Isolation, characterization and in silico analysis of 3-Hydroxy-3-methylglutaryl-coenzyme A reductase (HMGR) gene from Andrographis paniculata (Burm. f) Nees

Srinath, Mote; Bindu, Byreddi Bhavani Venkata; Shailaja, Ayeti; Giri, Charu Chandra

doi:10.1007/s11033-019-05172-0

Cited by 9 publications

(7 citation statements)

References 47 publications

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“…In our previous report, we found ApHMGR of the MVA pathway was highly expressed in roots than the stem and leaves. It was identified that HMGR expression was higher in roots (2.5 fold) followed by stem (twofold) and leaf tissues (1.5 fold) [34]. Hence, we could draw a conclusion that DXS from MEP and HMGR from MVA pathway have prominent roles for the production of andrographolide and other DLs.…”

Section: Discussionmentioning

confidence: 81%

“…Despite the intensive study of genes and proteins contributing for diterpene biosynthesis in A. paniculata plant, a meager information is available for the isolation and characterization of the MEP pathway genes. However, in our laboratory, we have ongoing comprehensive research programs to decipher both MVA and MEP pathway genes using genomic and proteomic approaches [34,38]. In the present study, we have isolated, cloned, and carried out in silico functional characterization of DXS gene from A. paniculata for the first time.…”

Section: Discussionmentioning

confidence: 98%

“…However, most of the genes in the MEP pathway have not been isolated and characterized as yet in A. paniculata particularly DXS. In our previous study, we have isolated and characterized ApHMGR gene from its cytosol counterpart MVA pathway and assessed its role in the DL biosynthesis [34].…”

Section: Introductionmentioning

confidence: 99%

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Molecular Cloning and Differential Gene Expression Analysis of 1-Deoxy-D-xylulose 5-Phosphate Synthase (DXS) in Andrographis paniculata (Burm. f) Nees

et al. 2020

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Andrographis paniculata 1-deoxy-D-xylulose-5-phosphate synthase (ApDXS) gene (GenBank Accession No MG271749.1) was isolated and cloned from leaves for the first time. Expression of ApDXS gene was carried out in Escherichia coli Rosetta cells. Tissue-specific ApDXS gene expression by quantitative RT-PCR (qRT-PCR) revealed maximum fold expression in the leaves followed by stem and roots. Further, the differential gene expression profile of Jasmonic acid (JA)-elicited in vitro adventitious root cultures showed enhanced ApDXS expression compared to untreated control cultures. A. paniculata 3-hydroxy-3-methylglutaryl-coenzyme A reductase (ApHMGR) gene expression was also studied where it was up-regulated by JA elicitation but showed lower expression compared to ApDXS. The highest expression of both genes was found at 25 µm JA elicitation followed by 50 µm. HPLC data indicated that the transcription levels were correlated with increased andrographolide accumulation. The peak level of andrographolide accumulation was recorded at 25 μM JA (9.38-fold) followed by 50 µM JA (7.58-fold) in elicitation treatments. The in silico generated ApDXS 3D model revealed 98% expected amino acid residues in the favored and 2% in the allowed regions of the Ramachandran plot with 92% structural reliability. Further, prediction of conserved domains and essential amino acids [Arg (249, 252, 255), Asn (307) and Ser (247)] involved in ligand/ inhibitor binding was carried out by in silico docking studies. Our present findings will generate genomic information and provide a blueprint for future studies of ApDXS and its role in diterpenoid biosynthesis in A. paniculata.

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Section: Discussionmentioning

confidence: 81%

Section: Discussionmentioning

confidence: 98%

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Molecular Cloning and Differential Gene Expression Analysis of 1-Deoxy-D-xylulose 5-Phosphate Synthase (DXS) in Andrographis paniculata (Burm. f) Nees

et al. 2020

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“…Overexpression of the DXS gene enhances the production of terpenoidal secondary metabolites in homologous as well as heterologous plant systems (Jadaun et al, 2017). In general, at least one copy of HMGR has been identified and isolated in several plants, including Andrographis paniculata , Vitis vinifera , and Pogostemon cablin (Srinath et al, 2020; Zhang et al, 2019; Zheng et al, 2021). Overexpression of the HMGR gene from Hevea brasiliensis in transgenic tobacco plants enhanced the accumulation of total sterols up to six‐fold (Schaller et al, 1995).…”

