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Research articleBiotechnology and Bioengineering (BB) | Enzyme, Protein Engineering, and Metabolic Engineering
Purification and Characterization of Anabaena flos-aquae Phenylalanine Ammonia-Lyase as a Novel Approach for Myristicin BiotransformationAsmaa Arafa 1, Afaf Abdel- Ghani 1, Samih El-Dahmy 1, Sahar Abdelaziz 1, Yassin El-Ayouty 2 and Ashraf El-Sayed 2*
1Department of Pharmacognosy, Faculty of Pharmacy, Zagazig University, Zagazig 44519, Egypt, 2Enzymology and Fungal Biotechnology Lab (EFBL), Botany and Microbiology Department, Faculty of Science, Zagazig University, Zagazig 44519, EgyptReceived: August 6, 2019; Accepted: September 27, 2019
J. Microbiol. Biotechnol. 2020; 30(4): 622-632
Published April 28, 2020
Copyright © The Korean Society for Microbiology and Biotechnology.
AbstractPhenylalanine 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 30oC, 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
KeywordsMMDA, Phenylalanine ammonia-Lyase, purification, myristicin, properties, MMDA
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