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Three trehalose synthetic pathways in the acarbose-producing Actinoplanes sp. SN223/29 and evidence for the TreY role in biosynthesis of component C

Abstract

In this study, three trehalose gene clusters, treX-Y-Z, tpS1, and treS, of the acarbose-producing strain, Actinoplanes sp. SN223/29, have been identified. In particular, five trehalose synthetic genes were sequenced and characterized in detail. They were cloned and expressed in Escherichia coli BL21(DE3)pLysS using the His-tag vector pET19b. The recombinant proteins were purified by Ni2+-nitrilotriacetic acid agarose affinity chromatography, and their functions were characterized biochemically. Both the maltooligosyltrehalose synthase (TreY–TreZ) pathway and the trehalose synthase (TreS) pathway have maximum activity at 40°C and at pH 7.5 and 7.0, respectively, in 100-mM phosphate buffer. Meanwhile, the trehalose-6-phosphate synthase (TpS1) showed maximum activity at 35°C and at pH 7.5 in 100 mM Tris–HCl. As a cofactor candidate, Mg2+ enhanced the activities of all three trehalose synthetic reactions significantly. TreY produced component C from acarbose by its proposed isomerase activity, but TreS did not. This study suggests that the mutation of treY can improve acarbose production by repressing component C production. Based on the data obtained in this study, a model for component C production in Actinoplanes sp. SN 223/29 is proposed.

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Acknowledgements

The authors thank Bayer AG (Wuppertal, Germany) for providing the strain Actinoplanes sp. SN223/29 and Professor Piepersberg from the University of Wuppertal (Germany) for the λ-EMBL3 phage library of Actinoplanes sp. SN223/29. This work was supported by a grant (Code # 20050301034449) from BioGreen 21 Program, Rural Development Administration, Republic of Korea.

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Correspondence to Joo-Won Suh.

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Lee, J., Hai, T., Pape, H. et al. Three trehalose synthetic pathways in the acarbose-producing Actinoplanes sp. SN223/29 and evidence for the TreY role in biosynthesis of component C. Appl Microbiol Biotechnol 80, 767 (2008). https://doi.org/10.1007/s00253-008-1582-3

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Keywords

  • Actinoplanes sp.
  • TreX-Y-Z
  • TpS1
  • TreS
  • Trehalose biosynthesis
  • Acarbose
  • Component C