JPS61236801A - Novel branched alpha-cyclodextrin and its preparation - Google Patents
Novel branched alpha-cyclodextrin and its preparationInfo
- Publication number
- JPS61236801A JPS61236801A JP60077597A JP7759785A JPS61236801A JP S61236801 A JPS61236801 A JP S61236801A JP 60077597 A JP60077597 A JP 60077597A JP 7759785 A JP7759785 A JP 7759785A JP S61236801 A JPS61236801 A JP S61236801A
- Authority
- JP
- Japan
- Prior art keywords
- cyclodextrin
- alpha
- maltotriosyl
- pullulanase
- maltotriose
- 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.)
- Granted
Links
- HFHDHCJBZVLPGP-RWMJIURBSA-N alpha-cyclodextrin Chemical compound OC[C@H]([C@H]([C@@H]([C@H]1O)O)O[C@H]2O[C@@H]([C@@H](O[C@H]3O[C@H](CO)[C@H]([C@@H]([C@H]3O)O)O[C@H]3O[C@H](CO)[C@H]([C@@H]([C@H]3O)O)O[C@H]3O[C@H](CO)[C@H]([C@@H]([C@H]3O)O)O3)[C@H](O)[C@H]2O)CO)O[C@@H]1O[C@H]1[C@H](O)[C@@H](O)[C@@H]3O[C@@H]1CO HFHDHCJBZVLPGP-RWMJIURBSA-N 0.000 title claims abstract description 25
- 229920001450 Alpha-Cyclodextrin Polymers 0.000 title claims abstract description 22
- 229940043377 alpha-cyclodextrin Drugs 0.000 title claims abstract description 22
- 238000002360 preparation method Methods 0.000 title abstract 2
- 108090000637 alpha-Amylases Proteins 0.000 claims abstract description 14
- DBTMGCOVALSLOR-UHFFFAOYSA-N 32-alpha-galactosyl-3-alpha-galactosyl-galactose Natural products OC1C(O)C(O)C(CO)OC1OC1C(O)C(OC2C(C(CO)OC(O)C2O)O)OC(CO)C1O DBTMGCOVALSLOR-UHFFFAOYSA-N 0.000 claims abstract description 8
- RXVWSYJTUUKTEA-UHFFFAOYSA-N D-maltotriose Natural products OC1C(O)C(OC(C(O)CO)C(O)C(O)C=O)OC(CO)C1OC1C(O)C(O)C(O)C(CO)O1 RXVWSYJTUUKTEA-UHFFFAOYSA-N 0.000 claims abstract description 8
- FYGDTMLNYKFZSV-UHFFFAOYSA-N mannotriose Natural products OC1C(O)C(O)C(CO)OC1OC1C(CO)OC(OC2C(OC(O)C(O)C2O)CO)C(O)C1O FYGDTMLNYKFZSV-UHFFFAOYSA-N 0.000 claims abstract description 8
- FYGDTMLNYKFZSV-BYLHFPJWSA-N β-1,4-galactotrioside Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@H]1O[C@@H]1[C@H](CO)O[C@@H](O[C@@H]2[C@@H](O[C@@H](O)[C@H](O)[C@H]2O)CO)[C@H](O)[C@H]1O FYGDTMLNYKFZSV-BYLHFPJWSA-N 0.000 claims abstract description 8
- 238000006243 chemical reaction Methods 0.000 claims description 13
- 238000004519 manufacturing process Methods 0.000 claims description 3
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 abstract description 7
- 239000000758 substrate Substances 0.000 abstract description 6
- 239000002537 cosmetic Substances 0.000 abstract description 2
- 230000000694 effects Effects 0.000 abstract description 2
- 235000013305 food Nutrition 0.000 abstract description 2
- 238000000926 separation method Methods 0.000 abstract description 2
- 238000009833 condensation Methods 0.000 abstract 1
- 230000005494 condensation Effects 0.000 abstract 1
- 239000003814 drug Substances 0.000 abstract 1
- 229940079593 drug Drugs 0.000 abstract 1
- 239000000463 material Substances 0.000 abstract 1
- LRHPLDYGYMQRHN-UHFFFAOYSA-N N-Butanol Chemical compound CCCCO LRHPLDYGYMQRHN-UHFFFAOYSA-N 0.000 description 15
- 229920000858 Cyclodextrin Polymers 0.000 description 14
- 239000000243 solution Substances 0.000 description 12
- 239000000126 substance Substances 0.000 description 10
- BDERNNFJNOPAEC-UHFFFAOYSA-N propan-1-ol Chemical compound CCCO BDERNNFJNOPAEC-UHFFFAOYSA-N 0.000 description 9
- 108090000790 Enzymes Proteins 0.000 description 8
- 102000004190 Enzymes Human genes 0.000 description 8
- 238000004816 paper chromatography Methods 0.000 description 7
- 238000000034 method Methods 0.000 description 6
- 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 5
- 239000008103 glucose Substances 0.000 description 5
- 125000002791 glucosyl group Chemical group C1([C@H](O)[C@@H](O)[C@H](O)[C@H](O1)CO)* 0.000 description 5
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 4
