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Production of optically active amino acids and their derivatives by microbial hydantoinase

微生物乙内酰脲酶生产光学活性氨基酸及其衍生物

基本信息

批准号：
02660113
负责人：
SHIMIZU Sakayu
金额：
$ 1.28万
依托单位：
Kyoto University
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (C)
财政年份：
1990
资助国家：
日本
起止时间：
1990 至 1991
项目状态：
已结题

项目摘要

(1)A novel ATP-dependent amidohydrolase(AIP-hydantoinase), which catalyzes hydrolysis of various 5-substituted hydantoins and ATP to the corresponding N-carbamoyl-L-amino acids, ammonia and carbon dioxide, and ADP and inorganic phosphate, respectively, was purified and crystallized from cells of a soil isolated bacterium PseudoMonas putida 77. the enzyme was found to catalyze the hydrolysis of several nucleoside triphosphates not only in the presence but also in the absence of amide substrate. The kinetic data suggested the conformational change of the enzyme after the binding of amide substrate. Such binding seemed to be absolutely necessary for the hydrolysis of nucleoside triphosphate. The enzyme was shown to be useful for the synthesis of various optically active N-carbamoyl-L-amino acids and for the determination of serum or urine creatine.(2)A new N-carbamoyl-D-amino acid amidohydrolase, which shows high specificity toward N-carbamoyl-D-p-hydroxyphenylglycine, was isolated from a Gram negative soil isolated bacterium and characterized in some details. The enzyme was shown to be highly thermostable and useful catalyst for the practical production of D-p-hydroxyphenylglycine.(3)A novel aromatic-aryl amidase which hydrolyzes various aromatic aryl-acyl amities, was found in a soil isolated Pseudomonase putida Sc-2. The enzyme was shown to be useful for optical resolution of various arcxuatic amines.(4)A new enzyme, nucleoside odidase which catalyzes oxidation of various nucleosides to the corresponding nucleoside 5'-carboxylic acids with concomitant production of hydrogenperoxide, was found in a soil isolated bacterium. The enzyme was seemed to have very close relation to the metabolism of hydantoin-pyrimidine metabolism.

(1)A从土壤分离细菌PseudoMonasputida 77的细胞中纯化并结晶了一种新的ATP依赖性酰胺水解酶（AIP-乙内酰脲酶），其催化各种5-取代乙内酰脲和ATP分别水解为相应的N-氨基甲酰基-L-氨基酸、氨和二氧化碳以及ADP和无机磷酸盐。发现该酶不仅在酰胺底物存在下而且在酰胺底物不存在下催化几种核苷三磷酸的水解。动力学数据表明酶与酰胺底物结合后构象发生了变化。这种结合似乎是三磷酸核苷水解所必需的。该酶可用于合成各种光学活性的N-氨基甲酰基-L-氨基酸，并可用于血清或尿肌酸的测定。(2)A从一株革兰氏阴性土壤分离菌中分离到一种新的N-氨甲酰-D-氨基酸酰胺水解酶，该酶对N-氨甲酰-D-对羟基苯甘氨酸具有高度的专一性。该酶具有很高的热稳定性，可用于D-对羟基苯甘氨酸的实际生产。(3)A从土壤中分离的恶臭假单胞菌Sc-2中发现了一种新的芳香族-芳基酰胺酶，它能水解各种芳香族芳基-酰基酰胺。该酶被证明是有用的光学拆分各种arxuatic胺。(4)A从土壤中分离的一种细菌中发现了一种新的酶--核苷二酸酶，它能催化各种核苷氧化成相应的核苷5 ′-羧酸，并同时产生过氧化氢。该酶与乙内酰脲-嘧啶代谢密切相关。