High temperature glycerolysis widely used in industrial scale for production of food-based emulsifiers has many disadvantages. On the other hand, high cost, unavailability, low activity and high reaction time of enzymes limit the use in industrial scale glycerolysis reactions for the production of food-based emulsifiers (i.e. monoglycerides). The goal of this project is to develop an efficient and optimal glycerolys is process for production of monoglycerides. This study found that optimal production through enzymatic glycerolysis was achieved with 15 wt% loading of Fermase CALB 10000 and oil to glycerol molar ratio was optimized at 1:5. A tert-butyl alcohol system was developed carefully with evaluation of more than 10 organic solvents. The suitable co-solvent addition was 1:50 molar ratio with respect to oil. In the above-optimized condition, the conversion of monoglyceride was 50-60% at 2-3 h and 70-80% at 5 to 6 h of reaction time. The reaction time was optimized at 5 h. Subsequent to the following reaction was found that, there is no drastic reduction of enzyme activity and could be reused for next reaction. Due to this behaviour, we recommend Fermase 10000 as an effective and low cost enzyme for future glycerolys is reaction in industrial scale. (C) 2013 Elsevier B.V. All rights reserved.