公开(公告)号：US5169758A

公开(公告)日：1992-12-08

申请号：US561358

申请日：1990-08-01

申请人： Stephan Fischer ,  Barbel Wurst ,  Hans-Otto Beutler ,  Georg-Burkhard Kresse ,  Herwig Brunner

发明人： Stephan Fischer ,  Barbel Wurst ,  Hans-Otto Beutler ,  Georg-Burkhard Kresse ,  Herwig Brunner

IPC分类号： C12N9/02 ,  C12N9/96 ,  C12Q1/26

CPC分类号： C12N9/0036 ,  C12N9/96 ,  C12Q1/26 ,  C12Y107/01002

摘要： The present invention provides a stabilized, NAD(P)H-dependent, soluble nitrate reductase of the assimilatory type, characterized by a molecular weight of about 90,000 D in the case of electrophoresis in the presence of sodium dodecyl sulphate and a residual activity after 3 weeks at 35.degree. C. of more than 60%, obtainable by preparing a suspension of the comminuted starting material in tris/tartaric acid buffer (pH 7-8.5), adding soluble polyethyleneimine thereto in such an amount that 10 to 20% of the nitrate reductase activity passes into the precipitation obtained, separating off the precipitate and working up the supernatant according to conventional biochemical methods of fractionation in the above-mentioned digestion buffer, dialyzing the enzyme solution obtained and lyophilizing in a zwitterionic buffer.

摘要翻译： 本发明提供了一种稳定的NAD（P）H依赖的可溶性硝酸还原酶，其特征在于在十二烷基硫酸钠存在下电泳时的分子量约为90,000D，并且在3 通过制备在三元/酒石酸缓冲液（pH7-8.5）中粉碎的起始材料的悬浮液，可以将其中的可溶性聚乙烯亚胺加入到其中，其量为10至20％ 硝酸还原酶活性进入所得沉淀物，根据常规的上述消化缓冲液中的分级分离方法分离出沉淀物并除去上清液，透析所得酶解液并在两性离子缓冲液中冻干。