Expression of salt-induced 2-Cys peroxiredoxin from Oryza sativa increases stress tolerance and fermentation capacity in genetically engineered yeast Saccharomyces cerevisiae

Abstract

Peroxiredoxins (Prxs), also termed thioredoxin peroxidases (TPXs), are a family of thiol-specific antioxidant enzymes that are critically involved in cell defense and protect cells from oxidative damage. In this study, a putative chloroplastic 2-Cys thioredoxin peroxidase (OsTPX) was identified by proteome analysis from leaf tissue samples of rice (Oryza sativa) seedlings exposed to 0.1 M NaCl for 3 days. To investigate the relationship between the OsTPX gene and the stress response, OsTPX was cloned into the yeast expression vector p426GPD under the control of the glyceraldehyde-3-phosphate dehydrogenase (GPD1) promoter, and the construct was transformed into Saccharomyces cerevisiae cells. OsTPX expression was confirmed by semi-quantitative reverse transcription-polymerase chain reaction and western blot analyses. OsTPX contained two highly conserved cysteine residues (Cys114 and Cys236) and an active site region (FTFVCPT), and it is structurally very similar to human 2-Cys Prx. Heterologous OsTPX expression increased the ability of the transgenic yeast cells to adapt and recover from reactive oxygen species (ROS)-induced oxidative stresses, such as a reduction of cellular hydroperoxide levels in the presence of hydrogen peroxide and menadione, by improving redox homeostasis. OsTPX expression also conferred enhanced tolerance to tert-butylhydroperoxide, heat shock, and high ethanol concentrations. Furthermore, high OsTPX expression improved the fermentation capacity of the yeast during glucose-based batch fermentation at a high temperature (40 °C) and at the general cultivation temperature (30 °C). The alcohol yield in OsTPX-expressing transgenic yeast increased by approximately 29 % (0.14 g g(-1)) and 21 % (0.12 g g(-1)) during fermentation at 40 and 30 °C, respectively, compared to the wild-type yeast. Accordingly, OsTPX-expressing transgenic yeast showed prolonged cell survival during the environmental stresses produced during fermentation. These results suggest that heterologous OsTPX expression increases acquired tolerance to ROS-induced oxidative stress by improving cellular redox homeostasis and improves fermentation capacity due to improved cell survival during fermentation, especially at a high temperature.

Enhanced fermentative performance under stresses of multiple lignocellulose-derived inhibitors by overexpression of a typical 2-Cys peroxiredoxin from Kluyveromyces marxianus

“…There were no obvious differences in growth between two strains under low concentration (0.2 mM). An elevated concentration of 3 mM could inhibit the growth of the control strain 423 significantly, while strain TPX1 had been less affected, which was in line with the results reported by Kim et al [29]. Consequently, an enhanced tolerance of S. cerevisiae 280 with a normally functioning KmTPX1 gene provided a good foundation for further analysis and validation of other possible functions.…”

“…2a), while the growth of control strain 423 had been greatly repressed on plates with 0.3 g/L of formic acid, 1.5 g/L of acetic acid, or 1.0 g/L of furfural, which was 1–2 gradients less than the strain TPX1 in the serial dilution assay. These results have been supported by a variety of similar oxidoreductases that might be contributed to the increased tolerance of S. cerevisiae to the inhibitors [8–15, 29]. However, despite sharing a similar mechanism of toxicity with furfural, the tolerance of TPX1 strain to 5-HMF was only slightly improved compared with the control strain.

Fig.

…”

“…However, when 3 mM of H 2 O 2 was added to the plates, TPX1 achieved a much better growth status than the strain 423, which indicated that the gene KmTPX1 might increase the tolerance of S. cerevisiae to H 2 O 2 . Similarly, a kind of 2-Cys Prxs from Oryza sativa [29] and the thioredoxin from Endocarpon pusillum [30] both have been proved to enhance the tolerance of S. cerevisiae to peroxides, including H 2 O 2 and 2-methyl-1,4-naphthoquinone (MD).

Fig. 2Effects of KmTPX1 overexpression on the growth behavior under oxidative stress.

…”

Understanding the Mechanism of Thermotolerance Distinct From Heat Shock Response Through Proteomic Analysis of Industrial Strains of Saccharomyces cerevisiae

“…S4C, its deletion strain no longer produced ethanol, implying the dominant role of ADH1 in ethanol biosynthesis. The evident growth advantage of ⌬adh1 could be mainly attributed to abolished synthesis of ethanol which is known to suppress cell growth and glucose uptake (33)(34)(35)(36)(37). However, glucose consumption was also reduced in ⌬adh1, suggesting possibly lowered activity of glycolysis (Fig.…”

Overexpression of 2-Cys Prx Increased Salt Tolerance of Photosystem II in Tobacco

“…Molecular verification and physiological testing was then conducted on the 2Cys Prx-overexpression tobacco. Kim et al (2013) transferred the 2-Cys Prx gene from rice (Oryza sativa) into Saccharomyces cerevisiae and found that the heterologous expression of 2-Cys Prx in S. cerevisiae increased the oxidation-reduction balance in S. cerevisiae 40°C, indicating that 2-Cys Prx effectively regulates the reactive oxygen balance in organisms. H2O2 in plant cells is the intermediate product in the process of water splitting and oxygen liberation.…”