Molecular cloning and expression of a glycosyltransferase from Bacillus subtilis for modification of morin and related polyphenols
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To characterize glycosyltransferases from Bacillus subtilis ATCC 6633 and investigate their substrate specificity towards plant polyphenols.
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.
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.
KeywordsBacillus subtilis Glycosyltransferase Glycosylation Morin Polyphenols
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).
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