Stabilization of Lysozyme by Benzyl Alcohol: Surface Tension and Thermodynamic Parameters Monu Kumari Goyal, Ipsita Roy, Aeshna Amin, Uttam Chand Banerjee,

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Presentation transcript:

Stabilization of Lysozyme by Benzyl Alcohol: Surface Tension and Thermodynamic Parameters Monu Kumari Goyal, Ipsita Roy, Aeshna Amin, Uttam Chand Banerjee, Arvind Kumar Bansal Journal of Pharmaceutical Sciences Volume 99, Issue 10, Pages 4149-4161 (October 2010) DOI: 10.1002/jps.22129 Copyright © 2010 Wiley-Liss, Inc. Terms and Conditions

Figure 1 Enzyme activity oflysozyme (5 mgmL−1) in the presence of preservatives at 75°C; phenol (A), benzalkonium chloride (BKC) (B), benzyl alcohol (BA) (C) (reproduced from Goyal et al.8) and m-cresol (D) (data are represented as Mean ± SD, n = 3). Samples were withdrawn at regular interval of 20 min up to 2 h. Journal of Pharmaceutical Sciences 2010 99, 4149-4161DOI: (10.1002/jps.22129) Copyright © 2010 Wiley-Liss, Inc. Terms and Conditions

Figure 2 Effect of pH on mean particle size (A) and turbidity measurements (B) of lysozyme (5 mgmL−1) heated at 75°C in the presence of 2% benzyl alcohol. A: Inset shows the expanded view of mean particle size at pH 9.0 and pH 7.0 (data are represented as Mean ± SD, n = 3). Control sample precipitated at higher pH and clogged the 100 nm filters hence particle size was not measured for control sample at higher pH (pH 7 and 9). B: The turbidity was measured at 600 nm as a function of time incubation. (Data are represented as Mean ± SD, n = 3.) Journal of Pharmaceutical Sciences 2010 99, 4149-4161DOI: (10.1002/jps.22129) Copyright © 2010 Wiley-Liss, Inc. Terms and Conditions

Figure 3 Effect of benzyl alcohol on surface tension of water (A) and protein solution (B) using drop shape analysis method at room temperature (data are represented as Mean ± SD, n = 6). Journal of Pharmaceutical Sciences 2010 99, 4149-4161DOI: (10.1002/jps.22129) Copyright © 2010 Wiley-Liss, Inc. Terms and Conditions

Figure 4 Effect of benzyl alcohol on guanidine hydrochloride induced unfolding of lysozyme at room temperature. All the samples were equilibrated overnight at room temperature in the presence of guanidine hydrochloride (0–7 M). Journal of Pharmaceutical Sciences 2010 99, 4149-4161DOI: (10.1002/jps.22129) Copyright © 2010 Wiley-Liss, Inc. Terms and Conditions

Figure 5 Representative data for fractions unfolded (fu) of lysozyme in the presence of benzyl alcohol as a function of temperature calculated from data obtained using circular dichroism. Each transition curve was measured three times and converted into fractions of unfolded protein using the Eq. (2). Journal of Pharmaceutical Sciences 2010 99, 4149-4161DOI: (10.1002/jps.22129) Copyright © 2010 Wiley-Liss, Inc. Terms and Conditions

Figure 6 Schematic representation of mechanism of interaction of lysozyme and benzyl alcohol. Journal of Pharmaceutical Sciences 2010 99, 4149-4161DOI: (10.1002/jps.22129) Copyright © 2010 Wiley-Liss, Inc. Terms and Conditions