多聚糖奈瑟菌中淀粉蔗糖酶基因的克隆与特性分析：线性α-1,4-葡聚糖的产生
Cloning and characterization of the gene for amylosucrase from Neisseria polysaccharea: production of a linear alpha-1,4-glucan.
作者信息
Büttcher V, Welsh T, Willmitzer L, Kossmann J
机构信息
Institut für Genbiologische Forschung GmbH Berlin, Germany.
出版信息
J Bacteriol. 1997 May;179(10):3324-30. doi: 10.1128/jb.179.10.3324-3330.1997.
Abstract
The gene for the amylosucrase from Neisseria polysaccharea (ATCC 43768) was cloned by use of a functional expression system in Escherichia coli XL1-Blue. The deduced amino acid sequence of the protein has homology to the sequences of the alpha-amylase class of enzymes, with the highest similarities being found to the sequences of the trehalose synthase from Pimelobacter sp. strain R48 (17) and amylomaltase from Thermotoga maritima (11). However, the regions of highest homology within the alpha-amylase class of enzymes, which are essential for the catalytic activity, are only scarcely found in the sequence of amylosucrase. By using the enzyme isolated from culture supernatants of transformed E. coli cells, it is possible to synthesize linear alpha-1,4-glucans from sucrose, indicating that the enzyme is not capable of producing alpha-1,6-glycosidic linkages on its own.
摘要
利用大肠杆菌XL1 - Blue中的功能表达系统克隆了多糖奈瑟菌（ATCC 43768）的支链淀粉酶基因。该蛋白质推导的氨基酸序列与α -淀粉酶类酶的序列具有同源性，与皮氏杆菌属菌株R48的海藻糖合酶序列（17%）和海栖热袍菌的淀粉麦芽糖酶序列（11%）相似度最高。然而，在α -淀粉酶类酶中对催化活性至关重要的最高同源区域，在支链淀粉酶序列中仅很少发现。通过使用从转化大肠杆菌细胞的培养上清液中分离出的酶，有可能从蔗糖合成线性α - 1,4 -葡聚糖，这表明该酶自身不能产生α - 1,6 -糖苷键。
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