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

Richard P, Viljanen K, Penttilä M - AMB Express (2015)

Bottom Line: This was suggested for the reason that the over-expression of PAD1 resulted in increased tolerance toward cinnamic acid, up to 0.6 mM.The overexpression of PAD1 and FDC1 resulted also in increased activity with the hydroxycinnamic acids ferulic acid, p-coumaric acid and caffeinic acid.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.

View Article: PubMed Central - PubMed

Affiliation: VTT Technical Research Centre of Finland Ltd, Tietotie 2, P.O. box 1000, 02044 VTT Espoo, Finland.

ABSTRACT
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.

No MeSH data available.


Related in: MedlinePlus

Trans-cinnamic acid is converted in a decarboxylation reaction to styrene.
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Fig1: Trans-cinnamic acid is converted in a decarboxylation reaction to styrene.

Mentions: The yeast Saccharomyces cerevisiae shows some resistance toward cinnamic acid and the gene that confers this resistance was identified as the PAD1 (phenylacrylic acid decarboxylase) gene (Clausen et al. 1994). The yeast can convert cinnamic acid to styrene (Figure 1) but also sorbic acid to 1,3-pentadiene however with a lower rate. The deletion of PAD1 resulted in the inability to convert cinnamic or sorbic acid suggesting that the Pad1p is active with both substrates (Stratford et al. 2007). Overexpression of the PAD1 gene resulted in strains with increased resistance to cinnamic and ferulic acid (Larsson et al. 2001). A PAD1 homologue is also present in the mold Aspergillus niger where it confers resistance to cinnamic acid and sorbic acid (Plumridge et al. 2008). Recently it was shown in S. cerevisiae that not only the PAD1 but the PAD1 and the FDC1 are essential for cinnamic acid, ferulic acid or coumaric acid resistance. Deletions in either of these two genes resulted in drastic reduced ability to decarboxylate these acids (Mukai et al. 2010). A possible interpretation of this observation was that the Pad1p and the Fdc1p are two essential subunits of a protein complex. The Pad1p and the Fdc1p are however believed to be in different cellular compartments. The Pad1p has a mitochondrial targeting sequence and the protein is located in the mitochondria (Huh et al. 2003) whereas the Fdc1p is a cytosolic protein.Figure 1


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

Richard P, Viljanen K, Penttilä M - AMB Express (2015)

Trans-cinnamic acid is converted in a decarboxylation reaction to styrene.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4384992&req=5

Fig1: Trans-cinnamic acid is converted in a decarboxylation reaction to styrene.
Mentions: The yeast Saccharomyces cerevisiae shows some resistance toward cinnamic acid and the gene that confers this resistance was identified as the PAD1 (phenylacrylic acid decarboxylase) gene (Clausen et al. 1994). The yeast can convert cinnamic acid to styrene (Figure 1) but also sorbic acid to 1,3-pentadiene however with a lower rate. The deletion of PAD1 resulted in the inability to convert cinnamic or sorbic acid suggesting that the Pad1p is active with both substrates (Stratford et al. 2007). Overexpression of the PAD1 gene resulted in strains with increased resistance to cinnamic and ferulic acid (Larsson et al. 2001). A PAD1 homologue is also present in the mold Aspergillus niger where it confers resistance to cinnamic acid and sorbic acid (Plumridge et al. 2008). Recently it was shown in S. cerevisiae that not only the PAD1 but the PAD1 and the FDC1 are essential for cinnamic acid, ferulic acid or coumaric acid resistance. Deletions in either of these two genes resulted in drastic reduced ability to decarboxylate these acids (Mukai et al. 2010). A possible interpretation of this observation was that the Pad1p and the Fdc1p are two essential subunits of a protein complex. The Pad1p and the Fdc1p are however believed to be in different cellular compartments. The Pad1p has a mitochondrial targeting sequence and the protein is located in the mitochondria (Huh et al. 2003) whereas the Fdc1p is a cytosolic protein.Figure 1

Bottom Line: This was suggested for the reason that the over-expression of PAD1 resulted in increased tolerance toward cinnamic acid, up to 0.6 mM.The overexpression of PAD1 and FDC1 resulted also in increased activity with the hydroxycinnamic acids ferulic acid, p-coumaric acid and caffeinic acid.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.

View Article: PubMed Central - PubMed

Affiliation: VTT Technical Research Centre of Finland Ltd, Tietotie 2, P.O. box 1000, 02044 VTT Espoo, Finland.

ABSTRACT
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.

No MeSH data available.


Related in: MedlinePlus