- 108010073178 Glucan 1,4-alpha-Glucosidase Proteins 0.000 description 4
- 102100022624 Glucoamylase Human genes 0.000 description 4
- JUJWROOIHBZHMG-UHFFFAOYSA-N Pyridine Chemical compound C1=CC=NC=C1 JUJWROOIHBZHMG-UHFFFAOYSA-N 0.000 description 4
- 238000004587 chromatography analysis Methods 0.000 description 4
- 229940097362 cyclodextrins Drugs 0.000 description 4
- 239000000203 mixture Substances 0.000 description 4
- HFHDHCJBZVLPGP-UHFFFAOYSA-N schardinger α-dextrin Chemical compound O1C(C(C2O)O)C(CO)OC2OC(C(C2O)O)C(CO)OC2OC(C(C2O)O)C(CO)OC2OC(C(O)C2O)C(CO)OC2OC(C(C2O)O)C(CO)OC2OC2C(O)C(O)C1OC2CO HFHDHCJBZVLPGP-UHFFFAOYSA-N 0.000 description 4
- SQGYOTSLMSWVJD-UHFFFAOYSA-N silver(1+) nitrate Chemical compound [Ag+].[O-]N(=O)=O SQGYOTSLMSWVJD-UHFFFAOYSA-N 0.000 description 4
- 239000007974 sodium acetate buffer Substances 0.000 description 4
- ZCYVEMRRCGMTRW-UHFFFAOYSA-N 7553-56-2 Chemical compound [I] ZCYVEMRRCGMTRW-UHFFFAOYSA-N 0.000 description 3
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 description 3
- 239000001116 FEMA 4028 Substances 0.000 description 3
- 229920001218 Pullulan Polymers 0.000 description 3
- 239000004373 Pullulan Substances 0.000 description 3
- WHGYBXFWUBPSRW-FOUAGVGXSA-N beta-cyclodextrin Chemical compound OC[C@H]([C@H]([C@@H]([C@H]1O)O)O[C@H]2O[C@@H]([C@@H](O[C@H]3O[C@H](CO)[C@H]([C@@H]([C@H]3O)O)O[C@H]3O[C@H](CO)[C@H]([C@@H]([C@H]3O)O)O[C@H]3O[C@H](CO)[C@H]([C@@H]([C@H]3O)O)O[C@H]3O[C@H](CO)[C@H]([C@@H]([C@H]3O)O)O3)[C@H](O)[C@H]2O)CO)O[C@@H]1O[C@H]1[C@H](O)[C@@H](O)[C@@H]3O[C@@H]1CO WHGYBXFWUBPSRW-FOUAGVGXSA-N 0.000 description 3
- 235000011175 beta-cyclodextrine Nutrition 0.000 description 3
- 229960004853 betadex Drugs 0.000 description 3
- 238000004128 high performance liquid chromatography Methods 0.000 description 3
- 229910052740 iodine Inorganic materials 0.000 description 3
- 239000011630 iodine Substances 0.000 description 3
- 239000000843 powder Substances 0.000 description 3
- 235000019423 pullulan Nutrition 0.000 description 3
- 239000002904 solvent Substances 0.000 description 3
- 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 description 2
- 241000588915 Klebsiella aerogenes Species 0.000 description 2
- GUBGYTABKSRVRQ-PICCSMPSSA-N Maltose Natural products O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@@H]1O[C@@H]1[C@@H](CO)OC(O)[C@H](O)[C@H]1O GUBGYTABKSRVRQ-PICCSMPSSA-N 0.000 description 2
- 241000235527 Rhizopus Species 0.000 description 2
- QAOWNCQODCNURD-UHFFFAOYSA-N Sulfuric acid Chemical compound OS(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 description 2
- WQZGKKKJIJFFOK-VFUOTHLCSA-N beta-D-glucose Chemical compound OC[C@H]1O[C@@H](O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-VFUOTHLCSA-N 0.000 description 2
- GUBGYTABKSRVRQ-QUYVBRFLSA-N beta-maltose Chemical compound OC[C@H]1O[C@H](O[C@H]2[C@H](O)[C@@H](O)[C@H](O)O[C@@H]2CO)[C@H](O)[C@@H](O)[C@@H]1O GUBGYTABKSRVRQ-QUYVBRFLSA-N 0.000 description 2
- 238000006482 condensation reaction Methods 0.000 description 2
- 239000013078 crystal Substances 0.000 description 2
- 238000000921 elemental analysis Methods 0.000 description 2
- GDSRMADSINPKSL-HSEONFRVSA-N gamma-cyclodextrin Chemical compound OC[C@H]([C@H]([C@@H]([C@H]1O)O)O[C@H]2O[C@@H]([C@@H](O[C@H]3O[C@H](CO)[C@H]([C@@H]([C@H]3O)O)O[C@H]3O[C@H](CO)[C@H]([C@@H]([C@H]3O)O)O[C@H]3O[C@H](CO)[C@H]([C@@H]([C@H]3O)O)O[C@H]3O[C@H](CO)[C@H]([C@@H]([C@H]3O)O)O[C@H]3O[C@H](CO)[C@H]([C@@H]([C@H]3O)O)O3)[C@H](O)[C@H]2O)CO)O[C@@H]1O[C@H]1[C@H](O)[C@@H](O)[C@@H]3O[C@@H]1CO GDSRMADSINPKSL-HSEONFRVSA-N 0.000 description 2
- 229940080345 gamma-cyclodextrin Drugs 0.000 description 2
- 239000007788 liquid Substances 0.000 description 2
- 238000000655 nuclear magnetic resonance spectrum Methods 0.000 description 2
