JPH0481433B2 - - Google Patents
Info
- Publication number
- JPH0481433B2 JPH0481433B2 JP60219707A JP21970785A JPH0481433B2 JP H0481433 B2 JPH0481433 B2 JP H0481433B2 JP 60219707 A JP60219707 A JP 60219707A JP 21970785 A JP21970785 A JP 21970785A JP H0481433 B2 JPH0481433 B2 JP H0481433B2
- Authority
- JP
- Japan
- Prior art keywords
- lactose
- culture
- galactosyllactose
- present
- microorganism
- 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 - Lifetime
Links
- 244000005700 microbiome Species 0.000 claims description 22
- GUBGYTABKSRVRQ-QKKXKWKRSA-N Lactose Natural products OC[C@H]1O[C@@H](O[C@H]2[C@H](O)[C@@H](O)C(O)O[C@@H]2CO)[C@H](O)[C@@H](O)[C@H]1O GUBGYTABKSRVRQ-QKKXKWKRSA-N 0.000 claims description 17
- 239000008101 lactose Substances 0.000 claims description 16
- 238000004519 manufacturing process Methods 0.000 claims description 12
- OWEGMIWEEQEYGQ-UHFFFAOYSA-N 100676-05-9 Natural products OC1C(O)C(O)C(CO)OC1OCC1C(O)C(O)C(O)C(OC2C(OC(O)C(O)C2O)CO)O1 OWEGMIWEEQEYGQ-UHFFFAOYSA-N 0.000 claims description 10
- 241000228143 Penicillium Species 0.000 claims description 7
- 241000223259 Trichoderma Species 0.000 claims description 6
- 241000223260 Trichoderma harzianum Species 0.000 claims description 6
- 241000228212 Aspergillus Species 0.000 claims description 5
- 240000006439 Aspergillus oryzae Species 0.000 claims description 5
- 235000002247 Aspergillus oryzae Nutrition 0.000 claims description 5
- 240000000064 Penicillium roqueforti Species 0.000 claims 1
- 235000002233 Penicillium roqueforti Nutrition 0.000 claims 1
- 239000002609 medium Substances 0.000 description 8
- 238000000034 method Methods 0.000 description 8
- 239000000203 mixture Substances 0.000 description 7
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 6
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 5
- 150000001720 carbohydrates Chemical class 0.000 description 5
- 235000014633 carbohydrates Nutrition 0.000 description 5
- 239000001965 potato dextrose agar Substances 0.000 description 5
- 239000000843 powder Substances 0.000 description 4
- 241000894006 Bacteria Species 0.000 description 3
- 230000002550 fecal effect Effects 0.000 description 3
- 239000000706 filtrate Substances 0.000 description 3
- 239000001963 growth medium Substances 0.000 description 3
- 230000001737 promoting effect Effects 0.000 description 3
- 239000002689 soil Substances 0.000 description 3
- 238000004809 thin layer chromatography Methods 0.000 description 3
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 3
- ATYSZLKTHMZHJA-UXLSSDPBSA-N (3R,4R,5S,6R)-6-(hydroxymethyl)-2-[(3R,4S,5R,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl]-5-[(2S,3R,4S,5R,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl]oxyoxane-2,3,4-triol Chemical compound C1([C@H](O)[C@@H](O)[C@@H](O)[C@H](O1)CO)C1(O)[C@H](O)[C@@H](O)[C@H](O[C@H]2[C@H](O)[C@@H](O)[C@@H](O)[C@H](O2)CO)[C@H](O1)CO ATYSZLKTHMZHJA-UXLSSDPBSA-N 0.000 description 2
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 2
- 241000186000 Bifidobacterium Species 0.000 description 2
- 241000233866 Fungi Species 0.000 description 2
- 230000001580 bacterial effect Effects 0.000 description 2
- 102000005936 beta-Galactosidase Human genes 0.000 description 2
- 108010005774 beta-Galactosidase Proteins 0.000 description 2
- 235000020256 human milk Nutrition 0.000 description 2
- 210000004251 human milk Anatomy 0.000 description 2
- 239000013028 medium composition Substances 0.000 description 2
