Sulfolobus acidocaldarius uses a complex trehalose metabolism for salt stress response involving a novel TPS/TPP pathway.

The Crenarchaeon Sulfolobus acidocaldarius has been described to synthesize trehalose via the maltooligosyltrehalose synthase (TreY) and maltooligosyltrehalose trehalohydrolase (TreZ) pathway and the trehalose glycosyltransferring synthase (TreT) pathway has been predicted. Deletion mutant analysis of single and double deletion strains of ΔtreY and ΔtreT in S. acidocaldarius revealed that next to these two pathways a third, novel trehalose biosynthesis pathway is operative in vivo: the trehalose-6-phosphate (T6P) synthase/T6P phosphatase (TPS/TPP) pathway. In contrast to known TPS proteins, which belong to the GT20 family, the S. acidocaldarius TPS belongs to the GT4 family establishing a new function within this group of enzymes. This novel GT4-like TPS turned out to be mainly present in the Sulfolobales. The ΔtreY/ΔtreT/Δtps triple mutant of S. acidocaldarius lacking the ability to synthesize trehalose showed no altered phenotype under standard conditions or heat stress, but was unable to grow under salt stress. Accordingly, in the wild type strain a significant increase of intracellular trehalose formation was observed under salt stress. Quantitative real-time PCR showed a salt stress mediated induction of all three trehalose synthesizing pathways. This demonstrates in Archaea that trehalose plays an essential role for growth under high salt conditions.ImportanceThe metabolism and function of trehalose as compatible solute was not well understood in Archaea. This combined genetic and enzymatic approach at the interface of microbiology, physiology and microbial ecology gives important insights into survival under stress, adaptation to extreme environments, and the role of compatible solutes in Archaea. Here we unravelled the complexity of trehalose metabolism and present a comprehensive study on trehalose function in stress response in S. acidocaldarius This sheds light on the general microbiology and the fascinating metabolic repertoire of Archaea involving many novel biocatalysts such as glycosyltransferases with great potential in biotechnology.