JPH02311490A - L-ascorbic acid derivative and its production - Google Patents
L-ascorbic acid derivative and its productionInfo
- Publication number
- JPH02311490A JPH02311490A JP13355089A JP13355089A JPH02311490A JP H02311490 A JPH02311490 A JP H02311490A JP 13355089 A JP13355089 A JP 13355089A JP 13355089 A JP13355089 A JP 13355089A JP H02311490 A JPH02311490 A JP H02311490A
- Authority
- JP
- Japan
- Prior art keywords
- ascorbic acid
- acid derivative
- galactosidase
- lactose
- production
- 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
- 150000000996 L-ascorbic acids Chemical class 0.000 title claims description 26
- 238000004519 manufacturing process Methods 0.000 title claims description 6
- CIWBSHSKHKDKBQ-JLAZNSOCSA-N Ascorbic acid Chemical compound OC[C@H](O)[C@H]1OC(=O)C(O)=C1O CIWBSHSKHKDKBQ-JLAZNSOCSA-N 0.000 claims abstract description 30
- 229960005070 ascorbic acid Drugs 0.000 claims abstract description 24
- 239000002211 L-ascorbic acid Substances 0.000 claims abstract description 15
- 235000000069 L-ascorbic acid Nutrition 0.000 claims abstract description 13
- 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 abstract description 13
- 102000005936 beta-Galactosidase Human genes 0.000 claims abstract description 13
- 108010005774 beta-Galactosidase Proteins 0.000 claims abstract description 13
- 239000008101 lactose Substances 0.000 claims abstract description 13
- 239000000126 substance Substances 0.000 claims abstract description 7
- 239000000203 mixture Substances 0.000 claims abstract description 4
- GUBGYTABKSRVRQ-XLOQQCSPSA-N Alpha-Lactose 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](O)[C@H]1O GUBGYTABKSRVRQ-XLOQQCSPSA-N 0.000 claims abstract description 3
- 230000001580 bacterial effect Effects 0.000 claims description 12
- 239000002537 cosmetic Substances 0.000 abstract description 3
- 235000013305 food Nutrition 0.000 abstract description 3
- 239000003814 drug Substances 0.000 abstract description 2
- 229940079593 drug Drugs 0.000 abstract 1
- 239000000463 material Substances 0.000 abstract 1
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 9
- 239000011668 ascorbic acid Substances 0.000 description 9
- 229930182830 galactose Natural products 0.000 description 9
- 238000006243 chemical reaction Methods 0.000 description 8
- 239000007795 chemical reaction product Substances 0.000 description 7
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 7
- 240000006439 Aspergillus oryzae Species 0.000 description 6
- 235000002247 Aspergillus oryzae Nutrition 0.000 description 6
- 239000012153 distilled water Substances 0.000 description 5
- 230000000694 effects Effects 0.000 description 5
- 238000000034 method Methods 0.000 description 5
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 4
- 102000004190 Enzymes Human genes 0.000 description 4
- 108090000790 Enzymes Proteins 0.000 description 4
- 235000010323 ascorbic acid Nutrition 0.000 description 4
- -1 ascorbic acid phosphate ester Chemical class 0.000 description 4
- 239000000047 product Substances 0.000 description 4
- 150000001875 compounds Chemical class 0.000 description 3
- 238000001644 13C nuclear magnetic resonance spectroscopy Methods 0.000 description 2
- NLXLAEXVIDQMFP-UHFFFAOYSA-N Ammonia chloride Chemical compound [NH4+].[Cl-] NLXLAEXVIDQMFP-UHFFFAOYSA-N 0.000 description 2
- 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 2
- 241000222025 Hamamotoa singularis Species 0.000 description 2
- 229910019142 PO4 Inorganic materials 0.000 description 2
- XSQUKJJJFZCRTK-UHFFFAOYSA-N Urea Chemical compound NC(N)=O XSQUKJJJFZCRTK-UHFFFAOYSA-N 0.000 description 2
- 239000002253 acid Substances 0.000 description 2
- 238000005119 centrifugation Methods 0.000 description 2
- 239000003153 chemical reaction reagent Substances 0.000 description 2
- 238000004440 column chromatography Methods 0.000 description 2
- 238000012258 culturing Methods 0.000 description 2
- 238000001914 filtration Methods 0.000 description 2
- 239000008103 glucose Substances 0.000 description 2
