Phenoloxidase activation, antimicrobial, and antibiofilm properties of β-glucan binding protein from Scylla serrata crab hemolymph

Mani Divya, Baskaralingam Vaseeharan, Mahalingam Anjugam, Arokiadhas Iswarya, Sivashanmugam Karthikeyan, Palaniyandi Velusamy, Marimuthu Govindarajan, Naiyf S. Alharbi, Shine Kadaikunnan, Jamal M. Khaled, C. Vágvölgyi

Research output: Contribution to journalArticle

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Abstract

In this study, we purified β-GBP from hemolymph of Scylla serrata crabs using affinity chromatography. The purified S. serrata β-GBP (Ss-β-GBP) had 100 kDa molecular mass in the SDS-PAGE. MALDI-TOF/TOF analysis was conducted, revealing that the purified 100 kDa protein had 96% similarity with β-GBP of Astacus leptodactylus. Ss-β-GBP was characterized using high-performance liquid chromatography (HPLC), X-ray diffraction (XRD) analysis, circular dichroism (CD) and Fourier transform infrared (FTIR) spectroscopy, which confirmed the structure of the Ss-β-GBP. The purified Ss-β-GBP was functionally analyzed by yeast agglutination and phagocytic reaction assays. Moreover, the PO enhancing ability of Ss-β-GBP was evidenced through PO activity. Specifically, the antibacterial activity of the Ss-β-GBP against Gram-positive (Enterococcus faecalis and Staphylococcus aureus) and Gram-negative (Escherichia coli and Pseudomonas aeruginosa) bacteria was evaluated by determining its minimum inhibitory concentration (MIC) < 60 μg/ml for all tested species. Furthermore, the antibiofilm efficacy of Ss-β-GBP at 50 and 100 μg/ml was outlined using light microscopy and confocal laser scanning microscopy (CLSM). Bacterial viability assays also outlined the dose-dependent activity of Ss-β-GBP based on the ratio of live/dead bacterial cells. The results of this study revealed that crab-borne Ss-β-GBP might be widely used to suppress the growth of pathogenic bacteria.

Original languageEnglish
Pages (from-to)864-873
Number of pages10
JournalInternational Journal of Biological Macromolecules
Volume114
DOIs
Publication statusPublished - Jul 15 2018

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Monophenol Monooxygenase
Hemolymph
Assays
Bacteria
Chemical activation
Microbial Viability
Affinity chromatography
Astacoidea
Enterococcus faecalis
Matrix-Assisted Laser Desorption-Ionization Mass Spectrometry
Agglutination
High performance liquid chromatography
Dichroism
Molecular mass
Microbial Sensitivity Tests
Fourier Transform Infrared Spectroscopy
Circular Dichroism
Affinity Chromatography
Confocal Microscopy
X-Ray Diffraction

Keywords

  • Arthropods
  • Crabs
  • Encapsulation
  • Live and dead assay
  • SDS-PAGE
  • β-Glucan binding protein

ASJC Scopus subject areas

  • Structural Biology
  • Biochemistry
  • Molecular Biology

Cite this

Phenoloxidase activation, antimicrobial, and antibiofilm properties of β-glucan binding protein from Scylla serrata crab hemolymph. / Divya, Mani; Vaseeharan, Baskaralingam; Anjugam, Mahalingam; Iswarya, Arokiadhas; Karthikeyan, Sivashanmugam; Velusamy, Palaniyandi; Govindarajan, Marimuthu; Alharbi, Naiyf S.; Kadaikunnan, Shine; Khaled, Jamal M.; Vágvölgyi, C.

In: International Journal of Biological Macromolecules, Vol. 114, 15.07.2018, p. 864-873.

Research output: Contribution to journalArticle

Divya, M, Vaseeharan, B, Anjugam, M, Iswarya, A, Karthikeyan, S, Velusamy, P, Govindarajan, M, Alharbi, NS, Kadaikunnan, S, Khaled, JM & Vágvölgyi, C 2018, 'Phenoloxidase activation, antimicrobial, and antibiofilm properties of β-glucan binding protein from Scylla serrata crab hemolymph', International Journal of Biological Macromolecules, vol. 114, pp. 864-873. https://doi.org/10.1016/j.ijbiomac.2018.03.159
Divya, Mani ; Vaseeharan, Baskaralingam ; Anjugam, Mahalingam ; Iswarya, Arokiadhas ; Karthikeyan, Sivashanmugam ; Velusamy, Palaniyandi ; Govindarajan, Marimuthu ; Alharbi, Naiyf S. ; Kadaikunnan, Shine ; Khaled, Jamal M. ; Vágvölgyi, C. / Phenoloxidase activation, antimicrobial, and antibiofilm properties of β-glucan binding protein from Scylla serrata crab hemolymph. In: International Journal of Biological Macromolecules. 2018 ; Vol. 114. pp. 864-873.
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AU - Karthikeyan, Sivashanmugam

AU - Velusamy, Palaniyandi

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AU - Alharbi, Naiyf S.

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