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Purification, characterization of an entomopathogenic fungal lectin from Purpureocillium lilacinum and its involvement in pathogenesis leading to mycotic keratitis

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Abstract

A lectin PCL, from Purpureocillium lilacinum a saprophytic, filamentous fungus was purified from the crude extract of the mycelia using 70% ammonium sulphate precipitation followed by affinity chromatography on mucin-Sepharose 4 B column. PCL is a monomer with an apparent molecular mass of 18.5 kDa as revealed by SDS-PAGE under both reducing and non-reducing conditions. PCL is a blood group non-specific lectin and has highest affinity towards chitin, mucin, asialomucin, fetuin with a MIC of 0.15 µg/mL and also recognizes l-fucose, galactose, lactose, n-acetyl galactosamine, hyaluronic acid. PCL is stable up to 60 °C and within the pH range 4–8. To understand its role in pathogenesis, effect of PCL was evaluated on human corneal epithelial cells (HCECs). PCL showed strong glycan mediated binding to HCECs and PCL showed proinflammatory response at lower concentrations by stimulating secretion of IL-6, 8. In contrast PCL at higher concentrations revealed opposite effect of HCECs growth inhibition. All these results collectively support the involvement of PCL in mediating host pathogen interactions possibly leading to pathogenesis. In addition, considering the entomopathogenic effect of Purpureocillium lilacinum, PCL may be attributed for this beneficiary effect, which needs to be explored.

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Funding

SRI thanks the Council of Scientific & Industrial Research (CSIR, No. 27 (0350)/19/EMR-II dated 17th May 2019) and N Jagadeesh thanks the CSIR for fellowship under CSIR-SRF (No. 09/101(0056)/2019/EMR-I).

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Authors and Affiliations

  1. Department of Studies in Biochemistry, Karnatak University, Dharwad, 580003, India

    Narasimhappagari Jagadeesh, Supreeth S. Kulkani, Vishwanath B. Chachadi & Shashikala R. Inamdar

  2. Prof. Brien Holden Eye Research Centre, LV Prasad Eye Institute, Hyderabad, India

    Sanhita Roy

Authors
  1. Narasimhappagari Jagadeesh
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  2. Supreeth S. Kulkani
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  3. Vishwanath B. Chachadi
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  4. Sanhita Roy
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  5. Shashikala R. Inamdar
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Contributions

All authors contributed to the study. NJ: Performed experiments and draft of the manuscript; SSK and NJ: purification of lectin; VBC: procuring the fungal culture and manuscript; SR: HCECs and manuscript; SRI: Study conception, design, data interpretation, final manuscript and laboratory facilities. All authors have read and approved the final manuscript.

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Correspondence to Shashikala R. Inamdar.

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Communicated by Erko Stackebrandt.

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Jagadeesh, N., Kulkani, S.S., Chachadi, V.B. et al. Purification, characterization of an entomopathogenic fungal lectin from Purpureocillium lilacinum and its involvement in pathogenesis leading to mycotic keratitis. Arch Microbiol 204, 673 (2022). https://doi.org/10.1007/s00203-022-03243-8

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  • DOI: https://doi.org/10.1007/s00203-022-03243-8

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