Refolding of β-Stranded Class I Chitinases of Hippophae rhamnoides Enhances the Antifreeze Activity during Cold Acclimation
Figure 4
Analysis of antifreeze activity in HrCHT1a and HrCHT1b purified from cold acclimated and non-acclimated seedlings.
(A) Upper panel-ice crystal morphology, analysed by phase contrast microscope attached with nanoliter osmometer. Lower Panel-Ice recrystallization inhibition assays as observed by sucrose sandwich splat assay. BSA was used a negative control which did not exhibit antifreeze activity. Magnification bar in ice crystal morphology shows 10 µm, while in splat assay it represents 50 µm. Error bars show standard deviation from five replicates. (B) Ice crystal morphologies showing different stages of formation of flower shaped ice crystal with 5 mg/mL HrCHT1a.