JPS6394987A - Novel production of beta-d-galactopyranosyl-(1-6)-beta-d-galactopyranosyl-(1-4)-d-fructose - Google Patents
Novel production of beta-d-galactopyranosyl-(1-6)-beta-d-galactopyranosyl-(1-4)-d-fructoseInfo
- Publication number
- JPS6394987A JPS6394987A JP23932186A JP23932186A JPS6394987A JP S6394987 A JPS6394987 A JP S6394987A JP 23932186 A JP23932186 A JP 23932186A JP 23932186 A JP23932186 A JP 23932186A JP S6394987 A JPS6394987 A JP S6394987A
- Authority
- JP
- Japan
- Prior art keywords
- galactopyranosyl
- lactulose
- fructose
- aqueous solution
- beta
- 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.)
- Pending
Links
- 238000004519 manufacturing process Methods 0.000 title claims description 24
- JZBCIZZDHIDCRE-VPUALBSKSA-N (3S,4R,5R)-1,3,5,6-tetrahydroxy-4-[(2S,3R,4S,5R,6R)-3,4,5-trihydroxy-6-[[(2R,3R,4S,5R,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl]oxymethyl]oxan-2-yl]oxyhexan-2-one Chemical compound [C@@H]1([C@H](O)[C@@H](O)[C@@H](O)[C@H](O1)CO)OC[C@@H]1[C@@H]([C@@H]([C@H]([C@@H](O1)O[C@@H]([C@@H](C(CO)=O)O)[C@H](O)CO)O)O)O JZBCIZZDHIDCRE-VPUALBSKSA-N 0.000 title 1
- JCQLYHFGKNRPGE-FCVZTGTOSA-N lactulose Chemical compound OC[C@H]1O[C@](O)(CO)[C@@H](O)[C@@H]1O[C@H]1[C@H](O)[C@@H](O)[C@@H](O)[C@@H](CO)O1 JCQLYHFGKNRPGE-FCVZTGTOSA-N 0.000 claims abstract description 21
- 229960000511 lactulose Drugs 0.000 claims abstract description 21
- PFCRQPBOOFTZGQ-UHFFFAOYSA-N lactulose keto form Natural products OCC(=O)C(O)C(C(O)CO)OC1OC(CO)C(O)C(O)C1O PFCRQPBOOFTZGQ-UHFFFAOYSA-N 0.000 claims abstract description 21
- 108010005774 beta-Galactosidase Proteins 0.000 claims abstract description 13
- 239000007864 aqueous solution Substances 0.000 claims abstract description 10
- 229930182830 galactose Natural products 0.000 claims abstract description 3
- 102000005936 beta-Galactosidase Human genes 0.000 claims abstract 3
- 238000006276 transfer reaction Methods 0.000 claims description 2
- 239000000243 solution Substances 0.000 abstract description 16
- 238000006243 chemical reaction Methods 0.000 abstract description 15
- 108090000790 Enzymes Proteins 0.000 abstract description 13
- 102000004190 Enzymes Human genes 0.000 abstract description 13
- 239000007788 liquid Substances 0.000 abstract description 7
- 238000010438 heat treatment Methods 0.000 abstract description 5
- 239000000047 product Substances 0.000 abstract description 3
- 238000000746 purification Methods 0.000 abstract description 3
- 150000003839 salts Chemical class 0.000 abstract description 3
- 239000012467 final product Substances 0.000 abstract description 2
- 230000001737 promoting effect Effects 0.000 abstract description 2
- 150000001875 compounds Chemical class 0.000 abstract 2
- 239000000470 constituent Substances 0.000 abstract 1
- 238000001035 drying Methods 0.000 abstract 1
- 230000002779 inactivation Effects 0.000 abstract 1
- 239000004615 ingredient Substances 0.000 abstract 1
- 229940068140 lactobacillus bifidus Drugs 0.000 abstract 1
- 238000000926 separation method Methods 0.000 abstract 1
- 230000009466 transformation Effects 0.000 abstract 1
- 239000005715 Fructose Substances 0.000 description 14
- 229940088598 enzyme Drugs 0.000 description 12
- 239000007787 solid Substances 0.000 description 11
- 102100026189 Beta-galactosidase Human genes 0.000 description 10
- 238000000034 method Methods 0.000 description 8
- 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 7
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 7
- 239000000126 substance Substances 0.000 description 7
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 6
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 6
- KRKNYBCHXYNGOX-UHFFFAOYSA-N citric acid Chemical compound OC(=O)CC(O)(C(O)=O)CC(O)=O KRKNYBCHXYNGOX-UHFFFAOYSA-N 0.000 description 6
- 230000000694 effects Effects 0.000 description 6
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 6
- RFSUNEUAIZKAJO-ARQDHWQXSA-N Fructose Chemical compound OC[C@H]1O[C@](O)(CO)[C@@H](O)[C@@H]1O RFSUNEUAIZKAJO-ARQDHWQXSA-N 0.000 description 5
- 108010059881 Lactase Proteins 0.000 description 5
- 239000003513 alkali Substances 0.000 description 5
- 229940116108 lactase Drugs 0.000 description 5
