Effect of overexpression of proline dehydrogenase on high saline adaptation through proline utilization in Escherichia coli
Effect of overexpression of proline dehydrogenase on high saline adaptation through proline utilization in Escherichia coli
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脯氨酸脱氢酶过表达对大肠杆菌利用脯氨酸对高盐适应的影响
DOI：
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发表时间：
2013
期刊：
影响因子：
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通讯作者：
S. Nagata
中科院分区：
文献类型：
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作者：
H. Sasaki;A. Oshima;A. Ishida;S. Nagata
分析了Pro脱氢酶PUTA的表达量对大肠杆菌细胞在高盐条件下适应机制的影响。将来源于pUTA基因的pUTA基因导入大肠杆菌BL21-PUTA细胞，获得pUTA高效表达菌株BL21-PUTA。添加基因表达诱导剂IPTG诱导PUTA转录，并在1MNaC l条件下观察到高转录。大肠杆菌BL21-puta在Davis微量培养基中的生长在1MNaC l条件下受到抑制，但添加L-脯氨酸可显著恢复其生长。另一方面，加入异丙基甘油三酯后，细胞在L-脯氨酸存在下的生长恢复受到抑制。在1MNaC l条件下，无论是哪种菌株，细胞内都能迅速积累L-脯氨酸。另一方面，经IPTG处理的BL21-puta细胞内L-脯氨酸浓度低于BL21细胞。这些数据表明，PUTA的表达量影响了大肠杆菌对L-脯氨酸对高盐度的适应。
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关键词：大肠杆菌，高盐适应，脯氨酸脱氢酶，过表达
The effect of the amount of proline dehydrogenase PutA expression on the mechanism of adaptation of Escherichia coli cells under high salinity was analyzed. The PutA gene derived from E. coli expression plasmid was introduced into E. coli cell, and PutA high expression strain BL21-PutA was developed. Transcription of the PutA was induced by the addition of gene expression inducer IPTG, and high transcription was observed under 1 M NaCl condition. Growth of E. coli BL21-PutA in a Davis minimal medium was inhibited under 1 M NaCl condition, but it was remarkably recovered by the addition of L-proline. On the other hand, the recovery of growth in the presence of L-proline was inhibited when IPTG was added. Under 1 M NaCl condition, L-proline was quickly accumulated in cells, regardless of the strains. On the other hand, intracellular concentration of L-proline in BL21-PutA cells cultured with IPTG was lower than BL21 cells. These data suggested that the amount of the expression of PutA affected the adaptation of high salinity with L-proline in E. coli cells.
Key words: Escherichia coli, high salinity adaptation, proline dehydrogenase PutA, overexpression
影响因子：
3
作者：
H. Sasaki;Ai Takaki;A. Oshima;A. Ishida;S. Nagata
通讯作者：
H. Sasaki;Ai Takaki;A. Oshima;A. Ishida;S. Nagata
高盐条件下脯氨酸和钾对大肠杆菌K-12细胞活性刺激的影响
DOI：
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发表时间：
2005
期刊：
Biosci.Biotech.Biochem. In press
影响因子：
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作者：
Kojima;M.;M.Tokumaru;K.Fujiki;K.Hayashi;他1名;Shinichi Nagata
通讯作者：
Shinichi Nagata