Ohnishi H, Nishida T, Yoshida A, Kamio Y, Izaki K
Department of Agricultural Chemistry, Faculty of Agriculture, Tohoku University, Sendai, Japan.
Biochem Biophys Res Commun. 1991 Apr 15;176(1):321-7. doi: 10.1016/0006-291x(91)90927-y.
The nucleotide sequence of pnl gene encoding pectin lyase (PNL; EC4.2.2.10)from Erwinia carotovora Er was determined. The structural gene of pnl consisted of 942 base pairs. An open reading frame that could encode a 33,700 dalton polypeptide consisting 314 amino acids was assigned. The molecular size of the polypeptide predicted from the amino acid composition was close to the value of PNL determined in E.carotovora Er. The nucleotide sequence of the 5'-flanking region showed the presence of the consensus sequence of ribosome binding site, Pribnow box and the RNA polymerase recognition site in E.carotovora and Escherichia coli. Between the presumed Pribnow box and the ribosome binding site, two pairs of inverted repeats were found. By comparing the predicted amino acid sequences of pnl, several reported bacterial pectate lyases and Aspergillus niger pectin lyase, short regions of homology were found despite the different substrate specificities of these enzymes.
测定了胡萝卜软腐欧文氏菌(Erwinia carotovora Er)中编码果胶裂解酶(PNL;EC4.2.2.10)的pnl基因的核苷酸序列。pnl的结构基因由942个碱基对组成。确定了一个开放阅读框,其可编码由314个氨基酸组成的33700道尔顿的多肽。根据氨基酸组成预测的多肽分子大小与在胡萝卜软腐欧文氏菌Er中测定的PNL值接近。5'-侧翼区的核苷酸序列显示在胡萝卜软腐欧文氏菌和大肠杆菌中存在核糖体结合位点、普里布诺框和RNA聚合酶识别位点的共有序列。在推测的普里布诺框和核糖体结合位点之间,发现了两对反向重复序列。通过比较pnl预测的氨基酸序列、几种已报道的细菌果胶酸裂解酶和黑曲霉果胶裂解酶,尽管这些酶的底物特异性不同,但仍发现了短的同源区域。