Purohit K, McFadden B A
J Bacteriol. 1977 Jan;129(1):415-21. doi: 10.1128/jb.129.1.415-421.1977.
Electrophoretically homogeneous ribulose-1,5-bisphosphate (RuBP) carboxylase was obtained from autotropically grown Hydrogenomonas eutropha by sedimentation of the 105,000 X g supernatant in a discontinuous sucrose gradient and by ammonium sulfate fractionation followed by another sucrose gradient centrifugation. The molecular weight of the enzyme determined by light scattering was 490,000 +/- 15,000. The enzyme could be dissociated by sodium dodecyl sulfate into three types of subunits, and the molecular weights (+/- 10%) could be measured. There were two species of large subunits, L and L' (molecular weight 56,000 and 52,000, respectively) and one species of small subunits (molecular weight, 15,000). The mole ratio of L to L' was 5:3, and the overall mole ratio of the small to large subunits was 1.08. The simplest quaternary structure of the enzyme is L5L'3S8. The enzyme contained RuBP oxygenase activity as evidenced by the O2-dependent production of phosphoglycolate and 3-phosphoglyceric acid in equimolar quantities from RuBP.
通过在不连续蔗糖梯度中对105,000×g上清液进行沉降，以及硫酸铵分级分离，随后再进行一次蔗糖梯度离心，从自养生长的真养产碱菌中获得了电泳纯的核酮糖-1,5-二磷酸（RuBP）羧化酶。通过光散射测定的该酶分子量为490,000±15,000。该酶可被十二烷基硫酸钠解离成三种类型的亚基，并可测定其分子量（±10%）。有两种大亚基，L和L'（分子量分别为56,000和52,000）以及一种小亚基（分子量为15,000）。L与L'的摩尔比为5:3，小亚基与大亚基的总摩尔比为1.08。该酶最简单的四级结构是L5L'3S8。该酶含有RuBP加氧酶活性，这可通过RuBP以等摩尔量产生磷酸乙醇酸和3-磷酸甘油酸依赖于氧气得到证明。
