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KMID : 1094720210260060956
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Biotechnology and Bioprocess Engineering
2021 Volume.26 No. 6 p.956 ~ p.967
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Production of the Minor Ginsenoside F2 from the PPD-mix-type Major Ginsenosides Using a Novel Recombinant Glycoside Hydrolase from Novosphingobium aromaticivorans
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Siddiqi Muhammad Zubair
Park Hye-Yoon
Kim Ga-Ryun
Cui Chang-Hao
Jo Young-Jun
Kim Sun-Chang
Im Wan-Taek
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Abstract
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Background: The minor ginsenosides such as F2, C-K, Rh2, and Rg3 make up less than 1% of the total ginseng extract; however, many studies have shown that ginsenoside F2 has anti-cancer and antioxidant effects and improves dementia and atopic dermatitis. Therefore, the enhanced production of this minor ginsenoside is a promising approach for the pharmaceutical industry. In this study, we found and cloned a novel glycoside hydrolase gene for the gram-scale production of ginsenoside F2.
Methods and Results: Novosphingobium aromaticivorans, which was isolated from deep-terrestrial-subsurface sediments, has shown ginsenoside-converting ability. From this bacterium, a novel glycoside hydrolase, named BglNar, was found that can efficiently biotransform the protopanaxatriol (PPD-mix-type) major ginsenosides (Rb1, Rb2 Rc, and Rd) into the minor ginsenoside F2. This enzyme was cloned, expressed in Escherichia coli BL21, and characterized. The BglNar comprises 439 amino acid and belongs to the glycoside hydrolase family 1. The Km value of p-nitrophenyl-β-D-glucopyranoside was 9.06 ± 0.28 and the Vmax value was 24.0 ± 0.34 μmol/min/mg of protein. For gram-scale production of minor ginsenoside F2, crude PPD-mix-type ginsenosides (2 g/400 mL) were treated with BglNar and 1.14 g of F2 with final purity of 82.5 ± 1.3%, was obtained after purification using a column packed with HP20 resin.
Conclusion: Our preliminary data demonstrate that the gram production of ginsenoside F2 using a recombinant enzyme will enhance the health benefits of Panax ginseng. This is the first study describing the gram-scale production of F2 from PPD-mix using a single novel ginsenoside-transforming β-glucosidase of the GH family 1. Significance and Impact of Study: In most of the previous studies, researchers have been used the combination of enzymes for production of F2, while in this study we found the possibility of gram-scale production of F2 from PPD-mix-type major ginsenosides using a single recombinant enzyme, thus simplifying the production and reducing the cost.
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KEYWORD
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biotransformation, β-glucosidase, recombinant enzyme, ginsenoside F2, Novosphingobium aromaticivorans
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