- 238000007796 conventional method Methods 0.000 description 4
- 239000006188 syrup Substances 0.000 description 4
- 235000020357 syrup Nutrition 0.000 description 4
- 229930091371 Fructose Natural products 0.000 description 3
- 239000003925 fat Substances 0.000 description 3
- 229920001542 oligosaccharide Polymers 0.000 description 3
- 150000002482 oligosaccharides Chemical class 0.000 description 3
- 239000000843 powder Substances 0.000 description 3
- 238000012360 testing method Methods 0.000 description 3
- NWUYHJFMYQTDRP-UHFFFAOYSA-N 1,2-bis(ethenyl)benzene;1-ethenyl-2-ethylbenzene;styrene Chemical compound C=CC1=CC=CC=C1.CCC1=CC=CC=C1C=C.C=CC1=CC=CC=C1C=C NWUYHJFMYQTDRP-UHFFFAOYSA-N 0.000 description 2
- 241000186000 Bifidobacterium Species 0.000 description 2
- RFSUNEUAIZKAJO-VRPWFDPXSA-N D-Fructose Natural products OC[C@H]1OC(O)(CO)[C@@H](O)[C@@H]1O RFSUNEUAIZKAJO-VRPWFDPXSA-N 0.000 description 2
- 101000993347 Gallus gallus Ciliary neurotrophic factor Proteins 0.000 description 2
- 239000003957 anion exchange resin Substances 0.000 description 2
- 229910052799 carbon Inorganic materials 0.000 description 2
- 238000004440 column chromatography Methods 0.000 description 2
- 239000000203 mixture Substances 0.000 description 2
- 102000004169 proteins and genes Human genes 0.000 description 2
- 108090000623 proteins and genes Proteins 0.000 description 2
- 239000002994 raw material Substances 0.000 description 2
- 239000013076 target substance Substances 0.000 description 2
- NMELTECMHKKXLF-YNMYPSOUSA-N (3s,4r,5r)-3,4,5,6-tetrahydroxy-1-[(2r,3r,4s,5r,6r)-3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl]oxyhexan-2-one Chemical compound OC[C@@H](O)[C@@H](O)[C@H](O)C(=O)CO[C@@H]1O[C@H](CO)[C@H](O)[C@H](O)[C@H]1O NMELTECMHKKXLF-YNMYPSOUSA-N 0.000 description 1
- 241001655328 Bifidobacteriales Species 0.000 description 1
- 241001562081 Ikeda Species 0.000 description 1
- 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 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- 238000007259 addition reaction Methods 0.000 description 1
- 239000006227 byproduct Substances 0.000 description 1
- 239000003054 catalyst Substances 0.000 description 1
- 238000005341 cation exchange Methods 0.000 description 1
- 239000003729 cation exchange resin Substances 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 239000011248 coating agent Substances 0.000 description 1
- 238000000576 coating method Methods 0.000 description 1
- 238000000354 decomposition reaction Methods 0.000 description 1
- 230000007423 decrease Effects 0.000 description 1
- 238000010828 elution Methods 0.000 description 1
- 230000002255 enzymatic effect Effects 0.000 description 1
- 238000006911 enzymatic reaction Methods 0.000 description 1
- 238000001914 filtration Methods 0.000 description 1
- 238000001641 gel filtration chromatography Methods 0.000 description 1
- 238000002523 gelfiltration Methods 0.000 description 1
- 238000004128 high performance liquid chromatography Methods 0.000 description 1
- 238000005342 ion exchange Methods 0.000 description 1
- 239000003456 ion exchange resin Substances 0.000 description 1
- 229920003303 ion-exchange polymer Polymers 0.000 description 1
- 150000002500 ions Chemical class 0.000 description 1
- 238000006317 isomerization reaction Methods 0.000 description 1
- 239000008101 lactose Substances 0.000 description 1
- 150000002597 lactoses Chemical class 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 230000035484 reaction time Effects 0.000 description 1
- 230000008707 rearrangement Effects 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 239000011347 resin Substances 0.000 description 1
- 229920005989 resin Polymers 0.000 description 1
- 238000003756 stirring Methods 0.000 description 1
Landscapes
- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
Description
【発明の詳細な説明】
〔産業上の利用分野〕
本発明はビフィズス菌増殖促進活性を有するβ−Ｄ−ガ
ラクトピラノシル−（ｌ→６）−β−Ｄ−ガラクトピラ
ノシル−（１→４）−〇−フラクトース（Ｃ，ＣＦ）を
ラクチュロースから製造する新規な方法に関する。Detailed Description of the Invention [Industrial Application Field] The present invention provides β-D-galactopyranosyl-(l→6)-β-D-galactopyranosyl-(1 →4) -Relates to a new method for producing -0-fructose (C, CF) from lactulose.
