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α-L-rhamnosidase from Penicillium tardum and Its Application for Biotransformation of Citrus Rhamnosides

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Abstract

Enzymatic deramnosylation of flavonoids is a convenient tool for improving the quality of citrus juices. α-L-rhamnosidase with a specific activity of 33.1 units/mg was isolated and characterized from the culture liquid of Penicillium tardum. The molecular weight of the enzyme was 95 kDa according to the data of gel filtration on Sepharose 6B and gel electrophoresis in SDS-PAGE. The pH optimum of the enzyme activity was 5.0, and the thermo optimum was 60 °C. Enzyme showed high stability in the temperature range of 45–50 and at 60–70 °C. It retained 80 to 50% of the initial activity for 90 min. The half-life of α-L-rhamnosidase at 70 °C increased twofold in the presence of 20–40% glycerol and 2.3-fold in the presence of 4 M sorbitol. The enzyme was completely inhibited in the presence of 10−3 M Ag+ and Cd2+ and approximately by 90% in the presence of Fe2+, Fe3+, and Al3+ ions. More than 60%, the enzyme activity was inhibited by Hg2+, Co2+, and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide methiodide. Activating effect of Ca2+ ions was also noted. Km and Vmax for the hydrolysis of p-nitrophenyl-α-L-rhamnopyranoside and naringin were 0.7 mM and 38.3 µM/min/mg and 1.34 mM and 43.7 µM/min/mg, respectively. Penicillium tardum α-L-rhamnosidase hydrolyzed naringin, neohesperidin, hesperidin, rutin, and narirutin at high rate, which allowed us to consider it as an effective tool for transformation of bioflavonoids in food industry.

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Funding

The work was funded by the National Academy of Science of Ukraine (No. 0113U001217).

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  1. Department of Biochemistry of Microorganisms, Zabolotny Institute of Microbiology and Virology, National Academy of Sciences of Ukraine, 154 Zabolotny st, Kyiv, 03143, Ukraine

    Nataliya Borzova, Olena Gudzenko & Lyudmila Varbanets

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  1. Nataliya Borzova
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Contributions

All authors contributed to the study conception and design. Material preparation, data collection, and analysis were performed by Nataliya Borzova, Olena Gudzenko, and Lyudmila Varbanets. The first draft of the manuscript was written by Nataliya Borzova, and all authors commented on previous versions of the manuscript. All authors read and approved the final manuscript.

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Correspondence to Nataliya Borzova.

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Borzova, N., Gudzenko, O. & Varbanets, L. α-L-rhamnosidase from Penicillium tardum and Its Application for Biotransformation of Citrus Rhamnosides. Appl Biochem Biotechnol 194, 4915–4929 (2022). https://doi.org/10.1007/s12010-022-04008-1

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