Abstract
Pleurotus tuoliensis, a kind of valuable and favorable edible mushroom in China, is always subjected to high environmental temperature during cultivation. In our previous study with P. tuoliensis, trehalose proved to be effective for tolerating heat stress. Trehalose-6-phosphate synthase (TPS; EC2.4.1.15) plays a key role in the biosynthesis of trehalose in fungi. In this study, a full-length of cDNA with 1,665 nucleotides encoding TPS (PtTPS) in P. tuoliensis was cloned. The PtTPS amino acid was aligned with other homologues and several highly conserved regions were analyzed. Thus, the TPS protein was expressed in Escherichia coli and purified by affinity chromatography to test its biochemical properties. The molecular mass of the enzyme is about 60 kDa and the optimum reaction temperature and pH is 30 °C and 7, respectively. The UDP-glucose and glucose-6-phosphate were the optimum substrates among all the tested glucosyl donors and acceptors. Metal cations like Mg2+, Co2+, Mn2+, Ni2+, K+, Ag+ stimulated PtTPS activity significantly. Metal chelators such as sodium citrate, citric acid, EDTA, EGTA and CDTA inhibited enzyme activity. Polyanions like heparin and chondroitin sulfate were shown to stimulate TPS activity.
Highlights
IntroductionPleurotus tuoliensis (previously named as P. eryngii var. tuoliensis) (Zhao et al, 2016) is a favored edible mushroom with rich nutritional and medicinal value in China
Pleurotus tuoliensis (Zhao et al, 2016) is a favored edible mushroom with rich nutritional and medicinal value in China
Exogenous addition of trehalose significantly reduced the oxidative damages resulted from heat stress to the cell membrane. These results indicated that both the trehalose and the Trehalose-6-phosphate synthase (TPS) gene in P. tuoliensis were important in tolerating heat stress
Summary
Pleurotus tuoliensis (previously named as P. eryngii var. tuoliensis) (Zhao et al, 2016) is a favored edible mushroom with rich nutritional and medicinal value in China. Tuoliensis) (Zhao et al, 2016) is a favored edible mushroom with rich nutritional and medicinal value in China. It is seasonally cultivated in agricultural facilities in some parts of China, and the spontaneous high environmental temperatures during summer usually affect its growth, development, and the quality and yield of the fruiting body. Trehalose is a stable and non-reducing disaccharide, which is widely spread in a variety of organisms (Elbein et al, 2003). At least five trehalose biosynthetic pathways have been reported in the organisms (Avonce et al, 2006).
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