JP2002330798A - Method for producing polysaccharide - Google Patents
Method for producing polysaccharideInfo
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- JP2002330798A JP2002330798A JP2001141827A JP2001141827A JP2002330798A JP 2002330798 A JP2002330798 A JP 2002330798A JP 2001141827 A JP2001141827 A JP 2001141827A JP 2001141827 A JP2001141827 A JP 2001141827A JP 2002330798 A JP2002330798 A JP 2002330798A
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- polysaccharide
- purified product
- producing
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- rice
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- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
Description
【０００１】[0001]
【発明の属する技術分野】本発明は、乳酸菌ラクトバチ
ラス・ケフィラノファシエンスを培養することで多糖を
製造する多糖製造方法に関する。TECHNICAL FIELD The present invention relates to a method for producing a polysaccharide by producing a polysaccharide by culturing a lactic acid bacterium Lactobacillus kefiranofaciens.
【０００２】[0002]
【従来の技術】近年の日本では、食生活の変化に伴って
高脂肪食品を摂取する機会が増え、高脂血症，動脈硬化
症などの成人病が増加している。これら成人病の予防と
して、摂取することで血圧降下作用，脂質代謝改善作
用，血糖上昇抑制作用などが期待できる多糖が注目され
ている。例えば、旧ソ連コーカサス地方原産の発酵乳ケ
フィールは、グルコースとがラクトースから構成された
粘質多糖ケフィランを含有しており、長寿につながる健
康飲料として東欧を中心に飲用されている。2. Description of the Related Art In recent years, in Japan, the chances of ingesting high-fat foods have increased with changes in eating habits, and adult diseases such as hyperlipidemia and arteriosclerosis have increased. As a prophylaxis against these adult diseases, polysaccharides, which can be expected to have a hypotensive effect, a lipid metabolism improving effect, a blood glucose elevation suppressing effect, and the like when ingested, have attracted attention. For example, fermented milk kefir native to the former Soviet Union Caucasus region contains a viscous polysaccharide kefiran composed of glucose and lactose, and is drunk mainly in Eastern Europe as a health drink leading to longevity.
【０００３】従来、このケフィランは、ラクトバチラス
・ケフィラノファシエンス（Ｌ．Ｋｅｆｉｒａｎｏｆａ
ｃｉｅｎｓ）を含む数種類の乳酸菌を利用して乳糖より
合成していた。上記乳酸菌が乳糖より生産するケフィラ
ンは、分岐を持つＧｌｃ−Ｇａｌ（−Ｇｌｃ）−Ｇａｌ
−Ｇａｌ−Ｇｌｃの繰り返し単位で、グルコース（Ｇｌ
ｃ）とガラクトース（Ｇａｌ）の構成比率がほぼ１：１
で、分子量が１００〜４００万の多糖類である。[0003] Conventionally, this kefiran has been used in L. kefiranofaciens (L. Kefiranofa).
ciens), and was synthesized from lactose using several types of lactic acid bacteria. Kefiran produced by the lactic acid bacterium from lactose is branched Glc-Gal (-Glc) -Gal.
-Gal-Glc repeating unit, glucose (Gl
c) and galactose (Gal) in a composition ratio of about 1: 1
And is a polysaccharide having a molecular weight of 1 to 4 million.
【０００４】[0004]
【発明が解決しようとする課題】以上説明したように、
近年では、成人病の予防が期待できる多糖が注目されて
おり、より多くの多糖製造方法が望まれている。本発明
は、以上のような問題点を解消するためになされたもの
であり、多糖を製造する新たな方法を提供することを目
的とする。As described above,
In recent years, polysaccharides that can be expected to prevent adult diseases have attracted attention, and more polysaccharide production methods are desired. The present invention has been made to solve the above problems, and has as its object to provide a new method for producing a polysaccharide.
【０００５】[0005]
【課題を解決するための手段】本発明の多糖製造方法
は、米を含む培地を用意し、培地で乳酸菌ラクトバチラ
ス・ケフィラノファシエンスを培養し、培養した後の培
養液中よりタンパク質と菌とを分離した分離液を作製
し、分離液より所定の分子量以下の物質を除去するケフ
ィランを含む精製物を得るものである。The polysaccharide production method of the present invention comprises preparing a medium containing rice, culturing lactic acid bacterium Lactobacillus kefiranofaciens in the medium, and culturing the protein and the bacterium from the culture solution after culturing. To obtain a purified product containing kefiran that removes substances having a predetermined molecular weight or less from the separated solution.
