Potential of Good?s buffers to inhibit denaturation of myofibrillar protein upon freezing

The current systematic study sought to examine the potential use of three Good's buffers (MES, MOPS and HEPES) in inhibiting myofibrillar protein (MFP) denaturation induced by acidity changes. The highest degree of acidity variation was found in the center and bottom of large bottles due to the freeze-concentration effect. Good's buffer tended to basify during freezing, and it could prevent the crystallization of sodium phosphate (NaP) buffer. Acidification upon freezing Na-P disrupted the natural conformation of MFP and induced the formation of large proteins aggregates with tight packing. The 15 mM MES, 20 mM MOPS, and 30 mM HEPES were respectively added to neutralize the strong acidity drop induced by freezing 20 mM Na-P, and all of them significantly improved the stability of the MFP conformation (P < 0.05). This work is not only critical to meet the growing demand for protein, but also groundbreaking for broadening the applicability of Good's buffers in the food industry.