A novel trehalose-synthesizing glycosyltransferase from Pyrococcus horikoshii: Molecular cloning and characterization

Ryu, Soo In; Park, Cheon Seok; Cha, Jaeho; Woo, Eui-Jeon; Lee, Soo Bok

doi:10.1016/j.bbrc.2005.01.149

Cited by 64 publications

(54 citation statements)

References 20 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…While some of these organisms can have two or even three pathways for trehalose synthesis, most bacteria as well as plants and animals have only the Tps/Tpp pathway (2,38,39). The TreS pathway is found almost exclusively in bacteria, and the TreT pathway is present mainly in hyperthermophilic bacteria and archaea (7,17,29,33). Occasionally, more than one copy of those genes exists in the same organism: Ralstonia solanacearum, for example, has two tps genes; Thermoanaerobacter tengcongensis has two treP genes; and Methanoculleus marisnigri has three copies of treT.…”

Section: Discussionmentioning

confidence: 99%

“…The TreT from R. xylanophilus catalyzes a reaction similar to that of the homofunctional enzymes from Thermococcus litoralis and Pyrococcus horikoshii, including the hydrolysis of trehalose with ADP as a cosubstrate (resembling the former), but it does not have the trehalase activity of the latter (29,33). Interestingly, the TreT from Thermoproteus tenax could catalyze only the formation and not the hydrolysis of trehalose (17).…”

Section: Discussionmentioning

confidence: 99%

“…With the exception of hyperthermophilic bacteria and a few deltaproteobacteria, treT is not found in most bacterial genomes. This gene is also present in a few euryarchaeal genomes but is frequent in those from hyperthermophilic crenarchaeotes (17,29,33). Such a scattered distribution suggests that treT has been involved in lateral gene transfer events and that R. xylanophilus may have acquired it from hyperthermophiles (8,31).…”

Section: Discussionmentioning

confidence: 99%

“…A less common and recently discovered pathway involves a trehalose glycosyltransferring synthase (TreT) that converts NDP-glucose and glucose into trehalose. This enzyme has been examined only in the hyperthermophilic archaea Thermococcus litoralis, Pyrococcus horikoshii, and Thermoproteus tenax (17,29,33).…”

mentioning

confidence: 99%

See 3 more Smart Citations

A Unique Combination of Genetic Systems for the Synthesis of Trehalose in Rubrobacter xylanophilus : Properties of a Rare Actinobacterial TreT

et al. 2008

View full text Add to dashboard Cite

Trehalose is the primary organic solute inRubrobacter xylanophilus under all conditions tested, including those for optimal growth. We detected genes of four different pathways for trehalose synthesis in the genome of this organism, namely, the trehalose-6-phosphate synthase (Tps)/trehalose-6-phosphate phosphatase (Tpp), TreS, TreY/TreZ, and TreT pathways. Moreover, R. xylanophilus is the only known member of the phylum Actinobacteria to harbor TreT. The Tps sequence is typically bacterial, but the Tpp sequence is closely related to eukaryotic counterparts. Both the Tps/Tpp and the TreT pathways were active in vivo, while the TreS and the TreY/TreZ pathways were not active under the growth conditions tested and appear not to contribute to the levels of trehalose observed. The genes from the active pathways were functionally expressed in Escherichia coli, and Tps was found to be highly specific for GDP-glucose, a rare feature among these enzymes. The trehalose-6-phosphate formed was specifically dephosphorylated to trehalose by Tpp. The recombinant TreT synthesized trehalose from different nucleoside diphosphate-glucose donors and glucose, but the activity in R. xylanophilus cell extracts was specific for ADP-glucose. The TreT could also catalyze trehalose hydrolysis in the presence of ADP, but with a very high K m . Here, we functionally characterize two systems for the synthesis of trehalose in R. xylanophilus, a representative of an ancient lineage of the actinobacteria, and discuss a possible scenario for the exceptional occurrence of treT in this extremophilic bacterium.

show abstract

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

mentioning

confidence: 99%

See 2 more Smart Citations

A Unique Combination of Genetic Systems for the Synthesis of Trehalose in Rubrobacter xylanophilus : Properties of a Rare Actinobacterial TreT

et al. 2008

View full text Add to dashboard Cite

show abstract

“…The recently discovered fourth pathway involves trehalose glycosyltransferring synthase (TreT), which catalyzes the reversible formation of trehalose from ADP-glucose and glucose. TreT has been characterized in hyperthermophilic bacteria (37,39).Many bacterial species possess a single pathway; for example, Escherichia coli (48) and Salmonella enterica (24) synthesize trehalose using the OtsAB pathway. In E. coli, the otsBA…”

mentioning

confidence: 99%

Trehalose Biosynthesis in Rhizobium leguminosarum bv. trifolii and Its Role in Desiccation Tolerance