【０００６】上記発明において、培地は、米を糖化した
米糖化液を含み、また、培地は、酵母エキスを含む。上
記発明において、所定の分子量以下の物質の除去は、限
外ろ過により行えばよい。[0006] In the above invention, the medium contains a rice saccharified solution obtained by saccharifying rice, and the medium contains a yeast extract. In the above invention, the removal of the substance having a predetermined molecular weight or less may be performed by ultrafiltration.
【０００７】[0007]
【発明の実施の形態】以下、本発明の実施の形態につい
て図を参照して説明する。始めに、本実施の形態におけ
る、多糖製造方法について説明する。まず、内容積が５
００ｌ程度のタンクを用意し、このタンク内で、米の粉
３６ｋｇを水２１６ｌに懸濁させ、これに乳酸２．８ｋ
ｇを加えて１２１℃にして３０分間攪拌することで乳化
させる。次いで、乳化させた液にペプシンを添加し、含
まれているタンパク質を加水分解する（ｐＨ１．８，５
０℃，４時間）。この後、加水分解した液に、グルコア
ミラーゼを添加して糖化し（ｐＨ４．５，５０℃，１時
間）、米糖化液（約２７０ｌ）を作製する。Embodiments of the present invention will be described below with reference to the drawings. First, a method for producing a polysaccharide according to the present embodiment will be described. First, if the internal volume is 5
A tank of about 00 l is prepared, and 36 kg of rice flour is suspended in 216 l of water in this tank, and 2.8 k of lactic acid is added thereto.
Then, the mixture is added to g and stirred at 121 ° C. for 30 minutes to emulsify. Next, pepsin is added to the emulsified liquid to hydrolyze the contained protein (pH 1.8, 5).
0 ° C, 4 hours). Thereafter, glucoamylase is added to the hydrolyzed liquid to saccharify it (pH 4.5, 50 ° C., 1 hour) to prepare a saccharified rice liquid (about 270 l).
【０００８】つぎに、上記タンク内の米糖化液に、酵母
エキス４ｋｇ（１％）と、「Ｔｗｅｅｎ８０」４００ｍ
ｌ（０．１％）と、リン酸水素二カリウム８００ｇ
（０．２％）と、酢酸ナトリウム２ｋｇ（０．５％）
と、クエン酸三アンモニウム８００ｇ（０．２％）と、
硫酸マグネシウム８０ｇ（０．０２％）と、硫酸マンガ
ン２０ｇ（０．００５％）と、寒天６０ｇ（０．０１
％）と、水１３０リットルを加え、これらの培地組成
で、乳酸菌ラクトバチラス・ケフィラノファシエンスを
培養する。なお、この場一組成において、米糖化液とし
て、グルコースが１０％、ポリペプトンが０．３５％含
まれていることになる。Next, 4 kg (1%) of yeast extract and 400 m of “Tween 80” were added to the rice saccharified solution in the tank.
1 (0.1%) and 800 g of dipotassium hydrogen phosphate
(0.2%) and 2 kg of sodium acetate (0.5%)
And 800 g (0.2%) of triammonium citrate,
80 g (0.02%) of magnesium sulfate, 20 g (0.005%) of manganese sulfate, and 60 g (0.01%) of agar
%) And 130 liters of water, and the lactic acid bacterium Lactobacillus kefiranofaciens is cultured in these medium compositions. In addition, in this in-situ composition, 10% of glucose and 0.35% of polypeptone are contained as a rice saccharified solution.
【０００９】上記培地における乳酸菌ラクトバチラス・
ケフィラノファシエンスの培養条件は、温度３３℃，攪
拌速度３０ｒｐｍ，ｐＨ５．０とする。なお、ｐＨの調
整には、水酸化ナトリウム（５Ｎ）を用いる。この培養
を７日間続け、培養液１６３０ｍｇ／リットルを得た。
７日間の培養の後、得られた培養液より、つぎに示す分
離・精製を行い、生成物を得た。まず、得られた培養液
に、当量の水を加え、かつ、ＨＣｌ（５Ｎ）でｐＨ１．
０に調整し、３０分間攪拌することで、培養液中のタン
パク質を凝固・沈殿させる。この後、水酸化ナトリウム
（５Ｎ）でｐＨを調整し（ｐＨ８．５）、また液温を５
０℃程度として多糖類が溶解しやすい状態として３０分
間攪拌する。The lactic acid bacteria Lactobacillus in the above medium
The culture conditions of Kefiranofaciens are a temperature of 33 ° C., a stirring speed of 30 rpm, and a pH of 5.0. Note that sodium hydroxide (5N) is used for pH adjustment. This culture was continued for 7 days to obtain a culture solution (1630 mg / liter).
