Sharma Eti, Pandey Saurabh, Gaur A K
Department of Molecular Biology and Genetic Engineering, College of Basic Sciences and Humanities, G. B. Pant University of Agriculture and Technology, Pantnagar, 263145, Uttarakhand, India.
Plant Molecular Biology Lab, The International Centre for Genetic Engineering and Biotechnology ICGEB, New Delhi, India.
3 Biotech. 2016 Jun;6(1):89. doi: 10.1007/s13205-016-0405-y. Epub 2016 Mar 7.
The 3-hydroxy-3-methyl glutaryl CoA reductase (HMGR) is the key enzyme of mevalonate pathway in plants. A partial genomic DNA fragment encoding HMGR conserved domain (denoted as AbHMGR) is isolated from Aconitum balfourii Stapf. It comprises 871 bp encoding 290 amino acids. In silico analysis reveals that it had extensive similarities to other plant HMGR gene. Domain analysis of AbHMGR showed two highly conserved NADPH and HMG CoA domains. Docking study predicted inhibitor, substrate and cofactor binding sites in the protein. Expression analysis revealed that AbHMGR is similarly expressed in all tested tissues with differential pattern. The highest expression was found in leaf tissue. However, fold expression in root and shoot tissue was almost similar. Enzyme activity of HMGR was found to be much higher in leaf tissue as compared to other tissues. The highest aconitine content (0.015 %) was obtained in root tissues. Our data laid a foundation for further investigation of HMGR role in Aconitum balfourii.
3-羟基-3-甲基戊二酰辅酶A还原酶（HMGR）是植物甲羟戊酸途径中的关键酶。从滇西乌头（Aconitum balfourii Stapf）中分离出一个编码HMGR保守结构域的基因组DNA片段（记为AbHMGR）。它包含871个碱基对，编码290个氨基酸。电子分析表明，它与其他植物HMGR基因有广泛的相似性。对AbHMGR的结构域分析显示有两个高度保守的NADPH和HMG CoA结构域。对接研究预测了该蛋白中抑制剂、底物和辅因子的结合位点。表达分析表明，AbHMGR在所有测试组织中均有表达，且模式不同。在叶组织中表达量最高。然而，根和茎组织中的表达倍数几乎相似。与其他组织相比，叶组织中HMGR的酶活性要高得多。根组织中乌头碱含量最高（0.015%）。我们的数据为进一步研究HMGR在滇西乌头中的作用奠定了基础。
