NDLI: Isolation, purification and functional characterization of glucansucrase from probiotic Lactobacillus plantarum DM5

Content Provider	Springer Nature Link
Author	Das, Deeplina Goyal, Arun
Copyright Year	2014
Abstract	The probiotic Lactobacillus plantarum DM5 isolated from fermented beverage Marcha of Sikkim was explored for its ability to produce extracellular glucansucrase during fermentation. The strain L. plantarum DM5 displayed glucansucrase activity of 2.71 U/ml (0.48 U/mg) at 27 °C under static condition. The medium compositions for glucansucrase production were optimized and it was found that K$_{2}$HPO$_{4}$ (2.5 %, w/v), yeast extract (2.5 %, w/v), Tween 80 (0.6 %, v/v), and sucrose (5 %, w/v) enhanced the activity by 20 %, 22 %, 68 % and 230 %, respectively. Glucansucrase was purified using polyethylene glycol 400 and 1500 fractionation followed by gel filtration that gave specific activity of 18.7 U/mg with 40-fold purification. Purified enzyme exhibited maximum activity at 30 °C and pH 5.4. Zymogram analysis of purified enzyme confirmed the presence of glucosyltransferase of approximately 148 kDa. The V$_{m}$ and K$_{m}$ of purified glucansucrase for sucrose as substrate was 19.6 μmoles/mg/min and 4.5 mM. Divalent cations Mg$^{2+}$, Ca$^{2+}$ and Co$^{2+}$ enhanced glucansucrase activity by 16 %, 18 % and 19 %, respectively, whereas Hg$^{2+}$ and Mn$^{2+}$ decreased the enzyme activity by 81 % and 79 %, respectively, when assayed in presence of sucrose. Among stabilizers, dextran T-40, PEG 6000, PEG 8000, glutaraldehyde, glycerol, Tween 80 and acetonitrile; Tween 80 provided maximum stabilization with half-life of 86 days at –20 °C. The overall biochemical characterization reveals a promising novel glucansucrase that can compensate for the increasing demand of glucan as viscosifier and stabilizer in food industry.
Starting Page	1715
Ending Page	1724
Page Count	10
File Format	PDF
ISSN	15904261
Journal	Annals of Microbiology
Volume Number	64
Issue Number	4
e-ISSN	18692044
Language	English
Publisher	Springer Berlin Heidelberg
Publisher Date	2014-01-28
Publisher Place	Berlin, Heidelberg
Access Restriction	Subscribed
Subject Keyword	Lactobacillus plantarum Probiotic Glucansucrase Glucan Marcha Tween 80 Microbiology Microbial Genetics and Genomics Microbial Ecology Fungus Genetics Medical Microbiology Applied Microbiology
Content Type	Text
Resource Type	Article
Subject	Applied Microbiology and Biotechnology

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