Typical of the Caatinga scrublands of northeastern Brazil, the umbu, or Brazilian plum, is known for its rich nutritional properties. It is high in vitamin C and juicy, and the mature fruit in particular contains a number of volatile components. In this region of Brazil, it is widely consumed, both raw and processed, in the form of pulp, jelly, candy and ice cream. A group of Brazilian and Swiss scientists recently concluded a study that revealed new properties of the fruit, which is round in shape, has a velvety skin and a slightly tart flavor. They found that the plum (Spondias tuberosa) is rich in phenolic compounds with antioxidant activity, which makes it a potential raw material for cosmetics that revitalize aging skin, such as anti-wrinkle creams or products for sagging skin. Two of the substances the study identified are new.
Coordinated by Vanderlan da Silva Bolzani, a pharmacologist and professor at the Chemistry Institute of São Paulo State University (IQ-Unesp) in Araraquara, the research was a collaboration with the University of Geneva (Unigen), in Switzerland, and the Center for Innovation and Pre-clinical Testing (CIEnP), a private non-profit company based in Florianópolis (Santa Catarina State). “The study provides the first complete documentation on isolating the compounds of umbu pulp, which contains antioxidant and rejuvenating properties for the skin,” notes Maria Luiza Zeraik, who worked on the team as a postdoctoral fellow. She is currently a professor in the Chemistry Department of the Center for Exact Sciences at Londrina State University (UEL) in the state of Paraná. “An important aspect of our study is to promote technological innovation of economic value for the northeastern region,” says Zeraik. The umbuzeiro, or Brazilian plum tree, is important in the Caatinga because it bears fruit during the dry season and provides a source of income for the local population.
The study was funded by the Swiss and Brazilian governments with financing from FAPESP through a postdoctoral research grant awarded to Zeraik, in addition to a National Biodiversity Research System (Sisbiota) project, a National Council for Scientific and Technological Development (CNPq) program in partnership with the Foundation. Starting in 2014, the study included a portfolio of projects of the Center for Research and Innovation in Biodiversity and Drug Discovery (CIBFar), one of the FAPESP Research, Innovation and Dissemination Centers (RIDC), coordinated by Glaucius Oliva of the Physics Institute of the University of São Paulo (IFSC-USP) in São Carlos.
Two patent applications were filed in Brazil and abroad. They have to do with the process of extraction and isolation of compounds present in umbu pulp related to their antioxidant properties and the inhibition of acetylcholinesterase, an enzyme that promotes connections (synapses) between neurons. “The substances related to acetylcholinesterase could, in the future, lead to a drug or food supplement to treat memory loss, a condition commonly found in the elderly,” says Bolzani.
Focus on biodiversity
The findings on the umbu were part of a larger study aimed at investigating the properties of 22 fruits belonging to Brazilian biodiversity with a view toward their potential use in the cosmetics or food industry. This project was part of a bilateral agreement between Brazil and Switzerland known as the Brazilian Swiss Joint Research Programme (BSJRP), coordinated on the Brazilian side by CNPq. The project was initiated in 2011 and completed in 2014 at IQ-Unesp, which was responsible for the initial chemical and biological screening of native or endemic fruits of northern and northeastern Brazil. Besides umbu, others on the study list include platonia, siriguela, mangaba, pitomba and the hog-plum, among others.
“The process of preparing the fruit samples, extracting chemical components by the usual analytical methods and conducting preliminary chemical tests of the extracts were done in our NuBBE laboratory [Center for Natural Products Bioassays, Biosynthesis and Eco-physiology, a green seal laboratory that prohibits the use of chlorinated solvents and other petroleum derivatives in many stages of extraction and purification],” notes Bolzani, who is also a member of the steering committee of the Biota-FAPESP program, which aims to map and analyze biodiversity in São Paulo and evaluate the possibilities for sustainable exploitation of plants and other organisms with economic potential. “All parts of the fruit [peels, pulp, seeds] were analyzed, resulting in more than 100 extracts. We separated some very active extracts from these, and the umbu pulp proved to be an excellent one with which to start the research.” Other fruits—whose names the group would like to keep confidential—also presented activities of interest and will be studied later.
