Protein Biomarkers of Bovine Defective Meats at a Glance: Gel-Free Hybrid Quadrupole-Orbitrap Analysis for Rapid Screening.

Enrique Sentandreu,Miguel A Sentandreu,Olga Pardo,Mamen Oliván,Noelia Aldai,Claudia Fuente-García,Vicent Yusà,Núria León

doi:10.1021/acs.jafc.1c02016

Abstract

An understanding of biological mechanisms that could be involved in the stress response of animal cattle prior to slaughter is critical to create effective strategies aiming at the production of high-quality meat. The sarcoplasmic proteome of directly extracted samples from normal and high ultimate pH (pHu) meat groups was studied through a straightforward gel-free strategy supported by liquid chromatography hybrid quadrupole-Orbitrap high-resolution mass spectrometry (LC-HRMS) analysis. A stepped proteomic pipeline combining rapid biomarker hunting supported by qualitative protein Mascot scores followed by targeted label-free peptide quantification revealed 26 descriptors that characterized meat groups assayed. The functional study of the proposed biomarkers suggested their relevant role in metabolic, chaperone/stress-related, muscle contractility/fiber organization, and transport activities. The efficiency, flexibility, rapidity, and easiness of the methodology proposed can positively contribute to the creation of innovative proteomic alternatives addressing meat quality assessment.

Highlights

Consumers are currently worried about ethics of food production concerning the implementation of animal welfare policies, greatly influencing their final decision on product selection.[1]
Pre-slaughter stress (PSS) is one of the most relevant issues among different conditioning factors related to animal care that can greatly affect the quality of meat, causing the occurrence of defective dark, firm, and dry (DFD) meat that is normally characterized by an ultimate pH ≥ 6.0
Original liquid chromatography hybrid quadrupole-Orbitrap high-resolution mass spectrometry (LC-HRMS) and Mascot generic format data files generated in this study are freely available at http://hdl.handle.net/10261/228237

Summary

Introduction

Consumers are currently worried about ethics of food production concerning the implementation of animal welfare policies, greatly influencing their final decision on product selection.[1]. Food authorities consider that pHu values higher than 6.0 at 24 h postmortem are intimately associated with PSS animals[3] and DFD meats.[4] early detection of high pHu meats in the food chain is critical for the industry[5] since defective raw material causes significant economic losses.[6] the value of pHu assessment is compromised since high values do not necessarily guarantee the presence of true DFD meats,[6] requiring new diagnostic strategies. The efficiency of proteomic research for the hunting of predictive high pHu protein biomarkers stands up as a novel approach to discriminate meat from normal and PSS animals.[7−11] results achieved to date strongly relied on gel-based methodologies that provided an excellent resolving power while still having important constraints such as the limited number of identified protein biomarkers and inaccurate quantitative results. The application of such approaches can be hindered by current trends in green analytical chemistry concerning the use of hazardous and nonsustainable chemicals (i.e., acrylamide) and excessive energy consumption by merging two protein purification steps.[12]

Objectives

Methods

Results