Purification, characterization and antioxidant activity of dextran produced by Leuconostoc pseudomesenteroides from homemade wine
Introduction
Exopolysaccharides (EPSs) have attracted increasing attention from researchers because of their medical, industrial, or food applications (Ibarburu et al., 2015). Many different microorganisms can be used to synthesize EPS, such as bacteria, algae, yeasts, and fungi (Chen, Liu, Xiao, Huang, & Liu, 2016; He et al., 2014; Miao et al., 2014; Qu, Li, Zhang, Zeng, & Fu, 2016; Tian, Zhao, Zeng, Zhang, & Zheng, 2016). Among all the microbial EPSs, lactic acid bacteria (LAB) EPS has been the most diffusely studied especially for the production of fermented food, attribute to their ability to generate lactic acid and they are generally recognized as safe (GRAS) status (Rühmkorf et al., 2012).
Depending on the chemical composition, EPSs are classified as homopolysaccharides (HoPSs) and heteropolysaccharides (HePSs) (De Vuyst & Degeest, 1999). Based on linkage bonds and nature of monomeric units, the HoPSs can be classified as four groups: (i) dextran with α-(1→6) glycosidic linkages (Van Cleve, Schaefer, & Rist, 1956), (ii) mutan with α-(1→3) glycosidic linkages (Kralj et al., 2004), (iii) alternan with alternating α-(1→6) and α-(1→3) glycosidic linkages (CôTÉ & Robyt, 1982), and (iv) reuteran with α-(1→4) and α-(1→6) glycosidic linkages (Feng et al., 2018). Among these EPS, dextran is α-glucan that contain consecutive α-(1→6) linkages in the main chain and α-(1→2), α-(1→3), or α-(1→4) branch linkages have been regarded as the most promising food improvers (Moon, Lee, Taniguchi, Miyamoto, & Kimura, 2001; Shukla, Shi, Maina, Juvonen, & Goyal, 2014). LAB EPS plays a crucial role in improving the rheology, texture, mouth feel of fermented food formulations and conferring beneficial physiological effects on human health, such as antitumour activity, immunomodulating bioactivity and anticarcinogenecity (Ibarburu et al., 2015; Patel, Majumder, & Goyal, 2012; Shao et al., 2014). The various characteristics of EPSs are not only because of their chemical structure, but also related to their composition, molecular mass, chain conformation and water-soluble (Inturri et al., 2017). Identifying structures of EPS is important to investigate its bioactivities and health benefits. While, there were only few reports about antioxidant activities of EPS from Leuconostoc.
Leuconostoc species are the primary producers of the dextran that have multipurpose uses (Ibarburu et al., 2015). However, the existing researches mainly focused on Leu. mesenteroides (Du, Xing, Zhou, & Han, 2017; Du, Xing, Yang et al., 2017) and Leu. citreum (Yang et al., 2018b, 2015). There is little information relating to the dextran from Leu. pseudomesenteroides which is relatively unknown and few studies have been reported the dextran with high production. In order to enrich the source of EPS-producing strains, identify structural and biochemical characterizations of EPS and explore the potential applications in the food, cosmetic, pharmaceuticals and other fields. In this study, we aim to isolate and identify an EPS produced by Leu. pseudomesenteroides from homemade wine, and to investigate the structural characteristics of EPS via high-performance size-exclusion chromatography (HPSEC), fourier-transform infrared (FT-IR) spectra, and nuclear magnetic resonance (NMR) spectroscopy. In addition, the physico-chemical properties including water solubility, water holding capacity, emulsifying activity, thermodynamic stability and antioxidant activity of the EPS were investigated.
Section snippets
Screening and identification of EPS-producing LAB
The sample of homemade wine was collected and stored at 4 °C. The sample was subjected to serial dilution. Each dilution was spread on MRS agar medium (50 g/L glucose, 10 g/L tryptone, 5 g/L yeast extract, 10 g/L beef extract, 2 g/L K2HPO4, 5 g/L anhydrous sodium acetate, 2 g/L ammonium citrate, 0.58 g/L MgSO4·7H2O, 0.25 g/L MnSO4·H2O, 1 mL/L Tween 80, 18 g/L agar, pH 6) and incubated at 30 °C for 36 h. The slimy and mucoid colony was inoculated to fermentation medium. According to the
Isolation and identification of EPS-producing strain
A total of 4 isolates of strains were able to synthesis EPS with sucrose. A highest EPS-producing bacterium, numbered as DRP-5, which produced 43.11 ± 3.08 g/L of 50 g/L sucrose within 36 h. DRP-5 was Gram-positive short rods with cell ranging between 0.6 and 1.1 μm in length and 0.3 and 0.7 μm in diameter. The pure colonies were slightly convex, slimy, regularly edged, and cream in color. The bacterium was identified as Leu. pseudomesenteroides according to physical-chemical tests,
Conclusion
In this study, an EPS-producing strain isolated from homemade wine was identified as Leu. pseudomesenteroides. The EPS produced by Leu. pseudomesenteroides DRP-5 was purified and its properties were analyzed. After the ethanol extraction, the purification of EPS was carried out by TCA deproteinization with Sephadex G-100 chromatography. The purified EPS had a molecular mass of 6.23 × 106 Da. The results of GC, FT-IR, 1H and 13C NMR spectral analysis confirmed that the EPS is a highly linear
Acknowledgements
This work was financially supported by the financial aid from the National Science-Technology Support Program of China (2015BAD16B01).
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These authors contributed equally to this work and share the first authorship.