After culturing for 7 days, the following culture was subjected to the following separation and purification to obtain a product. First, an equivalent amount of water was added to the obtained culture solution, and the mixture was adjusted to pH 1 with HCl (5N).
The protein in the culture solution is coagulated and precipitated by adjusting to 0 and stirring for 30 minutes. Thereafter, the pH was adjusted with sodium hydroxide (5N) (pH 8.5), and the liquid temperature was adjusted to 5%.
The mixture is stirred at about 0 ° C. for 30 minutes so that the polysaccharide is easily dissolved.
【００１０】この後、遠心分離装置により培養液より凝
固タンパク質や、ラクトバチラス・ケフィラノファシエ
ンスの菌体を遠心分離（８０００Ｇ，１０分，５０℃）
する。遠心分離した後、上澄みを取り出し、これを５０
℃の状態で分画分子量６０００で限外ろ過し、濾紙内に
残った残分を凍結乾燥して本実施の形態における精製物
（多糖）の粉末を約６００ｇ得る。Thereafter, the coagulated protein and the cells of Lactobacillus kefiranofaciens are separated from the culture solution by a centrifugal separator (8000 G, 10 minutes, 50 ° C.).
I do. After centrifugation, the supernatant is removed and
Ultrafiltration is performed at a molecular weight cut-off of 6000 at ℃, and the residue remaining in the filter paper is lyophilized to obtain about 600 g of the powder of the purified product (polysaccharide) in the present embodiment.
【００１１】つぎに、以下に示すことにより、得られた
精製粉末の分析を行った。まず、得られた精製物粉末の
５０％エタノール沈殿を、「ＤＥＡＥ ｓｅｐｈａｄｅ
ｘ Ａ−５０」で分画する。分画の結果は、図１に示
す。次いで、上記分画における素通り画分（図１のＦｒ
１５〜１８）に対して同容量のエタノールを加え、得ら
れた沈殿を遠心分離によって回収し、エタノールで洗浄
する。この後、洗浄したものを、「ＴＯＹＯＰＥＡＲＬ
ＨＷ６５」により分画する。この分画の結果は、図２に
示す。Next, the obtained purified powder was analyzed as described below. First, 50% ethanol precipitation of the obtained purified product powder was referred to as “DEAE sephade”.
x A-50 ". The results of the fractionation are shown in FIG. Next, the flow-through fraction (Fr in FIG. 1)
15-18), the same volume of ethanol is added, and the resulting precipitate is collected by centrifugation and washed with ethanol. After that, the washed product is referred to as "TOYOPEARL"
HW65 ". The result of this fractionation is shown in FIG.
【００１２】つぎに、上記分画した結果得られた精製多
糖の単糖組成を分析する。まず、精製多糖に、濃度２Ｍ
のトリフルオル酢酸（ＣＦ3ＣＯＯＨ）を加え，加水分
解（１００℃・５分）する。このことにより得られた加
水分解物を、水素ホウ酸ナトリウムで還元し、この後、
無水酢酸とピリジンを加えてアセチル化する。このアセ
チル化したものを、ガスクロマトグラフィー装置（ＧＣ
−ＭＳ−ＱＰ５０５０Ａ：島津製作所製）で分析する。
この分析では、カラムは、島津製作所製のＣＢＰ−５
（０．２０ｍｍ×２５ｍ）を用い、分離した各成分を質
量分析により検出する。この分析の結果、精製した多糖
の単糖組成は、グルコース（１．０）とガラクトース
（１．４）であることが判明した。Next, the monosaccharide composition of the purified polysaccharide obtained as a result of the above fractionation is analyzed. First, the purified polysaccharide was added at a concentration of 2M.
Of trifluoroacetic acid (CF 3 COOH) and hydrolyze (100 ° C., 5 minutes). The hydrolyzate obtained by this is reduced with sodium hydrogen borate, and then
Acetic anhydride and pyridine are added for acetylation. This acetylated product is subjected to gas chromatography (GC
-MS-QP5050A: manufactured by Shimadzu Corporation).
