Highly sensitive potassium (K)-doped carbon nanotube (CNT) and polypyrrole (PPy) nanocomposite membrane-based enzyme field effect transistor (ENFET) has been fabricated on indium tin oxide (ITO) for detection of cholesterol. P-type graphene has been deposited as substrate on ITO glass electrochemically. N-type graphene has been deposited in source and drain regions. Zirconium dioxide (ZrO2) has been deposited on the channel region as gate insulator. K/PPy/CNT composite has been deposited as sensing membrane on the top of ZrO2 layer; 1 mu l of cholesterol oxidase (ChOx) has been immobilized on K/PPy/CNT membrane via physical adsorption technique. The response of K/PPy/CNT/FET has been studied using Agilent 3458A digital multimeter in presence of phosphate buffer saline (PBS) of 50 mM, pH 7.0 and 0.9 % NaCl contained in a glass pot. During measurement, 20 mu l cholesterol solutions (0.5 to 25 mM) were poured into the pot through micropipette each time. It has been found that K/PPy/CNT/FET has linearly varied from 0.5 to 20 mM. The sensitivity of this FET has been found to be similar to 400 mu A/mM/mm(2) with regression coefficient (r) similar to 0.998. The proposed ENFET has response time of 1 s and stability up to 6 months. The experiment has been repeated 10 times, and only 2.0 % output variation has been observed. The limit of detection (LoD) and Michaelis-Menten constant (K (m)) were found to be similar to 1.4 and 2.5 mM, respectively. The results obtained in this work show negligible interference (3.7 %) with uric acid, glucose and urea.