- 235000015097 nutrients Nutrition 0.000 description 2
- 239000000047 product Substances 0.000 description 2
- 150000004044 tetrasaccharides Chemical class 0.000 description 2
- ISJHTNASRGGDRC-MWGHHZFTSA-N (4R)-3-acetyl-3,4-dihydroxy-4-[(1S,2R,3R)-1,2,3,4-tetrahydroxybutyl]hexane-2,5-dione Chemical compound C(C)(=O)[C@@](C(O)(C(C)=O)C(C)=O)(O)[C@@H](O)[C@H](O)[C@H](O)CO ISJHTNASRGGDRC-MWGHHZFTSA-N 0.000 description 1
- 229920001817 Agar Polymers 0.000 description 1
- 102000002464 Galactosidases Human genes 0.000 description 1
- 108010093031 Galactosidases Proteins 0.000 description 1
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 1
- 244000061456 Solanum tuberosum Species 0.000 description 1
- 235000002595 Solanum tuberosum Nutrition 0.000 description 1
- 239000008272 agar Substances 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 229910052799 carbon Inorganic materials 0.000 description 1
- 238000012258 culturing Methods 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 238000010828 elution Methods 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 210000000416 exudates and transudate Anatomy 0.000 description 1
- 239000000835 fiber Substances 0.000 description 1
- 238000001914 filtration Methods 0.000 description 1
- 235000013350 formula milk Nutrition 0.000 description 1
- 229930182830 galactose Natural products 0.000 description 1
- 239000008103 glucose Substances 0.000 description 1
- 238000011534 incubation Methods 0.000 description 1
- 238000001802 infusion Methods 0.000 description 1
- 238000003973 irrigation Methods 0.000 description 1
- 230000002262 irrigation Effects 0.000 description 1
- 230000011987 methylation Effects 0.000 description 1
- 238000007069 methylation reaction Methods 0.000 description 1
- 229910052757 nitrogen Inorganic materials 0.000 description 1
- 238000007747 plating Methods 0.000 description 1
- 235000008476 powdered milk Nutrition 0.000 description 1
- 238000004321 preservation Methods 0.000 description 1
- 235000021067 refined food Nutrition 0.000 description 1
- 229930187593 rose bengal Natural products 0.000 description 1
- AZJPTIGZZTZIDR-UHFFFAOYSA-L rose bengal Chemical compound [K+].[K+].[O-]C(=O)C1=C(Cl)C(Cl)=C(Cl)C(Cl)=C1C1=C2C=C(I)C(=O)C(I)=C2OC2=C(I)C([O-])=C(I)C=C21 AZJPTIGZZTZIDR-UHFFFAOYSA-L 0.000 description 1
- 229940081623 rose bengal Drugs 0.000 description 1
- STRXNPAVPKGJQR-UHFFFAOYSA-N rose bengal A Natural products O1C(=O)C(C(=CC=C2Cl)Cl)=C2C21C1=CC(I)=C(O)C(I)=C1OC1=C(I)C(O)=C(I)C=C21 STRXNPAVPKGJQR-UHFFFAOYSA-N 0.000 description 1
- 150000003839 salts Chemical class 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 241000894007 species Species 0.000 description 1
- 239000000758 substrate Substances 0.000 description 1
- 230000002194 synthesizing effect Effects 0.000 description 1
- 238000005918 transglycosylation reaction Methods 0.000 description 1
- 150000004043 trisaccharides Chemical class 0.000 description 1
- 238000005406 washing Methods 0.000 description 1
- 239000002023 wood Substances 0.000 description 1
Landscapes
- Preparation Of Compounds By Using Micro-Organisms (AREA)
Description
【発明の詳細な説明】
産業上の利用分野
本発明は、ビフイズス菌の増殖促進因子並びに
便性改善因子として利用される6′−ガラクトシル
ラクトースの製造方法に関する。DETAILED DESCRIPTION OF THE INVENTION Field of Industrial Application The present invention relates to a method for producing 6'-galactosyllactose, which is used as a growth promoting factor for Bifidobacterium and as a fecal quality improving factor.
