Generation of a Functionally Distinct Rhizopus oryzae Lipase through Protein Folding Memory.

Rhizopus oryzae lipase (ROL) has a propeptide at its N-terminus that functions as an intramolecular chaperone and facilitates the folding of mature ROL (mROL). In this study, we successfully generated a functionally distinct imprinted mROL (mROL^imp) through protein folding memory using a mutated propeptide. The mutated propeptide left its structural memory on mROL and produced mROL^imp that exhibited different substrate specificities compared with mROL^WT (prepared from the wild type propeptide), although the amino acid sequences of both mROLs were the same. mROL^imp showed a preference for substrates with medium chain-length acyl groups and, noticeably, recognized a peptidase-specific substrate. In addition, ROL^imp was more stable than mROL^WT. These results strongly suggest that proteins with identical amino acid sequences can fold into different conformations and that mutations in intramolecular chaperones can dynamically induce changes in enzymatic activity. [ABSTRACT FROM AUTHOR]