最近、ビフィズス菌の有用性が広く認識されるようにな
り、人間の体内におけるビフィズス菌の増殖促進活性を
有する物質について、多くノ研究が行なわれている。モ
してβ−Ｄ−ガラクトピラノシル−（１→６）−β−Ｄ
−ガラクトピラノシル−（ｌ→４）−Ｄ−フラクトース
にはそれの増殖促進活性があることが報告されている。Recently, the usefulness of Bifidobacteria has become widely recognized, and many studies have been conducted on substances that have the activity of promoting the growth of Bifidobacteria in the human body. β-D-galactopyranosyl-(1→6)-β-D
-Galactopyranosyl-(l→4)-D-fructose has been reported to have growth-promoting activity.
そして、その製造法としてガラクトシルラクトースのア
ルカリ異性化法（日本農芸化学会昭和６１年度大会、講
演要旨集、１８４頁、昭和６１年４月）が知られている
。As a method for producing it, the alkaline isomerization method of galactosyllactose (Japan Society of Agricultural Chemistry, 1988 Conference, Abstracts of Lectures, p. 184, April 1988) is known.
この方法は、ガラクトシルラクトースをアルカリ触媒下
で、ドブリー・ドブリュイン転移を行なわせて目的物質
を生成せしめるものであり、アルカリの添加、加熱反応
、添加アルカリイオンおよび反応副生物である着色物質
のイオン交換樹脂による除去、精製等の複雑な工程を要
し、またアルカリ添加反応時に分解する、例えば脂肪や
タンパク質等の物質の共存は不可能である欠点を有して
いる。This method generates the target substance by subjecting galactosyllactose to Dobrey-Debruyn rearrangement under an alkali catalyst, and involves the addition of an alkali, heating reaction, and ion exchange of the added alkali ions and the colored substance that is a reaction by-product. It requires complicated steps such as removal with a resin and purification, and also has the disadvantage that it is impossible to coexist with substances such as fats and proteins that are decomposed during the alkali addition reaction.
本発明者らは、前記従来法の欠点を改善し、より単純な
工程でβ−Ｄ−ガラクトピラノシル−（ｌ→６）−β−
Ｄ−ガラクトピラノシル−（ｌ→４）−Ｄ−フラクトー
スを製造する方法について研究を行なった。その結果、
ラクチュロース水溶液又はラクチュロースを含有する水
に液をβ−ガラクトシダーゼで処理することにより、簡
易に、かつ高収率にβ−Ｄ−ガラクトピラノシル−（ｌ
→６）−β−Ｄ−ガラクトピラノシル−（ｌ→４）−〇
−フラクトースを製造することが可能であることを見い
出し、本発明を完成した。The present inventors have improved the drawbacks of the conventional method and have achieved β-D-galactopyranosyl-(l→6)-β-
Research was conducted on a method for producing D-galactopyranosyl-(l→4)-D-fructose. the result,
By treating an aqueous lactulose solution or water containing lactulose with β-galactosidase, β-D-galactopyranosyl-(l
→6) It was discovered that it is possible to produce -β-D-galactopyranosyl-(l→4)-〇-fructose, and the present invention was completed.
