Schneider E, Freundlieb S, Tapio S, Boos W
Department of Biology, University of Konstanz, Federal Republic of Germany.
J Biol Chem. 1992 Mar 15;267(8):5148-54.
malS, the gene encoding the periplasmic alpha-amylase, is under the regulatory control of the MalT protein, the gene activator of the Escherichia coli maltose system. We sequenced the DNA region encoding malS and its control elements. malS consists of an open reading frame of 2,028 base pairs encoding a protein of 676 amino acids with a deduced molecular weight of 75,664 including a typical amino-terminal signal sequence of 17 amino acids. The deduced amino acid sequence of malS was compared with that of other proteins. We found homologies to alpha-amylases and a cyclodextrin glucanotransferase but not to beta-amylases. In addition, MalS showed significant homology to another maltodextrin-utilizing and MalT-dependent enzyme of E. coli, maltodextrin glucosidase (MalZ) but not to amylomaltase (MalQ), the major maltodextrin-degrading enzyme. Conserved regions that have been proposed to constitute enzymatically active sites in alpha-amylases are present in MalS. Two of these sequences can also be found in the amino terminus of the lambda-receptor, a maltodextrin-specific channel in the outer membrane. The nucleotide sequence of the control region of malS revealed MalT binding sites correctly spaced for the start of the malS transcript. At the position 219 base pairs upstream of malS, we found a divergently transcribed gene, bax, which has been recognized previously. Downstream of malS, after a 513-base pair intergenic region, lies the convergently transcribed gene avtA, which codes for the alanine/valine transaminase.
malS基因编码周质α-淀粉酶,它受MalT蛋白的调控,MalT蛋白是大肠杆菌麦芽糖系统的基因激活剂。我们对编码malS及其调控元件的DNA区域进行了测序。malS由一个2028个碱基对的开放阅读框组成,编码一个676个氨基酸的蛋白质,推导分子量为75664,包括一个17个氨基酸的典型氨基末端信号序列。将malS推导的氨基酸序列与其他蛋白质的序列进行了比较。我们发现它与α-淀粉酶和环糊精葡聚糖转移酶有同源性,但与β-淀粉酶没有同源性。此外,MalS与大肠杆菌另一种利用麦芽糊精且依赖MalT的酶——麦芽糊精葡糖苷酶(MalZ)有显著同源性,但与主要的麦芽糊精降解酶淀粉麦芽糖酶(MalQ)没有同源性。在α-淀粉酶中被认为构成酶活性位点的保守区域存在于MalS中。这些序列中的两个也可以在λ受体的氨基末端找到,λ受体是外膜中的一种麦芽糊精特异性通道。malS调控区域的核苷酸序列揭示了MalT结合位点,其间距适合malS转录本的起始。在malS上游219个碱基对处,我们发现了一个先前已被识别的反向转录基因bax。在malS下游,经过一个513个碱基对的基因间隔区后,是反向转录的基因avtA,它编码丙氨酸/缬氨酸转氨酶。