- UMJSCPRVCHMLSP-UHFFFAOYSA-N pyridine Natural products COC1=CC=CN=C1 UMJSCPRVCHMLSP-UHFFFAOYSA-N 0.000 description 2
- 239000002994 raw material Substances 0.000 description 2
- 229910001961 silver nitrate Inorganic materials 0.000 description 2
- 235000000346 sugar Nutrition 0.000 description 2
- 239000011800 void material Substances 0.000 description 2
- 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 1
- 229920000945 Amylopectin Polymers 0.000 description 1
- 241000193830 Bacillus <bacterium> Species 0.000 description 1
- 241000194110 Bacillus sp. (in: Bacteria) Species 0.000 description 1
- VYZAMTAEIAYCRO-UHFFFAOYSA-N Chromium Chemical compound [Cr] VYZAMTAEIAYCRO-UHFFFAOYSA-N 0.000 description 1
- 229920002527 Glycogen Polymers 0.000 description 1
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 1
- 238000000862 absorption spectrum Methods 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 239000007864 aqueous solution Substances 0.000 description 1
- 238000009835 boiling Methods 0.000 description 1
- 239000000872 buffer Substances 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 229910052804 chromium Inorganic materials 0.000 description 1
- 239000011651 chromium Substances 0.000 description 1
- 238000004440 column chromatography Methods 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 238000001816 cooling Methods 0.000 description 1
- 125000004122 cyclic group Chemical group 0.000 description 1
- 238000000354 decomposition reaction Methods 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 230000002255 enzymatic effect Effects 0.000 description 1
- 238000002523 gelfiltration Methods 0.000 description 1
- 150000004676 glycans Chemical class 0.000 description 1
- 229940096919 glycogen Drugs 0.000 description 1
- 238000010438 heat treatment Methods 0.000 description 1
- PNDPGZBMCMUPRI-UHFFFAOYSA-N iodine Chemical compound II PNDPGZBMCMUPRI-UHFFFAOYSA-N 0.000 description 1
- 239000003456 ion exchange resin Substances 0.000 description 1
- 229920003303 ion-exchange polymer Polymers 0.000 description 1
- 238000002955 isolation Methods 0.000 description 1
- 238000011068 loading method Methods 0.000 description 1
- 230000000873 masking effect Effects 0.000 description 1
- 244000005700 microbiome Species 0.000 description 1
- 229920001542 oligosaccharide Polymers 0.000 description 1
- 230000003287 optical effect Effects 0.000 description 1
- DHRLEVQXOMLTIM-UHFFFAOYSA-N phosphoric acid;trioxomolybdenum Chemical compound O=[Mo](=O)=O.O=[Mo](=O)=O.O=[Mo](=O)=O.O=[Mo](=O)=O.O=[Mo](=O)=O.O=[Mo](=O)=O.O=[Mo](=O)=O.O=[Mo](=O)=O.O=[Mo](=O)=O.O=[Mo](=O)=O.O=[Mo](=O)=O.O=[Mo](=O)=O.OP(O)(O)=O DHRLEVQXOMLTIM-UHFFFAOYSA-N 0.000 description 1
- 229920001282 polysaccharide Polymers 0.000 description 1
- 239000005017 polysaccharide Substances 0.000 description 1
- 238000000746 purification Methods 0.000 description 1
- 230000035484 reaction time Effects 0.000 description 1
- 230000001568 sexual effect Effects 0.000 description 1
- 230000003381 solubilizing effect Effects 0.000 description 1
- 241000894007 species Species 0.000 description 1
- 230000000087 stabilizing effect Effects 0.000 description 1
- 150000008163 sugars Chemical class 0.000 description 1
- 238000004809 thin layer chromatography Methods 0.000 description 1
Landscapes
- Medicinal Preparation (AREA)
- Cosmetics (AREA)
- Polysaccharides And Polysaccharide Derivatives (AREA)
- General Preparation And Processing Of Foods (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
Abstract
Description
【発明の詳細な説明】
a、産業上の利用分野
本発明は新規な分岐α−サイクロデキストリンおよびそ
の製造方法に関し、更に詳細には、マルトトリオシル−
a−サイクロデキストリンおよびその製造方法に関する
。DETAILED DESCRIPTION OF THE INVENTION a. Industrial Application Field The present invention relates to a novel branched α-cyclodextrin and a method for producing the same.
