JPS5828295A - Preparation of immobilized coenzyme - Google Patents
Preparation of immobilized coenzymeInfo
- Publication number
- JPS5828295A JPS5828295A JP56124920A JP12492081A JPS5828295A JP S5828295 A JPS5828295 A JP S5828295A JP 56124920 A JP56124920 A JP 56124920A JP 12492081 A JP12492081 A JP 12492081A JP S5828295 A JPS5828295 A JP S5828295A
- Authority
- JP
- Japan
- Prior art keywords
- nad
- protein
- transglutaminase
- coenzyme
- reaction
- 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
- 239000005515 coenzyme Substances 0.000 title claims abstract description 11
- BAWFJGJZGIEFAR-NNYOXOHSSA-N NAD zwitterion Chemical compound NC(=O)C1=CC=C[N+]([C@H]2[C@@H]([C@H](O)[C@@H](COP([O-])(=O)OP(O)(=O)OC[C@@H]3[C@H]([C@@H](O)[C@@H](O3)N3C4=NC=NC(N)=C4N=C3)O)O2)O)=C1 BAWFJGJZGIEFAR-NNYOXOHSSA-N 0.000 claims abstract description 35
- 229950006238 nadide Drugs 0.000 claims abstract description 32
- 108060008539 Transglutaminase Proteins 0.000 claims abstract description 15
- 102000004169 proteins and genes Human genes 0.000 claims abstract description 15
- 108090000623 proteins and genes Proteins 0.000 claims abstract description 15
- 102000003601 transglutaminase Human genes 0.000 claims abstract description 15
- 229930027945 nicotinamide-adenine dinucleotide Natural products 0.000 claims abstract description 3
- 238000000354 decomposition reaction Methods 0.000 claims description 6
- 238000004519 manufacturing process Methods 0.000 claims description 6
- 238000006243 chemical reaction Methods 0.000 abstract description 15
- 235000018102 proteins Nutrition 0.000 abstract description 11
- 239000005018 casein Substances 0.000 abstract description 9
- 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 abstract description 9
- 235000021240 caseins Nutrition 0.000 abstract description 9
- 239000000126 substance Substances 0.000 abstract description 9
- UXVMQQNJUSDDNG-UHFFFAOYSA-L Calcium chloride Chemical compound [Cl-].[Cl-].[Ca+2] UXVMQQNJUSDDNG-UHFFFAOYSA-L 0.000 abstract description 6
- 239000001110 calcium chloride Substances 0.000 abstract description 6
- 229910001628 calcium chloride Inorganic materials 0.000 abstract description 6
- 125000000404 glutamine group Chemical group N[C@@H](CCC(N)=O)C(=O)* 0.000 abstract description 4
- 238000000034 method Methods 0.000 abstract description 4
- 108010073771 Soybean Proteins Proteins 0.000 abstract description 2
- 238000000502 dialysis Methods 0.000 abstract description 2
- 230000003100 immobilizing effect Effects 0.000 abstract description 2
- 239000000203 mixture Substances 0.000 abstract description 2
- 235000019710 soybean protein Nutrition 0.000 abstract description 2
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 abstract description 2
- 239000012295 chemical reaction liquid Substances 0.000 abstract 1
- 239000000047 product Substances 0.000 description 6
- 239000000243 solution Substances 0.000 description 6
- 125000002252 acyl group Chemical group 0.000 description 4
- FBPFZTCFMRRESA-KVTDHHQDSA-N D-Mannitol Chemical compound OC[C@@H](O)[C@@H](O)[C@H](O)[C@H](O)CO FBPFZTCFMRRESA-KVTDHHQDSA-N 0.000 description 3
- 102000004190 Enzymes Human genes 0.000 description 3
- 108090000790 Enzymes Proteins 0.000 description 3
- 150000001875 compounds Chemical class 0.000 description 3
- 210000004185 liver Anatomy 0.000 description 3
- 102000007698 Alcohol dehydrogenase Human genes 0.000 description 2
- 108010021809 Alcohol dehydrogenase Proteins 0.000 description 2
- 241001465754 Metazoa Species 0.000 description 2
