Stabilization of dextransucrase from Leuconostoc mesenteroides NRRL B-640

Purama, Ravi Kiran; Agrawal, Mayur; Goyal, Arun

doi:10.1007/s12088-010-0057-2

Cited by 11 publications

(11 citation statements)

References 17 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The addition of 1mM CaCl 2 , MgCl 2 , FeSO 4 and CoCl 2 to the DS caused an improvement of enzyme activity by 6, 8, 14 and 12%, respectively. The slight enhancement of DS activity achieved with the addition of divalent ions has been observed in other studies [31,36,39]. This effect has been attributed to the stabilization effect of the divalent ions on the tertiary enzyme structure [36,40].…”

Section: Effect Of Salts and Edta On Dextransucrase Activitysupporting

confidence: 58%

Characterization of dextransucrase from Leuconostoc mesenteroides T3, water kefir grains isolate

Miljkovic¹,

Davidovic²,

Kralj

et al. 2017

Hem Ind

View full text Add to dashboard Cite

The production of dextransucrase (DS) by Leuconostoc mesenteroides T3, novel isolate from water kefir grain, was studied and optimized. Bacterial supernatant reached activity of 3.1 U/ml when the culture was grown at 23 °C and under static culture condition using classical Tsuchiya medium for DS production. The increase of sucrose concentration to 7% led to an increase of DS activity by 52% compared to the control. Medium with 2% beef extract and 1% yeast extract resulted in 4.52 U/ml, which was 47% higher than in the control (with 2% yeast extract). Finally, the increase of K 2 HPO 4 concentration from 2 to 3% resulted in the increased enzyme activity by 28%. Enzyme purified by polyethylene glycol 400 fractionation displayed maximum activity at 30 °C and pH 5.4. Zymogram analysis confirmed the presence of DS of approximately 180 kDa. The addition of divalent cations Ca 2+ , Mg 2+ , Fe 2+ and Co 2+ led to a minor increase of DS activity, while the addition of Mn 2+was the most prominent with 73% increase. These findings classify dextransucrase from Leuconostoc mesenteroides T3 as promising candidate for production of dextran, which has numerous applications in various industries.Lactic acid bacteria (LAB) are one of the main classes of microorganisms that are known to produce several industrially important biomolecules among which exopolysaccharides (EPS) have the widest range of uses [1]. Fermentation processes for EPS production are quite expensive due to purification costs, giving an advantage to the use of enzymes. Current challenges in the LAB enzymes production include both strain improvement and enhancement of enzyme production.Dextransucrase (DS) is an extracellular enzyme that belongs to the class of glucosyltransferases [2] and it is mainly produced by microorganisms belonging to the families Lactobacillaceae and Streptococcaceae, especially by the genera Lactobacillus, Leuconostoc and Streptococcus [3]. Enzyme DS catalyzes the transfer reaction of glucosyl residues from sucrose to dextran polymer chain and releases fructose [4,5]. In the presence of appropriate acceptor (glucose, maltose, isomaltose, etc.) this enzyme can also produce oligosaccharides [6].

show abstract

Section: Effect Of Salts and Edta On Dextransucrase Activitysupporting

confidence: 58%

Characterization of dextransucrase from Leuconostoc mesenteroides T3, water kefir grains isolate

Miljkovic¹,

Davidovic²,

Kralj

et al. 2017

Hem Ind

View full text Add to dashboard Cite

show abstract

“…Aqueous solutions of dextran (100 kDa), PEG-8000, glycerol, Tween-80 were added to dextansucrase solution of wild type and mutant (0.24 mg/ml, 18 U/mg specific activity and 0.3 mg/ml, 18.2 U/mg) in sodium acetate buffer, pH 5.4 to obtain the final concentrations of dextran (100 kDa, 10 μg/ml), glycerol (0.5%, v/v ), PEG 8000 (10 μg/ml) and Tween 80 (0.5%, v/v ). The enzyme with or without any stabilizers was incubated at 30 °C for 24 h and 4 °C for 15 days (Purama et al 2010 ). The aliquots (20 μl) were taken at indicated time intervals for the enzyme assay.…”

