えば３０分間～３時間であってよい。麹菌由来のプロテアーゼを用いたオキアミの破砕物の酵素処理を、ｐＨ３～４の条件下で行うのは、オキアミの破砕物を酵素処理することによって得られるオイル画分に含まれる脂質が、オキアミ自身が持つ脂質分解酵素によって分解されてしまうことを防ぐためである。なお、ｐＨの調整は、クエン酸、乳酸、酢酸、グルクロン酸、アスコルビン酸などの有機酸や、炭酸を用いて行えばよいが、中でもクエン酸を用いてｐＨの調整を行うことが、医薬品素材や食品素材などを製造するための安全性を確保する点で望ましい。クエン酸は、例えばオキアミの破砕物の水懸濁液１重量部に対して０．００１～０．０３重量部用いればよい。 The crushed krill thus obtained is suspended in water and enzymatically treated with a protease derived from Aspergillus oryzae under pH 3-4 conditions. The suspension of the crushed krill in water may be carried out, for example, by adding 0.3 to 0.7 parts by weight of water to 1 part by weight of the crushed krill and stirring. The koji mold-derived protease may be at least one selected from koji mold-derived acidic protease, neutral protease, and alkaline protease. ), neutral protease formulations such as Orientase OP (manufactured by HBI) and Sumiteam FP (manufactured by Shin Nihon Chemical Industry Co., Ltd.). For example, 0.001 to 0.03 parts by weight of protease derived from Aspergillus oryzae is used per 1 part by weight of crushed krill, and the crushed krill is subjected to enzymatic treatment at the optimum temperature. The optimum temperature for using Cochrase P granules as the koji mold-derived protease is 50 to 55°C. Treatment times may be, for example, 30 minutes to 3 hours. The enzymatic treatment of the crushed krill material using the krill-derived protease is performed under the condition of pH 3 to 4 because the lipids contained in the oil fraction obtained by enzymatically treating the crushed krill material are the krill itself. This is to prevent it from being decomposed by the lipolytic enzymes possessed by The pH may be adjusted using organic acids such as citric acid, lactic acid, acetic acid, glucuronic acid and ascorbic acid, or carbonic acid. It is desirable in terms of ensuring safety for manufacturing food materials and the like. Citric acid may be used, for example, in an amount of 0.001 to 0.03 parts by weight per 1 part by weight of the water suspension of crushed krill.
オキアミの破砕物を酵素処理した後、例えば処理液を７０～７５℃に加温して１０分間～１時間インキュベートすることでプロテアーゼの失活を行ってから、処理液に含まれるオイル画分を回収する。処理液からのオイル画分の回収は、例えば、室温環境下において、脱水型デカンタを用いて処理液を遠心分離することによってオイル画分を含む液分と固形分に分離した後、得られたオイル画分を含む液分を濃縮型デカンタを用いて遠心分離することによってオイル画分と水画分に分離することで行えばよい。 After enzymatically treating the crushed krill, for example, the treated solution is heated to 70 to 75° C. and incubated for 10 minutes to 1 hour to deactivate the protease, and then the oil fraction contained in the treated solution is extracted. to recover. The oil fraction is recovered from the treated liquid, for example, in a room temperature environment, by centrifuging the treated liquid using a dehydrated decanter to separate the oil fraction and the solid content, and then obtained. The liquid fraction containing the oil fraction may be separated into an oil fraction and a water fraction by centrifuging using a concentrated decanter.
最後に水画分から分離したオイル画分を乾燥することで、目的物であるオキアミ成分含有組成物を得る。オイル画分の乾燥は、例えばフリーズドライ法によって行えばよいが、スプレードライ法や熱風乾燥法などによって行うこともできる。目的物であるオキアミ成分含有組成物が固形物として得られた場合、顆粒状や粉末状に粉砕してもよい。 Finally, the oil fraction separated from the water fraction is dried to obtain the target krill component-containing composition. Drying of the oil fraction may be performed, for example, by a freeze-drying method, but can also be performed by a spray-drying method, a hot-air drying method, or the like. When the target krill component-containing composition is obtained as a solid, it may be pulverized into granules or powder.
