3-Hydroxy-3-methylglutaryl-coenzymeA reductase (HMGR), the first rate-limiting enzyme of Mevalonate (MVA) pathway was isolated from Andrographis paniculata (ApHMGR) and expressed in bacterial cells. Full length ApHMGR (1937 bp) was submitted to NCBI with accession number MG271748.1. The open reading frame (ORF) was flanked by a 31-bp 5′-UTR, 118-bp 3′-UTR and ApHMGR contained a 1787 bp ORF encoding protein of 595 amino acids. ApHMGR protein was approximately 64 kDa, with isoelectric point of 5.75. Isolated ApHMGR was cloned into pET102 vector and expressed in E. coli BL21 (DE 3) cells, and characterized by SDS-PAGE. HPLC analysis for andrographolide content in leaf, stem and root of A. paniculata revealed highest in leaf tissue. The expression patterns of ApHMGR in different plant tissues using qRT-PCR revealed high in root tissue correlating with HPLC data. Three dimensional (3D) structural model of ApHMGR displayed 90% of the amino acids in most favored regions of the Ramachandran plot with 93% overall quality factor. ApHMGR was highly conserved with plant specific N-terminal membrane domains and C-terminal catalytic regions. Phylogenetic analysis showed A. paniculata sharing common ancestor with Handroanthus impetiginosus. 3D model of ApHMGR was screened for the interaction with substrates NADPH, HMG CoA and inhibitor using Auto Dock Vina. In silico analysis revealed that full length ApHMGR had extensive similarities to other plant HMGRs. The present communication reports the isolation of full length HMGR from A. paniculata, its heterologous expression in bacterial cells and in silico structural and functional characterization providing valuable genomic information for future molecular interventions.

中文翻译：

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穿心莲中3-羟基-3-甲基戊二酰辅酶A还原酶（HMGR）基因的分离，鉴定和计算机分析

从穿心莲（Ap HMGR）中分离出甲羟戊酸（MVA）途径的第一个限速酶3-Hydroxy-3-methylglutaryl-coenzymeA还原酶（HMGR），并在细菌细胞中表达。全长Ap HMGR（1937 bp）已提交NCBI，登录号MG271748.1。开放阅读框（ORF）的侧翼是31-bp的5'-UTR，118-bp的3'-UTR，并且Ap HMGR包含编码595个氨基酸的1787bp的ORF。Ap HMGR蛋白约为64 kDa，等电点为5.75。将分离的Ap HMGR克隆到pET102载体中并在大肠杆菌中表达BL21（DE 3）细胞，并通过SDS-PAGE表征。在叶，茎和根的穿心莲内酯含量的HPLC分析穿心莲揭示叶组织中最高的。使用qRT-PCR在不同植物组织中Ap HMGR的表达模式显示与HPLC数据相关的根部组织较高。Ap HMGR的三维（3D）结构模型显示了Ramachandran图最喜欢的区域中90％的氨基酸，总质量因子为93％。Ap HMGR与植物特异的N端膜结构域和C端催化区域高度保守。系统发育分析表明穿心莲共享共同的祖先与Handroanthus impetiginosus。的3D模型使用Auto Dock Vina筛选Ap HMGR与底物NADPH，HMG CoA和抑制剂的相互作用。计算机分析表明，全长Ap HMGR与其他植物HMGR具有广泛的相似性。本来文报道了全长HMGR的分离，从胰腺曲霉，其在细菌细胞中的异源表达以及计算机模拟的结构和功能表征，为将来的分子干预提供了有价值的基因组信息。