Function of the prosequence for in vivo folding and secretion of active Rhizopus oryzae lipase in Saccharomyces cerevisiae

Citation
S. Takahashi et al., Function of the prosequence for in vivo folding and secretion of active Rhizopus oryzae lipase in Saccharomyces cerevisiae, APPL MICR B, 55(4), 2001, pp. 454-462
Citations number
42
Language
INGLESE
art.tipo
Article
Categorie Soggetti
Biotecnology & Applied Microbiology",Microbiology
Journal title
APPLIED MICROBIOLOGY AND BIOTECHNOLOGY
ISSN journal
0175-7598 → ACNP
Volume
55
Issue
4
Year of publication
2001
Pages
454 - 462
Database
ISI
SICI code
0175-7598(200105)55:4<454:FOTPFI>2.0.ZU;2-M
Abstract
The role of the prosequence of Rhizopus oryzae lipase (ROL) with a preprose quence was analyzed by an expression system using Saccharomyces cerevisiae. When the mature portion of ROL (mROL) fused to the pre-alpha -factor leade r sequence was expressed, secretion of active mROL was not observed. Howeve r, when mROL was synthesized together with the prosequence in trans (indivi dually and coincidentally), secretion of active mROL was observed. The resu lts indicate that the prosequence of ROL helped correct folding of mROL and its subsequent secretion from the yeast cells, and that physical linkage ( cis) of the prosequence to the mature region was not prerequisite. From the expression of the ROL mutants with deletions at the N-terminal end of the prosequence together with mROL in trans, the residues from 20 to 37 in the prosequence were essential for the secretion, and those from 38 to 57 were essential for the formation of the active ROL and might play a role as an i ntramolecular chaperone. The results using the fragment of the prosequence confirmed that these residues (20-57) were significant for in vivo folding and secretion of active mROL.