ガラクトシルラクトースは、乳糖にガラクトー
スが結合した３糖類の総称であり、そのうちの
6′−ガラクトシルラクトースは人乳中に含まれて
いて、育児用粉乳に添加してビフイズス菌の増殖
促進因子及び便性改善因子として利用されてい
る。 Galactosyllactose is a general term for trisaccharides in which galactose is bonded to lactose.
6'-galactosyllactose is contained in human milk, and is added to powdered milk for infants to be used as a growth promoting factor for bifidobacteria and a fecal quality improving factor.
従来の技術
従来、ガラクトシルラクトース（以下GLと略
記する）の製造法として、乳糖にアスペルギル
ス・オリーゼが生産するβ−ガラクトシダーゼを
作用させることにより合成する方法（特公昭58−
20266）が知られている。Conventional technology Conventionally, as a method for producing galactosyllactose (hereinafter abbreviated as GL), a method of synthesizing lactose by reacting β-galactosidase produced by Aspergillus oryzae (Japanese Patent Publication No. 1983-
20266) is known.
しかし、この方法ではβ−ガラクトシダーゼに
よる糖転移反応によつてGLを合成するため、少
くとも４種類のGLと４糖類及び５糖類が同時に
合成される。したがつて、人乳中に含まれる6′−
GLのみを合成することができないので、上述し
た４種類以上のGLと４糖類及び５糖類とから成
る混合物を育児用粉乳に添加することによつて、
該混合物中に含まれる6′−GLを利用しているの
が現状であり、6′−GLの有効な利用上の観点か
ら、6′−GLのみを合成し得る方法の開発が要望
されている。 However, in this method, GL is synthesized by transglycosylation reaction using β-galactosidase, so at least four types of GL, tetrasaccharides, and pentasaccharides are simultaneously synthesized. Therefore, the 6′− contained in human milk
Since it is not possible to synthesize GL alone, by adding a mixture consisting of the above-mentioned four or more types of GL and tetrasaccharides and pentasaccharides to infant milk powder,
Currently, 6'-GL contained in the mixture is used, and from the viewpoint of effective utilization of 6'-GL, there is a demand for the development of a method that can synthesize only 6'-GL. There is.
発明が解決しようとする問題点
本発明者は、上述した現状に鑑み、6′−GLの
みを効率よく製造し得る方法について検討した結
果、アスペルギルス属、ペニシリウム属、ならび
にトリコデルマ属に属するラクトース資化性菌
が、乳糖を基質として6′−GLのみを産出するこ
とを見出し、本発明をなすに至つた。Problems to be Solved by the Invention In view of the above-mentioned current situation, the present inventor investigated a method for efficiently producing only 6'-GL, and found that lactose assimilation of 6′-GL belonging to the genus Aspergillus, Penicillium, and Trichoderma The present inventors have discovered that 6'-GL is produced only by using lactose as a substrate, leading to the present invention.
すなわち、本発明は、微生物を利用して6′−
GLのみを効率よく製造し得る方法を提供するこ
とを目的とする。 That is, the present invention utilizes microorganisms to produce 6′-
The purpose is to provide a method for efficiently manufacturing only GL.
以下に本発明を詳しく説明する。 The present invention will be explained in detail below.
発明の構成
本発明の特徴は、アスペルギルス層（Aspergil
−lus）、ペニシリウム属（Penicillium）及びト
リコデルマ属（Trichoderma）に属する微生物
から成る群から選択される、乳糖から6′−ガラク
トシルラクトースの産生能を有する微生物を、乳
糖を含有する倍地中に培養し、得られた、培養物
から6′−ガラクトシルラクトースを分離、採取す
ることにある。Structure of the Invention The present invention is characterized by the Aspergillus layer (Aspergillus layer).
A microorganism capable of producing 6'-galactosyllactose from lactose, selected from the group consisting of microorganisms belonging to the genus Penicillium and Trichoderma, is cultured in a medium containing lactose. Then, 6'-galactosyllactose is separated and collected from the obtained culture.