- DFPAKSUCGFBDDF-UHFFFAOYSA-N Nicotinamide Chemical compound NC(=O)C1=CC=CN=C1 DFPAKSUCGFBDDF-UHFFFAOYSA-N 0.000 description 2
- 125000003277 amino group Chemical group 0.000 description 2
- 239000008280 blood Substances 0.000 description 2
- 210000004369 blood Anatomy 0.000 description 2
- 239000000872 buffer Substances 0.000 description 2
- 239000003638 chemical reducing agent Substances 0.000 description 2
- 239000003814 drug Substances 0.000 description 2
- 230000002255 enzymatic effect Effects 0.000 description 2
- 238000006911 enzymatic reaction Methods 0.000 description 2
- RWSXRVCMGQZWBV-WDSKDSINSA-N glutathione Chemical compound OC(=O)[C@@H](N)CCC(=O)N[C@@H](CS)C(=O)NCC(O)=O RWSXRVCMGQZWBV-WDSKDSINSA-N 0.000 description 2
- 235000010355 mannitol Nutrition 0.000 description 2
- 238000007254 oxidation reaction Methods 0.000 description 2
- 230000035484 reaction time Effects 0.000 description 2
- -1 that is Chemical compound 0.000 description 2
- LENZDBCJOHFCAS-UHFFFAOYSA-N tris Chemical compound OCC(N)(CO)CO LENZDBCJOHFCAS-UHFFFAOYSA-N 0.000 description 2
- QKNYBSVHEMOAJP-UHFFFAOYSA-N 2-amino-2-(hydroxymethyl)propane-1,3-diol;hydron;chloride Chemical compound Cl.OCC(N)(CO)CO QKNYBSVHEMOAJP-UHFFFAOYSA-N 0.000 description 1
- 229920000936 Agarose Polymers 0.000 description 1
- 102000009366 Alpha-s1 casein Human genes 0.000 description 1
- 108050000244 Alpha-s1 casein Proteins 0.000 description 1
- 241000283690 Bos taurus Species 0.000 description 1
- OYPRJOBELJOOCE-UHFFFAOYSA-N Calcium Chemical compound [Ca] OYPRJOBELJOOCE-UHFFFAOYSA-N 0.000 description 1
- 241000700199 Cavia porcellus Species 0.000 description 1
- 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 1
- GUBGYTABKSRVRQ-WFVLMXAXSA-N DEAE-cellulose Chemical compound OC1C(O)C(O)C(CO)O[C@H]1O[C@@H]1C(CO)OC(O)C(O)C1O GUBGYTABKSRVRQ-WFVLMXAXSA-N 0.000 description 1
- VGGSQFUCUMXWEO-UHFFFAOYSA-N Ethene Chemical compound C=C VGGSQFUCUMXWEO-UHFFFAOYSA-N 0.000 description 1
- 239000005977 Ethylene Substances 0.000 description 1
- 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 1
- 108010024636 Glutathione Proteins 0.000 description 1
- 235000010469 Glycine max Nutrition 0.000 description 1
- 244000068988 Glycine max Species 0.000 description 1
- XUJNEKJLAYXESH-REOHCLBHSA-N L-Cysteine Chemical compound SC[C@H](N)C(O)=O XUJNEKJLAYXESH-REOHCLBHSA-N 0.000 description 1
- 229930195725 Mannitol Natural products 0.000 description 1
- 108010064851 Plant Proteins Proteins 0.000 description 1
- QAOWNCQODCNURD-UHFFFAOYSA-L Sulfate Chemical compound [O-]S([O-])(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-L 0.000 description 1
- 239000007983 Tris buffer Substances 0.000 description 1
- 239000000370 acceptor Substances 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 238000003916 acid precipitation Methods 0.000 description 1
- 150000001298 alcohols Chemical class 0.000 description 1
- 150000001299 aldehydes Chemical class 0.000 description 1
- 235000001014 amino acid Nutrition 0.000 description 1
- 150000001413 amino acids Chemical class 0.000 description 1
- 235000021120 animal protein Nutrition 0.000 description 1
- 230000001580 bacterial effect Effects 0.000 description 1
- 229910052791 calcium Inorganic materials 0.000 description 1
- 239000011575 calcium Substances 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 238000004587 chromatography analysis Methods 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- XUJNEKJLAYXESH-UHFFFAOYSA-N cysteine Natural products SCC(N)C(O)=O XUJNEKJLAYXESH-UHFFFAOYSA-N 0.000 description 1
