Zeeman S C, Northrop F, Smith A M, Rees T
John Innes Centre, Norwich Research Park, Colney, Norfolk, UK.
Plant J. 1998 Aug;15(3):357-65. doi: 10.1046/j.1365-313x.1998.00213.x.
The aim of this work was to identify enzymes that participate in the degradation of transitory starch in Arabidopsis. A mutant line was isolated by screening leaves at the end of the night for the presence of starch. The mutant had a higher starch content than the wild-type throughout the diurnal cycle. This accumulation was due to a reduction in starch breakdown, leading to an imbalance between the rates of synthesis and degradation. No reduction in the activity of endo-amylase (alpha-amylase), beta-amylase, starch phosphorylase, maltase, pullulanase or D-enzyme could be detected in crude extracts of leaves of the mutant. However, native PAGE in gels containing amylopectin revealed that a starch-hydrolysing activity, putatively identified as an endo-amylase and present in wild-type chloroplasts, was absent or appreciably reduced in the mutant. This is the first time that a specific enzyme required for starch degradation has been identified in leaves.
这项工作的目的是鉴定参与拟南芥中暂时淀粉降解的酶。通过在夜间结束时筛选叶片中淀粉的存在来分离出一个突变株系。在整个昼夜周期中,该突变体的淀粉含量均高于野生型。这种积累是由于淀粉分解减少,导致合成与降解速率失衡。在突变体叶片的粗提物中未检测到内切淀粉酶(α-淀粉酶)、β-淀粉酶、淀粉磷酸化酶、麦芽糖酶、支链淀粉酶或D-酶的活性降低。然而,在含有支链淀粉的凝胶中进行的非变性聚丙烯酰胺凝胶电泳显示,一种假定为内切淀粉酶且存在于野生型叶绿体中的淀粉水解活性在突变体中缺失或明显降低。这是首次在叶片中鉴定出淀粉降解所需的特定酶。