Purification, characterization and antioxidant activity of dextran produced by Leuconostoc pseudomesenteroides from homemade wine

doi:10.1016/j.carbpol.2018.06.116

Carbohydrate Polymers

Volume 198, 15 October 2018, Pages 529-536

https://doi.org/10.1016/j.carbpol.2018.06.116 Get rights and content

Highlights

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An exopolysaccharide was produced by Leuconostoc pseudomesenteroides DRP-5.
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The exopolysaccharide was a highly linear dextran.
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The dextran showed moderate antioxidant activity in vitro experiments.
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The dextran has an excellent potential for use in the food industry.

Abstract

An exopolysaccharide (EPS) was produced by Leuconostoc pseudomesenteroides DRP-5 isolated from homemade wine. The EPS was obtained with ethanol extraction, which was further purified by chromatography of Sephadex G-100 to get a purified fraction. The monosaccharide composition of the EPS was glucose, and its molecular weight (Mw) was 6.23 × 10⁶ Da, as determined by gas chromatography (GC) and high-performance size-exclusion chromatography (HPSEC). Fourier transform infrared spectra (FT-IR) and nuclear magnetic resonance spectra (NMR) showed that the EPS was a linear glucan with α-(1→6)-linked glucosidic bonds. The water holding capacity (WHC), water solubility index (WSI) and emulsifying activity (EA) of DRP-5 EPS were 296.76 ± 18.93%, 98.62 ± 3.57% and 87.22 ± 2.18%, respectively. DRP-5 EPS have a higher degradation temperature of 278.36 °C, suggesting high thermal stability of the EPS. Also, DRP-5 EPS was found to have moderate 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical, hydroxyl radical, superoxide anion radical, 2,2'-Azinobis-(3-ethylbenzthiazoline-6-sulphonate) (ABTS) radical, Fe²⁺ scavenging activities and reducing power. All these characteristics suggest that DRP-5 EPS might have potential applications in the pharmaceutical, cosmetic and food industries.

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).

References (46)

Z. Ahmed et al.
Characterization of exopolysaccharide produced by Lactobacillus kefiranofaciens ZW3 isolated from Tibet kefir-Part II
Food Hydrocolloids
(2013)
Y.X. Chen et al.
Antioxidant activities of polysaccharides obtained from Chlorella pyrenoidosa via different ethanol concentrations
International Journal of Biological Macromolecules
(2016)
G.L. CôTÉ et al.
Isolation and partial characterization of an extracellular glucansucrase from Leuconostoc mesenteroides NRRL B-1355 that synthesizes an alternating (1→ 6), (1→ 3)-α-D-glucan
Carbohydrate Research
(1982)
L. De Vuyst et al.
Heteropolysaccharides from lactic acid bacteria
FEMS Microbiology Reviews
(1999)
J. Dou et al.
Purification, characterization and antioxidant activities of polysaccharides from thinned-young apple
International Journal of Biological Macromolecules
(2015)
R.P. Du et al.
Optimization, purification and structural characterization of a dextran produced by L. mesenteroides isolated from Chinese sauerkraut
Carbohydrate Polymers
(2017)
F. Feng et al.
Characterization of highly branched dextran produced by Leuconostoc citreum B-2 from pineapple fermented product
International Journal of Biological Macromolecules
(2018)
A.N. Hassan et al.
Microstructure and rheology of yogurt made with cultures differing only in their ability to produce exopolysaccharides
Journal of Dairy Science
(2003)
Y.L. He et al.
Purification, characterization and promoting effect on wound healing of an exopolysaccharide from Lachnum YM405
Carbohydrate Polymers
(2014)
I. Ibarburu et al.
Production and partial characterization of exopolysaccharides produced by two Lactobacillus suebicus strains isolated from cider
International Journal of Biological Macromolecules
(2015)

R. Inturri et al.

Chemical and biological properties of the novel exopolysaccharide produced by a probiotic strain of Bifidobacterium longum

Carbohydrate Polymers

(2017)

J.Y. Jiang et al.

Purification, structural characterization and in vitro antioxidant activity of a novel polysaccharide from Boshuzhi

Carbohydrate Polymers

(2016)

C.C. Li et al.

Microbiological, physicochemical and rheological properties of fermented soymilk produced with exopolysaccharide (EPS) producing lactic acid bacteria strains

LWT-Food Science and Technology

(2014)

J. Liu et al.

In vitro and in vivo antioxidant activity of exopolysaccharides from endophytic bacterium Paenibacillus polymyxa EJS-3

Carbohydrate Polymers

(2010)

Y. Liu et al.

Structural analysis and antioxidant activities of polysaccharide isolated from Jinqian mushroom

International Journal of Biological Macromolecules

(2014)

M. Miao et al.

Characterisation of a novel water-soluble polysaccharide from Leuconostoc citreum SK24.002

Food Hydrocolloids

(2014)

S.I. Moon et al.

Melt/solid polycondensation of l-lactic acid: An alternative route to poly (l-lactic acid) with high molecular weight

Polymer

(2001)

D.L. Qiao et al.

Extraction optimized by using response surface methodology, purification and preliminary characterization of polysaccharides from Hyriopsis cumingii

Carbohydrate Polymers

(2009)

Y. Qu et al.

Optimization of infrared-assisted extraction of Bletilla striata polysaccharides based on response surface methodology and their antioxidant activities

Carbohydrate Polymers

(2016)

C. Saravanan et al.

Isolation and characterization of exopolysaccharide from Leuconostoc lactis KC117496 isolated from idli batter

International Journal of Biological Macromolecules

(2016)

F.R. Seymour et al.

Determination of the structure of dextran by ¹³C-nuclear magnetic resonance spectroscopy

Carbohydrate Research

(1976)

F.R. Seymour et al.

Structural analysis of dextrans containing 4-O-α-D-glucosylated α-D-glucopyranosyl residues at the branch points, by use of ¹³C-nuclear magnetic resonance spectroscopy and gas-liquid chromatography-mass spectrometry

Carbohydrate Research

(1979)

L. Shao et al.

Partial characterization and immunostimulatory activity of exopolysaccharides from Lactobacillus rhamnosus KF5

Carbohydrate Polymers

(2014)

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¹: These authors contributed equally to this work and share the first authorship.

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Purification, characterization and antioxidant activity of dextran produced by Leuconostoc pseudomesenteroides from homemade wine

Highlights

Abstract

Introduction

Section snippets

Screening and identification of EPS-producing LAB

Isolation and identification of EPS-producing strain

Conclusion

Acknowledgements

Food Hydrocolloids

International Journal of Biological Macromolecules

Carbohydrate Research

FEMS Microbiology Reviews

International Journal of Biological Macromolecules

Carbohydrate Polymers

International Journal of Biological Macromolecules

Journal of Dairy Science

Carbohydrate Polymers

International Journal of Biological Macromolecules

Carbohydrate Polymers

Carbohydrate Polymers

LWT-Food Science and Technology

Carbohydrate Polymers

International Journal of Biological Macromolecules

Food Hydrocolloids

Polymer

Carbohydrate Polymers

Carbohydrate Polymers

International Journal of Biological Macromolecules

Carbohydrate Research

Carbohydrate Research

Carbohydrate Polymers