An effective biphase system accelerates hesperidinase-catalyzed conversion of rutin to isoquercitrin.
Isoquercitrin is a rare, natural ingredient with several biological activities that is a key precursor for the synthesis of enzymatically modified isoquercitrin (EMIQ). The enzymatic production of isoquercitrin from rutin catalyzed by hesperidinase is feasible; however, the bioprocess is hindered by low substrate concentration and a long reaction time. Thus, a novel biphase system consisting of [Bmim][BF4]:glycine-sodium hydroxide (pH 9) (10:90, v/v) and glyceryl triacetate (1:1, v/v) was initially established for isoquercitrin production. The biotransformation product was identified using liquid chromatography-mass spectrometry, and the bonding mechanism of the enzyme and substrate was inferred using circular dichroism spectra and kinetic parameters. The highest rutin conversion of 99.5% and isoquercitrin yield of 93.9% were obtained after 3 h. The reaction route is environmentally benign and mild, and the biphase system could be reused. The substrate concentration was increased 2.6-fold, the reaction time was reduced to three tenths the original time. The three-dimensional structure of hesperidinase was changed in the biphase system, which α-helix and random content were reduced and β-sheet content was increased. Thus, the developed biphase system can effectively strengthen the hesperidinase-catalyzed synthesis of isoquercitrin with high yield.
1] School of Biotechnology, Jiangsu University of Science and Technology, Zhenjiang. 212018, P R China [2] Sericultural Research Institute, Chinese Academy of Agricultural Sciences, Zhenjiang. 212018, P R China [3] School of the Environment and Safety Engineering, Jiangsu University, Zhenjiang. 212013, P R China.
Jun
J
Wang
1] School of Biotechnology, Jiangsu University of Science and Technology, Zhenjiang. 212018, P R China [2] Sericultural Research Institute, Chinese Academy of Agricultural Sciences, Zhenjiang. 212018, P R China.
An
A
Gong
1] School of Biotechnology, Jiangsu University of Science and Technology, Zhenjiang. 212018, P R China [2] Sericultural Research Institute, Chinese Academy of Agricultural Sciences, Zhenjiang. 212018, P R China.
Cai-Feng
CF
Yang
1] School of Biotechnology, Jiangsu University of Science and Technology, Zhenjiang. 212018, P R China [2] Sericultural Research Institute, Chinese Academy of Agricultural Sciences, Zhenjiang. 212018, P R China.
Qi
Q
Bao
1] School of Biotechnology, Jiangsu University of Science and Technology, Zhenjiang. 212018, P R China [2] Sericultural Research Institute, Chinese Academy of Agricultural Sciences, Zhenjiang. 212018, P R China.
Xin-Yi
XY
Shi
1] School of Biotechnology, Jiangsu University of Science and Technology, Zhenjiang. 212018, P R China [2] Sericultural Research Institute, Chinese Academy of Agricultural Sciences, Zhenjiang. 212018, P R China.
Bei-Bei
BB
Han
School of the Environment and Safety Engineering, Jiangsu University, Zhenjiang. 212013, P R China.
Xiang-Yang
XY
Wu
1] School of Biotechnology, Jiangsu University of Science and Technology, Zhenjiang. 212018, P R China [2] Sericultural Research Institute, Chinese Academy of Agricultural Sciences, Zhenjiang. 212018, P R China.
Fu-An
FA
Wu
D001711:Biotransformation; D002384:Catalysis; D006026:Glycoside Hydrolases / Q000737:chemistry* / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D011794:Quercetin / Q000031:analogs & derivatives* / Q000737:chemistry; D012431:Rutin / Q000737:chemistry*; D013696:Temperature
C016527:isoquercitrin; D012431:Rutin; D011794:Quercetin; D006026:Glycoside Hydrolases; C023307:hesperidinase