HUANGFU Jie, LI Qian, LI Jun, et al. Pathway analysis of recombinant Escherichia coli during protein production process under simulated microgravity[J]. CIESC Journal, 2015, (12): 4960-4965.
HUANGFU Jie, LI Qian, LI Jun, et al. Pathway analysis of recombinant Escherichia coli during protein production process under simulated microgravity[J]. CIESC Journal, 2015, (12): 4960-4965.DOI: 10.11949/j.issn.0438-1157.20150471.
对该重组大肠杆菌菌株在SMG和NG培养过程中加入IPTG诱导外源蛋白质表达4 h 后的全细胞蛋白质酶解产物
通过三重稳定同位素二甲基标记方法并结合质谱分离鉴定技术
在肽段水平上标记后进行差异定量蛋白质组学分析。结果表明:与NG对照相比
SMG条件下共有124个蛋白质表达水平发生显著变化
其中92个蛋白质上调
6个蛋白质下调。上调蛋白质集中在细胞压力胁迫、氨基酸代谢、翻译转录和碳代谢等途径
这些相关途径的蛋白质表达水平变化可以影响大肠杆菌生理代谢及外源蛋白质表达效率。
Abstract
Microgravity effects were reported to affect the physiological characteristics and perturbations at the molecular levels of the microorganisms. Our previous works reported that the recombinant β-glucuronidase (PGUS) expressing strain Escherichia coli BL21 (DE3)/pET28a-pgus grew faster and the efficiency of recombinant protein production was also enhanced under simulated microgravity (SMG) as compared with the normal gravity (NG). In this study
a quantitative proteomic analysis of triple stable isotope dimethyl labeling using mass spectrometry method to investigate the effects of SMG on the strain and the during recombinant protein production process was performed. A total of 124 differentially expressed proteins under SMG were identified as compared with NG. Among them
92 proteins were significantly up-regulated and 6 proteins were drastically down-regulated under SMG. The majority of the up-regulated proteins were involved with the metabolism of cell stress response
amino acid metabolism
translation
transcription and carbohydrate metabolic process. The results indicated that the differentially expressed levels of these proteins under SMG were beneficial to the synthesis of the heterologous protein expression of the recombinant Escherichia coli.