The present invention relates to a-cyclodextrin and a method for producing the same.
b、従来の技術
サイクロデキストリンはグルコース残基がα−1,4−
結合により環状に結合したオリゴ糖であって、グルコー
ス残基6個からなるα−サイクロデキストリン、7個か
らなるβ−サイクロデキストリン、8個からなるγ−サ
イクロデキストリンなどが一般に知6れている。b. Conventional technology cyclodextrin has glucose residues α-1,4-
Oligosaccharides bonded in a cyclic manner through bonds, such as α-cyclodextrin consisting of 6 glucose residues, β-cyclodextrin consisting of 7 glucose residues, and γ-cyclodextrin consisting of 8 residues, are generally known.
これらサイクロデキストリンは、その構造からそれぞれ
1分子につきα−サイクロデキストリンが6人、β−サ
イクロデキストリンが8Aおよびγ−サイクロデキスト
リンがIOAの空隙を内部に有しており、この空隙内部
は親油性領域となっているので各種の油性物質を取り込
むことができる。そのため、このような性質を利用して
■不安定物質の安定化■揮発性物質の保持■異臭のマス
キング■難・不溶性物質の可溶化など、種々の用途が考
えられている。Due to their structure, these cyclodextrins each have a void inside each molecule of 6 molecules for α-cyclodextrin, 8 A for β-cyclodextrin, and IOA for γ-cyclodextrin, and the inside of this void is a lipophilic region. Therefore, various oily substances can be taken in. Therefore, various uses are being considered by utilizing these properties, such as: 1) stabilizing unstable substances, 2) retaining volatile substances, 2) masking off-odors, and 4) solubilizing difficult to dissolve or insoluble substances.
しかしながら、α−サイクロデキストリンおよびβ−サ
イクロデキストリンは低温域(室温以下)での水に対す
る溶解度が低いことから、この点における改良が待たれ
ていた。However, since α-cyclodextrin and β-cyclodextrin have low solubility in water at low temperatures (below room temperature), improvements in this respect have been awaited.
C0発明が解決しようとする問題点
既に、a−サイクロデキストリンについてはグルコース
残基の06の位置にグルコースあるいはマルトースがf
f−1,6−結合により結合した分岐サイクロデキスト
リンが知られており、このも、 のは分岐の
ないものに比べて水への溶解度が高いことが知られてい
る。Problems to be solved by the C0 invention Already, in a-cyclodextrin, glucose or maltose is present at the 06 position of the glucose residue.
Branched cyclodextrins bonded by f-1,6-bonds are known, and these are also known to have higher solubility in water than those without branches.
本発明者らは、分岐a−サイクロデキスFリンの上述の
如き特性に着目し、種々研究を重ねたところ、プルラナ
ーゼの縮合反応を利用することにより、グルコース、マ
ルトースなどに代わりマルト)リオースを結合させるこ
とがで塾るとの知見を得、更に検討の結果、本発明に到
達したものである。The present inventors focused on the above-mentioned properties of branched a-cyclodextrin, and after conducting various studies, found that by utilizing the condensation reaction of pullulanase, it was possible to combine malto)liose instead of glucose, maltose, etc. The present invention was arrived at as a result of further study and the knowledge that this is a cram school.
d0問題点を解決するための手段
本発明は、マル))リオースとa−サイクロデキストリ
ンを含む混合物にプルラナーゼを作用させてマルトトリ
オシル−a−サイクロデキストリンを生成させ、生成し
たマルトトリオシル−a −サイクロデキストリンを反
応液から分離採取することからなる、マルトトリオシル
−a−サイクロマルトトリオシル−a−サイクロデキス
トリンに関するものである。Means for Solving the d0 Problems The present invention is directed to producing maltotriosyl-a-cyclodextrin by treating a mixture containing mal)) liose and a-cyclodextrin with pullulanase. - It relates to maltotriosyl-a-cyclomaltotriosyl-a-cyclodextrin, which consists of separating and collecting cyclodextrin from a reaction solution.
本発明により得られるマルトトリオシル−a −サイク
ロデキストリンは、6−0−a−マルトトリオシルサイ
クロヘキサオースともいい、分子式CS 4 HI。0
45 分子量1459で表わされる、下記の理化学的
性質を有する新規化合物である。Maltotriosyl-a-cyclodextrin obtained by the present invention is also called 6-0-a-maltotriosylcyclohexaose and has a molecular formula of CS4HI. 0
45 This is a new compound with a molecular weight of 1459 and the following physical and chemical properties.
1)比旋光度
[α]2D、’+171° (C=0.2 、 H2O
)2)薄層クロマトグラフィー
薄層板a DC−Fertigplatten Kie
selgel 60(メルク社製)
展開溶媒:(^)1−ブタノール:エタノール:水=5
:5:2
(B)1−プタノール:ピリジン二本
=6:4:3
Rf=0.1((^)の場合)
0.4((B)の場合)
本品は、ヨウ素溶液を用いた発色およびリンモリブデン
酸/硫酸を用いた発色により呈色され、ニスポットであ
ることが確認された。1) Specific rotation [α] 2D, '+171° (C=0.2, H2O
)2) Thin layer chromatography plate a DC-Fertigplatten Kie
selgel 60 (manufactured by Merck & Co.) Developing solvent: (^) 1-butanol: ethanol: water = 5
:5:2 (B) Two 1-butanol:pyridine = 6:4:3 Rf=0.1 (in the case of (^)) 0.4 (in the case of (B)) This product uses an iodine solution. The product was colored using phosphomolybdic acid/sulfuric acid and was confirmed to be Nispot.