- 235000018417 cysteine Nutrition 0.000 description 1
- 230000001419 dependent effect Effects 0.000 description 1
- VHJLVAABSRFDPM-QWWZWVQMSA-N dithiothreitol Chemical compound SC[C@@H](O)[C@H](O)CS VHJLVAABSRFDPM-QWWZWVQMSA-N 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- DNJIEGIFACGWOD-UHFFFAOYSA-N ethyl mercaptane Natural products CCS DNJIEGIFACGWOD-UHFFFAOYSA-N 0.000 description 1
- 238000005194 fractionation Methods 0.000 description 1
- 229960003180 glutathione Drugs 0.000 description 1
- BJHIKXHVCXFQLS-UYFOZJQFSA-N keto-D-fructose Chemical compound OC[C@@H](O)[C@@H](O)[C@H](O)C(=O)CO BJHIKXHVCXFQLS-UYFOZJQFSA-N 0.000 description 1
- 229940040511 liver extract Drugs 0.000 description 1
- 239000000594 mannitol Substances 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 235000013336 milk Nutrition 0.000 description 1
- 239000008267 milk Substances 0.000 description 1
- 210000004080 milk Anatomy 0.000 description 1
- 229960003966 nicotinamide Drugs 0.000 description 1
- 235000005152 nicotinamide Nutrition 0.000 description 1
- 239000011570 nicotinamide Substances 0.000 description 1
- BOPGDPNILDQYTO-NNYOXOHSSA-N nicotinamide-adenine dinucleotide Chemical class C1=CCC(C(=O)N)=CN1[C@H]1[C@H](O)[C@H](O)[C@@H](COP(O)(=O)OP(O)(=O)OC[C@@H]2[C@H]([C@@H](O)[C@@H](O2)N2C3=NC=NC(N)=C3N=C2)O)O1 BOPGDPNILDQYTO-NNYOXOHSSA-N 0.000 description 1
- 235000021118 plant-derived protein Nutrition 0.000 description 1
- 239000002244 precipitate Substances 0.000 description 1
- 239000002243 precursor Substances 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
- 238000011084 recovery Methods 0.000 description 1
- 238000006479 redox reaction Methods 0.000 description 1
- 238000005185 salting out Methods 0.000 description 1
- 239000007858 starting material Substances 0.000 description 1
- 238000006276 transfer reaction Methods 0.000 description 1
- DGVVWUTYPXICAM-UHFFFAOYSA-N β‐Mercaptoethanol Chemical compound OCCS DGVVWUTYPXICAM-UHFFFAOYSA-N 0.000 description 1
Landscapes
- Immobilizing And Processing Of Enzymes And Microorganisms (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
Abstract
Description
【発明の詳細な説明】
補酵素ニコチンアミドーアデニ／−ノヌクレオチド（Ｎ
ＡＤ番略す）をタンパク質物質に結合させて固定化した
ＮＡＤを製造する方法に関する。DETAILED DESCRIPTION OF THE INVENTION Coenzyme nicotinamide adeni/-nonucleotide (N
The present invention relates to a method for producing NAD in which NAD (abbreviated as AD) is bound to a protein substance and immobilized thereon.
ＮＡＤは、酸化還元反応に関係する酵素系において補酵
素として働き、極めて不安定な化合物で使用時、保存時
に変化しやす（、また高価なものである。従って、使用
したＮＡＤを反応系から分離、回収し再利用する工夫が
必要である。NAD acts as a coenzyme in the enzyme system involved in redox reactions, and is an extremely unstable compound that changes easily during use and storage (and is expensive. Therefore, it is necessary to separate the used NAD from the reaction system. , it is necessary to devise ways to collect and reuse them.
そこで、本発明者らは、ＮＡＤ？：固定化することを検
討した結果、トランスグルタミナーゼを使用すると首尾
よく目的を達成することな見出した。本発明は、この知
見に基づき完成されたもので、固定化された補酵素ＮＡ
Ｄの製法にかかり、トランスグルタミナーゼを作用させ
てタンノクク質又はタン／（’り質分解物にＮＡＤ，す
なわちニコチンアミド−アデニン−ジヌクレオチド類を
結合することを特徴とするものである。Therefore, the present inventors investigated NAD? : After considering immobilization, we found that the purpose could be successfully achieved by using transglutaminase. The present invention was completed based on this knowledge, and is based on the immobilized coenzyme NA.
The method for producing D is characterized in that NAD, that is, nicotinamide-adenine dinucleotides, is bound to protein or protein decomposition products by the action of transglutaminase.
