生物工程外文翻译--代谢控制工程提高大肠杆菌番茄红素产量的研究(编辑修改稿)

生物工程外文翻译--代谢控制工程提高大肠杆菌番茄红素产量的研究(编辑修改稿)内容摘要：

to control the rebinant cell flow path in order to adapt to the metabolic state of the cell. Power control rebinant cell path can potentially lead to the increase in production will hinder growth reduced to a minimum to reduce the formation of toxic byproducts. Gene expression rules to adapt to the physiological state is also very necessary for the success of gene therapy. Here we give examples of the application of this method to improve the yield of lycopene in E. coli. We chose as a typical example of lycopene is beneficial for human health because lycopene. Lycopene as an effective antioxidant, it has been proposed for the treatment of cancer and other degenerative diseases, therefore, has the position of the activity of lycopene and other carotenoids have been more and more people concern, and there is a large number of reports in the description of its products. Results and Discussion Rebinant cell path to build dynamics controller. The purpose of the power controller is used to control the flow of the path in order to facilitate the C and utilization of energy. Shown in Figure 1: Design of such a cell needs to identify the operating system: the controller processing sensory input of the sensor monitors the signal of the (1) signaling molecules reflection related to the metabolic state of (2) (3) (4) of the control valve (. : promote) regulate gene expression (5) of the control path speed limiting device. Identify the correct signal molecules and speed limits need to have a thorough understanding of the metabolic system. The rest of the ingredients adjustment system derived from two ponents, wherein the system includes a large number of geic adjusted molecule. Figure 1 power cyberic depicting the control engineering structure Figure The process to be controlled is the metabolic pathway from glucose to the product (lycopene) and some of the waste (for example: acetate). The standard variable (or signal) is the ACP, ACP provides signals to the rest of the flow into a waste. Feel molecule is a the NRI protein, it can bine DNA regulating transcription. The control valve is a promoter glnAp2 restricted process (LDI and PPS), it can be controlled in metabolic control system. The dashed box in FIG noted that the metabolic system to be controlled. Our initial task is to intracellular metabolic state of the signal molecules can be screened to identify one and one sensor able to monitor it. ACP can do for the metabolic state of signaling molecules, because he has been used as an indicator of glucose. In addition, it has also adjust the substandard as to identify two blend ponents, such as Che13 Pho14, and Ntr15 of. Enhance the level of ACP can be used as a good indicator. Accordingly, as shown in Figure 1 Our approach is the use of ACP as a signal molecule to control the expression of the enzyme in the desired path, but also to fully utilize an excessive amount of C flows and changing the direction of flow away from the toxic product acetate. In order to understand the ACP and control of gene expression, we use E. coli Ntr regulon (Figure 2). Ntr regulator allows cells to adapt to the lack of nitrogen status but its role in most biological reactor gone down, because adequate nitrogen environment. Ntr adjustment sub sensor called NRII, it is the the glnL the product of the gene, can be phosphorylated form of phosphate transferred to the NRI protein NRIP and then bined with the AP2 binding sites, translational start AP2. In NRII missing, NRI can respond to the ACP level. Design NRII missing loop control, NRI respond to the ACP level and target gene expression (Fig. 2A). In order to reconstruct such a control unit, limited area containing the NRI adhesive sites and nuclear glnAp2 promoter region, and the insert DNA fragment into a cloning vector. The NRI bonding sites overlap another by cAMPCRP regulated promoters glnAP1 of. Thus, the DNA fragment also contains glnAP1, but not cAMPCRP binding sites (Figure 2A). CAMPCRP activity, the glnAp2 activity is quite low, and contains the promoter construct a wildtype strain (JCL1595) to support this Classifying (Figure 2B). Dynamic control of a single gene expression Kiics controller described in FIG 2. (A) glnAp2 promoter NRII lack, ACPinduced glnAp2. NRI, NRIbinding sites。 glnAp2 core, the glnAp2 nuclear sequence。 RBS, glnA ribosome bonding sites。 ATG transformation begins. Note 35 and 10 region of that position of the two RNA polymerase, the glnAp2 promoter of RNA polymerase and the upstream contact. GlnAp2lacZ from a single copy (B, C) Gal activity of structure (B) cell density (at 550nm optical density) and (C) acetic acid in JCL1595 (glnL +) and JCL 1596glnL) was salt secretions . From the wildtype (D) gal activity activity in glucose aerobic growth period single copy PlaclacZ and strains VJS632 of growth kiics. Symbol (•) represents cell growth (■) on behalf of the gal activity (▽ ) on behalf of extracellular In order to test the expression as a product the power controller glnAp2 potential, we introduced glnAp2lacZ messenger RNA fusion technology into glnL through phage host strain (JCL1596) and measure the time course of gal activity. This strain contains glnL2020 allele. FIG 2 C sho。