Cloning and characterization of α-l-rhamnosidase from Chloroflexus aurantiacus and its application in the production of isoquercitrin from rutin
- 40 Downloads
This study was conducted to characterize recombinant α-l-rhamnosidase from Chloroflexus aurantiacus and apply the enzyme in the production of isoquercitrin from rutin.
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.
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.
KeywordsChloroflexus aurantiacus Rutin Isoquercitrin α-l-Rhamnosidase Enzymatic production
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).
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).
- Makino T, Kanemaru M, Okuyama S, Shimizu R, Tanaka H, Mizukami H (2013) Anti-allergic effects of enzymatically modified isoquercitrin (α-oligoglucosyl quercetin 3-O-glucoside), quercetin 3-O-glucoside, α-oligoglucosyl rutin, and quercetin, when administered orally to mice. J Nat Med 67:881–886. https://doi.org/10.1007/s11418-013-0760-5 CrossRefGoogle Scholar
- Vila-Real H, Alfaia AJ, Bronze MR, Calado AR, Ribeiro MH (2011) Enzymatic synthesis of the flavone glucosides, prunin and isoquercetin, and the aglycones, naringenin and quercetin, with selective α-l-rhamnosidase and β-D-glucosidase activities of naringinase. Enzyme Res 2011:692618. https://doi.org/10.4061/2011/692618 CrossRefPubMedCentralGoogle Scholar