The partnership with the University of Geneva, a major European center for research on natural products, had a budget of 173,400 Swiss francs (currently equal to R$700,000), divided between the governments of Brazil (35% of the total) and Switzerland.” We employed innovative methods of chemical characterization and detected, isolated and identified the chemical compounds present in umbu, responsible for inhibiting the activity of acetylcholinesterase, the target enzyme for the treatment of Alzheimer’s disease,” notes Brazilian pharmacologist Emerson Queiroz, a professor at Unigen’s School of Pharmaceutical Sciences in Switzerland. In vitro bioassays using the pure compounds were performed by professor Muriel Cuendet at the same university.
As part of the Swiss-Brazilian program, Zeraik, during her postdoctoral period supervised by Bolzani, spent nine months at the University of Geneva. While there she learned the principles of metabolomic study carried out by the group led by professors Jean-Luc Wolfender and Emerson Queiroz. Wolfender heads the Phytochemistry and Bioactive Natural Products Laboratory of the University and coordinates the bilateral project for the Swiss institution. “Metabolomic study is an advanced approach to chemical mapping that is ideal for quantifying all the natural products of an organism,” says Zeraik. “It is used to study all secondary metabolic compounds of a plant and, through these analyzes, we obtain a fingerprint, the plant’s metabolic identity, such as a panel of chemical substances present in the species.” Queiroz believes that training human resources and transferring knowledge and technology to Brazil are important aspects of the Brazil-Switzerland bilateral program.
In vitro tests
After the characterization done in Switzerland, the umbu extracts were standardized and sent to CIEnP in Florianópolis for proof of concept studies, an essential stage when the aim is further industrial cooperation with a view toward a potential product. “We do in vitro studies here on human skin cells—melanocytes and keratinocytes—to evaluate use of the product in developing anti-aging cosmetics,” says João Batista Calixto, the CEO of CIEnP and a former professor at the Federal University of Santa Catarina (UFSC). “We spent almost a year conducting about 30 tests, involving various enzymes and inflammatory mediators potentially responsible for aging of the skin.”
Funded by the government of Santa Catarina State, and the Ministries of Health (MS) and of Science, Technology and Innovation (MCTI), CIEnP was created two years ago with a mission to contribute to technological innovation in the pharmaceutical (human and veterinary drugs) and cosmetics sectors. Most of its projects are carried out in conjunction with the industrial sector. Research involving the umbu was the center’s first partnership with a university and the first proof of concept performed at CIEnP with a product of Brazilian biodiversity. According to Calixto, a standardized extract of umbu proved to be safe with acceptable levels of toxicity. “These results show that the fruit has the potential to be used as a cosmetic in the prevention of the inflammatory symptoms observed during the aging process,” he says. “Now we are looking for a company interested in producing and marketing this bioactive.”
In December 2015, Bolzani, Calixto and Zeraik received the Kurt Politzer Award for Technology, in the Researcher category, for the project “Sustainable use of the pulp of the umbu and hog-plum fruits: phenolic products of high added value for the cosmetic industry with anti-aging properties.” The recognition is awarded by the Brazilian Chemical Industry Association (Abiquim) to projects by companies and scientists that stimulate chemical research and innovation in
1. Prospecting for bioactive molecules and study of infra-specific variability in endophyte plants and microorganisms from the Cerrado and Caatinga. Contribution to knowledge and sustainable use of Brazilian biodiversity (Sisbiota) (nº 2010/52327-5); Grant Mechanism Biota Program; Principal Investigator Vanderlan Bolzani (Unesp); Investment R$ 552,668.55 and US$ 246,950.72
2. Natural products from plants of the Cerrado and Atlantic Forest, potential and useful models for identifying prototypes with oxidizing action on neutrophils and myeloperoxidase (MPO) enzyme (nº 2011/03017-6); Grant Mechanism Postdoctoral research grant (Maria Luiza Zeraik); Principal Investigator Vanderlan Bolzani (Unesp); Investment R$ 297,813.41
3. CIBFar – Center for Research and Innovation in Biodiversity and Drug Discovery (nº 2013/07600-3); Grant Mechanism Research, Innovation and Dissemination Centers (RIDC); Principal Investigator Glaucius Oliva (IFSC-USP); Investment R$ 21,485,493.35 (over four years)
ZERAIK, M.L., et al. Antioxidants, quinone reductase inducers and acetylcholinesterase inhibitors from Spondias tuberosa fruits. Journal of Functional Foods. V.21, pp. 396-405, Online. January 2016.