本発明の目的は、β−Ｄ−ガラクトピラノシル−（ｉ→
６）−β−Ｄ−ガラクトピラノシル−（１→４）−ｏ−
フラクトースを簡易に、かつ高収率に製造する方法を提
供することにある。The object of the present invention is to obtain β-D-galactopyranosyl-(i→
6) -β-D-galactopyranosyl-(1→4)-o-
An object of the present invention is to provide a method for producing fructose simply and with high yield.
本発明の他の目的は、ビフィズス菌増殖促進活性を有す
る有益なβ−Ｄ−ガラクトピラノシル−（ｌ→６）−β
−Ｄ−ガラクトピラノシル−（１→４） −〇−フラク
トースを従来知られていない新規な原料から製造する方
法を提供することにある。Another object of the present invention is to provide useful β-D-galactopyranosyl-(l→6)-β with bifidobacterial growth-promoting activity.
The object of the present invention is to provide a method for producing -D-galactopyranosyl-(1→4)-〇-fructose from a new, hitherto unknown raw material.
本発明は、ラクチュロース水溶液又はラクチュロースを
含有する水に液をβ−ガラクトシダーゼで処理し、ガラ
クトース転移反応を行なわせることを特徴とするβ−Ｄ
−ガラクトピラノシル−（ｌ→６）−β−Ｄ−ガラクト
ピラノシル−（１→４）−〇−フラクトースの製造法で
ある。The present invention is characterized in that a lactulose aqueous solution or water containing lactulose is treated with β-galactosidase to carry out a galactose transfer reaction.
This is a method for producing -galactopyranosyl-(l→6)-β-D-galactopyranosyl-(1→4)-〇-fructose.
次に本発明の方法について詳述する。 Next, the method of the present invention will be explained in detail.
高度に精製されたラクチュロースの水溶液又はラクチュ
ロースを一溝成成分として含有する物質の水溶液を使用
することができる。これらの水溶液のラクチュロース濃
度は５〜７０％（重量）である。Aqueous solutions of highly purified lactulose or of substances containing lactulose as one component can be used. The lactulose concentration of these aqueous solutions is 5-70% (by weight).
使用するβ−ガラクトシダーゼは市所品であり、特に高
度に精製されている必要はなく、粗酵素でも使用可能で
ある。酵素は液状でも、また固定化された状態でも使用
可能である。The β-galactosidase used is a commercially available product, and does not need to be particularly highly purified, and crude enzyme can also be used. Enzymes can be used in liquid or immobilized form.
β−ガラクトシダーゼによる処理は、２０〜５０°Ｃの
温度で、使用する酵素が失活しない温ｇ範囲で実施する
。またｐＨ１塩頂の皿類及び濃度等の条件は使用する酵
素の特性に合わせて調整するが、通常ｐＨ３〜８、塩類
０−０．５％（重量）である。The treatment with β-galactosidase is carried out at a temperature of 20 to 50°C, in a temperature g range in which the enzyme used is not inactivated. Conditions such as pH 1 salt top plate and concentration are adjusted according to the characteristics of the enzyme used, but usually the pH is 3 to 8 and the salt content is 0 to 0.5% (by weight).
ラクチュロースからのβ−Ｄ−ガラクトピラノシル−（
ｌ→６）−β−Ｄ−ガラクトピラノシル−（１→４）−
Ｄ−フラクトースの生成は、上記反応条件、および反応
時間あるいは通液速度に異なるので、その処理条件につ
いては、あらかじめ試験をした後、設定することが望ま
しい。β-D-galactopyranosyl-(
l→6)-β-D-galactopyranosyl-(1→4)-
Since the production of D-fructose depends on the above-mentioned reaction conditions, reaction time, or liquid passage rate, it is desirable to set the processing conditions after conducting a test in advance.
所定の処理時間後、酵素の分離、あるいは加熱による失
活を行なって酵素反応を停止させ、β−Ｄ−ガラクトピ
ラノシル−（ｌ→６）−β−Ｄ−ガラクトピラノシル−
（ｌ→４） −ｏ−フラクトース生成液を得る。After a predetermined treatment time, the enzyme is separated or inactivated by heating to stop the enzyme reaction, and β-D-galactopyranosyl-(l→6)-β-D-galactopyranosyl-
(l→4) -O-Fructose production liquid is obtained.