以上の方法によって製造される本発明のオキアミ成分含有組成物は、例えば、タンパク質を３０～６０重量％の割合で、脂質を２５～５０重量％の割合で、水分を１～１５重量％の割合で含むものであり、とりわけ、脂質として哺乳動物において細胞の機能保持などのために重要な役割を果たしているとされるプラズマローゲン（ｐｌａｓｍａｌｏｇｅｎ）に生体内で変換されることが知られている１－アルキルエーテル型リン脂質を多く含む（本発明のオキアミ成分含有組成物の１～５重量％の割合）点が特筆に値する。従って、本発明のオキアミ成分含有組成物は、医薬品素材や食品素材などとして健康の維持や増強のために用いることができる。 The krill component-containing composition of the present invention produced by the above method has, for example, a protein content of 30 to 60% by weight, a lipid content of 25 to 50% by weight, and a water content of 1 to 15% by weight. In particular, it is known to be converted in vivo to plasmalogen, which is said to play an important role in maintaining cell functions in mammals as a lipid 1- It is worth noting that it contains a large amount of alkyl ether type phospholipid (1 to 5% by weight of the krill component-containing composition of the present invention). Therefore, the krill component-containing composition of the present invention can be used as a pharmaceutical material, a food material, and the like for maintaining and enhancing health.
以下、本発明を実施例によって詳細に説明するが、本発明は以下の記載に限定して解釈されるものではない。 EXAMPLES The present invention will be described in detail below with reference to examples, but the present invention should not be construed as being limited to the following description.
実施例１：
三陸地方で漁獲されたイサダ（ツノナシオキアミ）の無加水冷凍ブロック（－５０℃保存のもの）１０ｔを－５℃で解凍し、市販のミートチョッパーを用いて粗く破砕した後、市販のマスコロイダーを用いてペースト状になるまで破砕した。得られたオキアミの破砕物１０ｔに５ｔの純水を加えて撹拌することで調製したオキアミの破砕物の水懸濁液１５ｔに、麹菌由来のプロテアーゼであるコクラーゼＰ顆粒（三菱ケミカルフーズ社製）５０ｋｇを加え、さらにクエン酸１５０ｋｇを加えることで液のｐＨを３～４に調整し、５０℃で酵素処理した。１時間後、処理液を７０℃に加温して３０分間インキュベートすることでプロテアーゼの失活を行った。処理液を室温まで冷却してから、市販の脱水型デカンタを用いて処理液を遠心分離することで、オイル画分を含む液分１３ｔを固形分から分離した。次に、得られたオイル画分を含む液分を市販の濃縮型デカンタを用いて遠心分離することで、オイル画分１．７ｔを水画分から分離した。得られたオイル画分（赤色ないし桃色のオイル状物）をフリーズドライした後、得られた固形物を粉砕することで、目的物である茶褐色の粉末状イサダ成分含有組成物３７４ｇを得た。こうして得られたイサダ成分含有組成物は、タンパク質を４４重量％の割合で、脂質を２８重量％の割合で、水分を１０重量％の割合で含むものであり（残部は炭化水素や灰分）、脂質の内訳は、中性脂質１６．４重量％、フォスファチジルエタノールアミン６．８重量％、フォスファチジルコリン４０．８重量％、ジアシルグリセロール１２．６重量％、遊離脂肪酸１６．７重量％、その他６．７重量％であって、イサダ成分含有組成物中に占める１－アルキルエーテル型リン脂質の割合は２．７重量％であった。なお、イサダ成分含有組成物に含まれる脂質の分析は、Ｂｌｉｇｈ－Ｄｙｅｒ法によって試料（イサダ成分含有組成物）０．２ｇから脂質を抽出して行った。抽出された脂質中の中性脂質、フォスファチジルエタノールアミン、フォスファチジルコリン、ジアシルグリセロール、遊離脂肪酸の割合は、１０ｍｇ／ｍＬの濃度でクロロホルムに溶解した脂質を１μＬ薄層クロマトグラフ－水素炎イオン化検出器（イアトロスキャン：ＬＳＩメディエンス社製）に供して測定した。薄層クロマトグラフは、クロロホルム／メタノール／水（４２：２４：２．５）で１次展開した後、ヘキサン／ジエチルエーテル（５０：３０）で２次展開することで、それぞれの脂質を分離した。１－アルキルエーテル型リン脂質は、抽出された脂質中の含量を高速液体クロマトグラフ（ＨＰＬＣ）を用いて定量し、試料中に占める割合を算出した。高速液体クロマトグラフは、０．３ｍｇ／ｍＬの濃度でクロロホルム／メタノール（２：１）に溶解した脂質の溶液３０μＬを分析サンプルとし、カラムはＬｉｃｈｒｏｓｐｈｅｒ １００ Ｄｉｏｌ ２５０ｘ４（Ａｇｉｌｅｎｔ社製）を用い、溶出液Ａとしてヘキサン／２－プロパノール／酢酸（８２：１７：１）０．０８％トリエチルアミン、溶出液Ｂとして２－プロパノール／水／酢酸（８５：１４：１）０．０８％トリエチルアミンを、溶出液Ａ９５％／溶出液Ｂ５％（２３分間グラジエント）溶出液Ａ６０％／溶出液Ｂ４０％（４分間グラジエント）溶出液Ａ１５％／溶出液Ｂ８５％（１分間ホールドの後に４分間グラジエント）溶出液Ａ９５％／溶出液Ｂ５％（１０分間ホールド）の条件で送液することによって行い、１－アルキルエーテル型リン脂質を分離し、分離した１－アルキルエーテル型リン脂質を光散乱検出器（ＳＯＦＴＡ社製）を用いて検出した。定量のための検量線作成には１－アルキルエーテル型リン脂質の標準品である１－ｈｅｘａｄｅｃｅｎｏｙｌ－２－ｏｌｅｏｙｌ－ｓｎ－ｇｌｙｃｅｒｏ－３－ｐｈｏｓｐｈｏｃｏｌｉｎｅ（フナコシ社製）を用いた。また、タンパク質はケルダール法、含水量は加熱乾燥法によってそれぞれ分析した。
Example 1:
Thaw 10 tons of non-hydrolyzed frozen block of Isada (horned krill) caught in the Sanriku region (stored at -50°C) at -5°C, crushed roughly using a commercially available meat chopper, and then crushed using a commercially available mass colloider. and crushed to a paste. To 15 tons of an aqueous suspension of the crushed krill prepared by adding 5 tons of pure water to 10 tons of the obtained crushed krill and stirring, was added Cochrase P granules (manufactured by Mitsubishi Chemical Foods Co., Ltd.), which is a protease derived from Aspergillus oryzae. 