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

• Published in: Journal of microbiology and biotechnology Vol. 30; no. 4; pp. 622 - 632
• Main Authors: Arafa, Asmaa M, Abdel-Ghany, Afaf E, El-Dahmy, Samih I, Abdelaziz, Sahar, El-Ayouty, Yassin, El-Sayed, Ashraf S A
• Format: Journal Article
• Language: English
• Published: Korea (South) The Korean Society for Microbiology and Biotechnology 28.04.2020; 한국미생물·생명공학회
• Subjects: 3,4-Methylenedioxyamphetamine - analogs & derivatives; 3,4-Methylenedioxyamphetamine - metabolism; Allylbenzene Derivatives; Bacterial Proteins - chemistry; Bacterial Proteins - isolation & purification; Bacterial Proteins - metabolism; Benzyl Compounds - metabolism; Biotransformation; Dioxolanes - metabolism; Dolichospermum flos-aquae - enzymology; Hydrogen-Ion Concentration; Molecular Structure; Molecular Weight; Phenylalanine Ammonia-Lyase - chemistry; Phenylalanine Ammonia-Lyase - isolation & purification; Phenylalanine Ammonia-Lyase - metabolism; Pyrogallol - analogs & derivatives; Pyrogallol - metabolism; Research article; Spirulina - enzymology; Substrate Specificity; Temperature; 생물학; myristicin; Phenylalanine ammonia-Lyase; MMDA; purification; properties
• ISSN: 1017-7825; 1738-8872
• DOI: 10.4014/jmb.1908.08009

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 and has been purified, comparatively characterized and its affinity to transform myristicin was assessed. The specific activity of purified PAL from (73.9 μmol/mg/min) and (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 and PAL. The biochemical properties of the purified PAL from both algal isolates were determined comparatively. The optimum temperature of and PAL for forward or reverse activity was reported at 30°C, while the optimum pH for PAL enzyme isolated from was 8.9 for forward and reverse activities, and 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.