酵素処理を終った反応液は、そのまま、あるいは適宜濃
縮して、又は乾燥して粉末化し、最終製品とすることが
できる。また必要に応じてβ−Ｄ−ガラクトビラノシル
−（ｌ→６）−β−Ｄ−ガラクトピラノシル−（ｌ→４
）−〇−フラクトース含量を高めるために精製を行なう
こともできる。The reaction solution that has been subjected to the enzyme treatment can be used as it is, or can be appropriately concentrated or dried to form a powder into a final product. In addition, β-D-galactopyranosyl-(l→6)-β-D-galactopyranosyl-(l→4
)-0- Purification can also be carried out to increase the fructose content.
ＷＪ製は種々の公知の方法で行なうことができるっ例え
ば、イオン交換樹脂あるいはゲル濾過剤によるカラムク
ロマトグラフ法、あるいは活性炭カラムによる吸傍、ア
ルコール水溶液による溶出法等である。WJ production can be carried out by various known methods, such as column chromatography using an ion exchange resin or gel filtration agent, absorption using an activated carbon column, and elution using an aqueous alcohol solution.
なお、得られたβ−Ｄ−ガラクトピラノシル−（１→６
）−β−Ｄ−ガラクトピラノシル−（１→４）−Ｄ−フ
ラクトースの測定は、活性炭カラムクロマトグラフ法あ
るいはゲル濾過法と、高速液体クロマトグラフ法を組み
合わせて分離し、測定することができる。Note that the obtained β-D-galactopyranosyl-(1→6
)-β-D-galactopyranosyl-(1→4)-D-fructose can be separated and measured using a combination of activated carbon column chromatography or gel filtration and high performance liquid chromatography. can.
次に本発明の方法によるβ−Ｄ−ガラクトピラノシル−
（１→６）−β−Ｄ−ガラクトピラノシル−（ｌ→４）
−Ｄ−フラクトースの製合と、従来法によるβ−Ｄ−ガ
ラクトピラノシル−（１→６）−β−Ｄ−ガラクトピラ
ノシル−（１→４）−Ｄ−フラクトースの製造とを比較
し、本発明の方法について詳述する。Next, β-D-galactopyranosyl-
(1→6)-β-D-galactopyranosyl-(l→4)
Comparison of the production of -D-fructose and the production of β-D-galactopyranosyl-(1→6)-β-D-galactopyranosyl-(1→4)-D-fructose by the conventional method The method of the present invention will now be described in detail.
（試＠ｌ）従来法によるβ−Ｄ−ガラクトピラノシル−
（１→６）−β−Ｄ−ガラクトピラノシル−（１→４）
−〇−フラクトースの製造：元勲５００１１を水１．０
Ｋｇに添加して加温溶解した後、クエン酸を用いてｐＨ
４，２に調整した。この乳糖溶液を４０℃に保持し、市
販の固定化ラクターゼ（スミラクト、住友化オニ業社製
、活性： ２，３００ＩＬＵ／ｇ） １００−を添加
し、３０分間バッチ式にて撹拌し、反応させた。所定時
間後、固定化ラクターゼを濾過により分離し、陽イオン
交換Ｍ脂及び陰イオン交換樹脂カラムに通液して、固形
分２０．４％（重量）、固形分中ガラクトシルラクトー
ス約１３％（重量）を含有する反応液２．２に９を得た
。(Trial @l) β-D-galactopyranosyl- by conventional method
(1→6)-β-D-galactopyranosyl-(1→4)
-〇- Production of fructose: Genkun 50011 and water 1.0
After adding it to Kg and dissolving it by heating, adjust the pH using citric acid.
Adjusted to 4.2. This lactose solution was maintained at 40°C, and commercially available immobilized lactase (SumiLact, manufactured by Sumitomo Kaonigyo Co., Ltd., activity: 2,300 ILU/g) 100- was added and stirred batchwise for 30 minutes to react. Ta. After a predetermined period of time, the immobilized lactase is separated by filtration and passed through a cation exchange M fat and anion exchange resin column to obtain a solid content of 20.4% (weight) and galactosyllactose in the solid content of approximately 13% (weight). ) was obtained in reaction solution 2.2 containing 9.
