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Trehalose biosynthesis in Thermus thermophilus RQ-1: biochemical properties of the trehalose-6-phosphate synthase and trehalose-6-phosphate phosphatase.
Extremophiles. 2005 Feb; 9(1):29-36.E

Abstract

The genes for trehalose synthesis in Thermus thermophilus RQ-1, namely otsA [trehalose-phosphate synthase (TPS)], otsB [trehalose-phosphate phosphatase (TPP)], and treS [trehalose synthase (maltose converting) (TreS)] genes are structurally linked. The TPS/TPP pathway plays a role in osmoadaptation, since mutants unable to synthesize trehalose via this pathway were less osmotolerant, in trehalose-deprived medium, than the wild-type strain. The otsA and otsB genes have now been individually cloned and overexpressed in Escherichia coli and the corresponding recombinant enzymes purified. The apparent molecular masses of TPS and TPP were 52 and 26 kDa, respectively. The recombinant TPS utilized UDP-glucose, TDP-glucose, ADP-glucose, or GDP-glucose, in this order as glucosyl donors, and glucose-6-phosphate as the glucosyl acceptor to produce trehalose-6-phosphate (T6P). The recombinant TPP catalyzed the dephosphorylation of T6P to trehalose. This enzyme also dephosphorylated G6P, and this activity was enhanced by NDP-glucose. TPS had an optimal activity at about 98 degrees C and pH near 6.0; TPP had a maximal activity near 70 degrees C and at pH 7.0. The enzymes were extremely thermostable: at 100 degrees C, TPS had a half-life of 31 min, and TPP had a half-life of 40 min. The enzymes did not require the presence of divalent cations for activity; however, the presence of Co2+ and Mg2+ stimulates both TPS and TPP. This is the first report of the characterization of TPS and TPP from a thermophilic organism.

Authors+Show Affiliations

Departamento de Bioquímica and Centro de Neurociências e Biologia Celular, Universidade de Coimbra, 3004-517 Coimbra, Portugal.No affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

15455210

Citation

Silva, Zélia, et al. "Trehalose Biosynthesis in Thermus Thermophilus RQ-1: Biochemical Properties of the Trehalose-6-phosphate Synthase and Trehalose-6-phosphate Phosphatase." Extremophiles : Life Under Extreme Conditions, vol. 9, no. 1, 2005, pp. 29-36.
Silva Z, Alarico S, da Costa MS. Trehalose biosynthesis in Thermus thermophilus RQ-1: biochemical properties of the trehalose-6-phosphate synthase and trehalose-6-phosphate phosphatase. Extremophiles. 2005;9(1):29-36.
Silva, Z., Alarico, S., & da Costa, M. S. (2005). Trehalose biosynthesis in Thermus thermophilus RQ-1: biochemical properties of the trehalose-6-phosphate synthase and trehalose-6-phosphate phosphatase. Extremophiles : Life Under Extreme Conditions, 9(1), 29-36.
Silva Z, Alarico S, da Costa MS. Trehalose Biosynthesis in Thermus Thermophilus RQ-1: Biochemical Properties of the Trehalose-6-phosphate Synthase and Trehalose-6-phosphate Phosphatase. Extremophiles. 2005;9(1):29-36. PubMed PMID: 15455210.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Trehalose biosynthesis in Thermus thermophilus RQ-1: biochemical properties of the trehalose-6-phosphate synthase and trehalose-6-phosphate phosphatase. AU - Silva,Zélia, AU - Alarico,Susana, AU - da Costa,Milton S, Y1 - 2004/09/29/ PY - 2004/04/26/received PY - 2004/08/04/accepted PY - 2004/9/30/pubmed PY - 2005/12/15/medline PY - 2004/9/30/entrez SP - 29 EP - 36 JF - Extremophiles : life under extreme conditions JO - Extremophiles VL - 9 IS - 1 N2 - The genes for trehalose synthesis in Thermus thermophilus RQ-1, namely otsA [trehalose-phosphate synthase (TPS)], otsB [trehalose-phosphate phosphatase (TPP)], and treS [trehalose synthase (maltose converting) (TreS)] genes are structurally linked. The TPS/TPP pathway plays a role in osmoadaptation, since mutants unable to synthesize trehalose via this pathway were less osmotolerant, in trehalose-deprived medium, than the wild-type strain. The otsA and otsB genes have now been individually cloned and overexpressed in Escherichia coli and the corresponding recombinant enzymes purified. The apparent molecular masses of TPS and TPP were 52 and 26 kDa, respectively. The recombinant TPS utilized UDP-glucose, TDP-glucose, ADP-glucose, or GDP-glucose, in this order as glucosyl donors, and glucose-6-phosphate as the glucosyl acceptor to produce trehalose-6-phosphate (T6P). The recombinant TPP catalyzed the dephosphorylation of T6P to trehalose. This enzyme also dephosphorylated G6P, and this activity was enhanced by NDP-glucose. TPS had an optimal activity at about 98 degrees C and pH near 6.0; TPP had a maximal activity near 70 degrees C and at pH 7.0. The enzymes were extremely thermostable: at 100 degrees C, TPS had a half-life of 31 min, and TPP had a half-life of 40 min. The enzymes did not require the presence of divalent cations for activity; however, the presence of Co2+ and Mg2+ stimulates both TPS and TPP. This is the first report of the characterization of TPS and TPP from a thermophilic organism. SN - 1431-0651 UR - https://www.unboundmedicine.com/medline/citation/15455210/Trehalose_biosynthesis_in_Thermus_thermophilus_RQ_1:_biochemical_properties_of_the_trehalose_6_phosphate_synthase_and_trehalose_6_phosphate_phosphatase_ L2 - https://dx.doi.org/10.1007/s00792-004-0421-4 DB - PRIME DP - Unbound Medicine ER -