RESUMO
Objective: To evaluate the neutralizing effects of flavonoids on snake venom toxicity by stand-alone and combinatorial approaches. Methods: Synthetic flavonoids were assessed, either individually or in combination with antivenom, for their neutralization of phospholipase A
RESUMO
Objective: To evaluate the neutralizing effects of flavonoids on snake venom toxicity by stand-alone and combinatorial approaches. Methods: Synthetic flavonoids were assessed, either individually or in combination with antivenom, for their neutralization of phospholipase A
RESUMO
Background: It was previously shown that the activity of a serine protease from a moderately halotolerant Bacillus aquimaris VITP4 strain is active in a wide range of pH and temperatures and could be modulated by the presence of the divalent metal ions
Objectives: In the present study, a quantitative analysis was done in order to explore the parameters that are contributing to the protease activity
Materials and Methods: Changes in the secondary structure of the enzyme was determined by circular dichroism analysis. The conditions for the optimal activity was investigated by Response Surface Methodology. Stability of the enzyme was determined by thermal inactivation experiments
Results: The initial one-factor-at-a-time experiments have indicated that the activity of the enzyme could be enhanced not only by the presence of low concentrations of NaCl but also by divalent metal ions, such as Ca[2+], Mn[2+] and Cu[2+]. A clear dependence of the activity to the secondary structure of the enzyme could be established using circular dichroism spectroscopy. In the next level of optimization, four factors; viz. pH, temperature, concentration of Ca[2+], and Mn[2+] were used to optimize the conditions required for the maximal activity of the enzyme by Response Surface Methodology, and the data could be explained using quadratic model. Under optimal condition of 43[degree]C, pH 8.0, 8.2 mM Ca[2+], and 4.3 mM Mn[2+] a 1.5 times enhancement in the enzyme activity could be achieved. The storage stability of the enzyme under these selected conditions has indicated a non-linear relation between the conditions for the enzymatic activity as well as stability. However, the condition for the maximal stability [267+/-18 min] has corresponded to that of the optimal conditions for the maximal activity
Conclusions: This study, for the first time, has explored the possibility of using statistical methods for identifying the optimal conditions for alkaline protease activity isolated from the halotolerant Bacillus aquimaris VITP4