一般に補酵素ＮＡＤ類を化学的に固定化する方法は知ら
れているが、ＮＡＤ類をトランスグルタミナーゼで酵素
的に固定化することは、゛これまで報告されていない。Although methods for chemically immobilizing coenzymes NAD are generally known, enzymatic immobilization of NAD with transglutaminase has not been reported so far.
本発明によれば、補酵素ＮＡＤの酵素的固定化により、
ＮＡＤの回収、再使用が容易になり、また、不安定なＮ
ＡＤを安定化した形で得ることができる。本発明で得ら
れた固定化ＮＡＤは、医薬品などの有用物質を生産する
酵素反応や有害物質を分解、除去する酵素反応において
、繰返し使用することができるから、工業上極めて有利
である。According to the present invention, by enzymatic immobilization of coenzyme NAD,
NAD can be easily recovered and reused, and unstable NAD can be easily recovered and reused.
AD can be obtained in a stabilized form. The immobilized NAD obtained in the present invention is industrially extremely advantageous because it can be used repeatedly in enzyme reactions for producing useful substances such as pharmaceuticals and for decomposing and removing harmful substances.
本発明ではＮＡＤ又は（及び）ＮＡＤ誘導体が用いられ
、ＮＡＤの誘導体としては、例えばアミノヘキシルＮＡ
Ｄ誘導体すなわち８−（６−アミノヘキシル）−ＮＡＤ
やＮ６−（（６−アミノヘキシル）カルバモイルメチル
）−ＮＡＤ等がある。In the present invention, NAD or (and) an NAD derivative is used, and examples of the NAD derivative include aminohexylNA
D derivative i.e. 8-(6-aminohexyl)-NAD
and N6-((6-aminohexyl)carbamoylmethyl)-NAD.
本発明で使用されるタンパク質又は夕／・クク質分解物
はＮＡＤを固定化する性能、すなわちグルタミン残基な
有するものならば制限はない。タンｔＲり質として例え
ば、カゼイ／、大豆タン・やり質など全ての動植物タン
パク質、菌体タンパク質が挙げられる。タンパク質分解
物としては上記の全てのタンパク質の分解物を例示する
ことができる。またグルタミン残基な含むアミノ酸の合
成４リマーも使用しつ る 。The protein or protein decomposition product used in the present invention is not limited as long as it has the ability to immobilize NAD, ie, has a glutamine residue. Examples of the protein protein include all animal and plant proteins such as casei, soybean tongue, and soybean protein, as well as bacterial cell proteins. Examples of protein decomposition products include all of the above-mentioned protein decomposition products. Synthetic 4-rimers of amino acids containing glutamine residues are also used.
トランスグルタミナーゼは、カルシウム依存性のアシル
転移反応を触媒する酵素であり、タンノック質又はタン
ノｅり質分解物中のグルタミン残基（アシル供与体）と
種々の化合物中のアミノ基（アシル供与体）と種々の化
合物中のアミン基（アシル受容体）との間にアミド結合
を形成させろ能力をもつ。トランスグルタミナーゼはコ
ネランらの方法〔Ｃｏｎｎｅ ］ １ａｎｅｔ ａｌ
Ｊ、Ｂｉｏｌ”、Ｃｈｅｍ、２４６．１０９３（１９７
１） 〕に従ってモノしモソトの肝臓から取得され、他
に牛などの血液からも得られる。Transglutaminase is an enzyme that catalyzes a calcium-dependent acyl transfer reaction, which involves glutamine residues (acyl donors) in decomposition products of tannock substances or tanno-elite substances and amino groups (acyl donors) in various compounds. and amine groups (acyl acceptors) in various compounds. Transglutaminase was determined using the method of Conne et al.
J. Biol”, Chem, 246.1093 (197
According to [1)], it is obtained from the liver of monosquids, and it can also be obtained from the blood of cows and other animals.
