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Biochemical and in silico molecular study of caffeic acid-O-methyltransferase enzyme associated with lignin deposition in tall fescue

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Abstract

Caffeic acid-O-methyltransferase (COMT), an important enzyme governing the process of lignification in plants, functions at the level of caffeic acid methylation along with 3-O-methylation of monolignol precursors. The present investigation was carried out to decipher the role of COMT in tall fescue lignification and to clone and characterize the COMT gene. The study on COMT activity variation at different growth stages of tall fescue exhibited a significant increase in activity over all the growth stages of tall fescue. A significant relative increase of 47.8% was observed from the first vegetative to reproductive stage. COMT activity exhibited a strong positive correlation with lignin content suggesting it to be an important enzyme of tall fescue lignification. Amplification and sequencing of tall fescue COMT gene resulted in an amplicon of size 1662 (Accession No.-MW442832) and an ORF of 346 amino acids. The deduced protein was hydrophobic, thermally stable and acidic with molecular formula C1679H2623N445O482S20, molecular mass 37.4 kDa and theoretical pI of 6.12. The protein possesses a conserved dimerization domain with a highly conserved SAM binding site. The COMT protein was found to be a homo-dimer with 1 catalytic SAH/SAM ligand per monomer interacting with 14 amino acid residues within 4 Å region.

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The data supporting the findings of this study are available from the corresponding author (SS) on request.

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Acknowledgements

Acknowledgement is due to Hon’ble Vice-Chancellor, Central Agricultural University, Imphal for granting study leave to Dr. Siddhartha Singh for PhD pursuance. Authors are also grateful to the Head, Department of Plant Pathology, College of Agriculture, CSK HPKV, Palampur for providing necessary facilities.

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This research did not receive any specific grant from funding agencies in the public, commercial, or not-for-profit sectors.

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SS: conceptualization; data curation; formal analysis; investigation; methodology; validation; writing original draft; writing & editing. NS: resources; supervision; writing & editing.

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Correspondence to Siddhartha Singh.

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Singh, S., Sharma, N. Biochemical and in silico molecular study of caffeic acid-O-methyltransferase enzyme associated with lignin deposition in tall fescue. Amino Acids 55, 1293–1304 (2023). https://doi.org/10.1007/s00726-022-03225-6

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  • DOI: https://doi.org/10.1007/s00726-022-03225-6

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