Enzymatic production of (R)-phenylacetylcarbinol by pyruvate decarboxylase from Zymomonas mobilis

• Published in: Biotechnology and bioprocess engineering 2008, 13(3), , pp.372-376
• Main Authors: Yun, H.D. (Yeungnam University, Gyeongsan, Republic of Korea), E-mail: [e-mail], Kim, B.G. (Seoul National University, Seoul, Republic of Korea)
• Format: Article
• Language: English
• Subjects: Biotechnology; biotransformation; carboligation; Chemistry; Chemistry and Materials Science; E coli; Escherichia coli; Industrial and Production Engineering; phenylacetylcarbinol; pyruvate decarboxylase; Yeasts; ZYMOMONAS MOBILIS; 생물공학
• ISSN: 1226-8372; 1976-3816
• DOI: 10.1007/s12257-008-0030-7

Herein, we synthesized (R)-phenylacetylcarbinol (PAC), a pharmaceutical intermediate for ephedrine. and pseudoephedrine, from benzaldehyde and pyruvate using a recombinant pyruvate decarboxylase (PDC) from Zymomonas mobilis. A whole cell reaction consisting of 30 mM benzaldehyde, 60 mM pyruvate, and a mutant PDC enzyme (PDC W329M; 1.6 mg DCW/mL produced 12.4 mM (R)-PAC and less than 0.3 mM benzyl alchohol in 15 h at 20℃, outperforming the crude enzyme extract reaction (1.2 mM (R)-PAC) and minimizing formation of benzyl alchohol, the major by-product of S. cerevisiae whole cell reaction. These observations suggested that recombinant E. coli whole cell reactions are more efficient than crude enzyme extract or yeast-based reactions. We also demonstrated that the E. coli whole cell reaction performed effectively without expensive thiamin diphosphate cofactor. Finally, whole cell reaction (8 mg DCW/mL) was carried out with 200 mM benzaldehyde, 400 mM pyruvate in 10 mL of 500 mM phosphate buffer (pH 6.5), and 72 mM (R)-PAC was produced with 36% conversion for 15 h.