CLONING AND SEQUENCING OF THE CDNA-ENCODING LIPASE-I FROM TRICHOSPORON FERMENTANS WU-C-12

A cDNA clone encoding extracellular lipase I (TFL I) from Trichosporon fermantans WU-C12 was isolated and characterized. The TFL I cDNA was isolated from a lambda gt10-based cDNA library using as a probe a 0.8 kb fragment, amplified by PCR with synthetic oligonucleotide correspon ding to the partial amino acid sequences of TFL I. The cDNA encodes a protein consisting of 563 amino acids containing a putative signal pep tide of 19 amino acids. The deduced amino acid sequence shares 99.5% o verall identity with that of lipase II (GCL II) from Geotrichum candid um ATCC 34614, whereas TFL I is a trimer enzyme and GCL II monomer. So uthern hybridization with the TFL I cDNA as a probe revealed that WU-C 12 contained two different lipase genes.