Isolation and characterization of a lectin-like chitinase from the hepatopancreas of freshwater prawn, Macrobrachium rosenbergii

Sonalina Sahoo; Mohan R Badhe; Anirban Paul; Pramoda Kumar Sahoo; Amol R Suryawanshi; Debabrata Panda; Bindu R Pillai; Snigdha Baliarsingh; Bharat Bhusan Patnaik; Jyotirmaya Mohanty

doi:10.1016/j.biochi.2023.09.025

Isolation and characterization of a lectin-like chitinase from the hepatopancreas of freshwater prawn, Macrobrachium rosenbergii

Biochimie. 2023 Sep 26:S0300-9084(23)00252-3. doi: 10.1016/j.biochi.2023.09.025. Online ahead of print.

Affiliations

¹ ICAR-Central Institute of Freshwater Aquaculture, Bhubaneswar, 751002, India.
² Institute of Life Sciences, Bhubaneswar, 751023, India.
³ P.G. Department of Biosciences and Biotechnology, Fakir Mohan University, Vyasa Vihar, Nuapadhi, Balasore, 756089, India.
⁴ P.G. Department of Biosciences and Biotechnology, Fakir Mohan University, Vyasa Vihar, Nuapadhi, Balasore, 756089, India; Department of Biology, College of Natural Sciences, Soonchunhyang University, Asan, Chungnam, 31538, South Korea; Korea Native Animal Resources Utilization Convergence Research Institute (KNAR), Soonchunhyang University, Asan, Chungnam, 31538, South Korea.
⁵ ICAR-Central Institute of Freshwater Aquaculture, Bhubaneswar, 751002, India. Electronic address: jmohantycifa@gmail.com.

PMID: 37769935
DOI: 10.1016/j.biochi.2023.09.025

Abstract

A lectin was isolated from the hepatopancreas of freshwater prawn, Macrobrachium rosenbergii by affinity chromatography using mucin-sepharose matrix. The purity of the isolated lectin was confirmed in native gradient PAGE that showed a single protein band of ∼37.9 kDa. In SDS-PAGE also one band of ∼43.3 kDa molecular weight was observed that indicated the protein to be a monomer. The band from the SDS-PAGE gel was identified through mass spectrometry as chitinase 1. The purified chitinase (50 μg/ml) hemagglutinated rabbit RBCs and, mucin and glucose inhibited hemagglutination with minimum concentrations of 0.1 mg/ml and 100 mM, respectively. Bacterial agglutination with Gram -ve Vibrio harveyi, Aeromonas sobria and Escherichia coli was also observed by this protein. Thus, chitinase 1 showed lectin-like properties besides its chitin hydrolytic activity. In western blot with hepatopancreas sample, rabbit antiserum against chitinase 1 cross-reacted to two additional proteins namely, chitinase 1C and obstructor E (a chitin-binding protein, CBP), besides its specific reactivity. An indirect ELISA was developed with the antiserum to quantify chitinases/CBP in hepatopancreas and serum samples of M. rosenbergii. The assay was used in samples from juvenile prawns following V. harveyi challenge. At 72 h post-challenge, significantly higher levels of chitinases/CBP were quantified in the hepatopancreas of the challenged group (1.8 ± 0.2 mg/g tissue) compared to the control (1.2 ± 0.1 mg/g tissue). This study suggests that the chitinase 1 protein with lectin-like properties is possibly induced at the protein level and can be putatively involved in the innate immune response of M. rosenbergii.

Keywords: Chitinase; ELISA; Hepatopancreas; Lectin; Macrobrachium rosenbergii.