In this analysis, the column was CBP-5 manufactured by Shimadzu Corporation.
(0.20 mm × 25 m), and each separated component is detected by mass spectrometry. As a result of this analysis, the monosaccharide composition of the purified polysaccharide was found to be glucose (1.0) and galactose (1.4).
【００１３】つぎに、上記分画により精製した多糖の分
子質量を、「ＧＣＰ−ＭＡＬＬＳ（ＡＳＴＲＡ４ ７
３．０４）」を用いて測定する（水溶媒０．７５ｍｌ／
ｍｉｎ，カラム：ＴＯＳＯＨ Ｇ６０００ＰＷＸＬ）。
測定の結果、上記分画により精製した多糖の分子質量
は、７８５０００ダルトンであった。なお、１ダルトン
は、１／（アボガドロ数）ｇである。[0013] Next, the molecular mass of the polysaccharide purified by the above fractionation was determined by the following method: "GCP-MALLS (ASTRA47
3.04) ”(aqueous solvent 0.75 ml /
min, column: TOSOH G6000PWXL).
As a result of the measurement, the molecular mass of the polysaccharide purified by the above fractionation was 785,000 dalton. One dalton is 1 / (Avocado number) g.
【００１４】また、上記多糖の１％水溶液の常温（２０
℃）における旋光度（ナトリウムＤ線、波長５８９ｎｍ
を使用）を測定し、比旋光度を求めたところ、＋６４．
５°であった。以上のことにより、本実施の形態におけ
る精製物には、グルコースとガラクトースを構成単位と
する多糖類（ケフィラン）が含まれていることが判る。Further, a 1% aqueous solution of the above-mentioned polysaccharide is at room temperature (20
° C) (sodium D line, wavelength 589 nm)
Was measured, and the specific rotation was determined to be +64.
5 °. From the above, it is understood that the purified product in the present embodiment contains a polysaccharide (kefiran) having glucose and galactose as constituent units.
【００１５】つぎに、本実施の形態における精製物の生
理活性を調査した。まず、血圧降下作用について調査し
た。血圧降下作用の調査では、供試動物としてラット
（ＳＨＲＳＰ／Ｉｚｍ）♂８週を用いた。また、試験グ
ループとして、上記精製物を与えない対照群（１０匹）
と、体重１ｋｇ当たり１００ｍｇの精製物を１日ごとに
投与したグループ（１０匹）と、体重１ｋｇ当たり３０
０ｍｇの精製物を１日ごとに投与したグループとの３グ
ループ（１０匹）を設定した。調査の方法は、まず、精
製物を蒸留水で各濃度に調整し、胃ゾンデを用いて対象
のグループに６０日間連続経口投与する。対照群には、
蒸留水を与える。投与開始前と、３０日後と６０日後と
に、各ラットの収縮期ならびに拡張期の血圧を各々測定
した。測定結果は、表１に示すとおりとなった。Next, the physiological activity of the purified product in the present embodiment was investigated. First, the hypotensive effect was investigated. In the investigation of the blood pressure lowering effect, rats (SHRSP / Izm) @ 8 weeks were used as test animals. As a test group, a control group not receiving the purified product (10 animals)
And a group (10 animals) receiving 100 mg of purified product per kg of body weight every day, and a group of 30 animals per kg of body weight.
Three groups (10 animals) were set, including a group to which 0 mg of the purified product was administered every day. First, the purified product is adjusted to each concentration with distilled water, and is orally administered to a group of subjects continuously for 60 days using a gastric probe. In the control group,
Give distilled water. Before the start of the administration, and 30 days and 60 days later, the systolic and diastolic blood pressures of each rat were measured. The measurement results were as shown in Table 1.
【００１６】[0016]
【表１】 [Table 1]
【００１７】この血圧降下作用の調査の結果、表１に示
すように、３０日目の測定において、精製物を投与した
グループは、精製物を投与しなかった対照群に比較し、
血圧の上昇が抑制されていることが判明した。表中の数
字は、平均値±標準誤差である。また、「student's-t
test」により有意差を検定したところ、精製物を３００
ｍｇずつ投与したグループでは、収縮期血圧，拡張期血
圧ともに有意差が認められた。また、６０日目の測定に
おいても、３０日目の測定結果と同様の傾向を示し、精
製物を投与したグループでは、収縮期血圧が有意に低い
値を示した。なお、精製物を投与したグループの血清の
分析を行ったところ、血清中の濃度が堅いと血圧を上昇
させる因子となる血清ＡＣＥ活性が、低い値を示した。As a result of the investigation of the blood pressure lowering effect, as shown in Table 1, in the measurement on the 30th day, the group to which the purified product was administered was compared with the control group to which the purified product was not administered,
It was found that the rise in blood pressure was suppressed. The numbers in the table are mean ± standard error. Also, "student's-t
test, the significant difference was determined.