- 229910052757 nitrogen Inorganic materials 0.000 description 2
- 239000010452 phosphate Substances 0.000 description 2
- 125000003808 silyl group Chemical group [H][Si]([H])([H])[*] 0.000 description 2
- PPASLZSBLFJQEF-RXSVEWSESA-M sodium-L-ascorbate Chemical compound [Na+].OC[C@H](O)[C@H]1OC(=O)C(O)=C1[O-] PPASLZSBLFJQEF-RXSVEWSESA-M 0.000 description 2
- 235000019187 sodium-L-ascorbate Nutrition 0.000 description 2
- 239000011755 sodium-L-ascorbate Substances 0.000 description 2
- 239000006228 supernatant Substances 0.000 description 2
- 241000228245 Aspergillus niger Species 0.000 description 1
- 241000894006 Bacteria Species 0.000 description 1
- 241000283690 Bos taurus Species 0.000 description 1
- 244000045232 Canavalia ensiformis Species 0.000 description 1
- 235000010520 Canavalia ensiformis Nutrition 0.000 description 1
- 229920002261 Corn starch Polymers 0.000 description 1
- 241001337994 Cryptococcus <scale insect> Species 0.000 description 1
- 241000196324 Embryophyta Species 0.000 description 1
- 241000588724 Escherichia coli Species 0.000 description 1
- 102000002464 Galactosidases Human genes 0.000 description 1
- 108010093031 Galactosidases Proteins 0.000 description 1
- DGAQECJNVWCQMB-PUAWFVPOSA-M Ilexoside XXIX Chemical compound C[C@@H]1CC[C@@]2(CC[C@@]3(C(=CC[C@H]4[C@]3(CC[C@@H]5[C@@]4(CC[C@@H](C5(C)C)OS(=O)(=O)[O-])C)C)[C@@H]2[C@]1(C)O)C)C(=O)O[C@H]6[C@@H]([C@H]([C@@H]([C@H](O6)CO)O)O)O.[Na+] DGAQECJNVWCQMB-PUAWFVPOSA-M 0.000 description 1
- 239000001888 Peptone Substances 0.000 description 1
- 108010080698 Peptones Proteins 0.000 description 1
- 241000222068 Sporobolomyces <Sporidiobolaceae> Species 0.000 description 1
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 description 1
- 229930006000 Sucrose Natural products 0.000 description 1
- WQZGKKKJIJFFOK-PHYPRBDBSA-N alpha-D-galactose Chemical compound OC[C@H]1O[C@H](O)[C@H](O)[C@@H](O)[C@H]1O WQZGKKKJIJFFOK-PHYPRBDBSA-N 0.000 description 1
- 235000019270 ammonium chloride Nutrition 0.000 description 1
- BFNBIHQBYMNNAN-UHFFFAOYSA-N ammonium sulfate Chemical compound N.N.OS(O)(=O)=O BFNBIHQBYMNNAN-UHFFFAOYSA-N 0.000 description 1
- 229910052921 ammonium sulfate Inorganic materials 0.000 description 1
- 235000011130 ammonium sulphate Nutrition 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 210000001557 animal structure Anatomy 0.000 description 1
- 239000003957 anion exchange resin Substances 0.000 description 1
- 239000003963 antioxidant agent Substances 0.000 description 1
- 230000003078 antioxidant effect Effects 0.000 description 1
- 235000006708 antioxidants Nutrition 0.000 description 1
- 239000007864 aqueous solution Substances 0.000 description 1
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 1
- 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 1
- 229940041514 candida albicans extract Drugs 0.000 description 1
- 239000004202 carbamide Substances 0.000 description 1
- 235000013877 carbamide Nutrition 0.000 description 1
- 229910052799 carbon Inorganic materials 0.000 description 1
- 239000005018 casein Substances 0.000 description 1
- BECPQYXYKAMYBN-UHFFFAOYSA-N casein, tech. Chemical compound NCCCCC(C(O)=O)N=C(O)C(CC(O)=O)N=C(O)C(CCC(O)=N)N=C(O)C(CC(C)C)N=C(O)C(CCC(O)=O)N=C(O)C(CC(O)=O)N=C(O)C(CCC(O)=O)N=C(O)C(C(C)O)N=C(O)C(CCC(O)=N)N=C(O)C(CCC(O)=N)N=C(O)C(CCC(O)=N)N=C(O)C(CCC(O)=O)N=C(O)C(CCC(O)=O)N=C(O)C(COP(O)(O)=O)N=C(O)C(CCC(O)=N)N=C(O)C(N)CC1=CC=CC=C1 BECPQYXYKAMYBN-UHFFFAOYSA-N 0.000 description 1
- 235000021240 caseins Nutrition 0.000 description 1
- 238000004587 chromatography analysis Methods 0.000 description 1
- 239000008120 corn starch Substances 0.000 description 1
- 229940099112 cornstarch Drugs 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 235000014113 dietary fatty acids Nutrition 0.000 description 1
- 230000007717 exclusion Effects 0.000 description 1
- 239000000284 extract Substances 0.000 description 1
- 229930195729 fatty acid Natural products 0.000 description 1
- 239000000194 fatty acid Substances 0.000 description 1
