Colloidal Stability & Conformational Changes in β-Lactoglobulin: Unfolding to Self-Assembly

International Journal of Molecular Sciences

Steven Blake²  Samiul Amin¹  Wei Qi²  Madhabi Majumdar²  E. Neil Lewis² 

[1] Malvern Instruments, 7221 Lee Deforest Drive, Suite 300, Columbia, MD 21046, USA;

关键词: dynamic light scattering;    differential scanning calorimetry;    Raman spectroscopy;    self-assembly;    protein aggregation;    protein unfolding;    microrheology;   

DOI  :  10.3390/ijms160817719

来源: mdpi

【 摘 要 】

A detailed understanding of the mechanism of unfolding, aggregation, and associated rheological changes is developed in this study for β-Lactoglobulin at different pH values through concomitant measurements utilizing dynamic light scattering (DLS), optical microrheology, Raman spectroscopy, and differential scanning calorimetry (DSC). The diffusion interaction parameter k_D emerges as an accurate predictor of colloidal stability for this protein consistent with observed aggregation trends and rheology. Drastic aggregation and gelation were observed at pH 5.5. Under this condition, the protein’s secondary and tertiary structures changed simultaneously. At higher pH (7.0 and 8.5), oligomerizaton with no gel formation occurred. For these solutions, tertiary structure and secondary structure transitions were sequential. The low frequency Raman data, which is a good indicator of hydrogen bonding and structuring in water, has been shown to exhibit a strong correlation with the rheological evolution with temperature. This study has, for the first time, demonstrated that this low frequency Raman data, in conjunction with the DSC endotherm, can be been utilized to deconvolve protein unfolding and aggregation/gelation. These findings can have important implications for the development of protein-based biotherapeutics, where the formulation viscosity, aggregation, and stability strongly affects efficacy or in foods where protein structuring is critical for functional and sensory performance.

【 授权许可】

CC BY   
© 2015 by the authors; licensee MDPI, Basel, Switzerland.