Characterization of a novel lipase from Bacillus sp. isolated from tannery wastes

Kinetics of a lipase isolated from Bacillus sp. was studied. The enzyme showed maximum activity at pH 9 and temperature 60ºC. The Michaelis constant (K M 0.31 µM) obtained from three different plots i.e., Lineweaver-Burk, Hanes-Wolf and Hofstee, was found to be lower than already reported lipases that confirmed higher affinity of the enzyme for its substrate p-NPL (p-nitrophenyl laurate). Vmax of the enzyme was found to be 7.6 µM/mL/min. Energy of activation calculated from Arrhenius plot was found to be 20.607 kJmol-1. Activation enthalpy (ΔH*) had negative trend and the value for the hydrolysis of p-NPL by the enzyme at optimum temperature was -2.748 kJmol-1 . Activation entropy (ΔS*) and free energy of activation (ΔG*) of the enzyme were found to be 1.468 Jmol-1K-1 and -3.237 kJmol-1, respectively at optimum temperature. Low value of Q10 (0.04788) shows high catalytic activity of the enzyme. Mn2+, Fe2+ and Mg2+ enhanced the lipase activity whereas Cu2+, Na+ and Co2+ inhibited the enzyme activity. However, the enzyme activity was not affected significantly by K+ ions. EDTA and SDS also significantly inhibited the lipase activity. Activity of the enzyme was increased in n-hexane while decreased with increase in concentration of acetone, chloroform, ethanol and isopropanol.

Characterization of a novel lipase from Bacillus sp. isolated from tannery wastes.

Kinetics of a lipase isolated from Bacillus sp. was studied. The enzyme showed maximum activity at pH 9 and temperature 60°C. The Michaelis constant (KM 0.31 mM) obtained from three different plots i.e., Lineweaver-Burk, Hanes-Wolf and Hofstee, was found to be lower than already reported lipases that confirmed higher affinity of the enzyme for its substrate p-NPL (p-nitrophenyl laurate). Vmax of the enzyme was found to be 7.6 µM/mL/min. Energy of activation calculated from Arrhenius plot was found to be 20.607 kJmol(-1). Activation enthalpy (ΔH*) had negative trend and the value for the hydrolysis of p-NPL by the enzyme at optimum temperature was -2.748 kJmol(-1). Activation entropy (ΔS*) and free energy of activation (ΔG*) of the enzyme were found to be 1.468 Jmol(-1)K(-1) and -3.237 kJmol(-1), respectively at optimum temperature. Low value of Q10 (0.04788) shows high catalytic activity of the enzyme. Mn(2+), Fe(2+) and Mg(2+) enhanced the lipase activity whereas Cu(2+), Na(+) and Co(2+) inhibited the enzyme activity. However, the enzyme activity was not affected significantly by K(+) ions. EDTA and SDS also significantly inhibited the lipase activity. Activity of the enzyme was increased in n-hexane while decreased with increase in concentration of acetone, chloroform, ethanol and isopropanol.

Functional relationship of ammonia with RNA, DNA, and protein in rat muscle.

The acid proteinases, neutral proteinases, protein, RNA, and DNA were estimated along with the ammonia content in muscle in rats subjected to experimental hyperammonemic states (acute and chronic) by the administration of ammonium acetate intraperitoneally. It was observed that there was a decrease in the activities of acid proteinases and neutral proteinases in muscle in acute as well as in chronic hyperammonemic states. A rise in protein content was observed in muscle under these conditions. No changes were seen in the DNA content while RNA showed a slight increase in muscle. These results were discussed with reference to the possible role of ammonia in lysosomal protein degradation as well as to its probable effect on protein synthesis through its action on transcription.