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Assessment of the denaturation of collagen protein concentrates using different techniques

  • Victor Perez-Puyana EMAIL logo , Francisco J. Ostos , Pilar López-Cornejo , Alberto Romero and Antonio Guerrero
Published/Copyright: May 24, 2019

Abstract

The use of collagen and gelatin in the field of regenerative medicine is widely extended. However, most of the studies in this topic are focused on the scaffolds’ properties, but only a few are related to the properties of the raw material used. The raw material analysis not only consists of a study of the composition, but also of the denaturation degree that can influence the processing and properties of the structure of the scaffold. Thus, the denaturation degree analysis of different collagen proteins was performed and assessed by the comparison of four different methods: differential scanning calorimetry (DSC), Fourier transform Infrared Spectroscopy (FTIR) and circular dichroism (CD) spectra and sulfhydryls content analysis. DSC measurements put forward a glass transition between 88°C and 95°C as well as from the FTIR measurements; the characteristic peaks for proteins are evidenced. However, from the sulfur content, only a small proportion of free sulfhydryls are present with respect to their total amount. In addition, CD spectra allow to estimate the secondary structure of the protein by the analysis of the α-helix and β-strand and also quantify the denaturation degree with the ‘positive/negative ratio’ (RPN) from the CD profiles, obtaining values in the range between 25% and 100%.

Acknowledgements

This work is part of a research project sponsored by the ‘Ministerio de Economía y Competitividad’ of the Spanish Government (Ref. CTQ2015-71164-P, MINECO/FEDER, UE). The authors gratefully acknowledge their financial support. The authors also thank the Characterization Service (CITIUS-Universidad de Sevilla) for providing full access and assistance to the Q20 (TA Instruments). The authors also acknowledge the University of Seville for the two VPPI-US grants of Victor M. Pérez Puyana and Francisco J. Ostos Marcos, funder ID: http://dx.doi.org/10.13039/100009042.

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Received: 2019-01-08
Accepted: 2019-05-12
Published Online: 2019-05-24
Published in Print: 2019-12-18

©2019 Walter de Gruyter GmbH, Berlin/Boston

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