Purification and biochemical characterization of a novel glucansucrase from Leuconostoc citreum B-2

Zhao, Bo; Du, Renpeng; Wang, Jingyue; Xu, Min; Han, Yongdian; Han, Xue

doi:10.1007/s10529-020-02881-6

Cited by 4 publications

(6 citation statements)

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“…Dextransucrase (Dsr) plays a decisive role in the structure of dextran. Studies show that Dsrs from different LABs have similar biochemical properties and structural characteristics [ 5 ]. However, these different Dsrs can form different glucan repeating units [ 6 ], which in turn affect the function of glucan.…”

Section: Introductionmentioning

confidence: 99%

Functional Identification of the Dextransucrase Gene of Leuconostoc mesenteroides DRP105

Zhou

2020

IJMS

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Leuconostoc mesenteroides DRP105 isolated from Chinese sauerkraut juice is an intensive producer of dextran. We report the complete genome sequence of Leu. mesenteroides DRP105. This strain contains a dextransucrase gene (dsr) involved in the production of dextran, possibly composed of glucose monomers. To explore the dextran synthesis mechanism of Leu. mesenteroides DRP105, we constructed a dsr-deficient strain derived from Leu. mesenteroides DRP105 using the Cre-loxP recombination system. The secondary structure prediction results showed that Leu. mesenteroides DRP105 dextransucrase (Dsr) was coded by dsr and contained 17.07% α-helices, 29.55% β-sheets, 10.18% β-turns, and 43.20% random coils. We also analyzed the dextran yield, monosaccharide change, organic acid, and amino-acid content of Leu. mesenteroides DRP105 and Leu. mesenteroides DRP105−Δdsr. The result showed that the lack of dsr changed the Leu. mesenteroides DRP105 sugar metabolism pathway, which in turn affected the production of metabolites.

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Section: Introductionmentioning

confidence: 99%

Functional Identification of the Dextransucrase Gene of Leuconostoc mesenteroides DRP105

Zhou

2020

IJMS

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“…A co‐pigmentation effect of lactic acid formed during fermentation on anthocyanins in BCJ was therefore investigated in this study and this anthocyanin stabilizing and/or color‐enhancement effect was revealed for the first time. Moreover, possible effects of exopolysaccharides such as kefiran, glucan, and cycloamylose on color and antioxidant activity of FBs were also discussed for the first time in the light of the data obtained from our study and previous studies in the literature 9‐13 …”

Section: Introductionmentioning

confidence: 53%

“…Thus, lower sucrose content in these samples could not have resulted from the invertase activity of yeasts. As LAB can use sucrose to produce a glucan by extracellular glucansucrase, 11 and the activity of this enzyme increases with the increase in pH from 3.0 to 5.0, 12 the main reason for the lowest sucrose content in the FBs at pH 4.50 and 4.35 could be the sucrose metabolisms of Lactobacillus spp. Moreover, the sucrose metabolism of LAB in the FBs also explained why small differences in the content of glucose and fructose in the samples were found despite large differences in the sucrose content of FBs.…”

Section: Resultsmentioning

confidence: 99%

“…The most important metabolites produced by kefir culture are lactic acid, exopolysaccharides (especially kefiran, glucan, and cycloamylose), acetic acid, glycerol, ethyl acetate, isoamyl acetate, ethyl hexanoate, ethyl octanoate, ethyl decanoate, isopentyl acetate, and phenethyl acetate. [9][10][11][12][13] Among these metabolites, lactic acid, exopolysaccharides, glucan and cycloamylose, in particular, can show significant effects on anthocyanins and other properties (such as texture, consistency, flavor and antioxidant activity) of FBs because of variable functional groups (e.g., hydroxyl and carboxyl) in their chemical structures. A co-pigmentation effect of lactic acid formed during fermentation on anthocyanins in BCJ was therefore investigated in this study and this anthocyanin stabilizing and/or color-enhancement effect was revealed for the first time.…”

