Cloning and sequence analysis of the lipase and lipase chaperone-encoding genes from Acinetobacter calcoaceticus RAG-1, and redefinition of a Proteobacterial lipase family and an analogous lipase chaperone family
Cloning and sequence analysis of the lipase and lipase chaperone-encoding genes from Acinetobacter calcoaceticus RAG-1, and redefinition of a Proteobacterial lipase family and an analogous lipase chaperone family
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DOI：
10.1016/s0378-1119(99)00026-8
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发表时间：
1999-04-16
期刊：
影响因子：
3.5
通讯作者：
Colwell, RR
中科院分区：
文献类型：
--
作者：
Sullivan, ER;Leahy, JG;Colwell, RR
对醋酸钙不动杆菌RAG-1的脂肪酶(LIPA)和脂肪酶伴侣(LipB)基因进行克隆和测序。这些基因是从基因组DNA文库中通过互补同一菌株的脂肪酶缺陷转座子突变体而分离出来的。在这个突变体和其他三个突变体中的转座子插入被定位到伴侣基因的一个单一位置。该脂肪酶及其伴侣蛋白的氨基酸序列分别编码313个氨基酸(32.5 kDa)和347个氨基酸(38.6 kDa)。该脂肪酶含有一个假定的前导序列，以及已知在其他脂肪酶中起催化作用的保守的Ser、His和Asp残基，在伴侣蛋白的N-末端区域发现了一个可能的跨膜疏水螺旋。系统发育比较表明，LIPA与醋酸钙杆菌BD413、霍乱弧菌El Tor和普通变形杆菌K80的脂肪酶同属假单胞菌和伯克霍尔德氏菌脂肪酶家族。这个新的家族，我们重新定义为第一组Protebacterial脂肪酶，根据整个序列的同源性和亚家族所特有的残基保守性，将其细分为四个亚家族。此外，LipB被发现是一个类似的脂肪酶伴侣家族的成员。我们建议将荧光假单胞菌和粘质沙雷氏菌产生的脂肪酶称为第二组变形杆菌脂肪酶，它们构成第二个序列家族。提供的证据支持这样的假设，即第一组和第二组家庭都是从共同血统和横向基因转移的组合进化而来的。(C)1999 Elsevier Science B.V.保留所有权利。
The genes encoding the lipase (LipA) and lipase chaperone (LipB) from Acinetobacter calcoaceticus RAG-1 were cloned and sequenced. The genes were isolated from a genomic DNA library by complementation of a lipase-deficient transposon mutant of the same strain. Transposon insertion in this mutant and three others was mapped to a single site in the chaperone gene. The deduced amino acid (aa) sequences for the lipase and its chaperone were found to encode mature proteins of 313 aa (32.5 kDa) and 347 aa (38.6 kDa), respectively. The lipase contained a putative leader sequence, as well as the conserved Ser, His, and Asp residues which are known to function as the catalytic triad in other Lipases, A possible trans-membrane hydrophobic helix was identified in the N-terminal region of the chaperone. Phylogenetic comparisons showed that LipA, together with the lipases of A. calcoaceticus BD413, Vibrio cholerae El Tor, and Proteus vulgaris K80, were members of a previously described family of Pseudomonas and Burkholderia lipases. This new family, which we redefine as the Group I Proteobacterial lipases, was subdivided into four subfamilies on the basis of overall sequence homology and conservation of residues which are unique to the subfamilies. LipB, moreover, was found to be a member of an analogous family of lipase chaperones. We propose that the lipases produced by P. fluorescens and Serratia marcescens, which comprise a second sequence family, be referred to as the Group II Proteobacterial lipases. Evidence is provided to support the hypothesis that both the Group I and Group II families have evolved from a combination of common descent and lateral gene transfer. (C) 1999 Elsevier Science B.V. All rights reserved.