- 125000002519 galactosyl group Chemical group C1([C@H](O)[C@@H](O)[C@@H](O)[C@H](O1)CO)* 0.000 description 1
- 125000002791 glucosyl group Chemical group C1([C@H](O)[C@@H](O)[C@H](O)[C@H](O1)CO)* 0.000 description 1
- 238000010438 heat treatment Methods 0.000 description 1
- IPCSVZSSVZVIGE-UHFFFAOYSA-M hexadecanoate Chemical compound CCCCCCCCCCCCCCCC([O-])=O IPCSVZSSVZVIGE-UHFFFAOYSA-M 0.000 description 1
- 238000004128 high performance liquid chromatography Methods 0.000 description 1
- 230000007062 hydrolysis Effects 0.000 description 1
- 238000006460 hydrolysis reaction Methods 0.000 description 1
- 230000003100 immobilizing effect Effects 0.000 description 1
- 150000002500 ions Chemical class 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 238000004811 liquid chromatography Methods 0.000 description 1
- 210000004185 liver Anatomy 0.000 description 1
- 235000013372 meat Nutrition 0.000 description 1
- 244000005700 microbiome Species 0.000 description 1
- 235000013379 molasses Nutrition 0.000 description 1
- 125000001477 organic nitrogen group Chemical group 0.000 description 1
- 239000001301 oxygen Substances 0.000 description 1
- 229910052760 oxygen Inorganic materials 0.000 description 1
- 235000019319 peptone Nutrition 0.000 description 1
- 229920001467 poly(styrenesulfonates) Polymers 0.000 description 1
- 238000000746 purification Methods 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
- 230000035484 reaction time Effects 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 238000010898 silica gel chromatography Methods 0.000 description 1
- 239000011734 sodium Substances 0.000 description 1
- 229910052708 sodium Inorganic materials 0.000 description 1
- 239000000243 solution Substances 0.000 description 1
- 239000005720 sucrose Substances 0.000 description 1
- 229940088594 vitamin Drugs 0.000 description 1
- 229930003231 vitamin Natural products 0.000 description 1
- 235000013343 vitamin Nutrition 0.000 description 1
- 239000011782 vitamin Substances 0.000 description 1
- 150000003722 vitamin derivatives Chemical class 0.000 description 1
- 238000005406 washing Methods 0.000 description 1
- 230000002087 whitening effect Effects 0.000 description 1
- 239000012138 yeast extract Substances 0.000 description 1
Landscapes
- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Saccharide Compounds (AREA)
- Food Preservation Except Freezing, Refrigeration, And Drying (AREA)
Abstract
Description
【発明の詳細な説明】
（産業上の利用分野）
本発明は、安定性に優れたＬ−アスコルビン酸誘導体及
びその製造法に関するものである。DETAILED DESCRIPTION OF THE INVENTION (Field of Industrial Application) The present invention relates to an L-ascorbic acid derivative with excellent stability and a method for producing the same.
（従来の技術）
Ｌ−アスコルビン酸は、必須ビタミンあるいは酸化防止
剤として、医薬品あるいは食品の分野において広く使用
されているばかりでなく、メラニン還元作用を有するこ
とから、美白化粧料としても利用されている化合物であ
る。(Prior Art) L-ascorbic acid is not only widely used as an essential vitamin or antioxidant in the pharmaceutical and food fields, but also as a whitening cosmetic because it has a melanin-reducing effect. It is a compound that has
しかし、このＬ−アスコルビン酸は、非常に不安定な物
質であり、熱、光、酸素等によって容易に分解や変性を
受けることが知られている。このように不安定なＬ−ア
スコルビン酸を安定化する試みは数多くなされており、
いくつかの誘導体が開発されている。例えば、アスコル
ビン酸リン酸エステル（特開昭５７−１４０７８９号公
報）。However, this L-ascorbic acid is a very unstable substance and is known to be easily decomposed or denatured by heat, light, oxygen, etc. Many attempts have been made to stabilize L-ascorbic acid, which is unstable in this way.
Several derivatives have been developed. For example, ascorbic acid phosphate ester (JP-A-57-140789).
グルコースが結合したグリコジルアスコルビン酸（特公
昭５８−５９２０号公報）、アスコルビン酸脂肪酸エス
テル（特開昭６２−８４０７２号公報）等を具体例とし
てあげることができる。Specific examples include glycodylascorbic acid bound to glucose (Japanese Patent Publication No. 58-5920), ascorbic acid fatty acid ester (Japanese Patent Application Laid-Open No. 62-84072), and the like.
これらのし−アスコルビン酸誘導体を製造するには９例
えばアスコルビン酸リン酸エステルの場合、原料として
シリルリン酸化された化合物を用い、この化合物のシリ
ル基を脱離することにより製造されている。For example, in the case of ascorbic acid phosphate ester, a silyl phosphorylated compound is used as a raw material and the silyl group of this compound is eliminated.