Section: Discussionmentioning

confidence: 99%

Transcriptome analysis reveals candidate genes related to steroid alkaloid biosynthesis in Fritillaria anhuiensis

Shan

Shi

et al. 2022

Physiologia Plantarum

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Fritillaria anhuiensis (F. anhuiensis), a traditional medicinal herb of the Liliaceae family, has various pharmacological effects, including anti‐tussive, anti‐asthmatic, and expectorant activities. Peimine, a steroidal alkaloid, is a major active constituent of F. anhuiensis. Although steroidal alkaloids of F. anhuiensis have been explored chemically and pharmacologically, the molecular basis of their biosynthesis remains unknown due to a lack of genomic and transcriptomic information. In this study, peimine was detected in bulbs, stems, and leaves of F. anhuiensis, with the highest content in the bulbs. We analyzed the transcriptomes of the bulbs, stems, and leaves of F. anhuiensis using an Illumina HiSeq 4000 platform. A total of 92,829 unigenes with a mean length of 804 base pairs (bp) and an N50 value of 1354 bp were assembled from a total of 30.56 Gb high‐quality reads. Based on the Kyoto Encyclopedia of Genes and Genomes annotation, 83 unigenes were found to be related to the steroid biosynthesis pathway, while 155 unigenes were related to the terpenoid backbone biosynthesis pathway. Numerous transcription factors are involved in the regulation of alkaloid biosynthesis. A putative steroidal alkaloid biosynthesis pathway in F. anhuiensis was constructed, providing a novel understanding of peimine biosynthesis. We performed extensive bioinformatics and phylogenetic analyses to investigate the functional characteristics and evolutionary history of cycloartenol synthase. This study is the first to report transcriptome data from F. anhuiensis and extends our knowledge of the molecular mechanisms of steroid alkaloid biosynthesis.

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“…FemB protein template was retrieved from the UniProt database (UniProt ID: P0A0A6.FEMB_STAAM) and confirmed with BLASTp, performed homology modeling using a server ITASSER, to obtain protein 3D structure [ 9 ]. This server generated five homology models and all of them were further subjected to SAVES 6.0 and RAMPAGE validations to determine the top best model with improved properties of stereochemistry, and quality [ 10 ].…”

Section: Homology Model Building Of Femb and Insilico ...mentioning

confidence: 99%

Molecular docking-based screening of methicillin-resistant Staphylococcus aureus FEM proteins with FDA-approved drugs

Sagurthi

2023

Bioinformation

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Antibiotic resistance stands as one of the most significant public health challenges in recent decades. FEM proteins are responsible for the synthesis of pentaglycine cross-bridge, a primary constituent of bacterial peptidoglycan polymer crosslinking during cell wall biosynthesis. Since they are necessary for bacterial survival and antibiotic resistance, they were considered as significant antibacterial targets. We report herein, the virtual screening and selection of FDA-approved drugs and their potent similar molecules as FEM protein inhibitors and analyzed for inhibiting affinity and their ADMET pharmacokinetic properties. This data provide a foundation for the development and optimization of structurally innovative antimicrobial drugs.

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Isolation, characterization and in silico analysis of 3-Hydroxy-3-methylglutaryl-coenzyme A reductase (HMGR) gene from Andrographis paniculata (Burm. f) Nees

Cited by 9 publications

References 47 publications

Molecular Cloning and Differential Gene Expression Analysis of 1-Deoxy-D-xylulose 5-Phosphate Synthase (DXS) in Andrographis paniculata (Burm. f) Nees

Molecular Cloning and Differential Gene Expression Analysis of 1-Deoxy-D-xylulose 5-Phosphate Synthase (DXS) in Andrographis paniculata (Burm. f) Nees

Transcriptome analysis reveals candidate genes related to steroid alkaloid biosynthesis in Fritillaria anhuiensis

Molecular docking-based screening of methicillin-resistant Staphylococcus aureus FEM proteins with FDA-approved drugs

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