Melting Temperature (Tm)

°C (Celsius)

Circular Dichroism (CD)

10 mM phosphate–citrate

30-80 °C


0.05  mg/mL

195–260 nm


Circular dichroism (CD) analysis was conducted on a JASCO-810 spectropolarimeter (Tokyo, Japan). Enzyme samples were diluted in 10 mM phosphate–citrate buffer (pH 3.8) to the final protein concentration of 0.05 mg/mL and then loaded to a 10 mm quartz cuvette for CD assay. The melting curve of the enzymes were collected in the far-ultraviolet (195–260 nm) spectrum at the temperature from 30 to 80 °C continuously. The Tm value were calculated from the CD using the Global 3 software.