問題点を解決するための手段
本発明において利用する微生物は、アスペルギ
ルス属またはペニシリウム属もしくはトリコデル
マ属に属するラクトース資化性菌であつて、アス
ペルギルス・オリゼ（Aspergillus oryzae）IFO
4135、ペニシリウム・ロツクフオルテ（Penicilli
−um roqueforti）IFO 4622ならびにトリコデル
マ・ハルジヤヌム（Trichoderma harzianum）
SBT 7045（微工研菌寄第8338号）を例示し得る。Means for Solving the Problems The microorganism used in the present invention is a lactose-assimilating bacterium belonging to the genus Aspergillus, Penicillium, or Trichoderma, and Aspergillus oryzae IFO
4135, Penicilli
−um roqueforti) IFO 4622 and Trichoderma harzianum
SBT 7045 (Feikoken Bibori No. 8338) can be exemplified.
トリコデルマは、ペニシリウム属とともに土壌
中に非常に多く分布する菌である。また、各種農
作物や加工食品の汚染菌、木材、繊維の腐朽菌と
しても知られている。 Trichoderma, together with the genus Penicillium, is a fungus that is widely distributed in soil. It is also known as a bacterium that contaminates various agricultural products and processed foods, and a bacterium that rots wood and fibers.
本発明で使用するトリコデルマ・ハルジヤヌム
は東北大農学部のキヤンパスの土壌を10％（Ｗ／
Ｖ）ラクトース溶液で洗浄（irrigation）し、
洗浄液を培養し、これを周知の土壌平板法によ
つて分離した。 Trichoderma harzianum used in the present invention was produced using 10% soil (W/
V) Irrigation with lactose solution, incubation of the washings and separation by the well-known soil plating method.
このカビはバレイシヨ・ブドウ糖寒天倍地
（PDA）、PDA＋ローズベンガル添加バレイシ
ヨ・ブドウ糖寒天培地（PDA＋Ｒ）で20〜30℃
で好湿度で発育する。集落の大きさは大で、色は
黄緑色乃至濃緑色を呈し、臭気がない。そして、
その表面の性状は綿状で湿性であり、浸出液がな
く、裏面が無色乃至淡黄色である。 This mold was grown at 20-30°C on potato-dextrose agar medium (PDA) or potato-dextrose agar medium supplemented with PDA + rose bengal (PDA + R).
It grows in good humidity. The size of the colony is large, the color is yellow-green to dark green, and there is no odor. and,
Its surface is cotton-like and moist, with no exudate, and its back surface is colorless to pale yellow.
菌糸は有隔壁があり無色である。分生子は球形
乃至要球形で方面が平滑である。分生子柄は無色
で気中菌色より直生する。 The hyphae are septate and colorless. Conidia are spherical to semi-spherical with smooth sides. Conidiophores are colorless and grow straight rather than aerial fungi.
これらの菌学的性質からみて、特に、分生子が
球形乃至要球形で裏面が平滑でするという点を考
慮してRifal，M.A.：Ａ revision of the genus
Trichoderma，Mycological Papers No.116.56
（1969）に従つてトリコデルマ・ハルジヤヌムに
属するものと同定した。 Considering these mycological properties, especially considering that the conidia are spherical or semi-spherical with a smooth back surface, Rifal, MA: A revision of the genus.
Trichoderma，Mycological Papers No.116.56
(1969), it was identified as belonging to Trichoderma harzianum.
これらの微生物は、いずれも公知の菌種であつ
て、上掲の各寄託番号で保管されていて入手可能
である。 All of these microorganisms are known species, and are stored and available under the above-mentioned deposit numbers.
本発明では、まず、上掲の各微生物を下記に示
す組成の基本培地を用いて前培地を行なつて微生
物を増殖させる。 In the present invention, first, each of the microorganisms listed above is precultured using a basic medium having the composition shown below to grow the microorganisms.