There was a significant difference in systolic blood pressure and diastolic blood pressure in the group administered mg. The measurement on the 60th day showed the same tendency as the measurement result on the 30th day, and the group to which the purified product was administered showed significantly lower systolic blood pressure. When the serum of the group to which the purified product was administered was analyzed, the serum ACE activity, which is a factor that increases blood pressure when the serum concentration was firm, showed a low value.
【００１８】つぎに、上記精製物による脂肪代謝改善作
用を調査した。この調査では、供試動物としてラット
（ＳＨＲＳＰ／Ｉｚｍ）♂８週を用いた。また、試験グ
ループとして、上記精製物を与えない対照群（１０匹）
と、体重１ｋｇ当たり１００ｍｇの精製物を１日ごとに
投与したグループ（１０匹）と、体重１ｋｇ当たり３０
０ｍｇの精製物を１日ごとに投与したグループとの３グ
ループ（１０匹）を設定した。Next, the effect of the purified product on improving fat metabolism was investigated. In this investigation, rats (SHRSP / Izm) @ 8 weeks were used as test animals. As a test group, a control group not receiving the purified product (10 animals)
And a group (10 animals) receiving 100 mg of purified product per kg of body weight every day, and a group of 30 animals per kg of body weight.
Three groups (10 animals) were set, including a group to which 0 mg of the purified product was administered every day.
【００１９】調査の方法は、まず、精製物を蒸留水で各
濃度に調整し、胃ゾンデを用いて対象のグループに１４
日間連続経口投与する。対照群には、蒸留水を与える。
また、ミルクカゼイン２０．００％，グラニュー糖５
６．７５％，粉末牛脂１０．００％，アビセルロース
４．００％，コレステロール１．００％，コール酸０．
２５％，ビタミン混合１．００％，ミネラル混合７．０
０％からなる飼料を、自由摂取可能な状態としておく。
１４日間の投与終了後、各マウスより採取した血清を分
析する。採取した血清について、表２に示すThe method of the survey is as follows. First, the purified product is adjusted to each concentration with distilled water, and 14 g
Oral administration for consecutive days. The control group receives distilled water.
In addition, milk casein 20.00%, granulated sugar 5
6.75%, powdered beef tallow 10.00%, avicellulose 4.00%, cholesterol 1.00%, cholic acid 0.
25%, 1.00% mixed with vitamins, 7.0 mixed with minerals
A feed consisting of 0% is kept in a state where it can be freely taken.
After 14 days of administration, the serum collected from each mouse is analyzed. Table 2 shows the collected serum.
【００２０】[0020]
【表２】 [Table 2]
【００２１】表２に示すように、脂質関連項目である総
脂質，総コレステロール，ＨＤＬ−コレステロール，Ｌ
ＤＬ−コレステロール，トリグリセライド，遊離脂肪
酸，β−リポ蛋白の分析結果では、対照群との比較にお
いて、精製物を投与した２つのグループに有効な結果が
認められた。また、「student's-t test」により有意差
を検定したところ、精製物を３００ｍｇ投与したグルー
プの、ＨＤＬ−コレステロール，遊離脂肪酸の２項目
で、有意差が確認された。As shown in Table 2, total lipids, total cholesterol, HDL-cholesterol, L
In the analysis results of DL-cholesterol, triglyceride, free fatty acid, and β-lipoprotein, effective results were observed in the two groups to which the purified product was administered, as compared with the control group. In addition, when a significant difference was tested by “student's-t test”, a significant difference was confirmed in two items of HDL-cholesterol and free fatty acid in the group to which 300 mg of the purified product was administered.