（発明が解決しようとする課題）
前記したように、各種のアスコルビン酸誘導体が開発さ
れているが、その安定性については未だ十分とはいえず
、しかもこれらの製造法も高価な試薬を必要とすること
、収率が低いこと１反応条件が極めて過酷であること等
９問題点が多かった。(Problems to be Solved by the Invention) As mentioned above, various ascorbic acid derivatives have been developed, but their stability is still not sufficient, and their production methods also require expensive reagents. There were many problems such as low yield, extremely harsh reaction conditions, etc.
（課題を解決するための手段）
本発明者らは、前記の課題を解決するために鋭意研究を
重ねた結果９次式で示される新規なＬ−アスコルビン酸
誘導体が極めて安定であるこきを見出し、また、この誘
導体を温和な条件で安価に収率よく合成することができ
ることを見出し１本発明を完成するに至った。(Means for Solving the Problems) As a result of extensive research in order to solve the above problems, the present inventors have discovered that a novel L-ascorbic acid derivative represented by the 9th formula is extremely stable. Furthermore, the inventors discovered that this derivative can be synthesized under mild conditions at low cost and in good yield, leading to the completion of the present invention.
すなわち、第１の発明は１式（１）
で示されるＬ−アスコルビン酸誘導体を要旨とするもの
である。That is, the gist of the first invention is an L-ascorbic acid derivative represented by formula (1).
また、第２の発明は、及びＬ−アスコルビン酸と、乳糖
又は乳糖含有物との混合物にβ−ガラクトシダーゼ又は
β−ガラクトシダーゼを含有する菌体を作用させること
を特徴とするＬ−アスコルビン酸誘導体の製造法を要旨
とするものである。Moreover, the second invention is an L-ascorbic acid derivative characterized in that β-galactosidase or bacterial cells containing β-galactosidase is allowed to act on a mixture of L-ascorbic acid and lactose or a lactose-containing substance. The gist is the manufacturing method.
本発明のＬ−アスコルビン酸誘導体は、前記の式（１）
で示される構造を持ち、化学名は、６−０−ガラクトピ
ラノシル−し−アスコルビン酸である。The L-ascorbic acid derivative of the present invention has the above formula (1)
The chemical name is 6-0-galactopyranosyl-ascorbic acid.
本発明のＬ−アスコルビン酸誘導体ハ、Ｌ−７スコルビ
ン酸と、乳糖又は乳糖含有物との混合物にβ−ガラクト
シダーゼ（Ｅ Ｃ３，２，１，２３） 又Ｌｔ β−ガ
ラクトシダーゼを含有する菌体を作用させて製造するこ
とができる。The L-ascorbic acid derivative of the present invention (3) Adding β-galactosidase (E C3,2,1,23) or Lt β-galactosidase-containing bacteria to a mixture of L-7scorbic acid and lactose or a lactose-containing substance. It can be manufactured by making it act.
本発明に用いられるβ−ガラクトシダーゼとしては、ア
スペルギルス オリーゼ（Ａｓｐｅｒｇ ｉ ｌ １ｕ
ｓｏｒｙａｚｅ） ＊ エシェリキア コリ （Ｂｓｃ
ｈｅｒｉｃｈｉａｃｏｌｉ） 、 アスペルギルス ニ
ガー（Ａｓｐｅｒｇｉｌ Ｉｕｓｎｉｇａｒ ）等の微
生物由来の酵素、牛肝臓等の動物臓器由来の酵素、ジャ
ック ビーンズ（Ｊａｃｋｂｅａｎｓ ）等の植物種子
由来の酵素があげられるが。As the β-galactosidase used in the present invention, Aspergillus oryzae (Aspergillus oryzae)
soryaze) * Escherichia coli (Bsc
Examples include enzymes derived from microorganisms such as P. herichiacoli and Aspergillus niger, enzymes derived from animal organs such as bovine liver, and enzymes derived from plant seeds such as jack beans.
反応収率の点１こおいてアスペルギルス オリーゼ（Ａ
ｓｐｅｒｇｉｌｌｕｓ ｏｒｙａｚｅ ）由来のβ−ガ
ラクトシダーゼが最も優れている。また、β−ガラクト
シダーゼを含有する菌体としては、リボマイセス（Ｌｉ
ｐｏｍｙｃｅｓ） ＮＫＤ −１４（微工研菌寄第８９
４８号）、スポロボロミセス シンギュラリス（Ｓｐｏ
ｒｏｂｏｌｏｍｙｃｅｓ ｓｉｎｇｕｌａｒｉｓ） Ａ
ＴＣＣ２４１９３、クリプトコツカス ローシンティ
１４２１等が使用できる。これらのうち、リボマイセス
（Ｌｉｐｏｍｙｃｅｓ） Ｎ Ｋ Ｄ −１４がＬ−ア
スコルビン酸へのガラクトースの転移作用が強く、最も
好ましい。Regarding reaction yield point 1, Aspergillus oryzae (A
β-galactosidase derived from Spergillus oryaze is the best. In addition, as a bacterial cell containing β-galactosidase, Ribomyces (Li
pomyces) NKD-14 (Microtechnical Laboratory No. 89
No. 48), Sporobolomyces singularis (Spo
robolomyces singularis) A
TCC24193, Cryptococcus lowcinti 1421, etc. can be used. Among these, Libomyces NK D-14 has a strong effect of transferring galactose to L-ascorbic acid and is most preferred.
