Evaluation of α-amylase expression and analysis of phytochemical in the leaf callus tissue of Rauvolfia serpentina (Linn.) Benth. Ex Kurz exposed to Cyclodextrin

Abstract

Rauvolfia serpentina (Linn.) Benth. Ex Kurz belongs to the family Apocynaceae. The objective of the present study was to establish an effective protocol for the regeneration of leaf explants from R. serpentina and to study the α-amylase expression and phytochemical profiling by gas chromatography and mass spectrometry (GC-MS). Further antibacterial activity was studied using silver, gold, and copper nanoparticles. The leaf explants were cultured on Murashige and Skoog (MS) medium containing Benzyl amino purine (BAP) (2.0 mg/L-1) and Naphthalene acetic acid (NAA) (1.0 mg/L-1) induced the formation of callus and expressed α-amylase with underexposed to Cyclodextrin. Explants growing on MS medium fortified with 2,4-Dichlorophenoxy acetic acid (2,4-D) (1.0 mg/L-1), NAA (1.0 mg/L-1) with and without BAP (2.0 mg/L-1) and showed a maximum concentration of protein on the 75th day. Synthesis of a-amylase enzyme was expressed 40 days old culture and were confirmatic by western, further Silver, gold, and copper nanoparticles were synthesized using the ethyl acetate extract of callus tissue and subjected to thin-layer chromatography (TLC), which resolved 5 bands. These five bands were characterized by Fourier Transform Infrared (FTIR) Spectroscopy and screened for antimicrobial and antioxidant activities. TLC band 4 alone showed inhibitory activity against both Gram-negative and positive bacteria and potent antioxidant activity. For the first time, α-amylase was found in the callus extract by SDS-PAGE and confirmed by Western blot. The fourth band of TLC from the ethyl acetate extract as well as silver and gold nanoparticles synthesized using this extract revealed pronounced antimicrobial and antioxidant activities. GC-MS analysis revealed 26 compounds, which included mainly the phytosterols and fatty acid esters.