Yamaguchi S, Mase T, Takeuchi K
Tsukuba Research Laboratories, Amano Pharmaceutical Co. Ltd., Ibaragi, Japan.
Gene. 1991 Jul 15;103(1):61-7. doi: 10.1016/0378-1119(91)90391-n.
A gene (mdlA) encoding mono- and diacylglycerol lipase (MDGL) from Penicillium camembertii U-150 has been cloned using a 0.9-kb DNA fragment, generated by mixed oligodeoxyribonucleotide (oligo)-primed polymerase chain reaction (PCR), as a probe. Comparison of the nucleotide sequence of the gene and its cDNA clone, obtained by PCR, revealed the presence of two short introns (56 and 53 bp). Two transcription start points (tsp) were localized by primer extension analysis at 37 and 30 bp upstream from the ATG start codon and were preceded by the canonical TATAAA and CAAT sequences. The deduced amino acid (aa) sequence corresponds to 305 aa including a putative signal peptide of 26 aa. Despite significant differences in substrate specificity, the primary structure of the mature region shows homology (29% and 40%) to the triacylglycerol lipases from Mucor miehei and Humicola lanuginosa. Furthermore, the three residues presumed to form the catalytic site, serine, aspartic acid and histidine, are conserved. Primary structure comparisons of MDGL and triacylglycerol lipases are shown.
利用通过混合寡脱氧核糖核苷酸（oligo）引发的聚合酶链反应（PCR）产生的0.9 kb DNA片段作为探针，克隆了来自卡门柏青霉U-150的编码单酰甘油和二酰甘油脂肪酶（MDGL）的基因（mdlA）。对该基因及其通过PCR获得的cDNA克隆的核苷酸序列进行比较，发现存在两个短内含子（56和53 bp）。通过引物延伸分析在ATG起始密码子上游37和30 bp处定位了两个转录起始点（tsp），并且在其之前有典型的TATAAA和CAAT序列。推导的氨基酸（aa）序列对应于305个氨基酸，包括一个26个氨基酸的假定信号肽。尽管底物特异性存在显著差异，但成熟区域的一级结构与米黑根毛霉和羊毛状腐质霉的三酰甘油脂肪酶具有同源性（分别为29%和40%）。此外，推测形成催化位点的三个残基，即丝氨酸、天冬氨酸和组氨酸，是保守的。展示了MDGL和三酰甘油脂肪酶的一级结构比较。
