Probing role of key residues in the divergent evolution of Yarrowia lipolytica lipase 2 and Aspergillus niger eruloyl esterase A

Abstract

Yarrowia lipolytica lipase 2 (YLLip2) and Aspergillus niger feruloyl esterase A (AnFaeA) are enzymes of similar structures but with different functions. They are both classified into the same homologous family in Lipase Engineering Database (LED). The major difference between the two enzymes is that YLLip2 exhibits interfacial activity while AnFaeA does not. In order to better understand the interfacial activation mechanisms of YLLip2, structure guided site-directed mutagenesis were performed, mutants were constructed, kinetics parameters and lipase properties were detected. Mutant enzymes showed enhanced catalytic efficiency towards p-nitrophenyl butyrin (pNPB) but their catalytic efficiency decreased towards p-nitrophenyl palmitate (pNPP), their catalysis behavior was more close to feruloyl esterase. Moreover, the mutant enzymes exhibited enhanced thermostability compared with their wild type. These results indicate that I100 and F129 are probably cut-off point of divergent functions between the two enzymes during evolution.

The effect of mutations in the lid region of Thermomyces lanuginosus lipase on interactions with triglyceride surfaces: A multi-scale simulation study

“…Existing experimental data suggest that amino acid substitutions within the lid region of the TL lipase influence interfacial activation and catalytic activity (Skjold-Jorgensen et al, 2016;Skjold-Jørgensen et al, 2016. Other studies of different lipases have demonstrated similar effects, highlighting the critical role of the lid region in enzyme activity and substrate specificity (Secundo et al, 2006;Wang et al, 2015;Yu et al, 2014). Here, CG and AT simulations were used to investigate how mutation of the lid region of TLL affects interfacial interactions with a natural substrate, as well as conformational dynamics at the interface and in solution.…”

Isolation, identification and characterization of soil bacteria for the production of ferulic acid through co-culture fermentation using banana stem waste

“…Feruloyl esterase or ferulic acid esterase (FAE) is known as the most important enzyme to cleave the feruloyl-polysaccharide. On the other hand, several enzymes have been observed to facilitate the production of FA [7,8].…”