Fig. 2: Through signal peptide shuffling identified novel UPO construct and their analysis of UV absorption spectra and pH profiles. | Communications Biology

Fig. 2: Through signal peptide shuffling identified novel UPO construct and their analysis of UV absorption spectra and pH profiles.

From: A modular two yeast species secretion system for the production and preparative application of unspecific peroxygenases

Fig. 2: Through signal peptide shuffling identified novel UPO construct and their analysis of UV absorption spectra and pH profiles.

A Golden Gate signal peptide shuffling was applied for the testing of described and putative UPO genes, and the two best signal peptide/UPO gene combinations are displayed. GmaUPO, MweUPO, MroUPO and CglUPO were screened in combination with a GFP11-tag. MthUPO and TteUPO were screened using the TwinStrep-GFP11 protein tag. UPO enzyme activity was determined by monitoring the conversion of 2,6-dimethoxyphenol (DMP) to coerulignone. The highest average fluorescence (split-GFP) and conversion values (DMP) within one enzyme panel were set to 100%, and the other values normalised accordingly. Data are mean ± s.d. of biological replicates (n ≥ 4). Corresponding primary data are displayed within the Source data file. B UV-Vis absorption spectra of the purified peroxygenases MroUPO, CglUPO, MthUPO and TteUPO in the wavelength range between 300 and 600 nm (measurement interval: 1 nm). C pH profiles of MroUPO, CglUPO, MthUPO and TteUPO catalysed enzymatic conversion of 5-nitro-1,3-benzodioxole (NBD) to 4-nitrocatechol. The highest mean activity of a respective enzyme was set to 100% and the other values normalised accordingly. Data are means ± s.d. of measurements performed in triplicates. Corresponding primary data are displayed within the Source data file.

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