JP2613097B2 - Lactoferrin sterilization method - Google Patents
Lactoferrin sterilization methodInfo
- Publication number
- JP2613097B2 JP2613097B2 JP63259311A JP25931188A JP2613097B2 JP 2613097 B2 JP2613097 B2 JP 2613097B2 JP 63259311 A JP63259311 A JP 63259311A JP 25931188 A JP25931188 A JP 25931188A JP 2613097 B2 JP2613097 B2 JP 2613097B2
- Authority
- JP
- Japan
- Prior art keywords
- lactoferrin
- iron
- ionic strength
- heating
- solution
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Expired - Fee Related
Links
Classifications
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23J—PROTEIN COMPOSITIONS FOR FOODSTUFFS; WORKING-UP PROTEINS FOR FOODSTUFFS; PHOSPHATIDE COMPOSITIONS FOR FOODSTUFFS
- A23J3/00—Working-up of proteins for foodstuffs
- A23J3/04—Animal proteins
- A23J3/08—Dairy proteins
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23J—PROTEIN COMPOSITIONS FOR FOODSTUFFS; WORKING-UP PROTEINS FOR FOODSTUFFS; PHOSPHATIDE COMPOSITIONS FOR FOODSTUFFS
- A23J1/00—Obtaining protein compositions for foodstuffs; Bulk opening of eggs and separation of yolks from whites
- A23J1/20—Obtaining protein compositions for foodstuffs; Bulk opening of eggs and separation of yolks from whites from milk, e.g. casein; from whey
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Biochemistry (AREA)
- Engineering & Computer Science (AREA)
- Food Science & Technology (AREA)
- Polymers & Plastics (AREA)
- Zoology (AREA)
- Health & Medical Sciences (AREA)
- Nutrition Science (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
- Peptides Or Proteins (AREA)
Description
【発明の詳細な説明】 産業上の利用分野 本発明は、種々の生理的活性を有し、薬理学的にも重
要な乳蛋白質であるラクトフェリンを、その生理的機能
性を維持した状態で加熱殺菌する方法に関する。The present invention relates to a method for heating lactoferrin, a milk protein having various physiological activities and also having important pharmacological properties, while maintaining its physiological functionality. It relates to a method of sterilization.
技術的背景 ラクトフェリンは、乳等の外分泌液中に存在する鉄結
合性の糖蛋白質であつて、病原性細菌に対する精菌作
用、白血球の分化調節作用及び殺菌力増強作用、リンパ
球の増殖作用及び鉄吸収調節作用等の種々の生理的活性
を有するため、栄養学的のみならず薬理学的にも重要な
乳蛋白質である。Technical backgroundLactoferrin is an iron-binding glycoprotein that is present in exocrine fluids such as milk, and has a bactericidal action against pathogenic bacteria, a regulatory action on leukocyte differentiation and a bactericidal action, a proliferation action on lymphocytes, Since it has various physiological activities such as an iron absorption regulating action, it is an important milk protein not only nutritionally but also pharmacologically.
これらラクトフェリンの生理的機能性は、主として、
ラクトフェリンの鉄結合力に由来しており、ラクトフェ
リンの三次元的立体構造が保持されている時のみ、鉄を
キレート結合する能力を発揮する。The physiological functions of these lactoferrins are mainly
It is derived from the iron binding power of lactoferrin and exerts the ability to chelate iron only when the three-dimensional steric structure of lactoferrin is maintained.
ところが、ラクトフェリンは、加熱によつて三次元的
立体構造が破壊されて沈澱する。実際、濃度の希薄な0.
5％程度であつても、60℃、15分間加熱しただけで、ラ
クトフェリンは沈澱するといわれている。したがつて、
ラクトフェリン水溶液から細菌を除くには、フィルター
除菌しか方法がなかつた。However, lactoferrin precipitates due to the destruction of its three-dimensional structure by heating. In fact, a concentration of 0.
It is said that lactoferrin precipitates when heated at 60 ° C. for 15 minutes even at about 5%. Therefore,
The only way to remove bacteria from the aqueous lactoferrin solution was to filter bacteria.