【００２２】つぎに、上記精製物による脂肪代謝改善作
用を調査した。この調査では、供試動物としてＫＫＡy
マウス♂８週を用いた。このマウスは、人のインスリン
非依存型糖尿病に類似した病体を呈する自然発症糖尿病
モデルマウスである。また、試験グループとして、上記
精製物を与えない対照群（１０匹）と、体重１ｋｇ当た
り１００ｍｇの精製物を１日ごとに投与したグループ
（１０匹）と、体重１ｋｇ当たり３００ｍｇの精製物を
１日ごとに投与したグループとの３グループ（１０匹）
を設定した。Next, the effect of improving fat metabolism by the purified product was examined. In this study, KKAy was used as a test animal.
Mice # 8 weeks were used. This mouse is a spontaneous diabetes model mouse that presents a pathology similar to human non-insulin dependent diabetes. As a test group, a control group (10 animals) not receiving the purified product, a group (100 animals) administered with 100 mg of the purified product per 1 kg of body weight every day, and a group of 300 mg / kg of the purified product were administered. 3 groups (10 animals) with the group administered daily
It was set.
【００２３】調査の方法は、まず、精製物を蒸留水で各
濃度に調整し、胃ゾンデを用いて対象のグループに３０
日間連続経口投与する。対照群には、蒸留水を与える。
投与開始前と、１５日後と３０日後とに、眼窩静脈叢よ
り採血し、血糖測定器を用いて採血した血清中の血糖値
を測定する。また、満腹時血糖の測定のための採血は、
測定日午前１０時に行う。空腹時血糖の測定のための採
血は、満腹時血糖測定のための採血を行った後で、１８
時間絶食させて測定日翌日午前１０時に行う。測定の結
果は、表３に示す。First, the purified product was adjusted to each concentration with distilled water, and 30 g was administered to the target group using a gastric probe.
Oral administration for consecutive days. The control group receives distilled water.
Before the start of administration, and 15 days and 30 days later, blood is collected from the orbital venous plexus, and the blood glucose level in the collected blood is measured using a blood glucose meter. In addition, blood sampling for measuring blood glucose at the time of satiety
It is performed at 10 am on the measurement day. The blood collection for the measurement of fasting blood glucose is performed after the blood collection for the measurement of the fasting blood glucose is performed.
It is fasted for 10 hours and performed at 10 am the day after the measurement. Table 3 shows the measurement results.
【００２４】[0024]
【表３】 [Table 3]
【００２５】表３に示すように、対照群が、試験期間を
通じて高血糖状態であったのに対し、本実施の形態にお
ける精製物を投与したグループでは、１５日目，３０日
目と血糖値の低下が認められた。また、「student's-t
test」により有意差を検定したところ、精製物を３００
ｍｇ投与したグループの３０日目の結果に、２４０．５
±１４．０ｍｇ／ｄｌと、有意に低い値を示した。As shown in Table 3, the control group was in a hyperglycemic state throughout the test period, while the group to which the purified product of the present embodiment was administered had blood glucose levels on the 15th and 30th days. Decreased. Also, "student's-t
test, the significant difference was determined.
240.5 results for the 30 mg group
The value was significantly lower at ± 14.0 mg / dl.
【００２６】最後に、精製物の安全性を調査した。ま
ず、ネズミチフス菌（ＴＡ９８，ＴＡ１００）を使用し
て、変異原性試験を行ったところ、細胞毒性は、精製物
１００ｍｇ／ｍｌで陰性であり、変異原性は、Ｓ９酵素
活性の有無によらず、精製物１００ｍｇ／ｍｌで陰性で
あった。また、急性毒性試験の結果は、「ＬＤ50＞１０
ｇ／ｋｇ」であった。これらのことから、本実施の形態
における精製物の高い安全性が確認された。Finally, the safety of the purified product was investigated. First, when a mutagenicity test was performed using Salmonella typhimurium (TA98, TA100), the cytotoxicity was negative at 100 mg / ml of the purified product, and the mutagenicity was determined regardless of the presence or absence of S9 enzyme activity. Was negative at 100 mg / ml of the purified product. In addition, the result of the acute toxicity test is “LD 50 > 10
g / kg ". From these, high safety of the purified product in the present embodiment was confirmed.
【００２７】[0027]
【発明の効果】以上説明したように、本発明では、米を
含む培地を用いて乳酸菌ラクトバチラス・ケフィラノフ
ァシエンスを培養することで、ケフィランを含む精製物
を得るようにしたので、多糖を製造する新たな方法が得
られるというすぐれた効果がある。As described above, in the present invention, a lactobacillus Lactobacillus kefiranofaciens is cultured in a medium containing rice to obtain a purified product containing kefiran. There is an excellent effect that a new method of performing the above can be obtained.
