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Purification and Characterization of a Novel α-L-Rhamnosidase from Papiliotrema laurentii ZJU-L07 and Its Application in Production of Icariin from Epimedin C.
- Source :
- Journal of Fungi; Jun2022, Vol. 8 Issue 6, p644, 17p
- Publication Year :
- 2022
-
Abstract
- Icariin is the most effective bioactive compound in Herba Epimedii. To enhance the content of icariin in the epimedium water extract, a novel strain, Papiliotrema laurentii ZJU-L07, producing an intracellular α-L-rhamnosidase was isolated from the soil and mutagenized. The specific activity of α-L-rhamnosidase was 29.89 U·mg<superscript>−1</superscript> through purification, and the molecular mass of the enzyme was 100 kDa, as assayed by SDS-PAGE. The characterization of the purified enzyme was determined. The optimal temperature and pH were 55 °C and 7.0, respectively. The enzyme was stable in the pH range 5.5–9.0 for 2 h over 80% and the temperature range 30–40 °C for 2 h more than 70%. The enzyme activity was inhibited by Ca<superscript>2+</superscript>, Fe<superscript>2+</superscript>, Cu<superscript>2+</superscript>, and Mg<superscript>2+</superscript>, especially Fe<superscript>2+</superscript>. The kinetic parameters of K<subscript>m</subscript> and V<subscript>max</subscript> were 1.38 mM and 24.64 μmol·mg<superscript>−1</superscript>·min<superscript>−1</superscript> using pNPR as the substrate, respectively. When epimedin C was used as a nature substrate to determine the kinetic parameters of α-L-rhamnosidase, the values of K<subscript>m</subscript> and V<subscript>max</subscript> were 3.28 mM and 0.01 μmol·mg<superscript>−1</superscript>·min<superscript>−1</superscript>, respectively. The conditions of enzymatic hydrolysis were optimized through single factor experiments and response surface methodology. The icariin yield increased from 61% to over 83% after optimization. The enzymatic hydrolysis method could be used for the industrialized production of icariin. At the same time, this enzyme could also cleave the α-1,2 glycosidic linkage between glucoside and rhamnoside in naringin and neohesperidin, which could be applicable in other biotechnological processes. [ABSTRACT FROM AUTHOR]
- Subjects :
- RESPONSE surfaces (Statistics)
MOLECULAR weights
NARINGIN
NEOLIGNANS
Subjects
Details
- Language :
- English
- ISSN :
- 2309608X
- Volume :
- 8
- Issue :
- 6
- Database :
- Complementary Index
- Journal :
- Journal of Fungi
- Publication Type :
- Academic Journal
- Accession number :
- 157764896
- Full Text :
- https://doi.org/10.3390/jof8060644