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Biotechnology Letters

, Volume 39, Issue 8, pp 1229–1235 | Cite as

Molecular cloning and expression of a glycosyltransferase from Bacillus subtilis for modification of morin and related polyphenols

  • Qianqian Wang
  • Yixiang Xu
  • Jiaqi Xu
  • Xudong Wang
  • Chen Shen
  • Yan Zhang
  • Xiufeng Liu
  • Boyang Yu
  • Jian ZhangEmail author
Original Research Paper

Abstract

Objectives

To characterize glycosyltransferases from Bacillus subtilis ATCC 6633 and investigate their substrate specificity towards plant polyphenols.

Results

Among the cloned and expressed six UDP-glycosyltransferases (BsGT1-6), BsGT-1 showed activity with a wide range of polyphenols: morin, quercetin, alizarin, rehin, curcumin and aloe emodin. The gene of BsGT-1 has an ORF of 1206 bp encoding 402 amino acids. The recombinant enzyme was purified to homogeneity by Ni–NTA affinity chromatograph, and its biochemical characteristics were identified by HPLC–UV/MS, 1H-NMR and 13C-NMR. BsGT-1 has an MW of approx. 46 kDa as indicated by SDS-PAGE; its activity was optimal at 40 °C and pH 8.5. The Km value of BsGT-1 towards morin was 110 μM.

Conclusions

BsGT-1 from B. subtilis was cloned. It had high catalytic capabilities towards polyphenols which would make it feasible for the structural modification of polyphenols.

Keywords

Bacillus subtilis Glycosyltransferase Glycosylation Morin Polyphenols 

Notes

Supporting information

Section 1—The structure elucidation of compounds 1a and 2a.

Section 2—The structure elucidation of compounds 3a, 4a and 6a.

Section 3—The structure elucidation of compound 5a.

Supplementary Table 1—The genebank accession numbers and the primers of BsGT1-6.

Supplementary Fig. 1—HPLC chromatogram of BsGT-1 reaction products of flavonols: (A) compound 1 with BsGT-1. (B) compound 2 with BsGT-1.

Supplementary Fig. 2—HPLC chromatogram of BsGT-1 reaction product of anthraquinone: (A) compound 3 with BsGT-1. (B) compound 4 with BsGT-1. (C) compound 6 with BsGT-1.

Supplementary Fig. 3—HPLC Chromatogram of reaction product of compound 5 with BsGT-1.

Supplementary Fig. 4—Enzymatic activity of BsGT-1 depending in pH (A) and temperature (B).

Supplementary material

10529_2017_2352_MOESM1_ESM.doc (1.2 mb)
Supplementary material 1 (DOC 1232 kb)

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Copyright information

© Springer Science+Business Media Dordrecht 2017

Authors and Affiliations

  1. 1.State Key Laboratory of Natural MedicinesChina Pharmaceutical UniversityNanjingChina
  2. 2.Jiangsu Key Laboratory of TCM Evaluation and Translational ResearchChina Pharmaceutical UniversityNanjingChina
  3. 3.Institute of Biotechnology for TCM Research, School of Traditional Chinese MedicineChina Pharmaceutical UniversityNanjingChina

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