50 kg was added, and 150 kg of citric acid was further added to adjust the pH of the liquid to 3 to 4, followed by enzymatic treatment at 50°C. After 1 hour, the treated solution was heated to 70° C. and incubated for 30 minutes to inactivate the protease. After cooling the treated liquid to room temperature, the treated liquid was centrifuged using a commercially available dewatering decanter to separate 13 t of the liquid portion containing the oil fraction from the solid content. Next, 1.7 t of the oil fraction was separated from the water fraction by centrifuging the resulting liquid containing the oil fraction using a commercially available concentrated decanter. After freeze-drying the obtained oil fraction (red to pink oily matter), the obtained solid matter was pulverized to obtain 374 g of the desired dark brown powdery Isada component-containing composition. The Isada component-containing composition thus obtained contains 44% by weight of protein, 28% by weight of lipid, and 10% by weight of water (the balance being hydrocarbons and ash), The breakdown of lipids is 16.4% by weight of neutral lipids, 6.8% by weight of phosphatidylethanolamine, 40.8% by weight of phosphatidylcholine, 12.6% by weight of diacylglycerol, and 16.7% by weight of free fatty acids. , and others 6.7% by weight, and the proportion of 1-alkyl ether type phospholipid in the Isada component-containing composition was 2.7% by weight. The lipid contained in the Isada component-containing composition was analyzed by extracting the lipid from 0.2 g of the sample (Isada component-containing composition) by the Bligh-Dyer method. The ratio of neutral lipids, phosphatidylethanolamine, phosphatidylcholine, diacylglycerol, and free fatty acids in the extracted lipids was determined by thin-layer chromatography-hydrogen flame of 1 μL of lipids dissolved in chloroform at a concentration of 10 mg/mL. Measurement was performed using an ionization detector (Iatroscan: manufactured by LSI Medience). In thin-layer chromatography, each lipid was separated by primary development with chloroform/methanol/water (42:24:2.5) followed by secondary development with hexane/diethyl ether (50:30). . The content of the 1-alkyl ether type phospholipid in the extracted lipid was quantified using high performance liquid chromatography (HPLC), and the ratio in the sample was calculated. In high-performance liquid chromatography, 30 μL of a lipid solution dissolved in chloroform/methanol (2:1) at a concentration of 0.3 mg/mL was used as an analysis sample, the column was Lichrospher 100 Diol 250x4 (manufactured by Agilent), and the eluent was Hexane/2-propanol/acetic acid (82:17:1) 0.08% triethylamine as A, 2-propanol/water/acetic acid (85:14:1) 0.08% triethylamine as eluent B, and eluent A95. %/5% eluate B (23 min gradient) 60% eluate A/40% eluate B (4 min gradient) 15% eluate A/85% eluate B (1 min hold followed by 4 min gradient) 95% eluate/eluate The 1-alkyl ether type phospholipid was separated by feeding under the condition of 5% liquid B (hold for 10 minutes), and the separated 1-alkyl ether type phospholipid was measured using a light scattering detector (manufactured by SOFTA). detected. 1-hexadecenoyl-2-oleoyl-sn-glycero-3-phosphocoline (manufactured by Funakoshi Co., Ltd.), which is a standard product of 1-alkyl ether type phospholipids, was used to prepare a calibration curve for quantification. The protein was analyzed by the Kjeldahl method, and the water content was analyzed by the heat drying method.
