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Objective: Although the extracellular polysaccharides have been analyzed in the previous period, the biochemical, enzymological characters and stimulation and inhibition effect on glucansucrase are not fully understood., Results: After three steps purification, salting out, DEAE-Sepharose and Sephadex G-75, the final specific activity was 264.84 U/mg protein with 4.31-fold. The SDS-PAGE analysis of fraction gave a single band 170.35 kDa in the stained gel. The active band was analyzed with LC-MS/MS to identify glucansucrase. The highest coverage rate of dextransucrase from Leu. citreum (ACY92456.2) was 55.60%, the results were speculated that the glucansucrase secreted from Leu. citreum B-2 may be a novel glucansucrase. The purified enzyme was optimally active at 20-30 °C and pH 6.0-8.0. Metal ions K + , Na + , Ca 2+ , Mn 2+ , Mg 2+ , and Cr + had an apparent stimulating effect on enzyme activity, especially in divalent ions Ca 2+ and Mn 2+ , the residual activities were higher than 200%. In a reverse, Hg + , acetonitrile, SDS, salt, and guanidine expressed inhibition effect on enzyme residual activity. The K M and V max were detected to be 4.82 mM and 0.97 U/mg, respectively., Conclusion: All these data collectively indicate that B-2 glucansucrase is a novel one, which have good properties and may applied to new food areas.

Sugarcane molasses is an agricultural by-product containing sucrose. In this study, the exopolysaccharide (M-EPS) produced by Leuconostoc citreum B-2 in molasses-based medium was characterized, optimized, and its application in set yogurt was investigated. The structure analysis, including gel permeation chromatography, Fourier transform infrared spectroscopy, and nuclear magnetic resonance, revealed that the M-EPS was a linear dextran composed of D-glucose units, which were linked by α-(1→6) glycosidic bonds with 19.3% α-(1→3) branches. The M-EPS showed a lower molecular weight than that produced from sucrose. The M-EPS was added into the set yogurt, and then the water holding capacity, pH, and microstructure of set yogurt were evaluated. Compared with the controls, the addition of M-EPS improved the water holding capacity and reduced the pH of set yogurt. Meanwhile, the structure of the three-dimensional network was also observed in the set yogurt containing M-EPS, indicating that M-EPS had a positive effect on the stability of set yogurt. The results provide a theoretical basis for the cost-effective utilization of sugarcane molasses. [ABSTRACT FROM AUTHOR]

Exopolysaccharide from Leuconostoc pseudomesenteroides XG5 (XG5 EPS) is a linear dextran that is built by glucose units via α-1,6 glycosidic bond. The primary objective of this study was to investigate the yield of XG5 EPS and its application in set yogurt. In laboratory scale, the culture conditions of XG5 EPS production were optimized using the L9 (33) orthogonal test. Here, the optimized yield of XG5 EPS was 26.02 g/L under the conditions of 100 g/L sucrose, initial pH 7.0, 25°C incubation, and 100 rpm for 36 h in a shaking flask. Based on the optimized parameters of laboratory scale, a pilot fed-batch fermentation was performed in a 50-L bioreactor with an adjusted agitation speed of 20 rpm. The XG5 EPS yield reached 40.07 g/L in fed-batch fermentation, which was 54% higher than that achieved in laboratory scale. In addition, XG5 EPS was added into set yogurt to investigate its effect on the stability of set yogurt. Our data demonstrated that the XG5 EPS improved the water-holding capacity, texture profile, and viscosity of set yogurt during cold storage compared with the controls. In particular, addition of 0.5% XG5 EPS increased the structure of 3-dimensional network of set yogurt, which eventually improved the physical stability of the set yogurt. Overall, this study provided new insights for exploring the pilot scale production and application of dextran. [Display omitted] [ABSTRACT FROM AUTHOR]