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Protection of lactoperoxidase activity with sugars during lyophilization and evaluation of its antibacterial properties.

Shariat SS, Jafari N, Tavakoli N, Najafi RB - Res Pharm Sci (2015 Mar-Apr)

Bottom Line: Combinations of disaccharides resulted in an increment in the stability of the enzyme, compared to the native enzyme.Combination of 200 mM trehalose and 200 mM sucrose were found most effective cryoprotectant in freeze-drying of LP.Antimicrobial efficacy of LP was more pronounced when 10(5) cfu/ml was used as compared to 10(6) cfu/ml.

View Article: PubMed Central - PubMed

Affiliation: Department of Clinical Biochemistry, School of Pharmacy and Pharmaceutical Sciences, Isfahan University of Medical Sciences, Isfahan, I.R. Iran.

ABSTRACT
The purpose of the present study was to compare the stabilizing effect of four disaccharides alone or in combination on the lactoperoxidase (LP) derived from bovine milk during lyophilization. Sucrose, lactose, maltose, and trehalose at different concentrations (5-500 mM) were used to compare their protective effects on LP activity. The activity of lyophilized and native LP enzyme was evaluated using the procedure of Schindler with slight modifications. The antibacterial activity of the lyophilized enzyme against Pseudomonas aeroginosa, Escherichia coli, and Staphylococcus aureus was also investigated using the antimicrobial effectiveness test. Trehalose at concentration of 500 mM was the most effective cryoprotectant in protecting the enzyme activity. It preserved LP activity for 40 days, while the native enzyme lost its activity after 6 days. Combinations of disaccharides resulted in an increment in the stability of the enzyme, compared to the native enzyme. Combination of 200 mM trehalose and 200 mM sucrose were found most effective cryoprotectant in freeze-drying of LP. The lyophilized LP decreased the growth rate of Ps.aeroginosa, E.coli, and S.aureus between up to 30.8% in 10(6) cfu/ml and 53.3% in 10(5) cfu/ml. Antimicrobial efficacy of LP was more pronounced when 10(5) cfu/ml was used as compared to 10(6) cfu/ml.

No MeSH data available.


Related in: MedlinePlus

The effect of different concentrations of trehalose on the stability of lactoperoxidase during 40 days after freeze-drying (n=3, mean ± SD).
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Figure 2: The effect of different concentrations of trehalose on the stability of lactoperoxidase during 40 days after freeze-drying (n=3, mean ± SD).

Mentions: For instance, sucrose (Fig. 1) at 300 mM and 500 mM concentrations showed similar effect on enzyme activity but increasing the concentrations from 100 to 300 mM improved the recovered activity considerably. Trehalose (Fig. 2) was found to be the most effective sugar in protecting the enzyme activity and at 500 mM concentration, 91 % of the original activity was recovered in the first day of lyophilization lasting for more than 41 days. Maltose (Fig. 3) and lactose (Fig. 4) showed similar effects on LP activitities. These disaccharides showed good stability at 100 and 300 mM concentrations on the first day of lyophilization but the activities were decreased profoundly after few days, whereas trehalose and sucrose at concentration of 100 mM could retain the LP activity for more than 35 days.


Protection of lactoperoxidase activity with sugars during lyophilization and evaluation of its antibacterial properties.

Shariat SS, Jafari N, Tavakoli N, Najafi RB - Res Pharm Sci (2015 Mar-Apr)

The effect of different concentrations of trehalose on the stability of lactoperoxidase during 40 days after freeze-drying (n=3, mean ± SD).
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4584454&req=5

Figure 2: The effect of different concentrations of trehalose on the stability of lactoperoxidase during 40 days after freeze-drying (n=3, mean ± SD).
Mentions: For instance, sucrose (Fig. 1) at 300 mM and 500 mM concentrations showed similar effect on enzyme activity but increasing the concentrations from 100 to 300 mM improved the recovered activity considerably. Trehalose (Fig. 2) was found to be the most effective sugar in protecting the enzyme activity and at 500 mM concentration, 91 % of the original activity was recovered in the first day of lyophilization lasting for more than 41 days. Maltose (Fig. 3) and lactose (Fig. 4) showed similar effects on LP activitities. These disaccharides showed good stability at 100 and 300 mM concentrations on the first day of lyophilization but the activities were decreased profoundly after few days, whereas trehalose and sucrose at concentration of 100 mM could retain the LP activity for more than 35 days.

Bottom Line: Combinations of disaccharides resulted in an increment in the stability of the enzyme, compared to the native enzyme.Combination of 200 mM trehalose and 200 mM sucrose were found most effective cryoprotectant in freeze-drying of LP.Antimicrobial efficacy of LP was more pronounced when 10(5) cfu/ml was used as compared to 10(6) cfu/ml.

View Article: PubMed Central - PubMed

Affiliation: Department of Clinical Biochemistry, School of Pharmacy and Pharmaceutical Sciences, Isfahan University of Medical Sciences, Isfahan, I.R. Iran.

ABSTRACT
The purpose of the present study was to compare the stabilizing effect of four disaccharides alone or in combination on the lactoperoxidase (LP) derived from bovine milk during lyophilization. Sucrose, lactose, maltose, and trehalose at different concentrations (5-500 mM) were used to compare their protective effects on LP activity. The activity of lyophilized and native LP enzyme was evaluated using the procedure of Schindler with slight modifications. The antibacterial activity of the lyophilized enzyme against Pseudomonas aeroginosa, Escherichia coli, and Staphylococcus aureus was also investigated using the antimicrobial effectiveness test. Trehalose at concentration of 500 mM was the most effective cryoprotectant in protecting the enzyme activity. It preserved LP activity for 40 days, while the native enzyme lost its activity after 6 days. Combinations of disaccharides resulted in an increment in the stability of the enzyme, compared to the native enzyme. Combination of 200 mM trehalose and 200 mM sucrose were found most effective cryoprotectant in freeze-drying of LP. The lyophilized LP decreased the growth rate of Ps.aeroginosa, E.coli, and S.aureus between up to 30.8% in 10(6) cfu/ml and 53.3% in 10(5) cfu/ml. Antimicrobial efficacy of LP was more pronounced when 10(5) cfu/ml was used as compared to 10(6) cfu/ml.

No MeSH data available.


Related in: MedlinePlus