{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,1,31]],"date-time":"2026-01-31T19:10:10Z","timestamp":1769886610516,"version":"3.49.0"},"reference-count":83,"publisher":"MDPI AG","issue":"3","license":[{"start":{"date-parts":[[2026,1,29]],"date-time":"2026-01-29T00:00:00Z","timestamp":1769644800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"CONACyT","award":["CB-2015\/258587"],"award-info":[{"award-number":["CB-2015\/258587"]}]},{"name":"CONACyT","award":["CB-CF-2022\/321009"],"award-info":[{"award-number":["CB-CF-2022\/321009"]}]},{"DOI":"10.13039\/501100003593","name":"CNPq","doi-asserted-by":"crossref","award":["PQ no. 305913\/2022-0"],"award-info":[{"award-number":["PQ no. 305913\/2022-0"]}],"id":[{"id":"10.13039\/501100003593","id-type":"DOI","asserted-by":"crossref"}]},{"DOI":"10.13039\/501100004263","name":"FAPERGS","doi-asserted-by":"crossref","award":["TO 24\/2551-0001467-5"],"award-info":[{"award-number":["TO 24\/2551-0001467-5"]}],"id":[{"id":"10.13039\/501100004263","id-type":"DOI","asserted-by":"crossref"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Molecules"],"abstract":"<jats:p>Conventional cancer therapies often harm healthy tissues due to their poor specificity, resulting in significant side effects that diminish patients\u2019 quality of life. Parasporins, a group of non-hemolytic parasporal proteins produced by Bacillus thuringiensis, are known for their selective cytotoxicity toward cancer cells. Typically, these proteins require activation through physical or biochemical treatments that fragment them into multiple peptides of varying sizes, which are then tested as mixtures, without purification, against cancer cell lines. In this study, a purification strategy that isolates the protein without prior activation and evaluates the resulting cytotoxic mechanism is proposed. The purification consists of four steps: (1) crystal solubilization with Laemmli buffer, (2) size-based separation via SDS-PAGE, (3) electroelution of the target protein from the gel, and (4) dialysis to remove the elution buffer. From the B. thuringiensis AX isolate, four proteins ranging from ~20 to 60 kDa were recovered, but only AX-2 displayed cytotoxic activity toward MCF-7 breast cancer cells, while remaining non-hemolytic and non-toxic to normal cells (erythrocytes, PBMCs, and MRC-5 fibroblasts). Thus, AX-2 qualifies as a parasporin. AX-2 induces apoptosis in MCF-7 breast cancer cells without generating oxidative stress, and the observed cell death appears to initiate at the plasma membrane rather than through intracellular pathways.<\/jats:p>","DOI":"10.3390\/molecules31030475","type":"journal-article","created":{"date-parts":[[2026,1,29]],"date-time":"2026-01-29T14:52:36Z","timestamp":1769698356000},"page":"475","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["AX-2: A Promising Non-Hemolytic Protein of Bacillus thuringiensis with Potent Selective Cytotoxicity Against Breast Cancer Cells"],"prefix":"10.3390","volume":"31","author":[{"ORCID":"https:\/\/orcid.org\/0009-0003-7973-9472","authenticated-orcid":false,"given":"Alain","family":"Cruz-Nolasco","sequence":"first","affiliation":[{"name":"Divisi\u00f3n de Estudios de Posgrado, Doctorado en Biotecnolog\u00eda, Universidad del Papaloapan, Circuito Central 200, Parque Industrial, Tuxtepec 68301, Oaxaca, Mexico"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8275-6044","authenticated-orcid":false,"given":"Miguel Angel","family":"Pe\u00f1a-Rico","sequence":"additional","affiliation":[{"name":"Centro de Investigaciones Cient\u00edficas, Instituto de Biotecnolog\u00eda, Universidad del Papaloapan, Circuito Central 200, Parque Industrial, Tuxtepec 68301, Oaxaca, Mexico"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Sibel