Abstract
The plant kingdom provides a large resource of natural products and various related enzymes are analyzed. The high catalytic activity and easy genetically modification of microbial enzymes would be beneficial for synthesis of natural products. But the identification of functional genes of target enzymes is time consuming and hampered by many contingencies. The potential to mine microbe-derived glycosyltransferases (GTs) cross the plant kingdom was assessed based on alignment and evolution of the full sequences and key motifs of target enzymes, such as Rhodiola-derived UDP-glycosyltransferase (UGT73B6) using in salidroside synthesis. The GTs from Bacillus licheniformis ZSP01 with high PSPG motif similarity were speculated to catalyze the synthesis of salidroside. The UGTBL1, which had similarity (61.4%) PSPG motif to UGT73B6, displayed efficient activity and similar regioselectivity. Highly efficient glycosylation of tyrosol (1 g/L) was obtained by using engineered E. coli harboring UGTBL1 gene, which generated 1.04 g/L salidroside and 0.99 g/L icariside D2. All glycosides were secreted into the culture medium and beneficial for downstream purification. It was the first report on the genome mining of UGTs from microorganisms cross the plant kingdom. The mining approach may have broader applications in the selection of efficient candidate for making high-value natural products.
Highlights
Natural products are a major source of current clinical drugs and are a substantial resource for the discovery of new drugs[1]
The available microorganism genome databases are increasing, previous gene identification methods of target proteins were mainly focused on biochemical studies and characterizations, and genome mining were largely based on the alignment of enzymes from similar genera and species
We propose that the mining of novel microbial biocatalysts for the production of natural products should be addressed through phylogenetic analysis of the complete sequences and key amino acid sequence motifs of enzymes generating secondary metabolites cross the plant kingdom
Summary
Natural products are a major source of current clinical drugs and are a substantial resource for the discovery of new drugs[1]. We propose that the mining of novel microbial biocatalysts for the production of natural products should be addressed through phylogenetic analysis of the complete sequences and key amino acid sequence motifs of enzymes generating secondary metabolites cross the plant kingdom. On the basis of alignment and analysis of complete sequences or conserved signature motifs, Bacillus licheniformis derived GTs with high PSPG motif similarity to UGT73B6 were mined according to the phylogenetic analysis.
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