文献类型：期刊文献

英文题名：Identification of a novel promoter gHp0169 for gene expression in Gluconobacter oxydans

作者：Shi, Lulu[1];Li, Kefei[1];Zhang, Huan[1];Liu, Xu[1];Lin, Jinping[1];Wei, Dongzhi[1]

机构：[1]E China Univ Sci & Technol, State Key Lab Bioreactor Engn, Shanghai 200237, Peoples R China

年份：2014

卷号：175

期号：1

起止页码：69

外文期刊名：JOURNAL OF BIOTECHNOLOGY

收录：;EI(收录号：20141117466365);WOS:【SCI-EXPANDED(收录号:WOS:000333090200012)】；

基金：This work was financially supported by the Natural Science Foundation of China (No. 21002029/B020706), National Key Basic Research Development Program of China ("973" Program, No. 2012CB721003), and the National High Technology Research and Development Program of China (No. 2012AA022201).

语种：英文

外文关键词：Gluconobacter oxydans; Promoter; gHp0169; Gene overexpression

摘要：Gluconobacter oxydans can perform rapid incomplete oxidation of many sugars, sugar polyols and alcohols, and this outstanding ability shows a great potential in industrial bioconversion. Improvements of these industrially important strains would boost their productivities of important metabolites. However, the shortage of molecular tools for homologous and heterologous gene expression has obviously hindered G. oxydans from further application. In this study, a putative promoter sequence (104 bp), designated as gHp0169, was isolated and characterized from the chromosome of G. oxydans DSM 2003. Within this promoter sequence, the typical motif, known as -35 and -10 sequences with a 19-bp spacing, was found. The availability and promoter strength of promoter gHp0169 were then evaluated, by insertion into the plasmid pBBR1MCS5 for expression of a green fluorescent protein (GFP) and a membrane-bound type II NADH dehydrogenase (NDH-2) of G. oxydans. In comparison with promoter G. oxydans_tufB, gHp0169 exhibited a stronger promoter activity of NDH-2, indicating its significant value of gene expression in G. oxydans. To promote the production of 2-keto-D-gluconic acid (2-KGA) from gluconic acid (GA) gHp0169 was attempted to equip the flavin-dependent gluconate-2-dehydrogenase (GA2DH) and successfully achieved its overexpression in G. oxydans DSM 2003. As a result, the space-time yield of 2-KGA was boosted up to 29.86 mM/h compared with 14.78 mM/h for the control, which corresponded to a yield of 98.3% (84% for control). Copyright (C) 2014 Elsevier B.V. All rights reserved.