Expression of Codon-Optimized Plant Glycosyltransferase UGT72B14 in Escherichia coli Enhances Salidroside Production
Result of the recombinant protein yield and salidroside production in vitro. (a) SDS-PAGE analysis of the recombinant protein expression and purification. Lane M: protein molecular weight markers. Lane 1: total protein in uninduced strain. Lane 2: total protein in induced strain. Lane 3: concentrated protein after being purified by Ni affinity chromatography and PD-10 column. Sample of 15 μL was used for SDS-PAGE analysis. Target protein is indicated by arrows. (b) Result of salidroside production catalysed by UGT72B14 in vitro. Protein yield data of the optimized UGT72B14 was obtained by purified protein after induction of 7 h initiated with OD600 = while that of the wild-type UGT72B14 was obtained by purified protein after induction of 9 h initiated with OD600 = . The recombinant enzyme catalysis reaction system (100 μL) contained 50 mM Tris-HCl (pH 7.5), 2 mM UDP-glucose, 250 μM tyrosol, and the enzyme protein of 0.2 mg, proceeded for 30 min at 30°C, and terminated with 200 μL MeOH. Data were presented as mean ± SD.
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