Part:BBa_K3753001

PAAS-Vanda

Vanda hybrid phenylacetaldehyde synthase (PAAS) is capable of transforming L-phenylalanine (L-phe) into phenylacetaldehyde by oxidative decarboxylation. Subsequently, phenylacetaldehyde can be reducted into phenylethanol, which possesses elegant, fascinating and long-lasting fragrance.

Characterization

Numerous experiments have shown that the growth of the modified strain was not significantly different from that of the wild type. Import genes had no significant effect on yeast growth and metabolism (Fig. 1). Because yeast contains Ehrlich pathway and other metabolic pathways, they can work together to produce 2-PE, Saccharomyces cerevisiae BY4741 wild-type contains a certain amount of 2-PE production (1.205g/L). After the introduction of exogenous paas gene, the 2-PE production has increased. Among Petunia PAAS BBa_K3753000, Vanda PAAS and Rosa PAAS BBa_K3753002, Petunia PAAS has the most notable effect. The 2-PE production of yeast cells carrying pRS426 plasmid with Petunia PAAS reached 1.570g/L, with Vanda PAAS (1.514g/L) and with Rosa PAAS (1.341g/L) following closely (Fig. 2).

Fig.1 Changes in WT, BY4741-pRS426-Petunia, BY4741-pRS426-Vanda, BY4741-pRS426-Rosa yeast biomass concentration over time. The OD600 of different strains cultures used for analysis of 2-PE was measured at the indicated time points (Samples 0h, 24h,48h,72h). Data represent means of triplicate cultures ± standard error.

Fig.2 Introduced plasmids with target gene paas elevates 2-PE levels. Yeast cells carrying pRS426 plasmid with Petunia PAAS , Vanda PAAS, Rosa PAAS or, as controls, the unrelated Saccharomyces cerevisiae BY4741 wild-type, were grown in fermentation media. Cultures analyzed sampling for 72h, followed by HPLC to determine 2-PE levels. Data represent means of triplicate culture±standard error.

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal NheI site found at 1033
  
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]