Analysis of secondary structure
For analysing the secondary structure conformational changes of the thermal and shear treated samples, far UV-CD spectroscopic measurement was performed using a Circular Dichroism spectrophotometer (JASCO, J-1500). A 0.1 cm path length quartz cuvette was used for this study. The spectra were collected in the range of 260-190 nm (Iannazzo et al. 2018; Militello et al. 2003). Samples were scanned as an average of 5 accumulations with a scan speed of 100 nm/min and at 0.1 nm of bandwidth. The spectrum of the buffer was used for the baseline correction prior the measurement of the samples. The machine CD signal value θ in millidegree (mdeg) has been converted to mean residue ellipticity value (MRE) in (deg cm2dmol-1 residue-1 ) to decipher the analysis in terms of residues in the protein chain (Johnson Jr 1988).