Section: Introductionmentioning

confidence: 99%

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Effects of fermentation time and pH on quality of black carrot juice fermented by kefir culture during storage

2021

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BACKGROUND The effects of fermentation time (17–48 h) and pH (3.37–4.50) on qualities of fermented beverages (FBs) produced from black carrot juice (BCJ) were monitored during storage at 4 °C for 20 weeks. RESULTS Fermentation and adjusting the pH level provided significant increases (up to 22%) in anthocyanin content and the absorbance value at λmax (Amax). Moreover, the stability of anthocyanins, color density, and Amax in FBs was somewhat higher than those in BCJ. Lactic acid showed a co‐pigmentation effect on cyanidin‐3‐galactoside‐xyloside‐glucoside‐sinapic acid and cyanidin‐3‐galactoside‐xyloside‐glucoside‐ferulic acid. Sucrose was degraded much faster at pH 4.50 (17 h) and 4.35 (48 h) than at lower pH levels. During storage, pH 4.35 caused a balanced distribution between counts of lactic acid bacteria (LAB) and yeasts, and antioxidant activity of all FBs increased. Fermented beverages at 4.35 and 3.90 were found to be more palatable by panelists. CONCLUSION We recommend FB production at pH 4.35 after 48 h fermentation due to the balanced distribution of probiotics, high color enhancement, and consumer preference. © 2021 Society of Chemical Industry.

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“…At the same time, there were no bands other than the target bands ( Figure 2), indicating that the purification with GEM particles was sufficient. In this experiment, the GEM purification method we choose saves the purification time and reduces the purification steps compared with the traditional Ni column purification method [40]. The traditional Ni column purification method requires the use of a large amount of imidazole to elute the enzyme from the Ni column [41].…”

Section: Expression and Purification Of Recombinant Fructan Sucrasementioning

confidence: 99%

A Fructan Sucrase Secreted Extracellular and Purified in One-Step by Gram-Positive Enhancer Matrix Particles

et al. 2021

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Fructan sucrase is a kind of biological enzyme that catalyzes the synthesis of fructan, and fructan is a polysaccharide product with important industrial application value. In this study, the Fructan sucrase gene of Bacillus subtilis was cloned to plasmid PET-28A-ACMA-Z, and three clones were obtained after the transformation of Escherichia coli BL21, namely BS-FF, BSO, and BS. The clones BS-FF and BSO secreted the recombinant enzymes outside the cells, while the clone BS expressed them inside the cells. The induction experiment results showed that the optimum IPTG concentration in the medium was 0.5 mM and 1.0 mM for clones BS-FF and BSO, respectively, while the incubation conditions were at 28 °C for 8 h. The recombinant fructan sucrase was purified one step using a material called GEM particles. The results indicated that 95.25% of fructan sucrase expressed by the clone BS-FF could be secreted into the extracellular area, and even 98.78% by the clone BSO. With the above purification system, the receiving rate of the recombinant enzyme for clones BS-FF and BSO was 97.70% and 84.99%, respectively. As for the bioactivity of recombinant fructan sucrase, the optimum temperature and pH were 50 °C and 5.6, respectively. The Km and Vmax of it were 33.96 g/L and 0.63 g/(L·min), respectively. The engineered strains with the high extracellular secretion of fructan sucrase were constructed, and a one-step method for the purification of the recombinant enzyme was established. The results might provide a novel selection for the enzymatic production of fructan on a large scale.

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Purification and biochemical characterization of a novel glucansucrase from Leuconostoc citreum B-2

Cited by 4 publications

References 40 publications

Functional Identification of the Dextransucrase Gene of Leuconostoc mesenteroides DRP105

Functional Identification of the Dextransucrase Gene of Leuconostoc mesenteroides DRP105

Effects of fermentation time and pH on quality of black carrot juice fermented by kefir culture during storage

A Fructan Sucrase Secreted Extracellular and Purified in One-Step by Gram-Positive Enhancer Matrix Particles

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