Soybean PM2 protein (LEA3) confers the tolerance of Escherichia coli and stabilization of enzyme activity under diverse stresses.
Late embryogenesis abundant (LEA) proteins are closely associated with the tolerance of diverse stresses in organisms. To elucidate the function of group 3 LEA proteins, the soybean PM2 protein (LEA3) was expressed in E. coli and the protective function of the PM2 protein was assayed both in vivo and in vitro. The results of a spot assay and survival ratio demonstrated that the expression of the PM2 protein conferred the tolerance to the E. coli recombinant for different temperature conditions (4, -20 or 50 degrees C) or high-salinity stresses (120 mmol/l MgCl(2) or 120 mmol/l CaCl(2)). In addition, it was demonstrated that the in vitro addition of the PM2 protein could prevent the lactate dehydrogenase (LDH) inactivation normally induced by freeze-thaw. In the 62 degrees C condition, the PM2 protein (1:5 mass ratio to LDH) effectively prevented the LDH thermo-denaturation by acting synergistically with trehalose (62.5 microg/ml), although the PM2 protein alone at this concentration showed little protective effect on LDH activity. Furthermore, the results showed that the PM2 protein could partially prevent the thermo-denaturation of the bacterial proteome after boiling for 2 min. Based on these results, we propose that the PM2 protein itself, or together with trehalose, conferred the tolerance to the E. coli recombinant against diverse stresses by protecting proteins and enzyme activity under low- or high- temperature conditions.
Liu
Yun
Y
Shenzhen University, People's Republic of China.
Zheng
Yizhi
Y
Zhang
Yuqin
Y
Wang
Weimao
W
Li
Ranhui
R
D003001:Cloning, Molecular; D003080:Cold Temperature; D004926:Escherichia coli / Q000235:genetics / Q000502:physiology*; D015870:Gene Expression; D006358:Hot Temperature; D007770:L-Lactate Dehydrogenase / Q000378:metabolism; D050296:Microbial Viability; D009997:Osmotic Pressure; D010940:Plant Proteins / Q000235:genetics / Q000502:physiology*; D054712:Salinity; D013025:Glycine max / Q000235:genetics / Q000502:physiology*; D013312:Stress, Physiological
D010940:Plant Proteins; C066246:late embryogenesis abundant protein, plant; D007770:L-Lactate Dehydrogenase