本発明の方法では夕／・クク質等とＮＡＤ類とを混合し
たものにトランスグルタミナーゼな作用させて酵素反応
を起させタンノック質等とＮＡＤ類を結合させる。一般
に、タン・クク質１〜５部′な水１０００部に混合した
液にＮＡＤ誘導体０．１〜１部、塩化カルシウム０．６
部、トランスグルタミナーゼ０．１〜０．２部を添加す
る。反応はｐｆｌ ６〜８．５、好ましくは７．５で、
温度２０〜４０℃、好ましくは３７℃前後で行う。反応
時間は１〜１０時間である。反応時間経過後、エチレン
、シアミン四酢酸１２部を加え反応を停止させる。反応
終了後、反応液を通常の方法例えば透析処理して目的物
を取得する。In the method of the present invention, a mixture of proteinaceous substances and NADs is treated with transglutaminase to cause an enzymatic reaction, thereby binding the proteinaceous substances and NADs. Generally, 0.1 to 1 part of NAD derivative and 0.6 parts of calcium chloride are mixed in 1000 parts of water with a tank-like quality of 1 to 5 parts.
1 part and 0.1 to 0.2 parts of transglutaminase are added. The reaction is carried out at a pfl of 6 to 8.5, preferably 7.5;
It is carried out at a temperature of 20 to 40°C, preferably around 37°C. Reaction time is 1 to 10 hours. After the reaction time has elapsed, ethylene and 12 parts of cyaminetetraacetic acid are added to stop the reaction. After the reaction is completed, the reaction solution is subjected to a conventional method such as dialysis treatment to obtain the target product.
還元剤、例えばシスティン、グルタチオン、メルカプト
エタノール、ジチオスレイトールなどはトランスグルタ
ミナーゼを活性化し、また安定化する。従って反応液に
還元剤を添加するのが望ましいが、必ずしも必須ではな
い０
好ましい実施態様の１例は下記のとおりである。Reducing agents such as cysteine, glutathione, mercaptoethanol, dithiothreitol, etc. activate and also stabilize transglutaminase. Therefore, it is desirable, but not necessarily essential, to add a reducing agent to the reaction solution.An example of a preferred embodiment is as follows.
トリス緩衝液（ｐ）１７．５ ）１００ｍＭ塩化カルシ
ウム ５ｍＭ
ＮＡＤ １ｍＭ
カゼイン １〜３ｖｑ／ｍｅトランスグ
ルタミナーゼ １８３μカ血６ｐＨ： ７．５、
反応温度 ３７℃
次に、タンパク質としてカゼインを例にとって、カゼイ
ン中の種々のタンＡり画分とＮＡＤとの結合量を検討し
た結果な述べる。Tris buffer (p) 17.5) 100mM calcium chloride 5mM NAD 1mM casein 1-3vq/me transglutaminase 183μ blood 6pH: 7.5,
Reaction temperature: 37°C Next, using casein as an example of a protein, the results of studying the amount of binding between various protein fractions in casein and NAD will be described.
この゛結合量は馬肝のアルコール脱水素酵素の補酵素活
性により測定され、測定結果はアセチル化αＳ１カゼイ
ン画分の場合が最も効率よくＮＡＤと結合することを示
している。The amount of this binding is measured by the coenzyme activity of horse liver alcohol dehydrogenase, and the measurement results show that the acetylated αS1 casein fraction binds to NAD most efficiently.
本発明で得られた固定化ＮＡＤを例えば医薬品等製造の
酸化反応に使用すると、ＮＡＤは還元型ＮＡＤ（ＮＡＤ
Ｈ）に変るが、ここに生じたＮＡＤＨ−タンパク質固定
化結合体は、塩化カルシウム等による塩析又は酸沈殿に
よって反応液から容易に分離、回収することが可能なの
で、ＮＡＤを有効に再使用することができる。この際の
回収率はほぼ１００チである。また、固定化ＮＡＤは遊
離のものに比し保存安定性に優れている。When the immobilized NAD obtained in the present invention is used, for example, in an oxidation reaction in the production of pharmaceuticals, NAD becomes reduced NAD (NAD
Shifting to H), the NADH-protein immobilized conjugate generated here can be easily separated and recovered from the reaction solution by salting out with calcium chloride etc. or acid precipitation, so NAD can be effectively reused. be able to. The recovery rate at this time is approximately 100 cm. Furthermore, immobilized NAD has better storage stability than free NAD.
次に、本発明を実施例によって説明する。Next, the present invention will be explained by examples.
実施例
〔トランスグルタミナーゼの製法〕
モルモット肝臓の抽出液を出発原料としてＤＥＡＥ−セ
ルロースによるクロマトグラフィー、ソロタミン硫酸分
画、アガロースによるグルクロマトグラフィーな行い、
トランスグルタミナーゼを精製した。２ｋｇの肝臓より
２００〜の！ｆｌ製酵素を得た。Examples [Production method of transglutaminase] Using guinea pig liver extract as a starting material, chromatography with DEAE-cellulose, solotamine sulfate fractionation, gluchromatography with agarose,
Transglutaminase was purified. 200 ~ from 2 kg of liver! The fl enzyme was obtained.
