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Ultrafast synthesis of isoquercitrin by enzymatic hydrolysis of rutin in a continuous-flow microreactor

Wang Jun (School of the Environment and Safety Engineering, Jiangsu University, Zhenjiang, P. R. China + School of Biotechnology, Jiangsu University of Science and Technology, Zhenjiang, P.R. China)
An Gong (Jiangsu University of Science and Technology, School of Biotechnology, Zhenjiang, P.R. China)
Gu Shuangshuang (Jiangsu University of Science and Technology, School of Biotechnology, Zhenjiang, P.R. China)
Cui Hongsheng (Jiangsu University of Science and Technology, School of Biotechnology, Zhenjiang, P.R. China)
Wu Xiangyang (Jiangsu University, School of the Environment and Safety Engineering, Zhenjiang, P.R. China)

Isoquercitrin is a rare flavonol glycoside with a wide range of biological activities and is a key synthetic intermediate for the production of enzymatically modified isoquercitrin. In order to establish an ultrafast bioprocess for obtaining isoquercitrin, a novel continuous flow biosynthesis of isoquercitrin using the hesperidinase-catalyzed hydrolysis of rutin in a glass-polydimethylsiloxane (PDMS) microreactor was first carried out. Using the developed microchannel reactor (200μm width, 50μm depth, and 2 m length) with one T-shaped inlet and one outlet, the maximum yield of isoquercitrin (98.6%) was achieved in a short time (40 min) under the following optimum conditions: rutin concentration at 1 g L-1, hesperidinase concentration at 0.1 g mL-1, reaction temperature at 40°C, and a flow rate at 2 μL min-1. The activation energy value Ea of the enzymatic reaction was 4.61 kJ mol-1, and the reaction rate and volumetric productivity were approximately 16.1-fold and 30% higher, respectively, than those in the batch reactor. Thus, the use of a continuous-flow microreactor for the enzymatic hydrolysis of rutin is an efficient and simple approach to achieve a relative high yield of isoquercitrin.

Keywords: biocatalysis, continuous flow, hesperidinase, isoquercitrin, microreactor