Abstract
The leghemoglobin protein (LegH) from soy (Glycine max) expressed in Pichia pastoris (LegH preparation, LegH Prep) imparts a meat-like flavor profile onto plant-based food products. The safety of LegH Prep was evaluated through a series of in vitro and in vivo tests. The genotoxic potential of LegH Prep was assessed using the bacterial reverse mutation assay (Ames test) and the in vitro chromosome aberration test. LegH Prep was nonmutagenic and nonclastogenic in each test, respectively. Systemic toxicity was assessed in a 28-day dietary study in male and female Sprague Dawley rats. There were no mortalities associated with the administration of LegH Prep. There were no clinical observations, body weight, ophthalmological, clinical pathology, or histopathological changes attributable to LegH Prep administration. There were no observed effects on male reproduction in this study, but the suggestion of a potential estrous cycle distribution effect in female rats prompted a second comprehensive 28-day dietary study in female Sprague Dawley rats. This study demonstrated that female reproductive parameters were comparable between rats treated with LegH Prep and concurrent control rats. These studies establish a no observed adverse effect level of 750 mg/kg/d LegH, which is over 100 times greater than the 90th percentile estimated daily intake. Collectively, the results of the studies presented raise no issues of toxicological concern with regard to LegH Prep under the conditions tested.
Highlights
Clinical pathology was preliminarily tested via Levene test[52] for homogeneity and via Shapiro-Wilk test[53] for normalcy followed by analysis of variance (ANOVA) followed with Dunnett test.[48,49]. The objective of this test was to determine the mutagenic potential of leghemoglobin protein (LegH) leghemoglobin protein preparation (Prep) using histidine-requiring strains of S. typhimurium (TA98, TA100, TA1535, and TA1537) and a tryptophan-requiring strain of E. coli (WP2 uvrA)
Leghemoglobin protein preparation was evaluated with and without an exogenous metabolic activation (S9 mix) at levels of 23.384, 74, 233.84, 740, 2338.4, 7400, 23,384 and 74,000 mg/plate, which corresponded to 1.58, 5.0, 15.8, 50, 158, 500, 1580, and 5000 mg/ plate of the characterizing component, LegH, with the high level being the standard limit for this test (Supplemental Table S2)
Replacing the myoglobin that catalyzes the unique flavor chemistry of meat derived from animals with LegH from soy opens an opportunity to develop plant-based meats that deliver to consumers the pleasure they demand from animal-derived meats, with a small fraction of the environmental impact
Summary
Western diets containing meat have a larger negative impact on the environment compared to more sustainable plant-based diets.[1,2,3,4] due to both social and personal reasons, many consumers are reluctant to reduce the amount of meat they eat.[2,5,6] To date, plant-based diets have been limited to small populations, such as consumers who follow vegetarian or vegan principles.[7,8,9] One potential way to catalyze widespread shift to more sustainable, plant-based diets is to create meat directly from plants that satisfies the tastes of meat consumers.[7,8,10] Achieving that goal would require products that recreate the sensory properties that people crave in meat, including texture, mouthfeel, taste, smell, and cooking experience, based on an understanding of the biochemical origins of meat sensory attributes.An investigation of the molecular mechanisms underlying the unique flavors and aromas of meat led to the discovery that heme is the critical catalyst of the chemical reactions that transform simple biomolecules into the complex array of odorants and flavor molecules that define the characteristic flavor profile of meat.[11]. Western diets containing meat have a larger negative impact on the environment compared to more sustainable plant-based diets.[1,2,3,4] due to both social and personal reasons, many consumers are reluctant to reduce the amount of meat they eat.[2,5,6] To date, plant-based diets have been limited to small populations, such as consumers who follow vegetarian or vegan principles.[7,8,9] One potential way to catalyze widespread shift to more sustainable, plant-based diets is to create meat directly from plants that satisfies the tastes of meat consumers.[7,8,10] Achieving that goal would require products that recreate the sensory properties that people crave in meat, including texture, mouthfeel, taste, smell, and cooking experience, based on an understanding of the biochemical origins of meat sensory attributes. Dixon D, Alison R, Bach U, et al Nonproliferative and proliferative lesions of the rat and mouse female reproductive system. The female rat reproductive cycle: a practical histological guide to staging.
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