J.","family":"Estrada-Escobedo","sequence":"additional","affiliation":[{"name":"Divisi\u00f3n de Estudios de Posgrado, Maestr\u00eda en Biotecnolog\u00eda, Universidad del Papaloapan, Circuito Central 200, Parque Industrial, Tuxtepec 68301, Oaxaca, Mexico"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Angel A.","family":"Ortela-Gregorio","sequence":"additional","affiliation":[{"name":"Divisi\u00f3n de Estudios de Posgrado, Maestr\u00eda en Biotecnolog\u00eda, Universidad del Papaloapan, Circuito Central 200, Parque Industrial, Tuxtepec 68301, Oaxaca, Mexico"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4844-8317","authenticated-orcid":false,"given":"Erick A.","family":"Juarez-Arellano","sequence":"additional","affiliation":[{"name":"Centro de Investigaciones Cient\u00edficas, Instituto de Qu\u00edmica Aplicada, Universidad del Papaloapan, Circuito Central 200, Parque Industrial, Tuxtepec 68301, Oaxaca, Mexico"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2319-8772","authenticated-orcid":false,"given":"Genaro","family":"V\u00e1zquez-Victorio","sequence":"additional","affiliation":[{"name":"Facultad de Ciencias, Universidad Nacional Aut\u00f3noma de M\u00e9xico, Circuito Exterior S\/N, Ciudad Universitaria, Ciudad de Mexico 04510, Mexico"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2474-4739","authenticated-orcid":false,"given":"Angelica S.","family":"Martinez-Ramirez","sequence":"additional","affiliation":[{"name":"Centro de Investigaciones Cient\u00edficas, Instituto de Biotecnolog\u00eda, Universidad del Papaloapan, Circuito Central 200, Parque Industrial, Tuxtepec 68301, Oaxaca, Mexico"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5659-159X","authenticated-orcid":false,"given":"Michele Rorato","family":"Sagrillo","sequence":"additional","affiliation":[{"name":"Graduate Program in Nanoscience, Franciscan University, Santa Maria 97010-032, RS, Brazil"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0533-3483","authenticated-orcid":false,"given":"Roberto C. Vianna","family":"Santos","sequence":"additional","affiliation":[{"name":"Oral Microbiology Laboratory, Universidade Federal de Santa Maria, Santa Maria 97065-060, RS, Brazil"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9864-3738","authenticated-orcid":false,"given":"Luz","family":"Camacho","sequence":"additional","affiliation":[{"name":"Laboratorio de Nutrici\u00f3n Experimental, Instituto Nacional de Pediatr\u00eda, Ciudad de Mexico 04530, Mexico"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5354-9900","authenticated-orcid":false,"given":"Nayeli G.","family":"Nieto-Vel\u00e1zquez","sequence":"additional","affiliation":[{"name":"Divisi\u00f3n de Investigaci\u00f3n, Hospital Ju\u00e1rez de M\u00e9xico, Ciudad de Mexico 07760, Mexico"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4124-5533","authenticated-orcid":false,"given":"A. Karin","family":"Navarro-Mtz","sequence":"additional","affiliation":[{"name":"Centro de Investigaciones Cient\u00edficas, Instituto de Biotecnolog\u00eda, Universidad del Papaloapan, Circuito Central 200, Parque Industrial, Tuxtepec 68301, Oaxaca, Mexico"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2026,1,29]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"21282","DOI":"10.1074\/jbc.M401881200","article-title":"A Bacillus thuringiensis crystal protein with selective cytocidal action to human cells","volume":"279","author":"Ito","year":"2004","journal-title":"J. Biol. Chem."},{"key":"ref_2","doi-asserted-by":"crossref","unstructured":"Aboul-Soud, M., Al-Amri, M., Kumar, A., Al-Sheikh, Y., Ashour, A., and El-Kersh, T. (2019). Specific cytotoxic effects of parasporal crystal proteins isolated from native Saudi Arabian Bacillus thuringiensis strains against cervical cancer cells. 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