Hydantoinase ( ec 3 . 5 . 2 ) hydrolyzes its substrate 5 ' - monosubstantial hydantoin to enantiomerical n - carbamyl - amino acids and in turn , the resulting product can be chemically or enzymatically converted into the corresponding optically active amino acids 海因酶( hydantoinase ， ec3 . 5 . 2 ) ，是一类催化海因、 5 ’ -单替代海因及其衍生物环酰胺键断裂的酰胺水解酶。
2.
To improve the production and activity of the enzyme , the hydantoin hydrolase gene ( hyuh ) was cloned into e . coli m15 and bl21 ( de3 ) by using vector pqe60 and pt221 respectively . two expression plasmids were thus constructed 用pcr技术从质粒puc18 - 169中扩增出l -乙内酰脲水解酶基因( hyuh )和n -氨甲酰基氨基酸水解酶基因( hyuc ) ，分别在大肠杆菌和毕赤酵母中实现了活性表达。
3.
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4.
Puc18 - 169 was a subcloned plasmid containing the whole operon sequence of l - hydantoinase from arthrobacter bt801 which can convert 5 - benzylhydantoin to l - phenylalanine . hydantoin hydrolase which is responsible for the ring opening of hydantoin is one of the components of hydantoin utility enzymes of arthrobacter bt801 . n - carbamoylase is a part of hydantoinase operon which can transform n - carbamoylamino acids into the corresponding amino acids 节杆菌( arthrobacter ) bt801是由军事医学科学院生物工程研究所保存的l -乙内酰脲酶产酶菌株， puc18 - 169是由本室构建的含有节杆菌bt801的l -乙内酰脲酶完整操纵子序列的亚克隆质粒。
5.
With the procedures , the overall recovery of enzymatic activity reached 20 % and the specific activity for substrate hydantoin was about 4 u / mg protein . the purification factor was about 4 . 7 folds with the purity about more than 95 % as estimated by sds - page analysis . d - hydantoinase gene from strain ss - ori was cloned to five different vectors to be five recombinant plasmids and in turn to transfer into five different e . coli strains , respectively 对其中产酶活性最高的一工程菌pexsec一hdt /云coljblz ] ( de3 )海因酶的表达条件进行了研究，目的蛋白的表达量约占总菌蛋白的20 % ，其酶活力为0 . 92u / ml ，约是原始菌株的d一海因酶表观活性的4 . 6倍。
6.
Hydantoinases which consist of hydantoin hydrolase , n - carbamoylase and sometimes hydantoin racemase can hydrolyze d , l - 5 - substituted hydantoin into corresponding optically active amino acids . therefore , the hydantoinases from micro - organisms are widely applied to the industrial production of natural and unnatural amino acids . microbial hydantoinases are considered as industrial enzymes from the origin of life 乙内酰脲酶是广泛存在于动物、植物和微生物的一类酶系，包括乙内酰脲水解酶( hydantoinhydrolase ， hyuh )和n -氨甲酰基氨基酸水解酶( n - carbamoylase ， hyuc ) ，有些情况下还包括乙内酰脲消旋酶( hydantoinracemase ， hyur ) 。
7.
The l - n - carbamoylase of arthrobacter bt801 , coded by the hyuc gene , is the rate - limiting and the only stereoselective enzyme . hydantoin hydrolase gene ( hyuh ) and n - carbamoylase gene ( hyu c ) were amplified from the plasmid of puc18 - 169 by pcr and , at the same time , activity expression was obtained in e . coli and pichia pastor is gs115 respectively 节杆菌bt801的乙内酰脲水解酶催化乙内酰脲类化合物水解开环， n -氨甲酰基氨基酸水解酶是该菌乙内酰脲酶系中惟一具有立体选择性的酶，也是整个反应体系的限速酶。
8.
In bioconversion of a d - amino acid and its derivative , the substrate , 5 ' - monosubstituted hydantoin is firstly transferred into an intermediate , d - carbamyl amino acid by d - hydantoinase , and in turn into an optically active d - amino acid by either d - carbamoylase or chemical , nano2 . nowadays , the bioconversion of d - amino acids has been industrialized by enzymes , so called one - step process D -型氨基酸及其衍生物的生物转化，经历两个反应历程，首先是在海因酶的作用下，催化5 -单替代海因形成氨甲酰类氨基酸中间物，后者再在酶或化学的作用下，生成相应的光学活性氨基酸或其衍生物。
