The goal of this study was to produce conjugated linoleic acid (CLA) monoacylglycerols (MAGs). A chemicoenzymatic process was employed starting from castor bean oil, which was chemically dehydrated and isomerized to obtain new modified triacylglycerols (TAGs) with a very high proportion of CLA fatty acids (>95%). Such TAGs were then used in enzymatic glycerolysis reaction using Candida antarctica B lipase at 55C in tert-butyl alcohol as a solvent. The effect of enzyme load and substrate molar ratio on the production yield of MAGs was evaluated and optimized using response surface methodology with a two-factor and three-level factorial design. A satisfactory linear correlation existed between experimental results and predicted values (R2 = 0.97 and R2adj. = 0.94). Using the contour plot generated from the model for MAG production yield, optimum parameters could be obtained corresponding to an 11/1 glycerol/TAG molar ratio and an enzyme load of 2% (% w/w of both substrates).