Generation of a functionally distinct Rhizopus oryzae lipase through protein folding memory

Abstract

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.