3)ペーパークロマトグラフィー
1−ブタノール:1−プロパノール二本=3 :5 :
4の展開溶媒を使用し、55℃で3回展開した。3) Paper chromatography 1-butanol: 2 bottles of 1-propanol = 3:5:
The sample was developed three times at 55° C. using the developing solvent No. 4.
本品は、ヨウ素溶液を用いた発色お上びグルコアミラー
ゼで前処理した後硝酸銀を用いた発色により呈色され、
1スボ−/ )であることが確認された。This product is colored using an iodine solution, pretreated with glucoamylase, and then colored using silver nitrate.
It was confirmed that the amount was 1 sbo/ ).
4)高速液体クロマトグラフィー
(条 件)
カラムサイズ二6φ×5011IIIl担体: LiC
hrosorb−NHz(メルク社製)溶媒:アセトニ
トリル二本=70:30流速: 2.Oml/a+in
検出器:示差屈折計ERC7520型(エルマ光学株式
会社製)
本品は上記条件で1ピークであった。4) High performance liquid chromatography (conditions) Column size 26φ x 5011III Support: LiC
hrosorb-NHZ (manufactured by Merck & Co.) Solvent: 2 acetonitrile = 70:30 flow rate: 2. Oml/a+in Detector: Differential refractometer model ERC7520 (manufactured by Elma Optical Co., Ltd.) This product had one peak under the above conditions.
5)溶解性 水に易溶、エタノールに難溶。5) Solubility Easily soluble in water, slightly soluble in ethanol.
6)性 状 水溶液は無色であり、粉末は白色。6) Sexual condition The aqueous solution is colorless and the powder is white.
7)赤外線吸収スペクトル
@i図参照
13)IC核磁気共鳴スペクトル
第2図参照
δ(D20) 68.7(1−8結合のC5)78.
5 (C,でα−1,6結合してているグルコースの0
4)
100.1 (1−6結合のCI)
9)α−サイクロデキストリンとマルトトリオースの構
成
プルラナーゼ(林原生物化学研究所製、結晶品)により
マルトトリオースとα−サイクロデキストリンに分解さ
れ、その構成比率をペーパークロマトグラフィーおよび
高速液体クロマトグラフィーで求めた結果、マル))リ
オース:α−サイクロデキストリン=1:1であった。7) Infrared absorption spectrum @ i see figure 13) IC nuclear magnetic resonance spectrum see figure 2 δ (D20) 68.7 (C5 of 1-8 bond) 78.
5 (0 of glucose bonded α-1,6 at C,
4) 100.1 (CI of 1-6 bond) 9) Composition of α-cyclodextrin and maltotriose Decomposed into maltotriose and α-cyclodextrin by pullulanase (manufactured by Hayashibara Biochemical Research Institute, crystal product), As a result of determining the composition ratio by paper chromatography and high performance liquid chromatography, it was found that liose:α-cyclodextrin=1:1.
プルラナーゼ(ノボ・インダストリー・ジャパン社製)
によりマルトトリオースとa−サイクロデキストリンに
分解され、その構成比率をペーパークロマトグラフィー
および高速液体りaマドグラフィーで求めた結果、マル
トトリオース:a−サイクロデキストリン=1:1であ
った。Pullulanase (manufactured by Novo Industry Japan)
It was decomposed into maltotriose and a-cyclodextrin, and the composition ratio thereof was determined by paper chromatography and high performance liquid atomography, and it was found that maltotriose: a-cyclodextrin = 1:1.
グルコアミラーゼ(生化学工業製、結晶品)により分解
した後、ペーパークロマトグラフィーで単離すると、グ
ルコース2モルとヨウ素発色で黄橙色を示す糖(グルコ
シル−a−サイクロデキストリン)が1モル得られた。After decomposition with glucoamylase (manufactured by Seikagaku Corporation, crystalline product) and isolation by paper chromatography, 2 moles of glucose and 1 mole of sugar (glucosyl-a-cyclodextrin), which shows a yellow-orange color when colored with iodine, were obtained. .
本発明によれば、かかるマルトトリオシル−a−サイク
ロデキストリンは次のごとくして製造される。According to the present invention, such maltotriosyl-a-cyclodextrin is produced as follows.
即ち、マルトトリオースとα−サイクロデキストリンを
含む基質濃度40〜80%溶液にプルラナーゼを所定量
加え、液の温度、pHなどを酵素の好適作用範囲に維持
して、1日〜6日間反応させ、マルトトリオシル−a−
サイクロデキストリンを生成し、次いで所望によりクロ
マトグラフィーなどの方法によって反応液から分離採取
することにより製造される。That is, a predetermined amount of pullulanase is added to a solution containing maltotriose and α-cyclodextrin with a substrate concentration of 40 to 80%, and the temperature and pH of the solution are maintained within the enzyme's preferred action range and allowed to react for 1 to 6 days. , maltotriosyl-a-
It is produced by producing cyclodextrin, and then, if desired, separating and collecting it from the reaction solution by a method such as chromatography.