次いでこの反応液２．０Ｋｇを３Ｉ！の活性炭カラムに
通液してオリゴ糖を吸着させた後、水洗し、１０％エタ
ノール６７！を通液した後、５０％エタノール６１でオ
リゴ糖画分を溶出し、濃縮乾燥して、ガラクトシルラク
トース約２９％（重量）を含有するオリゴ剪粉末２２９
を得た。Next, 2.0 kg of this reaction solution was mixed with 3I! After passing the liquid through an activated carbon column to adsorb oligosaccharides, it was washed with water and added with 10% ethanol 67! After passing through the liquid, the oligosaccharide fraction was eluted with 50% ethanol 61 and concentrated and dried to obtain an oligosaccharide powder 229 containing about 29% (weight) of galactosyllactose.
I got it.
次いでβ−Ｄ−ガラクトピラノシル−（ｌ→６）−β−
Ｄ−ガラクトピラノシル−（１→４）−り一フラクトー
スを得るために、このオリゴ着粉末２０ｇを水５０Ｉ！
に溶解し、１０％の水酸化ナトリウム溶液２ｍｌを添加
し、７５°Ｃで４５分間反応させた。反応時、溶液は次
第に褐変化した。所定時間経過扱、冷却し、陽イオン交
換ｍ脂および陰イオン交換樹脂カラムに通液して脱色精
製し、固形分８．８％（重量）のシロップ１７０ｇを得
た。このシロップは固形分中β−〇−ガラクトピラノシ
ル−（１→６）−β−Ｄ−ガラクトピラノシル−（１→
４）−Ｄ−フラクトースを約１・９％（重ｆ！ｋ）含有
し、反応前のガラクトシルラクトースよりのβ−Ｄ−ガ
ラクトピラノシル−（ｌ→６）−β−り一ガラクトビラ
ノシルー（１→４）−Ｄ−フラクトースの生成率は約４
・９％であった。Then β-D-galactopyranosyl-(l→6)-β-
To obtain D-galactopyranosyl-(1→4)-ri-fructose, 20 g of this oligo-coating powder was mixed with 50 l of water.
2 ml of 10% sodium hydroxide solution was added, and the mixture was reacted at 75°C for 45 minutes. During the reaction, the solution gradually turned brown. The mixture was treated for a predetermined period of time, cooled, and decolorized and purified by passing through a cation exchange resin column and an anion exchange resin column to obtain 170 g of syrup with a solid content of 8.8% (weight). This syrup contains β-〇-galactopyranosyl-(1→6)-β-D-galactopyranosyl-(1→
4) Contains about 1.9% (weight f!k) of -D-fructose, β-D-galactopyranosyl-(l→6)-β-ri-galactobyranosyl from galactosyllactose before reaction The production rate of rou(1→4)-D-fructose is approximately 4
・It was 9%.
（試料２）本発明の方法によるβ−Ｄ−ガラクトピラノ
シル−（１→６）−β−Ｄ−ガラクトピラノシル−（１
→４） −〇−フラクトースの製逍：実施例１と同じ方
法で製凸した。得られたシロップは固形分当り約５．３
％（重量）のβ−Ｄ−ガラクトピラノシル−（工→６）
−β−Ｄ−ガラクトピラノシル−（１→４）−Ｄ−フラ
クトースを含有し、ラクチュロースからのβ−Ｄ−ガラ
クトピラノシル−（ｌ→６）−β−Ｄ−ガラクトピラノ
シル−（１→４）−Ｄ−フラクトースの生成率は約７・
１％であった。(Sample 2) β-D-galactopyranosyl-(1→6)-β-D-galactopyranosyl-(1
→4) -〇-Fructose production: Convex production was performed in the same manner as in Example 1. The resulting syrup has a solid content of approximately 5.3
% (weight) of β-D-galactopyranosyl-(Engineering → 6)
-β-D-galactopyranosyl-(1→4)-containing D-fructose, β-D-galactopyranosyl-(1→6)-β-D-galactopyranosyl- from lactulose The production rate of (1→4)-D-fructose is approximately 7.
It was 1%.
本試験結果から明らかなように、従来法においては、ガ
ラクトシルラクトースを含有する溶液に水酸化ナトリウ
ムを添加し、反応させるために副分解が生じて反応疲は
褐変し、イオン交換翁脂で褐変物質を分離・精製する必
要があり、その固形分歩留りは落ちる（試料ｌでは７５
％）。また反応前のガラクトシルラクトースよりのβ−
Ｄ−ガラクトピラノシル−（１→６）−β−Ｄ−ガラク
トピラノシル−（１→４）−Ｄ−フラクトースの生成率
は約４．９％と低い。As is clear from the results of this test, in the conventional method, sodium hydroxide is added to a solution containing galactosyllactose, and due to the reaction, side decomposition occurs and reaction fatigue results in browning. It is necessary to separate and purify the solid content, and the solid content yield decreases (75
%). In addition, β- from galactosyllactose before reaction
The production rate of D-galactopyranosyl-(1→6)-β-D-galactopyranosyl-(1→4)-D-fructose is as low as about 4.9%.