Mcintyre¹,

Davies

Hore³

et al. 2007

Appl Environ Microbiol

View full text Add to dashboard Cite

Rhizobium leguminosarum bv. trifolii forms nitrogen-fixing root nodules on the pasture legume Trifolium repens, and T. repens seed is often coated with a compatible R. leguminosarum bv. trifolii strain prior to sowing. However, significant losses in bacterial viability occur during the seed-coating process and during storage of the coated seeds, most likely due to desiccation stress. The disaccharide trehalose is known to function as an osmoprotectant, and trehalose accumulation due to de novo biosynthesis is a common response to desiccation stress in bacteria. In this study we investigated the role of endogenous trehalose synthesis in desiccation tolerance in R. leguminosarum bv. trifolii strain NZP561. Strain NZP561 accumulated trehalose as it entered the stationary phase due to the combined actions of the TreYZ and OtsAB pathways. Mutants deficient in either pathway showed near-wild-type levels of trehalose accumulation, but double otsA treY mutants failed to accumulate any trehalose. The double mutants were more sensitive to the effects of drying, and their survival was impaired compared to that of the wild type when glass beads were coated with the organisms and stored at relative humidities of 5 and 32%. The otsA treY mutants were also less competitive for nodule occupancy. Gene expression studies showed that the otsA and treY genes were expressed constitutively and that expression was not influenced by the growth phase, suggesting that trehalose accumulation is controlled at the posttranscriptional level or by control of trehalose breakdown rates. Our results indicate that accumulated trehalose plays an important role in protecting R. leguminosarum bv. trifolii cells against desiccation stress and against stress encountered during nodulation.Rhizobium leguminosarum bv. trifolii forms nitrogen-fixing root nodules on the pasture legume Trifolium repens (white clover). Inoculation of legumes is an efficient and convenient way of introducing rhizobia into soil and subsequently into the rhizosphere of legumes. The seeds of many clover species (including white clover) are coated with compatible R. leguminosarum bv. trifolii strains prior to sowing. One of the main reasons for the ineffectiveness of legume inoculants in the field is the rapid death of rhizobia due to desiccation (12, 55). Significant losses in inoculant viability occur during the seedcoating process, which involves a drying stage, during storage of the coated seeds and after introduction into the field environment. These losses in bacterial viability are likely to be due mainly to desiccation stress.Bacteria accumulate osmoprotective compounds, referred to as compatible solutes or osmolytes, in response to osmotic or desiccation stress. Osmolytes can be obtained by uptake from the environment (exogenous) or through de novo biosynthesis (endogenous). De novo biosynthesis of trehalose, a nonreducing (␣,␣-1,1)-glucose disaccharide, is a common response to desiccation and osmotic stress in many bacteria (11,35,57). Trehalose has been shown to protec...

show abstract

A glycogen branching enzyme from Thermomonospora curvata: Characterization and its action on maize starch

et al. 2015

View full text Add to dashboard Cite

A gene (TcGBE) encoding a glycogen branching enzyme from Thermomonospora curvata was expressed in Escherichia coli and purified, and biochemically analyzed. The purified enzyme was approximately 84 kDa, and exhibited a maximum activity at 55°C and pH 8.5. The enzyme showed the highest specificity to amylose with a specific activity of 90.28 U/mg. The recombinant enzyme could act on maize starch, and starch treated with TcGBE was observed to have a lower amylose content and molecular weight. The number of short chains of amylopectin in TcGBEmodified starch with a degree of polymerization (DP) less than 13 increased from 20.19 to 32.59%, and was accompanied by a reduction in longer chains (DP > 25) from 25.58 to 10.03%. In addition, the medium length chains of amylopectin (13 < DP <25) also showed a slight increase from 54.23 to 58.38%. The 1 H NMR spectra revealed an increased ratio of a-1,6-to a-1, 4-glucosidic linkages from 4.1 to 10.3% following treatment with TcGBE for 10 h. Taken together, these results suggested that TcGBE could be used to generate a modified maize starch with a low amylose content and more branched amylopectin of low molecular weight but richer in short and intermediate chains.

show abstract

A novel trehalose-synthesizing glycosyltransferase from Pyrococcus horikoshii: Molecular cloning and characterization

Cited by 64 publications

References 20 publications

A Unique Combination of Genetic Systems for the Synthesis of Trehalose in Rubrobacter xylanophilus : Properties of a Rare Actinobacterial TreT

A Unique Combination of Genetic Systems for the Synthesis of Trehalose in Rubrobacter xylanophilus : Properties of a Rare Actinobacterial TreT

Trehalose Biosynthesis in Rhizobium leguminosarum bv. trifolii and Its Role in Desiccation Tolerance

A glycogen branching enzyme from Thermomonospora curvata: Characterization and its action on maize starch

Contact Info

Product

Resources

About