Micropropagation, myristicin production enhancement, and comparative GC-MS analysis of the n-hexane extracts of different organs of Daucus pumilus (Gouan), family Apiaceae.

AIM: This work aimed to study the somatic embryogenesis and organogenesis of endangered Daucus pumilus (Gouan) for the conservation of this plant and improving the production of secondary metabolites of medicinal value. MATERIALS AND METHODS: The callus formation and in vitro propagation of D. pumilus (Gouan) by using a different combination of naphthalene acetic acid and benzylaminopurine were established. Various embryogenic stages were tracked using scanning electron microscopy and light microscopy. The volatile constituents of the n-hexane extracts of D. pumilus (Gouan) that extracted by ultrasonic-assisted technique were analyzed by gas chromatography-mass spectrometry. RESULTS AND DISCUSSION: Somatic embryogenesis and organogenesis of endangered D. pumilus (Gouan) were established for the first time. Myristicin and elemicin were successfully increased during micropropagation to 70.89% and 2.19%, respectively. Furthermore, the induction of compounds such as 6-methoxymellein, eugenin, methyl behenate, and 1,6-dimethylnaphthalene was also detected. CONCLUSION: Commercially, this protocol decreases the dependence on wild medicinal plants, enhances the manufacturing of valuable phytochemicals to meet the great demands of the pharmaceutical industries, and acts as a mean for genetic transformation of this plant.

Purification and Characterization of Anabaena flos-aquae Phenylalanine Ammonia-Lyase as a Novel Approach for Myristicin Biotransformation.

Phenylalanine ammonia-lyase (PAL) catalyzes the reversible deamination of phenylalanine to cinnamic acid and ammonia. Algae have been considered as biofactories for PAL production, however, biochemical characterization of PAL and its potency for myristicin biotransformation into MMDA (3-methoxy-4, 5-methylenedioxyamphetamine) has not been studied yet. Thus, PAL from Anabaena flos-aquae and Spirulina platensis has been purified, comparatively characterized and its affinity to transform myristicin was assessed. The specific activity of purified PAL from S. platensis (73.9 µmol/mg/min) and A. flos-aquae (30.5 µmol/mg/min) was increased by about 2.9 and 2.4 folds by gel-filtration comparing to their corresponding crude enzymes. Under denaturing-PAGE, a single proteineous band with a molecular mass of 64 kDa appeared for A. flos-aquae and S. platensis PAL. The biochemical properties of the purified PAL from both algal isolates were determined comparatively. The optimum temperature of S. platensis and A. flos-aquae PAL for forward or reverse activity was reported at 30°C, while the optimum pH for PAL enzyme isolated from A. flos-aquae was 8.9 for forward and reverse activities, and S. platensis PAL had maximum activities at pH 8.9 and 8 for forward and reverse reactions, respectively. Luckily, the purified PALs have the affinity to hydroaminate the myristicin to MMDA successfully in one step. Furthermore, a successful method for synthesis of MMDA from myristicin in two steps was also established. Gas chromatography-mass spectrometry (GC-MS) analysis was conducted to track the product formation.