In vitro analysis of mushroom proteases that may tenderize beef
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Date
2019-05
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The Ohio State University
Abstract
Meat tenderness is an important characteristic that influences consumer purchasing decisions. Protease extracts from natural plant sources such as pineapple have exhibited broad proteolytic activity that can over-tenderize the meat and negatively affect texture and quality. Therefore, identification and evaluation of other proteases capable of tenderizing beef is necessary. Previously, mushrooms have been shown to enhance flavor and nutritional composition of meat dishes, as well as having beneficial antioxidant and health effects. Mushrooms also contain a variety of proteases that were analyzed in this study for their ability to proteolyze beef proteins using an in vitro model system. Eight mushroom varieties were tested including white button (white immature Agaricus bisporus), crimini (brown immature Agaricus bisporus), portobello (mature Agaricus bisporus), shitakke (Lentinula edodes), enoki (Flammulina velutipes), oyster (Pleurotus ostreatus), king trumpet (Pleurotus eryngii), and brown beech (Hypsizygus tessellatus). Mushrooms were homogenized in a 20 mM Tris buffer (pH 8.0), filtered, centrifuged, and combined with purified bovine myofibrils for incubation at 25°C. Samples were collected at 0, 30, 60, 240, and 1440 min. Myofibrillar proteins of each sample were solubilized and separated using SDS-PAGE. Densities of protein bands were compared between the time-points to determine which of the eight mushroom varieties proteolyzed myofibrillar proteins including actin and myosin. Mushroom proteolytic activity was also quantified with a standard casein assay. Pleurotus ostreatus, mature Agaricus bisporus, and Pleurotus eryngii had the greatest caseinolytic activity at 1.06 U/mL, 0.70 U/mL, and 0.68 U/mL respectively, while Flammulina velutipes had the least at 0.03 U/mL. These results support the possibility that mushroom proteases may be able to tenderize beef, forming the basis for future research trials.
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Keywords
in vitro, protease, mushroom, beef, tenderness