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<p>Shiitake mushroom (<i>Lentinula edodes</i>), widely recognized for its medicinal properties, the essential nutraceutical compound in this mushroom is a type of β-glucan called lentinan (LNT), and its triple helical conformation is crucial for its immunomodulatory and other pharmacological activities. In the present study, an attempt was carried out to screen biologically active form of LNT in 4 different metabotypes of <i>L. edodes,</i> along with optimizing efficient extraction for scale-up<i>.</i> The samples were collected from ICAR-NEH, Manipur, India and there genetic identification revealed 99.8–100 % similarity to known reference sequence and deposited in Genbank under the assigned accession numbers, LE-5 (PV460746.1), LE-12 (PX070480), LE-25 (PX070479), and LE-27 (PV366868.1). For screening LNT two extraction methods were used, alkali assisted (AAE) and sub-critical water extraction (SWE). The former one (AAE) revealed a bathochromic shift near 510&#xa0;nm upon reaction with congo red dye, suggesting it preserved the triple helical from and no such shift was observed in the SWE extracts. The molecular basis of this disruption in SWE was investigated through molecular dynamics (MD) simulation to understand the LNT-chitin interactions in aqueous medium, as chitin is naturally found in shiitake cell wall. The results demonstrated that chitin and LNT maintained structural stability (Root Mean Square Deviations (RMSD): 0.8&#xa0;Å for chitin; 1.3&#xa0;Å for LNT) and formed ~ 3 stable intermolecular hydrogen bonds, while both showed extensive hydrogen bonding with water (108 for chitin; 136 for LNT) which explains the resistance of the LNT–chitin scaffold to aqueous disruption and the inability of SWE to preserve the bioactive triple helix. This integrated experimental computational approach not only deepens mechanistic understanding of polysaccharide stability but also reflects an emerging trend in Indian scientific research, combining indigenous bio-resources, green technologies, and in silico modeling to create scalable, sustainable pathways for nutraceutical recovery.</p>

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Conformation-driven extraction strategies for lentinan: integrating subcritical water techniques and molecular simulations for high-value functional food development towards a sustainable bioeconomy

  • 期刊论文

  • 发表日期：

    2025年11月17日

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  • C. S. Keerthana,
  • Tripti Kundu,
  • Muskan Beura,
  • Suvarghya Chakraborty,
  • Vipin Verma,
  • Deeba Kamil,
  • Madhulika Gupta,
  • Veda Krishnan

摘要

Shiitake mushroom (Lentinula edodes), widely recognized for its medicinal properties, the essential nutraceutical compound in this mushroom is a type of β-glucan called lentinan (LNT), and its triple helical conformation is crucial for its immunomodulatory and other pharmacological activities. In the present study, an attempt was carried out to screen biologically active form of LNT in 4 different metabotypes of L. edodes, along with optimizing efficient extraction for scale-up. The samples were collected from ICAR-NEH, Manipur, India and there genetic identification revealed 99.8–100 % similarity to known reference sequence and deposited in Genbank under the assigned accession numbers, LE-5 (PV460746.1), LE-12 (PX070480), LE-25 (PX070479), and LE-27 (PV366868.1). For screening LNT two extraction methods were used, alkali assisted (AAE) and sub-critical water extraction (SWE). The former one (AAE) revealed a bathochromic shift near 510 nm upon reaction with congo red dye, suggesting it preserved the triple helical from and no such shift was observed in the SWE extracts. The molecular basis of this disruption in SWE was investigated through molecular dynamics (MD) simulation to understand the LNT-chitin interactions in aqueous medium, as chitin is naturally found in shiitake cell wall. The results demonstrated that chitin and LNT maintained structural stability (Root Mean Square Deviations (RMSD): 0.8 Å for chitin; 1.3 Å for LNT) and formed ~ 3 stable intermolecular hydrogen bonds, while both showed extensive hydrogen bonding with water (108 for chitin; 136 for LNT) which explains the resistance of the LNT–chitin scaffold to aqueous disruption and the inability of SWE to preserve the bioactive triple helix. This integrated experimental computational approach not only deepens mechanistic understanding of polysaccharide stability but also reflects an emerging trend in Indian scientific research, combining indigenous bio-resources, green technologies, and in silico modeling to create scalable, sustainable pathways for nutraceutical recovery.