これらの菌体を得るだめの条件としては、特に限定され
るものではないが、一般に乳糖を含む培地で培養するこ
とにより、ガラクトース転移作用の強い菌体が得られる
。また、炭素源としてグルコース、シヨ糖、廃糖蜜等を
用い、菌体を十分増殖させた後に乳糖を添加し、さらに
培養を続け。The conditions for obtaining these cells are not particularly limited, but in general, cells with a strong galactose transfer activity can be obtained by culturing in a medium containing lactose. In addition, glucose, sucrose, blackstrap molasses, etc. are used as a carbon source, and after the bacterial cells have sufficiently grown, lactose is added and the culture is continued.
β−ガラクトシダーゼを十分誘導させた後に菌体を遠心
、濾過等の通常用いられる方法により回収すれば、ガラ
クトース転移作用の強い菌体を得ることができる。培養
に用いる窒素源としては２例えば、ペプトン、カゼイン
、コーンステイープリカー、肉エキス、酵母エキス等の
有機窒素源や。If β-galactosidase is sufficiently induced and the bacterial cells are recovered by a commonly used method such as centrifugation or filtration, bacterial cells with a strong galactosyl transfer activity can be obtained. Nitrogen sources used for culture include organic nitrogen sources such as peptone, casein, cornstarch liquor, meat extract, and yeast extract.
硫安、塩化アンモニウム、尿素等の無機窒素源を用いる
ことができる。通常用いられる方法により培養後、菌体
を遠心、濾過等の方法により回収し。Inorganic nitrogen sources such as ammonium sulfate, ammonium chloride, urea, etc. can be used. After culturing by a commonly used method, the bacterial cells are collected by methods such as centrifugation and filtration.
洗浄後、そのまま反応に用いることができるし。After washing, it can be used for reaction as is.
さらには菌体を各種の固定化法により固定化することに
より使用することもできる。Furthermore, it can also be used by immobilizing bacterial cells by various immobilization methods.
本発明において１反応時のＬ−アスコルビン酸の濃度と
しては、０．１〜３０％（Ｗ／Ｖ）が適当であり、１〜
２０％（Ｗ／Ｖ）が好ましい。乳糖の濃度としては、１
〜４０％（Ｗ／Ｖ）が適当であり、５〜１０％（Ｗ／Ｖ
）が好ましい。また。In the present invention, the appropriate concentration of L-ascorbic acid during one reaction is 0.1 to 30% (W/V), and 1 to 30% (W/V).
20% (W/V) is preferred. The concentration of lactose is 1
~40% (W/V) is appropriate, and 5~10% (W/V)
) is preferred. Also.
反応時のｐＨとしては３〜９が適当であるが、アスペル
ギルス オリーゼ由来のβ−ガラクトシダーゼを用いる
場合は、ｐＨ５〜６が好ましく、リボマイセスＮＫＤ−
１４を用いる場合は、ｐＨ５゜５〜６．５が好ましい。A suitable pH during the reaction is 3 to 9, but when using β-galactosidase derived from Aspergillus oryzae, a pH of 5 to 6 is preferred;
When using No. 14, the pH is preferably 5.5 to 6.5.
次に２反応温度としては。Next, as for the second reaction temperature.
２０〜６０℃の範囲が適当であり、３０〜５０℃の範囲
が好ましい。さらに１反応時間としては。A range of 20 to 60°C is appropriate, and a range of 30 to 50°C is preferred. Furthermore, as one reaction time.
使用する酵素あるいは菌体量により適宜選べばよいが、
目的とするＬ−アスコルビン酸誘導体の生成量が最大に
なるような時間を選べばよい。It may be selected appropriately depending on the enzyme used or the amount of bacterial cells, but
The time may be selected such that the amount of production of the desired L-ascorbic acid derivative is maximized.
このような方法により生成するＬ−アスコルビン酸誘導
体は、シリカゲルカラムクロマトグラフィー、活性炭カ
ラムクロマトグラフィー、液体クロマトグラフィー等１
通常の分離手段で分離することができる。The L-ascorbic acid derivative produced by such a method can be processed by silica gel column chromatography, activated carbon column chromatography, liquid chromatography, etc.
It can be separated by conventional separation means.
（実施例） 次に１本発明を実施例により具体的に説明する。(Example) Next, one embodiment of the present invention will be specifically explained using examples.