一方、生理的機能性を付与した食品にラクトフェリン
を利用する上で、ラクトフェリンの殺菌工程は不可欠で
あり、例えば、食品衛生法の清涼飲料水製造基準には、
pH4.0以下の場合、65℃、10分間の加熱又はこれと同
等以上の効果を有する殺菌を、pH4.0以上の場合、85
℃、30分間の加熱又はこれと同等以上の効果を有する殺
菌をそれぞれ行なわければならない旨、明記されている
ので、フィルター除菌を行つたラクトフェリンは、清涼
飲料水等の素材として用いることができないという問題
があつた。On the other hand, in using lactoferrin in foods that have been given physiological functionality, a lactoferrin sterilization step is indispensable, for example, in the Soft Drinks Production Standards of the Food Sanitation Law,
In the case of pH 4.0 or lower, 65 ° C, 10 minutes of heating or sterilization having an effect equal to or more than this, in the case of pH 4.0 or higher, 85
℃, heating for 30 minutes or sterilization with an effect equal to or more than this must be performed, because it is clearly specified, lactoferrin subjected to filter sterilization can not be used as a material such as soft drinks There was a problem.
発明が解決しようとする課題 本発明は、叙上の状況に鑑みなされたものであつて、
種々の生理的機能性を発現する基本となる鉄結合能を保
持したままで、ラクトフェリン水溶液を加熱殺菌するた
めの方法を提供することを課題とする。Problem to be Solved by the Invention The present invention has been made in view of the above situation,
An object of the present invention is to provide a method for heat-sterilizing an aqueous lactoferrin solution while maintaining a basic iron binding ability for expressing various physiological functions.
課題を解決するための手段 本発明の特徴は、ラクトフェリン水溶液のイオン強度
Ｉを次式に示す条件下に調整した後、加熱殺菌すること
にある。Means for Solving the Problems The feature of the present invention lies in that the ionic strength I of the lactoferrin aqueous solution is adjusted under the conditions shown by the following formula, and then sterilized by heating.
log I≦−３ （Ｔ≧80℃） （Ｔは温度℃を表す。） 本発明でいうラクトフェリンは、未変性で鉄を結合す
る能力のあるものであれば、鉄を結合した形であつて
も、また、鉄を遊離している形であつてもよい。 log I ≦ −3 (T ≧ 80 ° C.) (T represents temperature ° C.) The lactoferrin in the present invention is an iron-bound form as long as it is unmodified and capable of binding iron. May also be in the form of free iron.
従来、ラクトフェリンの分離、精製方法として、イオ
ン変換樹脂を用いる方法（Gordon et al.,Biochim.Biop
hys.Acta,60:410〜411,1962）、ヘパリンアフィニテイ
クロマトグラフィを用いる方法（Blockberg et al.,FEB
S Lett.,109:180,1980）、抗ラクトフェリンモノクロー
ナル抗体固定化カラムを用いる方法（特開昭61−14520
0）、或いは、硫酸エステル化した担体を用いる方法
（特開昭62−88450）等が知られているが、未変性で鉄
を結合する能力のあるものであれば、どのような方法で
調製されたラクトフェリンであつてもよい。Conventionally, as a method for separating and purifying lactoferrin, a method using an ion conversion resin (Gordon et al., Biochim. Biop.
hys. Acta, 60: 410-411, 1962), a method using heparin affinity chromatography (Blockberg et al., FEB)
S Lett., 109: 180, 1980), a method using a column on which an anti-lactoferrin monoclonal antibody is immobilized (JP-A-61-14520).
0) Alternatively, a method using a sulfated carrier (Japanese Patent Application Laid-Open No. 62-88450) is known, but any method can be used as long as it is unmodified and has the ability to bind iron. Lactoferrin may be used.
加熱殺菌に用いるラクトフェリンは、溶液状のもので
あつても、或いは、粉末化されたものを溶解したもので
あつてもよい。ラクトフェリン水溶液の濃度は高濃度で
あつても構わない。The lactoferrin used for heat sterilization may be in the form of a solution or may be obtained by dissolving powdered one. The concentration of the lactoferrin aqueous solution may be high.
