The main purpose of this study was to evaluate the gills and gonads of Vimba vimba in terms of fatty acid composition, esterase activity, and esterase band patterns by native-PAGE method. In this context, C16:0 and C18:1n-9 were the most abundant fatty acids in the TLs, NLs, and the PLs fractions of both tissues. EPA (C20:5n-3) and DHA (C22:6n-3) levels were higher in the gills than in the gonads. MUFAs were the main fatty acid groups in the TLs, NLs, and PLs of both tissues. Gills had higher n-3/n-6 ratios when compared with the gonads, with the exception of the PLs. TLs of the gonads had the highest AI and TI values. A concentration of 1 mM Na-taurocholate increased the esterase activity of the gonads. Other chemicals having inhibition and surface active effects resulted in decreases in the esterase activities at concentrations of 1, 5, and 10 mM. In total, nine esterase bands were determined in both tissues. Four bands were shared by both the gills and gonads of V.vimba. These shared bands could be responsible for the similar esterase activities in both tissues. Gills and gonads of V. vimba have substantial amounts of fatty acids and esterase isozymes. More detailed studies are required to understand their industrial and nutritional uses.