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Enhanced Accumulation of Betulinic Acid in Transgenic Hairy Roots of Senna obtusifolia Growing in the Sprinkle Bioreactor and Evaluation of Their Biological Properties in Various Biological Models
Authors:Tomasz Kowalczyk  Przemysław Sitarek  Monika Toma  Patricia Rijo  Eva Domínguez-Martín  Irene Falcó  Gloria Sánchez  Tomasz Śliwiński
Affiliation:1. Department of Molecular Biotechnology and Genetics, University of Lodz, Banacha 12/16, 90-237 Lodz, Poland;2. Department of Biology and Pharmaceutical Botany, Medical University of Lodz, Muszynskiego 1 Łódź, 90-151 Lodz, Poland;3. Laboratory of Medical Genetics, Faculty of Biology and Environmental Protection, University of Lodz, Pomorska 141/143, 90-236 Lodz, Poland;4. CBIOS – Research Center for Biosciences & Health technologies, Universidade Lusófona de Humanidades e Tecnologias, Campo Grande 376, 1749-024 Lisboa, Portugal

iMed.ULisboa – Research Institute for Medicines, Faculdade de Farmácia da Universidade de Lisboa, Av. Prof. Gama Pinto, 1649-003 Lisboa, Portugal;5. iMed.ULisboa – Research Institute for Medicines, Faculdade de Farmácia da Universidade de Lisboa, Av. Prof. Gama Pinto, 1649-003 Lisboa, Portugal

Department of Biomedical Sciences, Faculty of Pharmacy, University of Alcalá, Campus universitario. Ctra. Madrid-Barcelona km. 33,600, 28805 Alcalá de Henares, Spain;6. Departament of Biotechnology, Institute of Agrochemistry and Food Technology (IATA-CSIC), Av. Agustín Escardino, 7, Paterna, 46980 Valencia, Spain;7. Department of Medical Biochemistry, Medical University of Lodz, 6/8 Mazowiecka Str, 92-215 Lodz, Poland

Abstract:Betulinic acid, which is found in transgenic roots of Senna obtusifolia (L.) H.S.Irwin & Barneby, is a pentacyclic triterpene with distinctive pharmacological activities. In this study, we report the differences in the content of betulinic acid and selected anthraquinones in transgenic S. obtusifolia hairy roots with overexpression of the PgSS1 gene (SOPSS2 line) and in transformed hairy roots without this genetic construct (SOA41 line). Both hairy root lines grew in 10 L sprinkle bioreactor. Additionally, the extracts obtained from this plant material were used for biological tests. Our results demonstrated that the SOPSS2 hairy root cultures from the bioreactor showed an increase in the content of betulinic acid (38.125 mg/g DW), compared to the SOA41 hairy root line (4.213 mg/g DW). Biological studies have shown a cytotoxic and antiproliferative effect on U-87MG glioblastoma cells, and altering the level of apoptotic proteins (Bax, p53, Puma and Noxa). Antimicrobial properties were demonstrated for both tested extracts, with a stronger effect of SOPSS2 extract. Moreover, both extracts showed moderate antiviral properties on norovirus surrogates.
Keywords:transgenic hairy roots  sprinkle bioreactor  gene expression  anticancer  antimicrobial  antiviral activity
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