title: A peculiar segmented flow microfluidics for isoquercitrin biosynthesis based on coupling of reaction and separation.
abstract: A segmented flow containing a buffer-ionic liquid/solvent in a micro-channel reactor was applied to synthesize isoquercitrin by the hesperidinase-catalyzed selective hydrolysis of rutin, based on a novel system of reaction coupling with separation. Within the developed microchannel reactor with one T-shaped inlet and outlet, the maximum isoquercitrin yield (101.7 ± 2.6%) was achieved in 20 min at 30 °C and 4 μL/min. Compared with a continuous-flow reactor, reaction rate was increased 4-fold due to a glycine-sodium hydroxide:[Bmim][BF4]/glycerol triacetate (1:1, v/v) system that formed a slug flow in microchannel and significantly increased mass transfer rates. The mass transfer coefficient significantly increased and exhibited a linear relationship with the flow rate. Hesperidinase could be efficiently reused at least 5 times, without losing any activity. The bonding mechanism and secondary structure of hesperidinase indicated that hesperidinase had a greater affinity to rutin at a production rate of 4 μL/min in this segmented flow microreactor.
authors.affiliation: School of Biotechnology & School of the Environment and Chemical Engineering, Jiangsu University of Science and Technology, Zhenjiang 212018, PR China.
authors.forename: An
authors.initials: A
authors.lastname: Gong
authors.affiliation: School of Biotechnology & School of the Environment and Chemical Engineering, Jiangsu University of Science and Technology, Zhenjiang 212018, PR China.
authors.forename: Shuang-Shuang
authors.initials: SS
authors.lastname: Gu
authors.affiliation: School of Biotechnology & School of the Environment and Chemical Engineering, Jiangsu University of Science and Technology, Zhenjiang 212018, PR China; Sericultural Research Institute, Chinese Academy of Agricultural Sciences, Zhenjiang 212018, PR China. Electronic address: wangjun@just.edu.cn.
authors.forename: Jun
authors.initials: J
authors.lastname: Wang
authors.affiliation: School of Biotechnology & School of the Environment and Chemical Engineering, Jiangsu University of Science and Technology, Zhenjiang 212018, PR China; Sericultural Research Institute, Chinese Academy of Agricultural Sciences, Zhenjiang 212018, PR China.
authors.forename: Sheng
authors.initials: S
authors.lastname: Sheng
authors.affiliation: School of Biotechnology & School of the Environment and Chemical Engineering, Jiangsu University of Science and Technology, Zhenjiang 212018, PR China; Sericultural Research Institute, Chinese Academy of Agricultural Sciences, Zhenjiang 212018, PR China.
authors.forename: Fu-An
authors.initials: FA
authors.lastname: Wu
mesh_terms: D055162:Biocatalysis; D019149:Bioreactors; D002942:Circular Dichroism; D006026:Glycoside Hydrolases / Q000737:chemistry / Q000378:metabolism; D007700:Kinetics; D044085:Microfluidics / Q000295:instrumentation / Q000379:methods*; D017433:Protein Structure, Secondary; D011794:Quercetin / Q000031:analogs & derivatives* / Q000096:biosynthesis; D059027:Recycling; D012212:Rheology; D012431:Rutin / Q000378:metabolism; D013696:Temperature; D014215:Triacetin / Q000302:isolation & purification
chemical_list: C016527:isoquercitrin; D012431:Rutin; D011794:Quercetin; D006026:Glycoside Hydrolases; C023307:hesperidinase; D014215:Triacetin
keywords: Biphasic catalysis; Enzymatic synthesis; Isoquercitrin; Micro-channel reactor; Segmented flow