実施例２：
麹菌由来のプロテアーゼとしてコクラーゼＰ顆粒（三菱ケミカルフーズ社製）のかわりにオリエンターゼＯＰ（エイチビィアイ社製）を用い、クエン酸のかわりに炭酸を用いてｐＨの調整を行うこと以外は実施例１と同様にして、目的物である粉末状イサダ成分含有組成物を得た。
Example 2:
Same as in Example 1 except that Orientase OP (manufactured by HBI Co., Ltd.) was used instead of Cochrase P granules (manufactured by Mitsubishi Chemical Foods Co., Ltd.) as a protease derived from Aspergillus oryzae, and carbonic acid was used instead of citric acid to adjust the pH. In the same manner, a target powdery Isada component-containing composition was obtained.
製剤例１：錠剤
以下の成分組成からなるオキアミ成分含有組成物を含む錠剤を自体公知の方法で製造した。
実施例１で得たイサダ成分含有組成物 １
乳糖 ８０
ステアリン酸マグネシウム １９（単位：重量％）
Formulation Example 1: Tablet A tablet containing a krill component-containing composition having the following component composition was produced by a method known per se.
Isada component-containing composition obtained in Example 1 1
Lactose 80
Magnesium stearate 19 (unit: weight%)
製剤例２：ビスケット
以下の成分組成からなるオキアミ成分含有組成物を含むビスケットを自体公知の方法で製造した。
実施例１で得たイサダ成分含有組成物 １
薄力粉 ３２
全卵 １６
バター １６
砂糖 ２４
水 １０
ベーキングパウダー １（単位：重量％）
Formulation Example 2: Biscuits A biscuit containing a krill component-containing composition having the following component composition was produced by a method known per se.
Isada component-containing composition obtained in Example 1 1
soft flour 32
Whole egg 16
Butter 16
sugar 24
water 10
Baking powder 1 (unit: weight%)
本発明は、医薬品素材や食品素材などとして用いることができるオキアミ成分含有組成物の製造方法を提供することができる点において産業上の利用可能性を有する。 INDUSTRIAL APPLICABILITY The present invention has industrial applicability in that it can provide a method for producing a krill component-containing composition that can be used as a pharmaceutical material or food material.
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|---|---|---|---|---|
| JP2005087017A (en) | 2003-09-12 | 2005-04-07 | T Hasegawa Co Ltd | Method for producing euphausiacea extract |
| US20130274496A1 (en) | 2011-02-11 | 2013-10-17 | Dae Duck Frd Co., Ltd | Krill oil and method for manufacturing the same |
| JP2015163607A (en) | 2014-01-29 | 2015-09-10 | 岩手県 | Acquisition method of hydroxy eicosapentaenoic acid |
| CN105542936A (en) | 2015-12-16 | 2016-05-04 | 江南大学 | Euphausia superba oil extraction method |
| JP2017122067A (en) | 2016-01-07 | 2017-07-13 | 株式会社マリン大王 | Oil of euphausia pacifica and method for producing the same |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2005087017A (en) | 2003-09-12 | 2005-04-07 | T Hasegawa Co Ltd | Method for producing euphausiacea extract |
| US20130274496A1 (en) | 2011-02-11 | 2013-10-17 | Dae Duck Frd Co., Ltd | Krill oil and method for manufacturing the same |
| JP2014509339A (en) | 2011-02-11 | 2014-04-17 | テドク・エフアールディー・カンパニー・リミテッド | Method for producing krill oil and krill oil produced by this method |
| JP2015163607A (en) | 2014-01-29 | 2015-09-10 | 岩手県 | Acquisition method of hydroxy eicosapentaenoic acid |
| CN105542936A (en) | 2015-12-16 | 2016-05-04 | 江南大学 | Euphausia superba oil extraction method |
| JP2017122067A (en) | 2016-01-07 | 2017-07-13 | 株式会社マリン大王 | Oil of euphausia pacifica and method for producing the same |
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