在固氮条件下生长的深红红螺菌中谷氨酸合酶的纯化及部分特性分析
Purification and partial characterization of glutamate synthase from Rhodospirillum rubrum grown under nitrogen-fixing conditions.
作者信息
Carlberg I, Nordlund S
机构信息
Department of Biochemistry, Arrhenius Laboratories for Natural Sciences, Stockholm University, Sweden.
出版信息
Biochem J. 1991 Oct 1;279 ( Pt 1)(Pt 1):151-4. doi: 10.1042/bj2790151.
Abstract
Glutamate synthase, a key enzyme in ammonia assimilation, has been purified from the photosynthetic bacterium Rhodospirillum rubrum. The purification procedure involves ion-exchange chromatography, affinity chromatography and gel filtration. The recovery in the procedure is high (62%) and the specific activity is 21 mumol of NADPH oxidized/min per mg. The enzyme is specific for its substrates, and no activity was demonstrated with NADH or NH4+ ions substituting for NADPH and glutamine respectively. The enzyme is composed of two dissimilar subunits with molecular masses of 53 and 152 kDa, and it is shown that Cl- ions have an effect on the aggregation of the enzyme. Km values for the substrates are: NADPH, 16 microM; 2-oxoglutarate, 10 microM; and glutamine, 65 microM. The enzyme is inhibited by amidotransferase inhibitors at micromolar concentrations. The role of the enzyme in the metabolic regulation of nitrogenase is discussed.
摘要
谷氨酸合酶是氨同化过程中的关键酶，已从光合细菌红螺菌中纯化出来。纯化过程包括离子交换色谱法、亲和色谱法和凝胶过滤法。该过程的回收率很高（62%），比活性为每毫克每分钟氧化21微摩尔NADPH。该酶对其底物具有特异性，用NADH或NH4+离子分别替代NADPH和谷氨酰胺时未表现出活性。该酶由两个分子量分别为53 kDa和152 kDa的不同亚基组成，并且表明Cl-离子对酶的聚集有影响。底物的Km值分别为：NADPH，16微摩尔；2-氧代戊二酸，10微摩尔；谷氨酰胺，65微摩尔。该酶在微摩尔浓度下会被酰胺转移酶抑制剂抑制。讨论了该酶在固氮酶代谢调节中的作用。
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