Efficient expression of mono- and diacylglycerol lipase gene from Penicillium camembertii U-150 in Aspergillus oryzae under the control of its own promoter.
The gene, mdlA, coding for mono- and diacylglycerol lipase from Penicillium camembertii U-150 was expressed efficiently in Aspergillus oryzae under the control of its own promoter. The gene product was secreted into the culture medium with a highest productivity of 1 g/liter and correctly processed at both N- and C-termini. KEX2-like processing was suggested to occur at the C-terminus in both A. oryzae and P. camembertii. Specific activity and substrate specificity of the purified recombinant protein were also almost the same to that of native protein but the extent of N-glycosylation in the recombinant protein was about half of that of the native protein. The presence of introns did not seem to affect the gene expression. The mdlA expression was induced by lipids and regulated transcriptionally in A. oryzae as well as P. camembertii. Promoter deletion analysis showed that the region between the positions at -382 and -554 bp from the translation initiation point was important to the higher expression of mdlA. The promoter sequence of mdlA was compared to that of the Geotrichum candidum lipase gene, which is also reported to be inducible by lipids, with three commonly observed oligonucleotide sequences.
Yamaguchi
S
S
Tsukuba Research Laboratories, Amano Pharmaceutical Co., Ltd., Ibaraki, Japan.
Takeuchi
K
K
Mase
T
T
Matsuura
A
A
D001236:Aspergillus oryzae; D017353:Gene Deletion; D015870:Gene Expression; D008055:Lipids / Q000494:pharmacology; D008071:Lipoprotein Lipase / Q000096:biosynthesis / Q000235:genetics*; D008994:Monoacylglycerol Lipases / Q000096:biosynthesis / Q000235:genetics*; D010407:Penicillium; D011401:Promoter Regions, Genetic; D011994:Recombinant Proteins / Q000096:biosynthesis; D014170:Transformation, Genetic
D008055:Lipids; D011994:Recombinant Proteins; D008994:Monoacylglycerol Lipases; D008071:Lipoprotein Lipase