In marine macroalgae, environmental stresses can induce major oxidative damage to cells. Aldehyde dehydrogenase (ALDH) is a key enzyme in the defense systems against oxidative damage and in protecting organisms from environmental stresses. In this study, we investigated changes in the proteome of Pyropia yezoensis (Bangiales) under two stress conditions, namely, hydrogen peroxide (H2O2) treatment and high temperature. Analysis of two-dimensional gels identified one spot that increased significantly following exposure to each of the stresses. MALDI-TOF spectrometric analysis of these spots identified three amino acid sequences, and these were used for a search of the EST database. This search identified a full-length cDNA clone encoding ALDH. Pyropia yezoensis ALDH (PyALDH) cDNA was 1515 bp in length and encoded a deduced polypeptide of 505 amino acids (57 kDa). We sub-cloned the ALDH gene into pET28a(+), and expression in Escherichia coli (Rosetta DE3) was used to determine the function in response to increased salinity and oxidative stress. Transgenic lines show improved tolerance following exposure to higher NaCl concentrations or to H2O2. An enzyme activity analysis of recombinant PyALDH showed that the most suitable reaction conditions were pH 7–7.5, 20–25 °C, and MnCl2 and ZnCl2 as activators. These results indicate that ALDH may play an important role in the ability of P. yezoensis to resist stress conditions.
Journal of Applied Phycology – Springer Journals
Published: Feb 6, 2018
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