本発明において用いられるプルラナーゼは、粘質多糖類
プルランのa−its−グルコシド結合を加水分解する
ほか、アミロペクチンやグリコーゲンのα−1,6−グ
ルコシド結合をも切断する能力を持つ酵素であり、主と
してエアロバクター・エアロデネス(A erobac
ter aerogenes)、パシラス・5p(Ba
cillus sp)などの微生物より得られる。これ
らプルラナーゼの使用量は、基質の品質あるいは反応の
実施形式などにより多少の違いはあるが、通常の場合、
a−サイクロデキストリン1グラム当たり10単位以上
用いられる。このプルラナーゼの酵素活性は次のごとき
方法により測定される。即ち、0.5%プルラン溶液(
プルランを50mM酢酸ナトリウム緩衝液、pH5,0
に溶解したもの)200μmに酵素液50μ!(同じ緩
衝液に溶解したもの)を加え、10分間、50℃で酵素
反応させる。反応後、反応液中に生成した還元糖をソモ
ギイーネルソン(Somogyi−Nelson)法で
測定する。酵素単位はこの条件で1分間に1μ5ole
のマル))リオースに相当する還元力を生成する酵素量
を1単位とする。The pullulanase used in the present invention is an enzyme that not only hydrolyzes the a-its-glucosidic bonds of the sticky polysaccharide pullulan, but also has the ability to cleave the α-1,6-glucosidic bonds of amylopectin and glycogen. Aerobacter aerobac
ter aerogenes), Pasillas 5p (Ba
It is obtained from microorganisms such as Cillus sp. The amount of pullulanase used varies slightly depending on the quality of the substrate and the method of reaction, but in normal cases,
10 or more units are used per gram of a-cyclodextrin. The enzymatic activity of this pullulanase is measured by the following method. That is, 0.5% pullulan solution (
Pullulan was added to 50mM sodium acetate buffer, pH 5.0.
Dissolved in ) 200μm and enzyme solution 50μ! (dissolved in the same buffer) and allowed to react with the enzyme at 50°C for 10 minutes. After the reaction, reducing sugars produced in the reaction solution are measured by the Somogyi-Nelson method. The enzyme unit is 1 μ5 ole per minute under these conditions.
)) The amount of enzyme that generates the reducing power equivalent to liose is defined as 1 unit.
本発明において原料として眉いちれるマル))リオース
およOa−サイクロデキストリンは、いずれも市販の製
品をそのまま用いることができるが、生成物の分離精製
の手数を考えると純度の高いものを用いるのが有利であ
る。これらマル))リオースお上りa−サイクロデキス
トリンの使用量は、α−サイクロデキストリンに対して
通常マル))リオース2〜7倍量、好ましくは3〜4倍
量用いられる。また、溶液の濃度は、本発明の方法がプ
ルラナーゼの縮合反応を利用するものである関係上、一
般的に原料基質の濃度が高いほど好ましく、従って、本
発明における基質濃度は50〜80%で使用することが
好ましい。In the present invention, both liose and Oa-cyclodextrin, which are used as raw materials, can be used as they are as commercially available products. is advantageous. The amount of a-cyclodextrin used is usually 2 to 7 times, preferably 3 to 4 times, the amount of α-cyclodextrin. In addition, since the method of the present invention utilizes the condensation reaction of pullulanase, it is generally preferable that the concentration of the raw material substrate is higher. Therefore, the concentration of the substrate in the present invention is 50 to 80%. It is preferable to use
本発明の方法においては、反応はプルラナーゼの作用条
件に適合させて実施される。従って、反応温度、pHな
どは用いられる酵素の種M(起源)によって差はあるが
、一般に40〜70℃、pH4,0〜6.0で行なうの
が好ましい。In the method of the invention, the reaction is carried out in a manner adapted to the operating conditions of pullulanase. Therefore, although the reaction temperature, pH, etc. vary depending on the species M (origin) of the enzyme used, it is generally preferable to carry out the reaction at 40 to 70°C and pH 4.0 to 6.0.
生成したマルトトリオシル−α−サイクロデキストリン
を反応液から分離するには、例えばトヨパールHW−4
O8を用いたカラムクロマトグラフィーあるいはワット
マン17クロムによるペーパークロマトグラフィーを用
いることにより容易に行うことができるが、工業的には
特にコスト上の理由からイオン交換樹脂クロマトグラフ
ィー、大量ゲルろ過分離性などを用いるのが有利である
。To separate the generated maltotriosyl-α-cyclodextrin from the reaction solution, for example, Toyopearl HW-4
This can be easily carried out using column chromatography using O8 or paper chromatography using Whatman 17 chromium, but from an industrial perspective, ion exchange resin chromatography, large-scale gel filtration, etc. It is advantageous to use
e0発明の効果
本発明により得られる新規な分岐a−サイクロデキスト
リンは、公知のa−サイクロデキストリンと同程度の強
い抱接力を有し、かつ、その溶解性において格段に優れ
ているので、医薬品、食品、化粧品その他一般の化学工
業分野でのサイクロデキストリンの用途開発に寄与する
ところが大きい。e0 Effects of the Invention The novel branched a-cyclodextrin obtained by the present invention has a strong adhesion force comparable to that of known a-cyclodextrins, and has much better solubility, so it can be used as a pharmaceutical, This will greatly contribute to the development of applications for cyclodextrins in food, cosmetics, and other general chemical industry fields.
r、実施例
次に実施例を示し、本発明を更に詳細かつ具体
)J的に説明する。r, Examples Next, examples will be shown to further explain the present invention in detail and concretely.