前培養のための基本培地組成：
乳糖 30ｇ
K2HPO4 １ｇ
MgSO4・7H2O 0.5ｇ
NaNO3 ２ｇ
KCl 0.5ｇ
FeSO4 0.01ｇ
水 １
PH 6.0
次いで、微生物を上記前培養のための培地中で
増殖させたものを下記に示す組成の基本培地を用
いて本培養を行なつて6′−GLを生産する。 Basic medium composition for preculture: Lactose 30g K 2 HPO 4 1g MgSO 4・7H 2 O 0.5g NaNO 3 2g KCl 0.5g FeSO 4 0.01g Water 1 PH 6.0 Then, microorganisms were added to the above medium for preculture. 6'-GL is produced by carrying out main culture using a basic medium having the composition shown below.
本培養のための基本培地組成：
乳糖 150ｇ
K2HPO4 １ｇ
MgSO4・7H2O 0.5ｇ
NaNO3 ２ｇ
KCl 0.5ｇ
FeSO4 0.01ｇ
水 １
PH ４〜９
なお、上記各培地の使用に当つては、微生物が
生育上利用し得る他の炭素源、窒素源及び無機
塩、さらにはその他の栄養分を適宜添加して含有
させることができる。 Basic medium composition for main culture: Lactose 150g K 2 HPO 4 1g MgSO 4・7H 2 O 0.5g NaNO 3 2g KCl 0.5g FeSO 4 0.01g Water 1 PH 4-9 In addition, when using each of the above media may contain other carbon sources, nitrogen sources, inorganic salts, and other nutrients that can be used by microorganisms for growth, as well as other nutrients, as appropriate.
因に、本発明で利用する上記微生物を保存する
には下記に示す培地を用いるとよい。 Incidentally, in order to preserve the above-mentioned microorganisms used in the present invention, it is preferable to use the culture medium shown below.
保存培養培地組成：
バレイシヨ浸出液 200ｇ
グルコース 20ｇ
寒天 15ｇ
水 １
本発明において、上記微生物を利用して6′−
GLを産生するには、該微生物を前述のようにし
て増殖させたものを、上記本培養のための培地に
接種してPH４〜９、20〜40℃、好ましくは30℃前
後で通常５〜15日間振とう培養を行なう。Preservation culture medium composition: Potato infusion 200g Glucose 20g Agar 15g Water 1 In the present invention, 6'-
To produce GL, the microorganisms grown as described above are inoculated into the medium for the main culture described above and incubated at pH 4-9, 20-40°C, preferably around 30°C, usually 5-5°C. Perform shaking culture for 15 days.
参考として、アスペルギルス・オリゼ
IFO4135、ペニシリウム・ロツクフオルテ
IFO4622及びトリコデルマ・ハルジヤヌム
SBT7045（微工研菌寄第8338号）の各微生物の10
日間の培養におけるPHと温度条件が6′−GLの産
生量に及ぼす影響を調べた結果を添付の第１図
（ａ及びｂ）に示す。 For reference, Aspergillus oryzae
IFO4135, Penicillium rockfuorte
IFO4622 and Trichoderma harzianum
10 of each microorganism in SBT7045 (Feikoken Bibori No. 8338)
The results of examining the effects of PH and temperature conditions on the production amount of 6'-GL during the 1-day culture are shown in the attached Figures 1 (a and b).
第１図にみられるとおり、各微生物とも培養時
のPH４〜９及び温度20〜40℃において6′−GLの
産生量が高くなることが認められる。 As seen in FIG. 1, it is recognized that the production amount of 6'-GL increases for each microorganism at a pH of 4 to 9 and a temperature of 20 to 40°C during culture.
本発明に従つて、上記微生物を上述のようにし
て培養して得られる培養液中には糖質として未反
応の乳糖と6′−GLのみが検出され、他の糖質の
産生はみられない。 According to the present invention, only unreacted lactose and 6'-GL were detected as carbohydrates in the culture solution obtained by culturing the microorganism as described above, and no production of other carbohydrates was observed. do not have.
上記培養液中の糖質を薄層クロマトグラフイー
により測定した結果を薄層クロマトパターンで示
すと添付の第２図のとおりである。 The results of measuring carbohydrates in the above culture solution by thin layer chromatography are shown in the attached FIG. 2 as a thin layer chromatography pattern.
