Accumulation of a recombinant Aspergillus oryzae lipase artificially localized on the Bacillus subtilis cell surface

cutL cDNA encoding an extracellular lipase, L1, from Aspergillus oryzae was fused to the cell wall-binding domain (CWB) region of a plasmid, pHCB3R. S DS-polyacrylamide gel electrophoresis (PAGE) and zymography of proteins ext racted from the cell surface of Bacillus subtilis 168 harboring a fused lip ase plasmid (pHCB3RCL) revealed that the fused gene product, CWB-CutL, was localized in the B. subtilis cell wall and retained Lipase activity. B, sub tilis WASD (wprA sigD), recently used for the accumulation of CWB-LipB (the CWB protein fused with B. subtilis lipase B), was also a suitable host for the accumulation of CWB-CutL, the amount being 10% of the total proteins e xtracted from the cell surface.