Structural Study of Thermostable Ginsenoside β-Glucosidase from <i>Caldicellulosiruptor bescii</i>

Protopanaxadiol-type ginsenosides, the major bioactive components of <i>Panax ginseng</i>, exhibit diverse pharmacological activities, but suffer from low oral bioavailability due to poor water solubility and membrane permeability. Enzymatic deglycosylation has emerged as an effective st...

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Main Author: Jung-Min Choi
Format: Article
Language:English
Published: MDPI AG 2025-05-01
Series:Crystals
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Online Access:https://www.mdpi.com/2073-4352/15/5/447
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author Jung-Min Choi
author_facet Jung-Min Choi
author_sort Jung-Min Choi
collection DOAJ
description Protopanaxadiol-type ginsenosides, the major bioactive components of <i>Panax ginseng</i>, exhibit diverse pharmacological activities, but suffer from low oral bioavailability due to poor water solubility and membrane permeability. Enzymatic deglycosylation has emerged as an effective strategy to enhance their therapeutic potential; however, most glucosidases lack sufficient thermostability for industrial applications. A β-glucosidase from the thermophilic bacterium <i>Caldicellulosiruptor bescii</i> (CbBGL) has demonstrated efficient conversion of major ginsenosides into compound K at elevated temperatures. In this study, the high-resolution crystal structure of CbBGL was determined at 1.9 Å. Structural analysis revealed that CbBGL adopts a classical (α/β)<sub>8</sub> TIM barrel fold and functions as a homodimer. Comparative studies with other glucosidases highlighted structural features contributing to its thermostability, including moderate B-factor distribution and a limited hydrogen bond network. Docking analyses revealed a narrow, inverted conical substrate-binding cleft, which imposes specific binding orientations and underlies the enzyme’s stepwise deglycosylation mechanism. These insights provide a structural basis for CbBGL’s thermal resilience and substrate specificity, offering a valuable platform for the rational engineering of glucosidases in ginsenoside bioconversion processes.
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spelling doaj-art-3741eb81d30d42ab8e8f6671a329861e2025-08-20T02:33:38ZengMDPI AGCrystals2073-43522025-05-0115544710.3390/cryst15050447Structural Study of Thermostable Ginsenoside β-Glucosidase from <i>Caldicellulosiruptor bescii</i>Jung-Min Choi0Department of Food Science and Biotechnology, Kyungsung University, Busan 48434, Republic of KoreaProtopanaxadiol-type ginsenosides, the major bioactive components of <i>Panax ginseng</i>, exhibit diverse pharmacological activities, but suffer from low oral bioavailability due to poor water solubility and membrane permeability. Enzymatic deglycosylation has emerged as an effective strategy to enhance their therapeutic potential; however, most glucosidases lack sufficient thermostability for industrial applications. A β-glucosidase from the thermophilic bacterium <i>Caldicellulosiruptor bescii</i> (CbBGL) has demonstrated efficient conversion of major ginsenosides into compound K at elevated temperatures. In this study, the high-resolution crystal structure of CbBGL was determined at 1.9 Å. Structural analysis revealed that CbBGL adopts a classical (α/β)<sub>8</sub> TIM barrel fold and functions as a homodimer. Comparative studies with other glucosidases highlighted structural features contributing to its thermostability, including moderate B-factor distribution and a limited hydrogen bond network. Docking analyses revealed a narrow, inverted conical substrate-binding cleft, which imposes specific binding orientations and underlies the enzyme’s stepwise deglycosylation mechanism. These insights provide a structural basis for CbBGL’s thermal resilience and substrate specificity, offering a valuable platform for the rational engineering of glucosidases in ginsenoside bioconversion processes.https://www.mdpi.com/2073-4352/15/5/447ginsenoside β-glucosidasethermostabilitycrystal structureenzymatic deglycosylation
spellingShingle Jung-Min Choi
Structural Study of Thermostable Ginsenoside β-Glucosidase from <i>Caldicellulosiruptor bescii</i>
Crystals
ginsenoside β-glucosidase
thermostability
crystal structure
enzymatic deglycosylation
title Structural Study of Thermostable Ginsenoside β-Glucosidase from <i>Caldicellulosiruptor bescii</i>
title_full Structural Study of Thermostable Ginsenoside β-Glucosidase from <i>Caldicellulosiruptor bescii</i>
title_fullStr Structural Study of Thermostable Ginsenoside β-Glucosidase from <i>Caldicellulosiruptor bescii</i>
title_full_unstemmed Structural Study of Thermostable Ginsenoside β-Glucosidase from <i>Caldicellulosiruptor bescii</i>
title_short Structural Study of Thermostable Ginsenoside β-Glucosidase from <i>Caldicellulosiruptor bescii</i>
title_sort structural study of thermostable ginsenoside β glucosidase from i caldicellulosiruptor bescii i
topic ginsenoside β-glucosidase
thermostability
crystal structure
enzymatic deglycosylation
url https://www.mdpi.com/2073-4352/15/5/447
work_keys_str_mv AT jungminchoi structuralstudyofthermostableginsenosidebglucosidasefromicaldicellulosiruptorbesciii