Functional characterization of phospholipase B enzyme from Giardia lamblia

Rituparna Sarkar; Sanjib Kumar Sardar; Ajanta Ghosal; Koushik Das; Yumiko Saito-Nakano; Shanta Dutta; Tomoyoshi Nozaki; Sandipan Ganguly

doi:10.1016/j.exppara.2023.108602

Functional characterization of phospholipase B enzyme from Giardia lamblia

Exp Parasitol. 2023 Oct:253:108602. doi: 10.1016/j.exppara.2023.108602. Epub 2023 Aug 22.

Authors

Rituparna Sarkar¹, Sanjib Kumar Sardar¹, Ajanta Ghosal¹, Koushik Das², Yumiko Saito-Nakano³, Shanta Dutta⁴, Tomoyoshi Nozaki⁵, Sandipan Ganguly⁶

Affiliations

¹ Division of Parasitology, ICMR-National Institute of Cholera and Enteric Diseases, Kolkata, India.
² Division of Parasitology, ICMR-National Institute of Cholera and Enteric Diseases, Kolkata, India; Department of Allied Health Sciences, School of Health Sciences and Technology, University of Petroleum and Energy Studies, Dehradun, India.
³ Department of Parasitology, National Institute of Infectious Diseases (NIID), Tokyo, Japan.
⁴ Division of Bacteriology, ICMR-National Institute of Cholera and Enteric Diseases (ICMR-NICED), Kolkata, India.
⁵ Department of Biomedical Chemistry, School of International Health, Graduate School of Medicine, The University of Tokyo, Tokyo, Japan.
⁶ Division of Parasitology, ICMR-National Institute of Cholera and Enteric Diseases, Kolkata, India. Electronic address: sandipanganguly@hotmail.com.

PMID: 37619808
DOI: 10.1016/j.exppara.2023.108602

Abstract

The microaerotolarent amitochondriate protozoan Giardia lamblia causes Giardiasis and produces a unique enzyme called Phospholipase B (PLB) in contrast to higher eukaryotes. The enzyme is produced upon induction with oxidative (H2O2) stress, thus leading to prostaglandin E2 (PGE2) production. It exists in dimeric form, and its molecular weight is 56 kDa. This PLB was extracellularly cloned in the pET21d vector. The ORF is 1620 bp (Genbank accession no. -OM939681) long and codes for a protein 539 amino acid long, with a 15 amino acid long amino-terminal signal peptide. The highest enzyme activity of PLB was identified at pH 7.5 and 35 °C. This specific enzyme was also active at 50 °C pH 10, but activity was low. We also analyzed the expression of PLB protein in G. lamblia, which was significantly induced under increased oxidative stress.

Keywords: Giardia lamblia; Oxidative stress; Phospholipase; Phospholipase B.

MeSH terms

Amino Acids
Giardia lamblia* / genetics
Giardiasis*
Humans
Hydrogen Peroxide
Lysophospholipase

Substances

Lysophospholipase
Hydrogen Peroxide
Amino Acids