実施例Ｉ
Ｌ−アスコルビン酸ナトリウム（石津製薬製特級試薬）
ｌＯｇ及び乳糖１０ｇを１００ｍ１の蒸留水に溶解して
、ｐＨを６．０に調整した。これにアスペルギルス オ
リーゼ（ＡｓｐｅｒｇｉｌｌＪｓ ｏｒｙｚａｅ）由来
のβ−ガラクトシダーゼ（シグマ社製、グレードＸＩ）
を８４０ユニット加えて、３０℃で４時間反応させて反
応物を得た。得られた反応物をバイオラッド（ＢＩＤ
Ｒ＾口）社製高速液体カラムクロマトグラフィー用力う
ムアミネックスイオンエクスクルージ：ｌン（ＡＭＩＮ
ＢＸ ＩＯＮ ＢＸＣＬＩＩＳＩＯＮ） ＨＰ Ｘ−８
７Ｈを用いて分析した。このとき、溶出液として０．０
Ｉ ＮのＨ２Ｓ Ｏ４を用い、流速０．６ｍｊ２／ｍ
ｉ ｎで反応物を溶出させた。また、そのときの検出
をＵＶ２，５で行った。Example I Sodium L-ascorbate (special grade reagent manufactured by Ishizu Pharmaceutical)
lOg and 10 g of lactose were dissolved in 100 ml of distilled water and the pH was adjusted to 6.0. In addition, β-galactosidase derived from Aspergillus oryzae (manufactured by Sigma, grade XI)
840 units of were added and reacted at 30°C for 4 hours to obtain a reaction product. The obtained reaction product was transferred to Bio-Rad (BID).
AMINEX Ion Exclusion for High Performance Liquid Column Chromatography manufactured by AMIN
BX ION BXCLIISION) HP X-8
Analyzed using 7H. At this time, 0.0
Using IN H2S O4, flow rate 0.6 mj2/m
The reaction was eluted in. Further, detection at that time was performed using UV2 and UV5.
その結果、３．６ｇのＬ−アスコルビン酸誘導体が生成
していた。このときの反応収率は、１９．８％であった
。As a result, 3.6 g of L-ascorbic acid derivative was produced. The reaction yield at this time was 19.8%.
次に、上記の反応物５０ｍｌを４５０ｍｌの活性炭カラ
ム（活性炭として和光紬薬製クロマトグラフィー用活性
炭を用いた）にかけ、流速１０ｍ ｌ／ｍｉｎで水溶出
させた。このとき、Ｌ−アスコルビン酸、Ｌ−アスコル
ビン酸誘導体の順に溶出してくるので、この溶出したＬ
−アスコルビン酸誘導体のみを集め、集めたＬ−アスコ
ルビン酸誘導体を２０ｍｊ！の陰イオン交換樹脂ダウエ
ックス（口ｏｗｅｘ） Ｓ Ａ Ｒカラム（ＣＩ−型）
にかけ、流速５ｙｎｊｉ’／ｍｉｎで６０ｍｌの蒸留水
を通液させたのち、１００ｍｊｉ！の０．０４Ｍ Ｎａ
Ｃｊ！−０，０ｌＮＨＣｌを通液し、目的のＬ−アスコ
ルビン酸誘導体を溶出させた。この溶出画分を濃縮乾固
し、１゜４ｇのＬ−アスコルビン酸誘導体を得た。この
ときの精製収率は、７７．８％であった。Next, 50 ml of the above reaction product was applied to a 450 ml activated carbon column (activated carbon for chromatography manufactured by Wako Tsumugi Pharmaceutical Co., Ltd. was used as the activated carbon) and eluted with water at a flow rate of 10 ml/min. At this time, L-ascorbic acid and L-ascorbic acid derivatives are eluted in this order, so this eluted L
- Collect only ascorbic acid derivatives and collect 20mj of the collected L-ascorbic acid derivatives! Anion exchange resin DOWEX SAR column (CI-type)
After applying 60ml of distilled water at a flow rate of 5ynji'/min, 100ml of distilled water was passed through the water at a flow rate of 5ynji'/min. 0.04M Na
Cj! -0,01 NHCl was passed through the solution to elute the desired L-ascorbic acid derivative. This eluted fraction was concentrated to dryness to obtain 1.4 g of L-ascorbic acid derivative. The purification yield at this time was 77.8%.
得られたＬ−アスコルビン酸誘導体を”Ｃ−ＮＭＲ分析
を行ったところ１次のような結果が得られた。When the obtained L-ascorbic acid derivative was subjected to C-NMR analysis, the following results were obtained.