一方、本発明の方法によれば、反応時の固形分歩留りは
９９％と高く、また反応前のラクチュロースよりのβ−
Ｄ−ガラクトピラノシル−（ｌ→６）−β−Ｄ−ガラク
トピラノシル−（１→４）−〇−フラクトースの生成率
は約７．＋９６と高い。On the other hand, according to the method of the present invention, the solid content yield during the reaction is as high as 99%, and β-
The production rate of D-galactopyranosyl-(l→6)-β-D-galactopyranosyl-(1→4)-〇-fructose is approximately 7. It is high at +96.
以上の結果から１本発明の方法によれば、単純な工程で
、固形分歩留りが良く、かつ生成率の高い状態でβ−Ｄ
−ガラクトピラノシル−（１→６）−β−Ｄ−ガラクト
ピラノシル−（ｌ→４）−り一フラクトースが得られる
ことは明らかである。From the above results, 1. According to the method of the present invention, β-D
It is clear that -galactopyranosyl-(1→6)-β-D-galactopyranosyl-(l→4)-ri-fructose is obtained.
実施例１
ラクチュロース３７％（重量）およびその池田類１３％
（重量）を含有し、固形分５０％（重量）のラクチュロ
ースシロップ１．ＯＫ９ （ｐＨ６，０）を４０°Ｃに
加温し、市販の固定化ラクターゼ（スミラクト、住方化
学工業社製、活性： ２，３００ＦＬＵ ／ 、Ｓ＋
） ５０−を充填したカラムに体積流速２０で通液し
、酵素処理液１．２に９を得た。Example 1 Lactulose 37% (weight) and its Ikeda type 13%
Lactulose syrup containing (by weight) and 50% solids (by weight) 1. OK9 (pH 6,0) was heated to 40°C, and commercially available immobilized lactase (SumiLact, manufactured by Sumikata Chemical Co., Ltd., activity: 2,300FLU/, S+
) The solution was passed through a column filled with 50- at a volumetric flow rate of 20 to obtain an enzyme-treated solution of 1.2 to 9.
この酵素処理液は、固形分含量４１．２％（重量）、ｐ
Ｈ６，０であり、固形分当り約５．３％（重重）のβ−
Ｄ−ガラクトピラノシル−（ｌ→６）−β−Ｄ−ガラク
トピラノシル−（１→４）−〇−フラクトースを含有し
ていた。This enzyme-treated solution had a solid content of 41.2% (weight), p
H6.0, and approximately 5.3% (by weight) of β-
It contained D-galactopyranosyl-(l→6)-β-D-galactopyranosyl-(1→4)-〇-fructose.
実■例２
３００ｇのラクチュロースを１．５ｇのＫ ＨＰＯを含
む水７００ｇに溶解し、クエン酸でｐＨを７．０に調整
し、４０°Ｃに保持した。次いで市販の液状ラクターゼ
（ＧＯＤＯ−ＹＮＬ 、合同酒精製、活性： １０．０
００ ｃＬｔｙ／ｇ） ２．８−を添加し、９０分撹
拌しながら保持してラクターゼ処理を行ない、のち８０
°Ｃ″Ｃ′ｌＯ分間加熱して酵２゛を失活した。Example 2 300 g of lactulose was dissolved in 700 g of water containing 1.5 g of K HPO, the pH was adjusted to 7.0 with citric acid, and the solution was maintained at 40°C. Next, commercially available liquid lactase (GODO-YNL, Joint Sake Refining Co., Ltd., activity: 10.0
00 cLty/g) 2.8- was added and kept stirring for 90 minutes to perform lactase treatment, and then
The enzyme 2' was inactivated by heating for 10 minutes.
得られた酵素処理液は、固形分当り約４．８％のβ−Ｄ
−ガラクトピラノシル−（ｌ→６）−β−Ｄ−ガラクト
ピラノシル−（１→４）−〇−フラクトースを含有して
いた。The resulting enzyme-treated solution contained approximately 4.8% β-D based on the solid content.
