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).