Cloning and expression of the gene encoding catalytic subunit of thermostable glucose dehydrogenase from Burkholderia cepacia in Escherichia coli
Cloning and expression of the gene encoding catalytic subunit of thermostable glucose dehydrogenase from Burkholderia cepacia in Escherichia coli
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DOI：
10.1016/s1570-9639(02)00534-4
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发表时间：
2003-02-21
影响因子：
3.2
通讯作者：
Sode, K
中科院分区：
文献类型：
--
作者：
Inose, K;Fujikawa, M;Sode, K
我们已经克隆了一个1620个核苷酸的基因编码的热稳定性葡萄糖脱氢酶（GDH）的催化亚基（α亚基）从洋葱伯克霍尔德氏菌。在α亚基的N-末端区域发现FAD结合基序。推测的α t亚基的一级结构与葡萄糖酸氧化酶的山梨醇脱氢酶（SDH）催化亚基和欧文氏菌（Erwiniacetylola）和柠檬泛菌（Pantoeacitrea）的2-酮基-D-葡萄糖酸脱氢酶（2KGDH）催化亚基的同源性约为48%。B的α t亚基。cepacia在大肠杆菌中以其活性水溶性形式表达，在70 ℃下显示最大的染料介导的GDH活性，保持高热稳定性。在编码18-kDa肽的α亚基的上游还发现了一个507个核苷酸的推定开放阅读框（ORF），命名为γ亚基。推测的γ亚基一级结构与G. oxydans和2KGDH。和P citrea。(C)2002 Elsevier Science B.V保留所有权利。
We have cloned a 1620-nucleotide gene encoding the catalytic subunit (alpha subunit) of a thermostable glucose dehydrogenase (GDH) from Burkholderia cepacia. The FAD binding motif was found in the N-terminal region of the alpha subunit. The deduced primary structure of the ot subunit showed about 48% identity to the catalytic subunits of sorbitol dehydrogenase (SDH) from Gluconobacter oxydans and 2-keto-D-gluconate dehydrogenases (2KGDH) from Erwinia herbicola and Pantoea citrea. The ot subunit of B. cepacia was expressed in Escherichia coli in its active water-soluble form, showing maximum dye-mediated GDH activity at 70 degreesC, retaining high thermal stability. A putative open reading frame (ORF) of 507 nucleotides was also found upstream of the a subunit encoding an 18-kDa peptide, designated as gamma subunit. The deduced primary structure of gamma subunit showed about 30% identity to the small subunits of the SDH from G. oxydans and 2KGDHs from E. herbicola and P citrea. (C) 2002 Elsevier Science B.V All rights reserved.