Pineapple Peel Extract as an Effective Substrate for Esterase Production from Bacillus subtilis E9

Esterase, belonging to hydrolase class of enzymes catalyzes the cleavage and formation of ester bonds. Esterase producing isolates E9 and E46, isolated from pineapple waste enriched soil were identified as Bacillus subtilis E9 and Bacillus sp. E46 respectively. Bacillus subtilis E9 with 10 U/mg esterase activity in basal media was further chosen for media optimization studies. Several factors including the effect of organic solvents and fruit peel extracts were studied by one factor at a time optimization method and statistical models. An enhanced enzyme production of 250.50 U/mg could be obtained under the optimal conditions of pH 6.5, incubation time 25 h and 1.8%v/v of acetone extract of pineapple peel. The four-stage purification improved the purity of the enzyme by 1.5-fold with 5.3% recovery and specific activity of 384 U/mg. The monomeric nature and the molecular weight (45 KDa) of the enzyme were determined by performing SDS PAGE and its activity was confirmed by zymogram analysis. The substrate specificity of the purified fraction exhibited a higher activity towards lower chain length esters, indicating the enzyme as esterase. The partially purified esterase showed an optimal temperature of 40 °C at an optimum pH of 7. K_m and V_max of the enzyme were 1.12 mM and 1.18 mM of released pNP · min^-1 respectively.