Abstract The neutral sphingomyelinase (nSMase) 1 homologue gene LsSMase was cloned from Laodelphax striatellus , a direct sap‐sucker and virus vector of gramineous plants, and expressed via a Bac to Bac baculovirus expression system. The LsSMase‐enhanced green fluorescent protein fusion protein was located in the endoplasmic reticulum in a similar manner to mammalian nSMase 1. The biochemical properties of LsSMase were determined in detail. The optimal pH and temperature for recombinant LsSMase were 8 and 37 °C, respectively. LsSMase was an Mg 2+ or Mn 2+ dependent enzyme, but different concentration of each were needed. The activity of LsSMase was significantly stimulated by Ethylene glycol bis(2‐aminoethyl ether)tetraacetic acid (EGTA), whereas it was inhibited by ethylenediaminetetraacetic acid. Millimolar concentrations of Zn 2+ completely inhibited LsSMase. The reducing agents dithiothreitol and β‐mercaptoethanol varied in their effects on activity. Phospholipids were not found to stimulate LsSMase.