正十二烷作为氧载体在重组大肠杆菌中增强富马酸酶活性的应用：细胞内微环境的作用
Application of n-dodecane as an oxygen vector to enhance the activity of fumarase in recombinant Escherichia coli: role of intracellular microenvironment.
作者信息
Zhang Sen, Song Ping, Li Shuang
机构信息
Nanjing University of Chinese Medicine, College of Pharmacy, Jiangsu Collaboration Innovation Center of Chinese Medical Resources Industrialization, Nanjing, People's Republic of China; National and Local Collaborative Engineering Center of Chinese Medicinal Resources Industrialization and Formulae Innovative Medicine, Nanjing, People's Republic of China.
Nanjing University of Technology, College of Biotechnology and Pharmaceutical Engineering, State Key Laboratory of Materials-Oriented Chemical Engineering, Nanjing, People's Republic of China.
出版信息
Braz J Microbiol. 2018 Jul-Sep;49(3):662-667. doi: 10.1016/j.bjm.2017.09.005. Epub 2018 Feb 2.
The effect of the intracellular microenvironment in the presence of an oxygen vector during expression of a fusion protein in Escherichia coli was studied. Three organic solutions at different concentration were chosen as oxygen vectors for fumarase expression. The addition of n-dodecane did not induce a significant change in the expression of fumarase, while the activity of fumarase increased significantly to 124% at 2.5% n-dodecane added after 9h induction. The concentration of ATP increased sharply during the first 6h of induction, to a value 7600% higher than that in the absence of an oxygen-vector. NAD/NADH and NADP/NADPH ratios were positively correlated with fumarase activity. n-Dodecane can be used to increase the concentration of ATP and change the energy metabolic pathway, providing sufficient energy for fumarase folding.
研究了在大肠杆菌中表达融合蛋白时，细胞内微环境在存在氧载体的情况下的影响。选择三种不同浓度的有机溶液作为富马酸酶表达的氧载体。添加正十二烷并未引起富马酸酶表达的显著变化，而在诱导9小时后添加2.5%正十二烷时，富马酸酶的活性显著提高至124%。在诱导的前6小时内，ATP浓度急剧增加，达到比不存在氧载体时高7600%的值。NAD/NADH和NADP/NADPH比率与富马酸酶活性呈正相关。正十二烷可用于提高ATP浓度并改变能量代谢途径，为富马酸酶折叠提供足够的能量。
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