The increasing use of petroleum plastics has caused environmental damage due to the degradation time of these materials. An alternative to petroleum plastics could be thermoplastic starch (TPS). However, thermoplastic starch does not exhibit satisfactory tensile properties. The mechanical properties of thermoplastic starch can be improved by adding sisal microfibers. Thus, the objective of this study was to evaluate the influence of different levels of glycerol and sisal microfibers on the thermal and tensile properties of thermoplastic corn starch composites. The microfibers were obtained via mechanical treatment followed by chemical treatment (alkaline treatment and bleaching). The films were obtained by the casting method using commercial corn starch and glycerol as a plasticizing agent, reinforced with sisal microfibers. Fourier transform infrared spectroscopy (FTIR) results revealed that the addition of microfibers did not change the chemical structure of the TPS matrix. The films from the samples with 18% glycerol and 10% microfibers had the highest value for the maximum tension, equal to 4.78 MPa. The thermal decomposition profile of TPS was not altered by the addition of microfibers. Our findings demonstrated the profound influence of glycerol and microfiber contents on the tensile properties of thermoplastic starch composites.