Protein Extraction from Porcine Myocardium Using Ultrasonication

Abstract

Porcine myocardium is regarded as a byproduct in slaughterhouses and is rarely used as a food source due to its unsuitability for processing and consumption. In this study, we sought to develop an efficient ultrasonication method to extract protein from porcine myocardium. Comparisons of protein yield using various ultrasonication conditions with porcine myocardium revealed that treatment with 0.2 M NaCl, with pH 8.0, at an extraction temperature of less than 40 °C and an amplitude of 60% to 80% was optimal, yielding an extraction rate of 90%. In addition, SDS-PAGE analysis showed that increasing the time interval for ultrasonication increased the presence of myosin heavy chain and actin protein content. Functional analysis of the physiological properties of the isolated proteins using an ATPase assay showed that Ca and Mg ATPase activity was virtually undetectable in the early stages of ultrasonic treatment and that the proteins denatured rapidly. An analysis of protein digestion also showed that the digestive capacity of proteins treated by ultrasonication methods was greater. These results demonstrate that the ultrasonication method is effective for high-yield protein extraction from cardiac myofibrils of porcine myocardium with low salt concentrations, low Ca and Mg ATPase activities, and high digestive capacities.

RSM Optimization for the Recovery of Technofunctional Protein Extracts from Porcine Hearts

“…As shown in the response surface (Figure 3a), higher solubility values correspond to the pH range from 6.5 to 8.5, and solubility improves with salt addition from 1.5 to 3% NaCl, due to an enhancement of myofibrillar proteins solubilization by salt. These results agree with studies on porcine cardiac proteins [15] or proteins extracted from other sources, i.e., porcine spleen [11] or mechanically separated turkey meat [31]. Additionally, ultrasonic treatments were claimed as useful to increase the effective extraction of protein from normal residual meat byproducts such as porcine myocardium without the need for high salt concentrations, as required in conventional extraction methods [15].…”

“…Soluble protein fractions are composed mainly of myofibrillar proteins, actin, and myosin [14,32]. All samples exhibited bands corresponding to heavy (H) and light (L) myosin chains (with a MW of 200-220 and 15-25 kDa, respectively) and bands around 40-50 kDa that could correspond to the globular monomeric form of actin (G-actin) (MW of 42 kDa) [15,33]. Bands in the range between 30 and 40 kDa may be assigned to the regulatory proteins associated with actin, troponin complex, and tropomyosin.…”

“…Some studies on this topic can be found in the recent literature. Kim et al (2017) tested the use of ultrasound-assisted extraction methods to extract proteins from myofibrils of porcine myocardium. They reported that sonicated samples produced higher protein extraction rates without the need for high salt concentrations [15].…”

“…Kim et al (2017) tested the use of ultrasound-assisted extraction methods to extract proteins from myofibrils of porcine myocardium. They reported that sonicated samples produced higher protein extraction rates without the need for high salt concentrations [15]. Tsermoula et al (2018) compared alkali (pH 11) and acid (pH 2) solubilization followed by isoelectric precipitation to prepare protein-rich extracts from bovine and porcine hearts.…”

Bromelain Kinetics and Mechanism on Myofibril from Golden Pomfret (Trachinotus blochii)

“…The SDS‐PAGE running buffer, sample loading buffer were purchased from Bio‐Rad Laboratories, Inc., Hercules, CA, USA. The protein standard (10 to 250 kDa) was used as the marker (Kim et al., ). Electrophoresis was performed on a Bio‐Rad Mini‐PROTEAN Tetra Cell at 80 V for approximately 100 min, followed by gel staining and distaining processes.…”