) Explain in a J-like manner.
実施例1゜
マル))リオース(日本食品化工KK製、純度99%)
i s、oOgとa−サイクロデキストリン(日本食品
化工KK製、純度98%)5.00gに、pH5,0,
50−M酢酸ナトリウム緩衝液7.701111を加え
沸騰浴中加熱溶解する。冷却後、これにバシラス* s
p (Bacillus sp)の耐熱性プルラナーゼ
(/ボ・インダストリー・ジャパン社製、200単位/
g)2.5gを加え、60℃で48時間反応させる。Example 1゜Mar)) Liose (manufactured by Nihon Shokuhin Kako KK, purity 99%)
To 5.00 g of i s, oOg and a-cyclodextrin (manufactured by Nihon Shokuhin Kako KK, purity 98%), pH 5.0,
Add 50-M sodium acetate buffer 7.701111 and dissolve by heating in a boiling bath. After cooling, add Bacillus*s
Heat-stable pullulanase of Bacillus sp. (manufactured by Bo Industry Japan, 200 units)
g) Add 2.5g and react at 60°C for 48 hours.
終了後、この反応液をトヨパールHW−4OSを充填し
たカラム(2,5X100cmX2本)によりデルろ過
クロマトグラフィーにかけて分離精製を行う。試料負荷
後24〜26時間後に溶出されてくる7ラクレヨンを集
め、ロータリーエバポレータで濃縮乾燥して、マルtト
リオシルーa−サイクロデキストリンの白色粉末264
7mgを得る。After completion, the reaction solution is subjected to Delfiltration chromatography using columns (2.5 x 100 cm x 2 columns) packed with Toyopearl HW-4OS for separation and purification. 7 Lacrayons eluted 24 to 26 hours after loading the sample were collected and concentrated and dried using a rotary evaporator to obtain a white powder of malt-triosil-a-cyclodextrin.
Get 7 mg.
−二のものの元素分析値は次の通りであった。The elemental analysis values of -2 were as follows.
元素分析値(Cs<L。Ol、)
実測値C=44.47%H=6.20%また、このもの
は277℃で分解した。Elemental analysis value (Cs<L.Ol,) Actual value C=44.47%H=6.20% Also, this product decomposed at 277°C.
次ニ、この粉末を、070%1−プロパツール■1−ブ
タノール:ピリジン:水= 6 :4 :3 ■1−ブ
タノール:1−プロパ7−ル:水= 3 :5 :4を
展開剤に用いてペーパー上に展開後、ヨウ素溶液を用い
る発色お上びグルコアミラーゼでペーパーを一度処理し
た後硝酸銀発色させたところ、1スポツトを与え単一物
質であることが確認された。Next, this powder was mixed with 070% 1-propanol ■1-butanol:pyridine:water = 6:4:3 ■1-butanol:1-propanol:water = 3:5:4 as a developing agent After development on paper using iodine solution, the paper was treated once with glucoamylase and then silver nitrate color was developed, which gave one spot and confirmed that it was a single substance.
次に、上記で得らへた物質5mgとエアロバクター〇エ
アロデネス(A erobacter aerogen
es)のプルラナーゼ(体厚生物化学研究所製、結晶品
40単位/a+g)1単位をpH6,0,50mM酢酸
ナトリウム緩衝液500μmに溶解し、30℃で一夜反
応させたところ、マル))リオースとa−サイクロデキ
ストリンを1:1の割合で生成することがペーパークロ
マトグラフィーおよび高速液体クロマトグラフィーによ
り確認された。Next, 5 mg of the dead substance obtained above and Aerobacter aerogenes were added.
When 1 unit of pullulanase (manufactured by Taishu Biochemistry Institute, crystal product 40 units/a+g) of es) was dissolved in 500 μm of pH 6, 0, 50 mM sodium acetate buffer and reacted overnight at 30°C, it was found that liose It was confirmed by paper chromatography and high performance liquid chromatography that a-cyclodextrin and a-cyclodextrin were produced in a 1:1 ratio.
更に、上記で得られた物質51とリゾプス・デ1/ ?
−(Rhizopus delemer)のグルコア
ミラーゼ(生化学工業製、結晶品30単位/ erg)
10単位をpH5,0,50oM酢酸ナトリウム緩衝
液に溶解し、30℃で一夜反応させたところ、グルコー
スとグルコシル−α−サイクロデキストリンを2:1の
割合で生成することがペーパークロマトグラフィーおよ
び高速液体クロマトグラフィーにより確認された。Furthermore, the substance 51 obtained above and Rhizopus de1/?
