Cloning and functional characterization of lignan glycosyltransferase gene IiUGT349 in Isatis indigotica

• Published in: Zhongguo zhongyao zazhi Vol. 47; no. 24; p. 6587
• Main Authors: Jiang, Yin-Yin, Tan, Yu-Ping, Sun, Shu-Fu, Tang, Jin-Fu
• Format: Journal Article
• Language: Chinese
• Published: China 01.12.2022
• Subjects: Cloning, Molecular; Glucosides - metabolism; Glycosyltransferases - metabolism; Isatis - chemistry; Isatis - genetics; Lignans - metabolism; Phylogeny; lignan glycosyltransferases; gene cloning; subcellular localization; Isatis tinctoria; bioinformatics analysis; protein expression; real-time fluorescence quantitative PCR; functional characterization
• ISSN: 1001-5302
• DOI: 10.19540/j.cnki.cjcmm.20220916.101

Based on the transcriptome data of Isatis indigotica, a total of 110 putative glycosytransferases were identified. Through prokaryotic expression and enzymic activity assay in vitro, a novel lignan glycosyltransferase gene was screened out and named IiUGT349, which catalyzed lariciresinol into lariciresinol-4-O-β-D-glucoside and lariciresinol-4'-O-β-D-glucoside. Bioinformatics analysis suggested that IiUGT349 contained an open reading frame(ORF) of 1 401 bp encoding a protein of 467 amino acids. A protein analysis indicated that IiUGT349 have a predecited molecular weight of 52.77 kDa and pI of 5.96. Phylogenetic analysis showed that IiUGT349 belonging to UGT90 family shared low amino acid sequence identity with the reported lignan glycosyltransferases, which may represent a novel type of lignan glycosyltransferases. Quantitative real-time PCR(qRT-PCR) analysis showed that IiUGT349 was expressed in roots, stems, young leaves and leaves, with the highest expression level in stems. Further biochemical analysis