Molecular cloning, inducible expression and antibacterial analysis of a novel i-type lysozyme (lyz-i2) in Pacific white shrimp, Litopenaeus vannamei.

Abstract

The full-length cDNA coding for a novel invertebrate (i-type) lysozyme was identified in Pacific white shrimp (Litopenaeus vannamei). The newly obtained L.vannamei lysozyme is similar to the Penaeus monodon i-type lysozyme 2, but it is distant from the known L.vannamei c-type lysozyme and i-type lysozyme 1 in protein sequence; therefore, it was defined as L.vannamei i-type lysozyme 2 (lyz-i2). Expression of L.vannamei lyz-i2 transcripts were ubiquitously detected in all tissues we selected, with the highest abundance observed in the hemolymph. Challenge with Vibrio harveyi might elicit L.vannamei lyz-i2 mRNA expression in the hepatopancreas, intestine, muscle, gill and hemolymph. In the themolymph, specifically, the stimulatory effects of Vibrio and lipopolysaccharide (LPS) on lyz-i2 transcript levels were durable and transient, respectively; while Polyinosinic:polycytidylic acid [Poly (I:C)] treatment did not affect lyz-i2 expression. L.vannamei lyz-i2 recombinant protein was generated in an Escherichia coli system. By lysoplate and turbidimetric assays, the L.vannamei lyz-i2 recombinant protein showed a broad spectrum of antimicrobial properties with high activities against Micrococcaceae lysodeikticus and various Vibrio species and relatively low activity against E.coli. In conclusion, L.vannamei lyz-i2 might be a potent antibacterial protein with a role in innate immunity in Penaeid shrimp.