- (Rhizopus delemer) glucoamylase (manufactured by Seikagaku Corporation, crystalline product 30 units/erg)
When 10 units were dissolved in pH 5, 0, 50oM sodium acetate buffer and reacted overnight at 30°C, paper chromatography and high performance liquid analysis revealed that glucose and glucosyl-α-cyclodextrin were produced in a 2:1 ratio. Confirmed by chromatography.
以上の結果から、上記の物質はマルトトリオシル−α−
サイクロデキストリンであることが確認された。From the above results, the above substance is maltotriosyl-α-
It was confirmed that it was cyclodextrin.
実施例2〜13
実施例1と同様な操作手順により、但し基質濃度゛、酵
素量、反応温度、反応時間を種々に変えて反応を行ない
、次表の結果を得た。Examples 2 to 13 Reactions were carried out in the same manner as in Example 1, but with various substrate concentrations, enzyme amounts, reaction temperatures, and reaction times, and the results shown in the following table were obtained.
第1図は、マルトトリオシル−α−サイクロデキストリ
ンの赤外線吸収スペクトルを示し、第2図は、マルトト
リオシル−α−サイクロデキストリンのI”C核磁気共
鳴スペクトルを示す。FIG. 1 shows the infrared absorption spectrum of maltotriosyl-α-cyclodextrin, and FIG. 2 shows the I''C nuclear magnetic resonance spectrum of maltotriosyl-α-cyclodextrin.
Claims (2)
プルラナーゼの存在下に反応させ、該反応液からマルト
トリオシル−α−サイクロデキストリンを分離、採取す
ることを特徴とするマルトトリオシル−α−サイクロデ
キストリンの製造方法。(2) Maltotriosyl-α-cyclodextrin, characterized in that maltotriose and α-cyclodextrin are reacted in the presence of pullulanase, and maltotriosyl-α-cyclodextrin is separated and collected from the reaction solution. manufacturing method.
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
JP60077597A JPS61236801A (en) | 1985-04-13 | 1985-04-13 | Novel branched alpha-cyclodextrin and its preparation |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
JP60077597A JPS61236801A (en) | 1985-04-13 | 1985-04-13 | Novel branched alpha-cyclodextrin and its preparation |
Publications (2)
Publication Number | Publication Date |
---|---|
JPS61236801A true JPS61236801A (en) | 1986-10-22 |
JPH044874B2 JPH044874B2 (en) | 1992-01-29 |
Family
ID=13638357
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
JP60077597A Granted JPS61236801A (en) | 1985-04-13 | 1985-04-13 | Novel branched alpha-cyclodextrin and its preparation |
Country Status (1)
Country | Link |
---|---|
JP (1) | JPS61236801A (en) |
Cited By (5)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JPS62116604A (en) * | 1985-11-18 | 1987-05-28 | Shokuhin Sangyo Baioriakutaa Syst Gijutsu Kenkyu Kumiai | Separation of glucosylcyclodextrin |
JPS63208510A (en) * | 1987-02-25 | 1988-08-30 | Sansho Seiyaku Kk | Melanization inhibitory drug for external use |
JPH01179697A (en) * | 1988-01-11 | 1989-07-17 | Nikken Chem Co Ltd | Production of branched cyclodextrin |
JPH0770499A (en) * | 1993-06-28 | 1995-03-14 | Color Fastener Kogyo Kk | Ink using branched cyclodextrin and printed form and production thereof |
US5925254A (en) * | 1995-03-17 | 1999-07-20 | Commissariat A L'energie Atomique | Process for separating sodium from aqueous effluents resulting from the reprocessing of spent nuclear fuel elements |
Citations (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JPS61212297A (en) * | 1985-03-19 | 1986-09-20 | Tokuyama Soda Co Ltd | Production of branched cyclodextrin |
-
1985
- 1985-04-13 JP JP60077597A patent/JPS61236801A/en active Granted
Patent Citations (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JPS61212297A (en) * | 1985-03-19 | 1986-09-20 | Tokuyama Soda Co Ltd | Production of branched cyclodextrin |
Cited By (5)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JPS62116604A (en) * | 1985-11-18 | 1987-05-28 | Shokuhin Sangyo Baioriakutaa Syst Gijutsu Kenkyu Kumiai | Separation of glucosylcyclodextrin |
JPS63208510A (en) * | 1987-02-25 | 1988-08-30 | Sansho Seiyaku Kk | Melanization inhibitory drug for external use |
JPH01179697A (en) * | 1988-01-11 | 1989-07-17 | Nikken Chem Co Ltd | Production of branched cyclodextrin |
JPH0770499A (en) * | 1993-06-28 | 1995-03-14 | Color Fastener Kogyo Kk | Ink using branched cyclodextrin and printed form and production thereof |
US5925254A (en) * | 1995-03-17 | 1999-07-20 | Commissariat A L'energie Atomique | Process for separating sodium from aqueous effluents resulting from the reprocessing of spent nuclear fuel elements |
Also Published As
Publication number | Publication date |
---|---|
JPH044874B2 (en) | 1992-01-29 |
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