In vitro gastrointestinal digestion of a peanut, soybean, guava and beet beverage supplemented with Lactobacillus rhamnosus GG
Graphical abstract
Introduction
The use of probiotics in products for health is increasing (Kechagia et al., 2013) using both dairy and non-dairy formulations (Martins, Ramos, Martins, & Leite Júnior, 2016). Consumers are trying to incorporate healthy foods into their routine (Bezerra, Araujo, Santos, & Correia, 2015), which has prompted the development of products with functional appeal, such as those containing probiotics. The global acceptability of these foods has been increasing recently due to a better understanding of their benefits (Espitia, Batista, Azeredo, & Otoni, 2016; Najgebauer-Lejko, 2014). According to Tripathi and Giri (2014), the advances in research on functional foods containing probiotics in developed countries have resulted in the incorporation of these microorganisms not only in dairy products but also in other foods (Martins et al., 2013; Mattila-Sandholm et al., 2002; Ranadheera, Vidanarachchi, Rocha, Cruz, & Ajlouni, 2017).
Fruit-based beverages are considered a good culture medium for probiotics (Martins et al., 2016) and have a positive image with bioactive components of their own. Traditionally, probiotic microorganisms have been added to fermented dairy products. However, the number of lactose-intolerant, milk-allergic, hypercholesterolemic and dyslipidemic individuals is growing (https://www.sciencedirect.com/science/article/pii/S2212429217307253; Panghal et al., 2018; Vasudha & Mishra, 2013) so non-dairy probiotic products are potentially useful. Thus, the introduction of these microorganisms into non-dairy matrices would allow their consumption by these individuals (Dias et al., 2018; Martins et al., 2015, 2016; Rivera-Espinoza & Gallardo-Navarro, 2010).
In addition to fruits, root-crop vegetables and legumes can also be used for probiotic beverages (Espirito-Santo et al., 2014; Santos, Libeck, & Schwan, 2014). Peanut is considered a functional food because it contains high amounts of B-complex vitamins, is rich in unsaturated fatty acids, and is a good source of vegetable protein, dietary fiber, antioxidant vitamins, minerals and some phytochemicals (Bansal, Mangal, Sharma, Yaday, & Gupta, 2016; Basode, Randolph, Hurley, & Ahmedna, 2012). Soybean is rich in quality proteins and has low saturated fat (Costa, Júnior, Rosa, Caliari, & Pimentel, 2017). So, peanuts and soybeans can be considered as good sources of vegetable protein (Bansal et al., 2016; Basode et al., 2012; Costa et al., 2017).
Beverages of fruits and vegetables have also been appreciated by consumers for allowing them to obtain new functional flavors. Guava is a widely consumed fruit in Brazil and contains, in addition to fiber and lycopene, high levels of minerals and vitamins A, C and B-complex (Tanwar, Andallu, & Chandel, 2014), and beet is a vegetable source of fiber and its juice is easily digestible (Ravichandran et al., 2013). These vegetables can also serve as substrates for probiotic cultures.
However, the use of probiotic microorganisms in foods requires that it be shown that the organisms can survive processing and storage, along with gastrointestinal transit (Bron & Kleerebezem, 2011; Papadimitriou et al., 2015). Initial testing can be done using in vitro studies simulating digestion. The objective of this study was to investigate the survival of Lactobacillus rhamnosus GG with gastrointestinal digestion in vitro in fermented mixed beverages of legumes (peanuts and soybean), guava and beet and to evaluate the products.
Section snippets
Peanut and soybean water-soluble extracts, guava and beet pulp and beverage preparation
Peanut extract was obtained according to the methodology described by Bechaut and Nail (1978), in which 1 kg of raw Brazilian peanut seeds, cultivar IAC red Tatu (Pereira, Cajuri, MG, Brazil) was immersed in 2 L of tap water containing 0.2% citric acid (Synth, Diadema, São Paulo, SP, Brazil) and held for 16–18 h at 26 °C. The liquid was drained and the peanuts were washed and ground for ~2 min at 18,000 rpm (Vithory TR 010, Curitiba Equipments, Catanduva, SP, Brazil) with 5 L of water. They
Survival of L. rhamnosus GG to gastrointestinal tract conditions simulated in vitro
The count of L. rhamnosus GG in probiotic mixed beverages did not differ (p ≥ 0.05), at the times evaluated, in the products after gastric stress at pH 2.0–2.5 (data not shown). MBPP reached the end of shelf life, in enteric phase II, with 6.34 Log CFU/mL and MBSP with 5.50 Log CFU/mL (Fig. 1). Although the count of the viable cell for enteric phases I and II were lower (p < 0.05) at all times of analysis, when compared to the gastric phase (Fig. 1), the beverages provided the consumer the
Conclusions
The soy or peanut blend, guava and beet were good as a carrier of L. rhamnosus GG as any beverage. They met the probiotic survival requirements in the test using in vitro bile salts as recommended by the FAO, with the quantities required to cross the gastrointestinal tract barriers and arrive viable in the simulated large intestine. L. rhamnosus GG remained viable during the shelf life of the mixed soybean or peanut beverages with counts >7.3 and 7.4 Log CFU/mL, respectively. Although the
CRediT authorship contribution statement
Shimeny Ramos Montanari: Conceptualization, Investigation, Formal analysis. Bruno Ricardo de Castro Leite Júnior: Validation, Writing - review & editing. Maurilio Lopes Martins: Project administration, Supervision, Writing - original draft. Afonso Mota Ramos: Validation, Writing - review & editing. Mirella Lima Binoti: Visualization, Investigation. Renata Cristina de Almeida Bianchini Campos: Investigation, Methodology. Eliane Maurício Furtado Martins: Conceptualization, Funding acquisition,
Declaration of competing interest
There is no conflict of interest from the authors. The authors are solely responsible for the content of this article.
Acknowledgements
The authors are thankful to the IF Sudeste MG, Rio Pomba Campus and the National Council for Scientific and Technological Development (CNPq, grant number 488348/2013-7) for financial support.
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