塩化カルシウム０．６２を溶解したトリス塩酸緩衝液（
ｐＨ７，５） １ ｌ中に、カゼインを３２およびアミ
ノヘキシルＮＡＤ誘導体０．８２を添加、溶解して反応
液とした。Tris-HCl buffer containing 0.62% calcium chloride (
pH 7.5) 32% casein and 0.82% aminohexyl NAD derivative were added and dissolved in 1 liter to prepare a reaction solution.
これにトランスグルタミナーゼを２００ｍｇ加え、３７
℃で８時間反応させた。Add 200 mg of transglutaminase to this and
The reaction was carried out at ℃ for 8 hours.
反応は０．４ Ａ Ｄ Ｔ Ａ（ｐＨ８，０＞ ２７
ｏｍｌを加えて停止させた。The reaction was 0.4 A D T A (pH 8, 0 > 27
oml was added to stop.
反応液を７０ｍＭＫＵを含む１０ｍＭイミタソール緩衝
液（ｐＨ６，０）に透析して、遊離のＮＡＤ誘導体を除
き、ＮＡＤ誘導体・カゼイン複合体２．８２を得た。The reaction solution was dialyzed against 10 mM imitasol buffer (pH 6.0) containing 70 mM KU to remove free NAD derivatives, yielding an NAD derivative/casein complex of 2.82.
使用例
Ｄ−フラクトースの前駆体Ｄ−マニトールを原料として
マニトールデヒドロケ９ナーゼと、実施例１で得たＮＡ
Ｄ誘導体・カゼイン複合体を共用して酸化反応させ、Ｄ
−フラクトースを得た。Usage example: D-fructose precursor D-mannitol dehydroke9ase using mannitol as a raw material and NA obtained in Example 1
By using the D derivative/casein complex and causing an oxidation reaction, D
- Fructose was obtained.
反応終了後、反応液に塩化カルシウムを最終濃度が４０
〜６０ｍＡになるようにカゼイン複合体を沈殿させて、
分離、回収した。After the reaction is complete, add calcium chloride to the reaction solution to a final concentration of 40
Precipitate the casein complex to ~60 mA,
Separated and collected.
この回収したＮＡＤＨ誘導体・カゼイン複合体はアルコ
ールデヒドロゲナーゼと共用して例えばアルデヒド類か
らアルコール類を得ることができる。The recovered NADH derivative/casein complex can be used together with alcohol dehydrogenase to obtain alcohols from aldehydes, for example.
特許出願人 雪印乳業株式会社 代理人 弁理士 土 居 三 部Patent applicant: Snow Brand Milk Products Co., Ltd. Agent Patent Attorney Third Department
Claims (1)
ンパク質分解物に補酵素ニコチンアミド−アデニン−ジ
ヌクレオチド類を結合することを特徴とする固定化補酵
素の製法。1. A method for producing an immobilized coenzyme, which comprises binding a coenzyme nicotinamide-adenine dinucleotide to a protein or protein decomposition product through the action of transglutaminase.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP56124920A JPS5828295A (en) | 1981-08-10 | 1981-08-10 | Preparation of immobilized coenzyme |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP56124920A JPS5828295A (en) | 1981-08-10 | 1981-08-10 | Preparation of immobilized coenzyme |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| JPS5828295A true JPS5828295A (en) | 1983-02-19 |
| JPH0145358B2 JPH0145358B2 (en) | 1989-10-03 |
Family
ID=14897402
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP56124920A Granted JPS5828295A (en) | 1981-08-10 | 1981-08-10 | Preparation of immobilized coenzyme |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JPS5828295A (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5834232A (en) * | 1996-05-01 | 1998-11-10 | Zymogenetics, Inc. | Cross-linked gelatin gels and methods of making them |
-
1981
- 1981-08-10 JP JP56124920A patent/JPS5828295A/en active Granted
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5834232A (en) * | 1996-05-01 | 1998-11-10 | Zymogenetics, Inc. | Cross-linked gelatin gels and methods of making them |
Also Published As
| Publication number | Publication date |
|---|---|
| JPH0145358B2 (en) | 1989-10-03 |
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