Section: Methodsmentioning

confidence: 99%

Dextransucrase from the mutant of Pediococcus pentosaceus (PPm) is more stable than the wild type

et al. 2011

Self Cite

View full text Add to dashboard Cite

A comparative study on both wild type and mutant of Pediococcus pentosaceus for dextransucrase activity, its stability, dextran synthesizing activity, antibiotic sensitivity and carbohydrate utilization was performed. The wild type P. pentosaceus had specific activity of 0.58 U/mg whereas the mutant showed that of 1.0 U/mg with 72% enhancement. The antibiogram of 27 antibiotics tested against mutant showed significant differences with 9 antibiotics when compared to wild type. In carbohydrate fermentation profile, trehalose, galactose, maltose, lactose and fructose are metabolized by both the strains, but weakly in case of mutant. Stabilization of purified dextransucrase from wild type and mutant with various stabilizers was studied at 30 and 4 °C. Both enzymes were more stable at 4 °C. Among various stabilizers such as dextran (100 kDa, 10 μg/ml), glycerol (0.5%, v/v), PEG 8000 (10 μg/ml) and Tween 80 (0.5%, v/v), Tween 80 provided maximum stabilization at 4 and 30 °C. The mutant showed better stabilization than that of the wild type at both 30 and 4 °C. The loss of activity at 30 °C after 24 h in wild type and mutant in the presence of Tween 80 was only 34 and 32%, respectively, whereas the loss of activity in control of wild type and mutant was 76 and 59%, respectively. After 15 days at 4 °C, the loss of activity in control of wild type and mutant in the presence of Tween 80 was only 15 and 8%, respectively, whereas at 30 °C, the loss of activity in control of wild type and mutant was 49 and 42% respectively. Half-life of the enzyme with Tween 80 was 28.5 and 33.5 h for wild type and mutant, respectively, at 30 °C and 52.1 and 106.6 days for wild type and mutant respectively, at 4 °C.

show abstract

“…Such external factors as temperature, fermentation time, pH value and sucrose concentration of medium might affect the activity and types of GTFs, but the individual differences among the strains are still the most important in the type's formation of glucosyltransferases (Cortezi et al, 2005). Kobayashi & Matsuda found L. mesenteroides NRRL B-1299 (ATCC 11449) could produce a soluble and two insoluble GTFs (Purama et al, 2010). Most of alternansucrases produced by L. mesenteroides B-1355 were ectoenzymes (Cote & Leathers, 2009).…”

Section: Discussionmentioning

confidence: 99%

“…Glucosyltransferases have always been considered as extracellular enzymes (Purama et al, 2010). N-terminal of protein molecules of GTFs has typical Grampositive bacterium signal peptide, the role of which is to lead GTFs precursor into secretion system.…”

Section: Introductionmentioning

confidence: 99%

Effects of differences between cell‐free and cell‐associated glucosyltransferases from Leuconostoc mesenteroides on gluco‐oligosaccharides structure

Wang

Sun

Gao

et al. 2015

Int J of Food Sci Tech

View full text Add to dashboard Cite

The gluco-oligosaccharide structures depended largely upon the glucosyltransferase (GTF) differences. The oligosaccharides with four polymerisation degrees only was produced by Leuconostoc mesenteroides 6055L that contain more cell-free GTFs, while the gluco-oligosaccharides with two polymerisation degrees could be yielded by strains PC13 and L4 that have high cell-related GTFs. Also, more gluco-oligosaccharides with a-1, 6 glycosidic link as the backbone, accompanied by the a-1, 4 glycosidic link as a branch structure was formed in the presence of cell-free enzymes. The oligosaccharide with a-1, 3 glycosidic link that connects to all of other branch structures was observed in the presence of cell-related enzymes. To produce gluco-oligosaccharides with a-1, 3 glycosidic link, the L. mesenteroides strains with high proportion of cell-related GTFs should be recommended. This is the first time to discuss the effects of the differences between the cell-free and the cell-associated GTFs produced by L. mesenteroides on oligosaccharide structure.

show abstract

Stabilization of dextransucrase from Leuconostoc mesenteroides NRRL B-640

Cited by 11 publications

References 17 publications

Characterization of dextransucrase from Leuconostoc mesenteroides T3, water kefir grains isolate

Characterization of dextransucrase from Leuconostoc mesenteroides T3, water kefir grains isolate

Dextransucrase from the mutant of Pediococcus pentosaceus (PPm) is more stable than the wild type

Effects of differences between cell‐free and cell‐associated glucosyltransferases from Leuconostoc mesenteroides on gluco‐oligosaccharides structure

Contact Info

Product

Resources

About