Overexpression of PAD1 and FDC1 results in significant cinnamic acid decarboxylase activity in Saccharomyces cerevisiae

• Published in: AMB Express Vol. 5; no. 1; p. 12
• Main Authors: Richard, Peter, Viljanen, Kaarina, Penttilä, Merja
• Format: Journal Article
• Language: English
• Published: Berlin/Heidelberg Springer Berlin Heidelberg 18.02.2015; Springer Nature B.V; BioMed Central Ltd
• Subjects: Biomedical and Life Sciences; Biotechnology; Life Sciences; Microbial Genetics and Genomics; Microbiology; Original; Original Article; Decarboxylase; Cinnamic acid; Styrene; PAD1; FDC1; S. cerevisiae
• ISSN: 2191-0855; 2191-0855
• DOI: 10.1186/s13568-015-0103-x

The S. cerevisiae PAD1 gene had been suggested to code for a cinnamic acid decarboxylase, converting trans -cinnamic acid to styrene. This was suggested for the reason that the over-expression of PAD1 resulted in increased tolerance toward cinnamic acid, up to 0.6 mM. We show that by over-expression of the PAD1 together with the FDC1 the cinnamic acid decarboxylase activity can be increased significantly. The strain over-expressing PAD1 and FDC1 tolerated cinnamic acid concentrations up to 10 mM. The cooperation of Pad1p and Fdc1p is surprising since the PAD1 has a mitochondrial targeting sequence and the FDC1 codes for a cytosolic protein. The cinnamic acid decarboxylase activity was also seen in the cell free extract. The activity was 0.019 μmol per minute and mg of extracted protein. The overexpression of PAD1 and FDC1 resulted also in increased activity with the hydroxycinnamic acids ferulic acid, p-coumaric acid and caffeinic acid. This activity was not seen when FDC1 was overexpressed alone. An efficient cinnamic acid decarboxylase is valuable for the genetic engineering of yeast strains producing styrene. Styrene can be produced from endogenously produced L-phenylalanine which is converted by a phenylalanine ammonia lyase to cinnamic acid and then by a decarboxylase to styrene.