Developing of improved methods for lactate detection has been increased due to its importance for clinical analysis, sports medicine, and the food industry [1, 2]. In this communication, we fabricated for the first time a highly sensitive and selective lactate biosensor based on RF sputtered NiO nanostructure FET. Due to excellent electron transfer capabilities besides high adsorption ability and electrocatalytic activity of the NiO nanostructure, it has been applied for LOH enzyme immobilization. Key feature to our biosensor design is easy and low-cost production of biosensing element achieved by developing metal oxide p-type semiconducting NiO thin film using reproducible RF sputtering technique, which results in high sensitivity, linear responses to lactate concentration and better selectivity. The fabricated FET devices exhibited p-channel behavior, with good electrical conductivity, and Ohmic contacts were formed with the source and drain electrodes. The FET based biosensor devices exhibited a novel strategy for lactate at attomolar level sensitivity and a linear dynamic range laM-lpM. High reproducibility and repeatability, with insignificant interference are major observed advantageous in this study. The sensor also has performances such as low cost, small size, and ease of operation, which is promising for making portable and disposable devices for routinely monitoring personal lactic acid levels. This simple yet versatile approach may open up a new route to the fabrication of various types of biosensor platforms based on metal oxide nanomaterials.