Cloning and sequencing of two chromosomal lipase genes from Geotrichum candidum.

Two chromosomal lipase genes (lip1 and lip2 encoding lipases I and II, respectively) were cloned from Geotrichum candidum by colony hybridization using lipase cDNAs as probes, and their nucleotide sequences were determined. Both genes contained 5'- and 3'-flanking regions in addition to their coding regions. The coding regions contained no intron, and there was 86% homology between their nucleotide sequences. However, no homology was observed between the flanking regions. The primer extension analysis showed that four transcription initiation sites were localized around 100-bp upstream from the ATG start codon of the lip2 gene. The putative promoter sequences of eukaryotic genes, CCAAT-----TATAAA, were found in the 5'-flanking regions of two lipase genes. These sequences might participate in the lipase induction by long-chain fatty acid. There were two inverted repeat sequences and a pyrimidine-rich sequence in the 5'-flanking region of lip2, but not in the 5'-flanking region of lip1. This specific structure of lip2 might reflect the higher transcriptional efficiency.