【図１】 「ＤＥＡＥ ｓｅｐｈａｄｅｘ Ａ−５０」
で分画した結果を示す特性図である。Fig. 1 "DEAE sephadex A-50"
FIG. 4 is a characteristic diagram showing the results of fractionation in FIG.
【図２】 「ＴＯＹＯＰＥＡＲＬＨＷ６５」により分画
した結果を示す特性図である。FIG. 2 is a characteristic diagram showing a result of fractionation by “TOYOPEARLHW65”.
───────────────────────────────────────────────────── フロントページの続き (72)発明者 大橋 茂 東京都世田谷区三軒茶屋１−16−19 大和 薬品株式会社内 (72)発明者 朱 霞 東京都世田谷区三軒茶屋１−16−19 大和 薬品株式会社内 (72)発明者 二宮 泰夫 東京都世田谷区三軒茶屋１−16−19 大和 薬品株式会社内 Ｆターム(参考） 4B064 AF12 CA02 CD19 CD22 CE06 DA10 4B065 AA30X BB18 BB26 BB29 BD15 BD18 BD22 CA22 CA41 CA44 ──────────────────────────────────────────────────続 き Continued on the front page (72) Inventor Shigeru Ohashi 1-16-19 Sangenjaya, Setagaya-ku, Tokyo Daiwa Pharmaceutical Co., Ltd. (72) Inventor Zhu 1-16-19 Sangenjaya, Setagaya-ku, Tokyo (72) Inventor Yasuo Ninomiya 1-16-19 Sangenjaya, Setagaya-ku, Tokyo Daiwa Pharmaceutical Co., Ltd.F-term (reference) 4B064 AF12 CA02 CD19 CD22 CE06 DA10 4B065 AA30X BB18 BB26 BB29 BD15 BD18 BD22 CA22 CA41 CA44
Claims (4)
ンスを培養し、 前記培養した後の培養液中よりタンパク質と菌とを分離
した分離液を作製し、 前記分離液より所定の分子量以下の物質を除去すること
でケフィランを含む精製物を得ることを特徴とした多糖
製造方法。1. A medium containing rice is prepared, a lactic acid bacterium Lactobacillus kefiranofaciens is cultured in the medium, and a separated solution in which proteins and bacteria are separated from the culture solution after the culturing is prepared. A method for producing a polysaccharide, characterized in that a purified product containing kefiran is obtained by removing a substance having a predetermined molecular weight or less from a separated solution.
する多糖製造方法。2. The method for producing a polysaccharide according to claim 1, wherein the medium contains a saccharified liquid of rice obtained by saccharifying rice.
おいて、 前記培地は、酵母エキスを含むことを特徴とする多糖製
造方法。3. The method for producing a polysaccharide according to claim 1, wherein the culture medium contains a yeast extract.
製造方法において、 前記所定の分子量以下の物質の除去は、限外ろ過により
行うことを特徴とする多糖製造方法。4. The method for producing a polysaccharide according to claim 1, wherein the substance having a predetermined molecular weight or less is removed by ultrafiltration.
Priority Applications (1)
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|---|---|---|---|
| JP2001141827A JP2002330798A (en) | 2001-05-11 | 2001-05-11 | Method for producing polysaccharide |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP2001141827A JP2002330798A (en) | 2001-05-11 | 2001-05-11 | Method for producing polysaccharide |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| JP2002330798A true JP2002330798A (en) | 2002-11-19 |
Family
ID=18988220
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP2001141827A Pending JP2002330798A (en) | 2001-05-11 | 2001-05-11 | Method for producing polysaccharide |
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| Country | Link |
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| JP (1) | JP2002330798A (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2008530034A (en) * | 2005-02-11 | 2008-08-07 | テクノロジー バイオラクティス インコーポレイティド | Use of Lactobacillus kefiranofaciens as probiotics and symbiotics |
-
2001
- 2001-05-11 JP JP2001141827A patent/JP2002330798A/en active Pending
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2008530034A (en) * | 2005-02-11 | 2008-08-07 | テクノロジー バイオラクティス インコーポレイティド | Use of Lactobacillus kefiranofaciens as probiotics and symbiotics |
| JP2013139462A (en) * | 2005-02-11 | 2013-07-18 | Technologies Biolactis Inc | Use of probiotics and lactobacillus kefiranofacience as synbiotic |
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