培養終了後の培養液から6′−GLを分離、採取
するには公知の手法を適用して行ない得る。例え
ば、上記培養液もしくはそれを濾別して得られた
瀘液を減圧下に濃縮して培養液中に残存する乳糖
を析出させて分離、除去した後、更にエシエリヒ
ア・コリ（Escherichia coli）由来のβ−ガラク
トシダーゼを作用させて培養液中に残存する乳糖
を分解し、ついで該培養液を活性炭のカラムに通
して6′−GLのみを吸着させ、この吸着した6′−
GLを20％エタノールで溶出させることにより、
精製した6′−GLが得られる。 6'-GL can be separated and collected from the culture solution after completion of culture by applying known techniques. For example, after concentrating the above culture solution or the filtrate obtained by filtering it under reduced pressure to precipitate, separate, and remove the lactose remaining in the culture solution, - Activate galactosidase to decompose the lactose remaining in the culture solution, then pass the culture solution through an activated carbon column to adsorb only 6'-GL, and this adsorbed 6'-
By eluting GL with 20% ethanol,
Purified 6′-GL is obtained.
このようにして得られた6′−GLの構造をメチ
ル化分析により調べた結果、２，３，４，６−テ
トラ−ｏ−メチル−１，５−ジ−ｏ−アセチル−
ガラクチトール、、２，３，４−トリ−ｏ−メチ
ル−１，５，６−トリ−ｏ−アセチル−ガラクチ
トール、２，３，６−ｏ−メチル−１，４，５−
トリ−アセチル−ソルビトールが検出されたこと
から、純粋な6′−ガラクトシルラクトースである
ことが確認された。 As a result of investigating the structure of 6'-GL thus obtained by methylation analysis, it was found that 2,3,4,6-tetra-o-methyl-1,5-di-o-acetyl-
Galactitol, 2,3,4-tri-o-methyl-1,5,6-tri-o-acetyl-galactitol, 2,3,6-o-methyl-1,4,5-
Since tri-acetyl-sorbitol was detected, it was confirmed that it was pure 6'-galactosyllactose.
叙上のように、本発明によると、微生物を利用
して乳糖から直接的に、且つ他の糖質生成物を伴
なうことなく、6′−GLのみを効率よく製造し得
るので、ビフイズス菌の増殖促進因子及び便性改
善因子として有用な6′−GLの製造上有益である
と言える。 As mentioned above, according to the present invention, only 6'-GL can be efficiently produced directly from lactose using microorganisms and without involving other carbohydrate products. It can be said that it is useful in the production of 6'-GL, which is useful as a bacterial growth promoting factor and a fecal quality improving factor.
以下に実施例を示して本発明を具体的に説明す
る。 EXAMPLES The present invention will be specifically described below with reference to Examples.
実施例 １
アスペルギルス・オリゼ（Aspergillus
oryzae）IFO 4135の菌株を前述した組成の前培
養培地100mlに接種し、30℃で５日間培養した。Example 1 Aspergillus oryzae
oryzae) IFO 4135 was inoculated into 100 ml of preculture medium having the composition described above, and cultured at 30°C for 5 days.
上述のように得られた前培養液100mlを、前述
した組成の本培養培地５に接種し、30℃で10日
間振とう培養した。得られた培養液を遠心分離し
て菌体を除去した後、減圧濃縮して培養濾液中の
乳糖を析出させて除去した。ついで、得られた培
養濾液を15cm×30cmの活性炭カラムに通じて6′−
GLを吸着させた後、５％エタノール３を通じ
て残存している乳糖を溶出除去した。ついで、20
％エタノール３を通じて得た溶出区分を減圧濃
縮した後、凍結乾燥して白色の6′−GLの粉末150
ｇを得た。 100 ml of the preculture solution obtained as described above was inoculated into main culture medium 5 having the composition described above, and cultured with shaking at 30° C. for 10 days. The obtained culture solution was centrifuged to remove bacterial cells, and then concentrated under reduced pressure to precipitate and remove lactose in the culture filtrate. Next, the obtained culture filtrate was passed through a 15 cm x 30 cm activated carbon column and 6′-
After adsorbing GL, remaining lactose was eluted and removed using 5% ethanol. Then, 20
The elution fraction obtained through 3% ethanol was concentrated under reduced pressure, and then lyophilized to obtain a white 6′-GL powder of 150%.
