Understanding how proteins fold into their native structure is a fundamental problem in biophysics, crucial for protein design. It has been hypothesized that the formation of a molten globule intermediate precedes folding to the native conformation of globular proteins; however, its thermodynamic properties are poorly known. We perform single-molecule pulling experiments of protein barnase in the range of 7 degrees C to 37 degrees C using a temperature-jump optical trap. We derive the folding free energy, entropy and enthalpy, and heat capacity change (Delta C-p = 1,050 +/- 50 cal/mol.K) at low ionic strength conditions. From the measured unfolding and folding kinetic rates, we also determine the thermodynamic properties of the transition state, finding a significant change in Delta C-p (similar to 90%) between the unfolded and the transition states. In contrast, the major change in enthalpy (similar to 80%) occurs between the transition and native states. These results highlight a transition state of high energy and low configurational entropy structurally similar to the native state, in agreement with the molten globule hypothesis.