１３Ｃ−ＮＭＲ（６値ｐｐｍ）
ガラクトースのＣ−１１０４，７２９
ガラクトースのＣ−２７２，５２６
ガラクトースのＣ−３７４，３４３
ガラクトースのＣ−４７０，４３４
ガラクトースのＣ−５７６，９２６
ガラクトースのＣ−６６２，８１Ｏ
Ｌ−アスコルビン酸のＣ−１１０４，８８６Ｌ−アスコ
ルビン酸のＣ−２１５７，０９４Ｌ−アスコルビン酸の
Ｃ−３１７４，９７５Ｌ−アスコルビン酸のＣ−４７７
，９４８Ｌ−アスコルビン酸のＣ−５７１，９４６Ｌ−
アスコルビン酸のＣ−６６８，９３４ま′た。得られた
Ｌ−アスコルビン酸誘導体をアスペルギルス オリーゼ
（＾ｓｐｅｒｇｉｌｌｕｓ ｏｒｙｚａｅ）由来のβ−
ガラクトシダーゼで加水分解することより、Ｌ−アスコ
ルビン酸とガラクトースが生成した。13C-NMR (6-value ppm) Galactose C-1104,729 Galactose C-272,526 Galactose C-374,343 Galactose C-470,434 Galactose C-576,926 Galactose C-662,81O L-ascorbic acid C-1104,886L-ascorbic acid C-2157,094L-ascorbic acid C-3174,975L-ascorbic acid C-477
,948L-Ascorbic acid C-571,946L-
C-668,934 of ascorbic acid. The obtained L-ascorbic acid derivative was mixed with β-derived from Aspergillus oryzae.
Hydrolysis with galactosidase produced L-ascorbic acid and galactose.
以上の結果から、上記で得られたＬ−アスコルビン酸誘
導体の構造が６−〇−ガラクトピラノシルーＬ−アスコ
ルビン酸であることが判明した。From the above results, it was revealed that the structure of the L-ascorbic acid derivative obtained above was 6-0-galactopyranosyl-L-ascorbic acid.
次に上記で得られた本発明品とＬ−アスコルビン酸及び
Ｉ、−丁スコルビルパルミテート（東京化成工業部）と
の加熱安定性の比較をｐＨ７で行った。このとき、各々
１％水溶液（ｐＨ７）で煮沸させ、経時的にサンプリン
グし、高速液体クロマトグラフィーにより定量した。Next, the heating stability of the product of the present invention obtained above and L-ascorbic acid and I,-di-scorbyl palmitate (Tokyo Kasei Kogyo Department) was compared at pH 7. At this time, each was boiled with a 1% aqueous solution (pH 7), sampled over time, and quantified by high performance liquid chromatography.
その結果を表１に示す。The results are shown in Table 1.
表１から１本発明品が極めて安定であることがわかる。It can be seen from Table 1 that the product of the present invention is extremely stable.
表 １
実施例２
Ｌ−７スコルビン酸ナトリウムｌＯｇ及び乳糖１０ｇを
１００ｍｊ？の蒸留水に溶解し、ｐＨ６，（１に調整し
た。これにリボマイセスＮＫＤ−１４（微工研菌寄第８
９４８号）湿菌体１０ｇを加えて４５℃で２４時間反応
させて反応物を得た。得られた反応物を遠心し、菌体除
去後の上澄みを実施例１と同様にして精製し、１．５ｇ
のＬ−アスコルビン酸誘導体を得た。Table 1 Example 2 1Og of sodium L-7 scorbate and 10g of lactose in 100mj? was dissolved in distilled water and adjusted to pH 6 (1).
No. 948) 10 g of wet bacterial cells were added and reacted at 45° C. for 24 hours to obtain a reaction product. The obtained reaction product was centrifuged, and the supernatant after removing the bacterial cells was purified in the same manner as in Example 1, and 1.5 g
An L-ascorbic acid derivative was obtained.
次に実施例１と同様にしてＬ−アスコルビン酸誘導体の
構造等を確認したところ、実施例１と全く同じものが得
られていた。Next, the structure etc. of the L-ascorbic acid derivative was confirmed in the same manner as in Example 1, and it was found that exactly the same product as in Example 1 was obtained.
実施例３
Ｌ−アスコルビン酸ナトリウム１０ｇ及び乳糖１０ｇを
１００ｍｊ！の蒸留水に溶解して、ｐＨを６．０に調整
した。これにスポロボロミセス シンジュラリスＡＴＣ
Ｃ２４１９３湿菌体１０ｇを加えて４０℃で２４時間反
応させて反応物を得た。Example 3 100mj of 10g of sodium L-ascorbate and 10g of lactose! of distilled water and the pH was adjusted to 6.0. In this, Sporobolomyces sindularis ATC
10 g of wet C24193 cells were added and reacted at 40°C for 24 hours to obtain a reaction product.
得られた反応物を遠心し、菌体除去後の上澄みを実施例
１と同様にして精製し、１．３ｇのＬ−アスコルビン酸
誘導体を得た。The obtained reaction product was centrifuged, and the supernatant after removing the bacterial cells was purified in the same manner as in Example 1 to obtain 1.3 g of L-ascorbic acid derivative.
