Mechanism of inhibition of rice bran lipase by polyphenols: a case study with chlorogenic acid and caffeic acid

ABSTRACT:  Rice bran is a byproduct obtained from the rice milling industry, and to arrest lipolysis caused by lipolytic enzyme, rice bran lipase (RBL) was inactivated by inhibitors such as polyphenols. This study describes the inhibition and interaction of enzyme with chlorogenic acid (CGA) and caffeic acid (CA). The inhibition of the enzyme was competitive in nature in both CGA and CA. The inhibition constant K _i of the reaction was found to be 1.8 and 1.5 μM for CGA and CA, respectively. Fluorescence emission measurements indicated a decrease in the fluorescence emission intensity and a red shift in the emission maximum as these ligands concentrations are increased, indicating the minor changes in the tryptophan environment and the effect of binding that is stronger in the case of CA compared to CGA with RBL. Far UV-circular dichroic data suggest that there are no significant changes in the conformation of the enzyme as a result of binding of CGA or CA. The instability of the enzyme in the presence of these polyphenols has been indicated by decrease in apparent thermal transition temperatures of the enzyme from a control value of 60 °C as revealed by thermal denaturation measurements. These results demonstrate that both CGA and CA are inhibitors of RBL and bind to the enzyme through both hydrogen and hydrophobic interaction in bringing about inhibition with minor structural alterations. These inactivation phenomena of polyphenols that act as inhibitors on RBL can be utilized to prevent oxidation of the rice bran oil.