本発明において重要なことは、ラクトフェリン水溶液
のイオン強度Ｉであつて、そのイオン強度Ｉが次式に示
す条件となるように水溶液を調整しなければならない。What is important in the present invention is the ionic strength I of the lactoferrin aqueous solution, and the aqueous solution must be adjusted so that the ionic strength I satisfies the condition shown in the following equation.
log I≦−３ （Ｔ≧80℃） （Ｔは温度℃を表す。） Ｔが60℃以下の場合、ラクトフェリンは変性しないの
で、特にイオン強度Ｉを調整する必要はないが、Ｔが60
℃以上の場合、上記式に示した条件からイオン強度Ｉが
外れると、ラクトフェリンの沈澱が生じたり、鉄結合能
が低下する。したがつて、塩濃度の高いラクトフェリン
水溶液を加熱殺菌する場合は、上記式に示した条件に見
合つたイオン強度Ｉとなるよう、脱塩して水溶液のイオ
ン強度を調整する必要がある。 log I ≦ −3 (T ≧ 80 ° C.) (T represents temperature ° C.) When T is 60 ° C. or less, lactoferrin is not denatured. Therefore, it is not particularly necessary to adjust the ionic strength I.
When the temperature is not lower than ℃, if the ionic strength I deviates from the condition shown in the above formula, lactoferrin precipitates or the iron binding ability decreases. Therefore, when heat sterilizing an aqueous solution of lactoferrin having a high salt concentration, it is necessary to adjust the ionic strength of the aqueous solution by desalting so that the ionic strength I meets the conditions shown in the above equation.
ラクトフェリン水溶液のイオン強度及び加熱温度と沈
澱生成又は鉄結合能との関係を示すと表１並びに表２の
とおりである。Tables 1 and 2 show the relationship between the ionic strength and heating temperature of the aqueous lactoferrin solution and the precipitation or iron binding ability.
鉄結合能の算出は、沈澱を除去した上清1mlに５％重
炭酸ソーダ水を１滴加えた時の465nmにおける吸光度O
DO、これに２％FeCl3・6H2O溶液100μを加えた時の46
5nmにおける吸光度をODFとし、同様に、未加熱のラクト
フェリンを用いて測定した▲▼Ｏ及び▲▼Ｆの
値から、次式に従つて行つた。 The iron binding ability was calculated by measuring the absorbance at 465 nm when 1 drop of 5% aqueous sodium bicarbonate was added to 1 ml of the supernatant from which the precipitate had been removed.
D O , 46% of which was added to 100% of a 2% FeCl 3 .6H 2 O solution.
The absorbance at 5nm and OD F, similarly, from the value of the measured ▲ ▼ O and ▲ ▼ F using unheated lactoferrin accordance connexion KoTsuta the following equation.
以下に実施例を示して本発明を具体的に説明する。 Hereinafter, the present invention will be described specifically with reference to examples.
実施例１ 凍結乾燥されたウシラクトフェリン粉末100gを脱イオ
ン水１に溶解した。これをフタ付容器に移して85℃温
湯中で30分間加熱殺菌した。加熱後、直ちに流水で室温
まで冷却した。沈澱物の生成は全く認められなかつた。
また、冷却後測定した鉄結合能は97％であつた。Example 1 100 g of lyophilized bovine lactoferrin powder was dissolved in deionized water 1. This was transferred to a container with a lid, and sterilized by heating in hot water at 85 ° C. for 30 minutes. After heating, it was immediately cooled to room temperature with running water. No formation of a precipitate was observed.
The iron binding ability measured after cooling was 97%.
実施例２ キトサンビーズに硫酸エステルを付与した担体に生脱
脂乳10トンを流し、結合したウシラクトフェリンを1M食
塩水で溶出した。溶出液を限外濾過装置（DDS 20）で脱
塩・濃縮を行い、蛋白質濃度5.0％のラクトフェリン溶
液20を得た。この液のイオン強度は原子吸光計でナト
リウムイオン濃度から計算した結果、４×10-4であつ
た。Example 2 10 tons of raw skim milk was flowed through a carrier obtained by adding a sulfate ester to chitosan beads, and bound bovine lactoferrin was eluted with 1M saline. The eluate was desalted and concentrated using an ultrafiltration device (DDS 20) to obtain a lactoferrin solution 20 having a protein concentration of 5.0%. The ionic strength of this solution was 4 × 10 −4 as a result of calculation from the sodium ion concentration by an atomic absorption spectrometer.
