Journal
JOURNAL OF PLANT BIOCHEMISTRY AND BIOTECHNOLOGY
Volume 30, Issue 2, Pages 317-325Publisher
SPRINGER INDIA
DOI: 10.1007/s13562-020-00591-8
Keywords
Rosmarinic acid synthase; Ocimum tenuiflorum; Cloning; Gene expression; Abiotic stress
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Funding
- ICAR-DMAPR, India
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Rosmarinic acid synthase (RAS) is a key enzyme involved in the biosynthesis of rosmarinic acid from l-Phenylalanine and l-Tyrosine. Its expression is influenced by various abiotic stresses, indicating a potential role in plant defense mechanisms.
Rosmarinic acid synthase (RAS) is a control point at which metabolites derived froml-Phenylalanine andl-Tyrosine are channeled for the biosynthesis of rosmarinic acid (RA).Ocimum tenuiflorumor holy basil has been utilized as a source of RA. To acquire a better understanding of the biosynthesis of this metabolite, isolation, and cloning ofO. tenuiflorumRAS (OtRAS) cDNA and its heterologous expression in bacterial cells was assessed. The full-length OtRAS (Accession No. MN542659) was 1278 bp in size encoding 425 amino acid long protein product. The subunit molecular mass of recombinant OtRAS was estimated to be similar to 47.5 kDa with a theoretical pI of 6.35. Sequence analysis indicated OtRAS belongs to BAHD superfamily proteins. Phylogenetic analysis revealed close evolutionary relatedness of OtRAS with RAS proteins reported in other lamiaceae members. Gene expression analysis showed that callus derived from leaves accumulated more transcripts of OtRAS than leaf and stem tissues derived from field-grown plants. OtRAS was up-regulated under flood and cold stress whereas down-regulated under drought and salt conditions. Further, three-dimensional structure of OtRAS showed two pseudo symmetrical domains harboring an active site in between these two domains. OtRAS active site comprised of conserved His41, His152, Trp363, and Lys391 residues. Overall, our results indicated the OtRAS regulation is a mechanism that probably depended on a variety of abiotic stresses.
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