A thermostable glycosyltransferase from Paenibacillus polymyxa NJPI29: recombinant expression, characterization, and application in synthesis of glycosides.

ABSTRACT: Glycosylation is a prominent biological mechanism, affecting the structural and functional diversity of many natural products. In this study, a novel thermostable uridine diphosphate-dependent glycosyltransferase gene PpGT1 was cloned from Paenibacillus polymyxa NJPI29 and recombinantly expressed in B. subtilis WB600. The purified PpGT1 had a molecular weight of 45 kDa, as estimated using SDS-PAGE. The PpGT1 could catalyze the glycosylation of vanillic acid, methyl vanillate, caffeic acid, cinnamic alcohol, and ferulic acid. Moreover, PpGT1 possessed good thermostability and retained 80% of its original activity even after 12 h of incubation at 45 °C. In addition, PpGT1 remained stable within a neutral to alkaline pH range as well as in the presence of metal ions. The synthesis of methyl vanillate 4-O-β-D-glucoside by purified PpGT1 reached a yield 3.58 mM in a system with pH 8.0, 45 °C, 12 mM UDP-Glc, and 4 mM methyl vanillate. 3D-structure-based amino acid sequence alignments revealed that the catalytic residues and C-terminated PSPG motif were conserved. These unusual properties indicated that PpGT1 is a candidate UGT for valuable natural product industrial applications.

Supplementary information

The online version contains supplementary material available at 10.1007/s13205-021-02855-z.

SUBMITTER: Chang S 

PROVIDER: S-EPMC8178423 | biostudies-literature | 2021 Jul

REPOSITORIES: biostudies-literature