-Galactopyranosyl-(l→6)-β-D-galactopyranosyl-(1→4)-〇-fructose was contained.
本発明によって謁せられる効果は、次のとおりである。 The effects achieved by the present invention are as follows.
（１）β−ガラクトシダーゼを作用させるだけの単純な
工程で目的物質が得られる。(1) The target substance can be obtained through a simple process of allowing β-galactosidase to act.
（２）酵素処理の時、生成物中より除去を必要とするア
ルカリの添加、あるいは褐変物質の生成がない。(2) During enzymatic treatment, there is no addition of alkali that needs to be removed from the product or generation of browning substances.
（３）酵素処理の時の固形分歩留りが高い。(3) High solids yield during enzyme treatment.
（４）酵素処理の時、タンパ列り脂肪等のラクチュロー
ス以外の物質が存在しても、高い生成率が得られるので
、未精製のラクチュロース原料から、目的とする物質を
製造できる。(4) During enzyme treatment, even if substances other than lactulose such as protein and fat are present, a high production rate can be obtained, so the desired substance can be produced from unpurified lactulose raw material.
Claims (4)
有する水溶液を、β−ガラクトシダーゼで処理し、ガラ
クトース転移反応を行なわせることを特徴とするβ−Ｄ
−ガラクトピラノシル−（１→６）−β−Ｄ−ガラクト
ピラノシル−（１→４）−Ｄ−フラクトースの新規な製
造法。(1) β-D characterized in that a lactulose aqueous solution or an aqueous solution containing lactulose is treated with β-galactosidase to perform a galactose transfer reaction.
- A novel method for producing galactopyranosyl-(1→6)-β-D-galactopyranosyl-(1→4)-D-fructose.
する水溶液が固定化されたβ−ガラクトシダーゼで処理
されることを特徴とする特許請求の範囲第１項に記載の
β−Ｄ−ガラクトピラノシル−（１→６）−β−Ｄ−ガ
ラクトピラノシル−（１→４）−Ｄ−フラクトースの新
規な製造法。(2) β-D-galactopyranosyl-(1→ 6) Novel method for producing -β-D-galactopyranosyl-(1→4)-D-fructose.
で行なわれることを特徴とする特許請求の範囲第１項又
は第２項のいずれかに記載のβ−Ｄ−ガラクトピラノシ
ル−（１→６）−β−Ｄ−ガラクトピラノシル−（１→
４）−Ｄ−フラクトースの新規な製造法。(3) β-D-galactopyranosyl-(1 →6)-β-D-galactopyranosyl-(1→
4) Novel method for producing -D-fructose.
のラクチュロースを含有していることを特徴とする特許
請求の範囲第１項ないし第３項のいずれかに記載のβ−
Ｄ−ガラクトピラノシル−（１→６）−β−Ｄ−ガラク
トピラノシル−（１→４）−Ｄ−フラクトースの新規な
製造法。(4) Lactulose aqueous solution at least 5% (by weight)
β- according to any one of claims 1 to 3, characterized in that the β-
A novel method for producing D-galactopyranosyl-(1→6)-β-D-galactopyranosyl-(1→4)-D-fructose.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP23932186A JPS6394987A (en) | 1986-10-09 | 1986-10-09 | Novel production of beta-d-galactopyranosyl-(1-6)-beta-d-galactopyranosyl-(1-4)-d-fructose |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP23932186A JPS6394987A (en) | 1986-10-09 | 1986-10-09 | Novel production of beta-d-galactopyranosyl-(1-6)-beta-d-galactopyranosyl-(1-4)-d-fructose |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| JPS6394987A true JPS6394987A (en) | 1988-04-26 |
Family
ID=17042976
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP23932186A Pending JPS6394987A (en) | 1986-10-09 | 1986-10-09 | Novel production of beta-d-galactopyranosyl-(1-6)-beta-d-galactopyranosyl-(1-4)-d-fructose |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JPS6394987A (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5861289A (en) * | 1990-03-16 | 1999-01-19 | Sigma-Tau Industrie Farmaceutiche Riunite S.P.A. | Heat-resistant β-galactosyltransferase, its production process and its use |
-
1986
- 1986-10-09 JP JP23932186A patent/JPS6394987A/en active Pending
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5861289A (en) * | 1990-03-16 | 1999-01-19 | Sigma-Tau Industrie Farmaceutiche Riunite S.P.A. | Heat-resistant β-galactosyltransferase, its production process and its use |
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