次に実施例１と同様にしてＬ−アスコルビン酸誘導体の
構造等を確認したところ、実施例１と全く同じものが得
られていた。Next, the structure etc. of the L-ascorbic acid derivative was confirmed in the same manner as in Example 1, and it was found that exactly the same product as in Example 1 was obtained.
（発明の効果）
本発明のＬ〜アスコルビン酸誘導体は、極めて安定性に
優れており、今まで安定性が悪かったために利用できな
かった医薬品１食品、化粧品等の分野で利用範囲が拡大
できる。(Effects of the Invention) The L-ascorbic acid derivative of the present invention has extremely excellent stability, and can be used in fields such as pharmaceuticals, foods, cosmetics, etc., which could not be used until now due to poor stability.
また１本発明によれば、この誘導体を温和な条件で安価
に収率よく合成することができる。Furthermore, according to the present invention, this derivative can be synthesized under mild conditions at low cost and with good yield.
Claims (2)
混合物にβ−ガラクトシダーゼ又はβ−ガラクトシダー
ゼを含有する菌体を作用させることを特徴とするＬ−ア
スコルビン酸誘導体の製造法。(2) A method for producing an L-ascorbic acid derivative, which comprises allowing β-galactosidase or a bacterial cell containing β-galactosidase to act on a mixture of L-ascorbic acid and lactose or a lactose-containing substance.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP13355089A JP2690779B2 (en) | 1989-05-25 | 1989-05-25 | L-ascorbic acid derivative and method for producing the same |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP13355089A JP2690779B2 (en) | 1989-05-25 | 1989-05-25 | L-ascorbic acid derivative and method for producing the same |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| JPH02311490A true JPH02311490A (en) | 1990-12-27 |
| JP2690779B2 JP2690779B2 (en) | 1997-12-17 |
Family
ID=15107436
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP13355089A Expired - Lifetime JP2690779B2 (en) | 1989-05-25 | 1989-05-25 | L-ascorbic acid derivative and method for producing the same |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JP2690779B2 (en) |
-
1989
- 1989-05-25 JP JP13355089A patent/JP2690779B2/en not_active Expired - Lifetime
Also Published As
| Publication number | Publication date |
|---|---|
| JP2690779B2 (en) | 1997-12-17 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| EP0096547B1 (en) | Process for preparing uridine diphosphate-n-acetylgalactosamine | |
| US4908311A (en) | Process for enzymatic preparation of cellooligosaccharides | |
| EP0075262B1 (en) | Process for the preparation of 5-ribo nucleotides | |
| EP0049801B1 (en) | Sucrose-mutase, immobilized sucrose-mutase and use of this immobilized sucrose-mutase for the production of isomaltulose(6-o-alpha-d-glucopyranosido-d fructose) | |
| US3930953A (en) | Glucose oxidase poor in catalase and process for obtaining it | |
| CA2604328C (en) | Purification method and production method for cellobiose | |
| Chow et al. | 4-Methylumbelliferyl 2-acetamido-2-deoxy-α-d-glucopyranoside, a fluorogenic substrate for N-acetyl-α-D-glucos-aminidase | |
| US3956337A (en) | Process for the preparation of D-gluconic-δ-lactam | |
| JPH02311490A (en) | L-ascorbic acid derivative and its production | |
| WO1991006663A1 (en) | Process for preparing ganglioside g¿m1? | |
| DE3626915A1 (en) | METHOD FOR THE ENZYMATIC SYNTHESIS OF A TRISACCHARID CONTAINING N-ACETYLNEURAMINE ACID | |
| SU1571047A1 (en) | Method of obtaining water-soluble oligosaccharides | |
| DE2337312B2 (en) | Process for the isolation and purification of dehydrogenases | |
| JP2001112496A (en) | Production of cellooligosaccharide | |
| JPH0662852A (en) | Fucoidanase | |
| JPH05176785A (en) | Production of arbutin | |
| JP3006615B2 (en) | Method for producing D-β-hydroxy amino acid | |
| JPS62289198A (en) | Novel process of producing hyaluronic acid | |
| JPS6024798B2 (en) | Moranoline derivatives and their production method | |
| JPS6120268B2 (en) | ||
| FR2459288A1 (en) | PROCESS FOR THE PREPARATION OF MILDIOMYCIN BY A MICROORGANISM OF THE GENUS STREPTOVERTICILLIUM IN A CULTURE CONTAINING AN N-METHYL COMPOUND | |
| WO1991006662A1 (en) | Process for preparing asialo g¿m1? | |
| JPH07274990A (en) | Method for purifying cyclic inulooligosaccharide | |
| DE2167034A1 (en) | Isolation of creatinine amido hydrolase and creatine - amidinohydrolase - from microorganisms, for use in clinical liver fun | |
| SU1255642A1 (en) | Method of producing polysaccharide solution from bacterial suspension |