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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 the lactoperoxidase system against E.coli in Muller-Hinton broth.
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Figure 5: The effect of the lactoperoxidase system against E.coli in Muller-Hinton broth.

Mentions: The microbiological effect of LP-S on the E.coli growth rate is shown in Fig. 5. Absorbances of samples in the presence and absence of LP-S at 580 nm after inoculation during 20 h are shown in this figure. Table 1 shows the percent reduction in the absorbances at 6 h time point in LP and non-LP containing tubes to show the effect of LP in delaying the growth rate of three microorganisms. It shows that the overall effect of LP in 105 cfu/ml suspension is greater than 106 cfu/ml one. As a result, the lower the initial inoculums, the higher the enzyme activity would be.


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 the lactoperoxidase system against E.coli in Muller-Hinton broth.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

Figure 5: The effect of the lactoperoxidase system against E.coli in Muller-Hinton broth.
Mentions: The microbiological effect of LP-S on the E.coli growth rate is shown in Fig. 5. Absorbances of samples in the presence and absence of LP-S at 580 nm after inoculation during 20 h are shown in this figure. Table 1 shows the percent reduction in the absorbances at 6 h time point in LP and non-LP containing tubes to show the effect of LP in delaying the growth rate of three microorganisms. It shows that the overall effect of LP in 105 cfu/ml suspension is greater than 106 cfu/ml one. As a result, the lower the initial inoculums, the higher the enzyme activity would be.

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