Evaluation of the genotoxic potential of protoporphyrin IX and the safety of a protoporphyrin IX‐rich algal biomass

Chlamydomonas reinhardtii is a nonpathogenic, nontoxigenic green algae used as a sustainable source of protein in foods. In order to mimic meat‐like qualities, a strain rich in protoporphyrin IX (PPIX), an endogenous heme/chlorophyll precursor, was developed using an evolution and selection strategy...

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Published in:Journal of applied toxicology Vol. 42; no. 7; pp. 1253 - 1275
Main Authors: Murbach, Timothy S., Glávits, Róbert, Moghadam Maragheh, Niloofar, Endres, John R., Hirka, Gábor, Goodman, Richard E., Lu, Guihua, Vértesi, Adél, Béres, Erzsébet, Pasics Szakonyiné, Ilona
Format: Journal Article
Language:English
Published: England Wiley Subscription Services, Inc 01.07.2022
Subjects:
Algae
Animals
Aquatic plants
Biocompatibility
Biomass
Chlamydomonas reinhardtii
Chlorophyll
Chromosome aberrations
Digestibility
DNA Damage
Genotoxicity
Heme
In vitro methods and tests
In vivo methods and tests
Mammals
Mammals - metabolism
Meat
meat substitute
Mutation
NOAEL
Organs
Pepsin
Protein sources
Proteins
Proteomics
Protoporphyrin
Protoporphyrin IX
Protoporphyrins - metabolism
Protoporphyrins - toxicity
Rats
Safety
Target recognition
Toxicity
toxicity
meat substitute
protoporphyrin IX
Chlamydomonas reinhardtii
safety
NOAEL
ISSN:0260-437X, 1099-1263, 1099-1263
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Summary:Chlamydomonas reinhardtii is a nonpathogenic, nontoxigenic green algae used as a sustainable source of protein in foods. In order to mimic meat‐like qualities, a strain rich in protoporphyrin IX (PPIX), an endogenous heme/chlorophyll precursor, was developed using an evolution and selection strategy, and investigations were carried out to evaluate the safety of the novel strain, C. reinhardtii (red), strain TAI114 (TAI114). Digestibility and proteomic evaluations were conducted to determine whether any potentially allergenic or toxic proteins occurred as the result of the mutation process. The genotoxic potential of pure PPIX was evaluated using a bacterial reverse mutation test, an in vitro mammalian chromosomal aberration test, and an in vivo mammalian micronucleus test. Finally, the novel TAI114 biomass was evaluated for general toxicity and identification of target organs in a 90‐day repeated‐dose oral toxicity study in rats. All proteins were rapidly degraded in pepsin at pH 2.0 suggesting low allergenic potential. The proteomic evaluation indicated that TAI114 biomass contains typical C. reinhardtii proteins. PPIX was unequivocally negative for genotoxic potential and no target organs or adverse effects were observed in rats up to the maximum feasible dose of 4000 mg/kg bw/day TAI114 biomass, which was determined to be the no‐observed‐adverse‐effect‐level (NOAEL). These results support the further development and risk characterization of TAI114 biomass as a novel ingredient for use in the meat analogue category of food. Protoporphyrin IX (PPIX) is an endogenous component of hemoglobin that can impart meat‐like qualities to non‐meat foods. An evolution and selection strategy was employed to develop a novel PPIX‐rich strain of the edible algae, Chlamydomonas reinhardtii. Before introducing the novel strain—TAI114—into food, preclinical safety studies were conducted and determined a low allergenic potential, lack of novel proteins, lack of genotoxic potential, and a subchronic no‐observed‐adverse‐effect‐level (NOAEL) in rats of 4000 mg/kg bw/day, the highest feasible dose.
Bibliography:Funding information
Richard E. Goodman, Guihua Lu, Adél Vértesi, Erzsébet Béres and Ilona Pasics Szakonyiné are co‐senior authors.
Triton Algae Innovations
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ISSN:0260-437X
1099-1263
1099-1263
DOI:10.1002/jat.4293