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Cloning and characterization of α-l-rhamnosidase from Chloroflexus aurantiacus and its application in the production of isoquercitrin from rutin

  • Kyung-Chul Shin
  • Min-Ju Seo
  • Deok-Kun Oh
  • Mi-Na Choi
  • Dae-Wook Kim
  • Yeong-Su KimEmail author
  • Chang-Su Park
Original Research Paper
  • 40 Downloads

Abstract

Objective

This study was conducted to characterize recombinant α-l-rhamnosidase from Chloroflexus aurantiacus and apply the enzyme in the production of isoquercitrin from rutin.

Results

The α-l-rhamnosidase from C. aurantiacus was cloned and expressed in Escherichia coli BL21 and purified as a soluble enzyme. α-l-rhamnosidase purified from C. aurantiacus has a molecular mass of approximately 105 kDa and is predicted to exist as a homodimer with a native enzyme of 200 kDa. The purified enzyme exhibited the highest specific activity for rutin among the reported isoquercitrin producing α-l-rhamnosidases and was applied in the production of isoquercitrin from rutin. Under the optimised conditions of pH 6.0, 50 °C, 0.6 U mL−1 α-l-rhamnosidase, and 30 mM rutin, α-l-rhamnosidase from C. aurantiacus produced 30 mM isoquercitrin after 2 h with a 100% conversion yield and productivity of 15 mM h−1.

Conclusions

We achieved a high productivity of isoquercitrin from rutin. Moreover, these results suggest that α-l-rhamnosidase from C. aurantiacus is an effective isoquercitrin producer.

Keywords

Chloroflexus aurantiacus Rutin Isoquercitrin α-l-Rhamnosidase Enzymatic production 

Notes

Acknowledgements

This research was supported by Basic Science Research Program through the National Research Foundation of Korea (NRF) funded by the Ministry of Education (Grant Number 2015R1D1A1A01059570).

Supporting information

Supplementary Figure 1—Structure of isoquercitrin.

Supplementary Figure 2—1H-NMR spectrum of isoquercitrin using NMR spectroscopy (800 MHz NMR).

Supplementary Figure 3—13C-NMR spectrum of isoquercitrin using NMR spectroscopy (800 MHz NMR).

Supplementary material

10529_2019_2648_MOESM1_ESM.docx (336 kb)
Supplementary material 1 (DOCX 336 kb)

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Copyright information

© Springer Nature B.V. 2019

Authors and Affiliations

  1. 1.Department of Bioscience and BiotechnologyKonkuk UniversitySeoulRepublic of Korea
  2. 2.Department of Food Science and TechnologyCatholic University of DaeguHayangSouth Korea
  3. 3.Plant Resource Industry DivisionBaekdudaegan National ArboretumBonghwagunRepublic of Korea

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