そこでプレート型殺菌機を用い、95℃の温度で４秒間
加熱殺菌し、７℃まで冷却した。沈澱物は認められず、
鉄結合能も98％であつた。Therefore, the plate was sterilized by heating at a temperature of 95 ° C. for 4 seconds using a plate-type sterilizer and cooled to 7 ° C. No precipitate was observed,
The iron binding ability was also 98%.
発明の効果 本発明の方法に従つて、ラクトフェリン水溶液を加熱
殺菌することにより、沈澱物を生成するということもな
く、また、殺菌されたラクトフェリンの鉄結合能も十分
維持された。Effect of the Invention According to the method of the present invention, the aqueous solution of lactoferrin is sterilized by heating without generating a precipitate, and the sterilized lactoferrin also has a sufficient iron-binding ability.
したがつて、ラクトフェリンの生理的機能を殆ど損な
うことなく、ラクトフェリン加熱殺菌し得ることがで
き、多様な生理的機能性を有するラクトフェリンを食品
素材として利用可能にするものであり、その産業上の効
果は計り知れない。Therefore, lactoferrin can be heat-sterilized without substantially impairing the physiological function of lactoferrin, and lactoferrin having various physiological functions can be used as a food material, and its industrial effects Is immeasurable.
Claims (1)
して、水溶液のイオン強度Ｉを次式に示す条件下に調整
した後、殺菌することを特徴とするラクトフェリンの殺
菌方法。 log I≦−３ （Ｔ≧80℃） （Ｔは温度℃を表す。）1. A method for sterilizing lactoferrin, which comprises sterilizing an aqueous lactoferrin solution after adjusting the ionic strength I of the aqueous solution under the conditions shown in the following formula. log I ≦ −3 (T ≧ 80 ° C.) (T indicates temperature ° C.)
Priority Applications (5)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP63259311A JP2613097B2 (en) | 1988-10-17 | 1988-10-17 | Lactoferrin sterilization method |
| DE8989119130T DE68903208T2 (en) | 1988-10-17 | 1989-10-14 | METHOD FOR THE THERMAL TREATMENT OF LACTOTRINE. |
| EP89119130A EP0364912B1 (en) | 1988-10-17 | 1989-10-14 | Method for thermally treating lactoferrin |
| NZ231038A NZ231038A (en) | 1988-10-17 | 1989-10-17 | Method of thermally treating aqueous lactoferrin |
| US07/422,489 US5116953A (en) | 1988-10-17 | 1989-10-17 | Method for thermally treating lactoferrin |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP63259311A JP2613097B2 (en) | 1988-10-17 | 1988-10-17 | Lactoferrin sterilization method |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| JPH02108629A JPH02108629A (en) | 1990-04-20 |
| JP2613097B2 true JP2613097B2 (en) | 1997-05-21 |
Family
ID=17332315
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP63259311A Expired - Fee Related JP2613097B2 (en) | 1988-10-17 | 1988-10-17 | Lactoferrin sterilization method |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JP2613097B2 (en) |
Families Citing this family (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2747935B2 (en) * | 1989-07-21 | 1998-05-06 | 雪印乳業株式会社 | Manufacturing method of iron-fortified beverage |
| JP3821981B2 (en) * | 1999-04-09 | 2006-09-13 | 森永乳業株式会社 | Sterilized milk containing unmodified lactoferrin and method for producing the same |
| JP4005103B2 (en) * | 2006-04-05 | 2007-11-07 | 森永乳業株式会社 | Bactericidal beverage containing unmodified lactoferrin and method for producing the same |
| JP2007175065A (en) * | 2007-03-28 | 2007-07-12 | Morinaga Milk Ind Co Ltd | Sterilized drink containing native lactoferrin, and method for producing the same |
| US11470858B2 (en) | 2018-03-12 | 2022-10-18 | Morinaga Milk Industry Co., Ltd. | Method for producing lactoferrin-containing aqueous solution |
-
1988
- 1988-10-17 JP JP63259311A patent/JP2613097B2/en not_active Expired - Fee Related
Non-Patent Citations (1)
| Title |
|---|
| Ｊｏｕｒｎａｌ ｏｆ Ｆｏｏｄ Ｓｃｉｅｎｃｅ，４８（１）（１９８３）（米）Ｐ．４２−４６，７０ |
Also Published As
| Publication number | Publication date |
|---|---|
| JPH02108629A (en) | 1990-04-20 |
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