I got g.
実施例 ２
ペニシリウム・ロツクフオルテIFO4622の菌株
を用いて実施例１に記載したと同様の方法によ
り、6′−GLの粉末50ｇを得た。Example 2 50 g of 6'-GL powder was obtained in the same manner as described in Example 1 using Penicillium lockforde IFO4622 strain.
実施例 ３
トリコデルマ・ハルジヤヌムSBT7045（微工研
菌第8338号）の菌株を用いて、実施例２に記載し
たと同様の方法により、6′−GLの粉末130ｇを得
た。Example 3 130 g of 6'-GL powder was obtained in the same manner as described in Example 2 using the strain of Trichoderma harzianum SBT7045 (Feikokenboku No. 8338).
添付の第１図（ａ及びｂ）は、本発明の方法に
おける微生物の培養条件と6′−ガラクトシルラク
トースの産生量との関係を示したものであり、第
２図は、本発明により培養して得られた培養液中
の糖質の薄層クロマトパターンを示したものであ
る。
Attached Figures 1 (a and b) show the relationship between the culture conditions of microorganisms and the production amount of 6'-galactosyllactose in the method of the present invention, and Figure 2 shows the relationship between microorganisms cultured according to the present invention and the amount of 6'-galactosyllactose produced. This figure shows the thin-layer chromatography pattern of carbohydrates in the culture solution obtained.
Claims (1)
ウム属（Penicillium）及びトリコデルマ属
（Trichoderma）に属する微生物から成る群から
選択される、乳糖から6′−ガラクトシルラクトー
スの産生能を有する微生物を乳糖を含有する培地
中に培養し、得られた培養物より6′−ガラクトシ
ルラクトースを分離、採取することを特徴とする
6′−ガラクトシルラクトースの製造法。 ２ 微生物がアスペルギルス・オリゼ
（Aspergillus oryzae）である特許請求の範囲第
１項記載の製造法。 ３ 微生物がペニシリウム・ロツクフオルテ
（Penicillium roqueforti）である特許請求の範囲
第１項記載の製造法。 ４ 微生物がトリコデルマ・ハルジヤヌム
（Trichoderma harzianum）である特許請求の
範囲第１項記載の製造法。[Scope of Claims] 1. A microorganism having the ability to produce 6'-galactosyllactose from lactose, which is selected from the group consisting of microorganisms belonging to the genus Aspergillus, Penicillium, and Trichoderma, is 6′-galactosyllactose is separated and collected from the resulting culture.
Method for producing 6′-galactosyllactose. 2. The production method according to claim 1, wherein the microorganism is Aspergillus oryzae. 3. The production method according to claim 1, wherein the microorganism is Penicillium roqueforti. 4. The production method according to claim 1, wherein the microorganism is Trichoderma harzianum.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP60219707A JPS6279791A (en) | 1985-10-02 | 1985-10-02 | Production of 6'-galactosyl lactose utilizing lactose-assimilating microorganism |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP60219707A JPS6279791A (en) | 1985-10-02 | 1985-10-02 | Production of 6'-galactosyl lactose utilizing lactose-assimilating microorganism |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| JPS6279791A JPS6279791A (en) | 1987-04-13 |
| JPH0481433B2 true JPH0481433B2 (en) | 1992-12-24 |
Family
ID=16739703
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP60219707A Granted JPS6279791A (en) | 1985-10-02 | 1985-10-02 | Production of 6'-galactosyl lactose utilizing lactose-assimilating microorganism |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JPS6279791A (en) |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR20200026274A (en) * | 2017-07-04 | 2020-03-10 | 가부시키가이샤 야쿠르트 혼샤 | Method for producing galactooligosaccharide |
-
1985
- 1985-10-02 JP JP60219707A patent/JPS6279791A/en active Granted
Also Published As
| Publication number | Publication date |
|---|---|
| JPS6279791A (en) | 1987-04-13 |
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