【正文】 contains BW18302/pPs706 cell juice (C) the time course of expression from BW18302/pPs706 (glnAp2pps) PPS. The lycopene path dynamics control When the avoid growth delay, the impetus provided by the glnAp2 control can be used to improve the level of expression of specific genes. Therefore, by adjusting the molecular rebinant cell path to improve the lycopene biosynthesis, this range can be expanded in E. coli from the path of isoprenoid. The isoprenoid E. coli pyruvate and G3P as a pioneer, as shown in Figure 4 A. Transformation in E. coli of a rebinant lycopene path, by expression of the gene. These genes dxs, gps, crtBI into the lowcopy plasmid pCL1920 construct the plasmid pCW9, while overexpression. In this path, GPS and IDI have been identified to control the flow to the final product. In addition, we have shown that phosphoenolpyruvate synthase (PPS gene product) control of pyruvate and G3P balance, thereby also control the flow of to isoprenoids path is. In order to control the carbon flow, we use the glnAp2 promoter to control the expression of GPS and IDI. These plasmids were individually introduced into containing pCW9 the the gln strain (BW18302). (FIG. 5A), as shown in p2IDI strain (glnAp2 IDI), in the glucosecontaining medium after 26 hours to produce 100 mg / l of lycopene. Strain containing Ptacof IDI (pTacIDI) the other hand, in the same strip Figure 4. Lycopene metabolic control engineering (A) with the genes dxs, gps, IDI and crtBI, G3P, pyruvate, 3 phosphate, IPP, Pyr, DMAPP, dimethyl phosphate, FPP, GPP, GGPP reconstruction the lycopene path (B) use the IDI and PPS dynamics control to change the metabolic flux to the the lycopene path strategy. Two types of dotted lines represent applied to the control circuit of the IDI and PPS () and carbon flow to changes in the path of lycopene (...). Glc, glucose。 Cells contain BW18302/pAROG juice, labeling gel runway the 39。 Metabolic control engineering to improve the production of E. coli lycopene William R. Farmer and James C. Liao * Department of Chemical Engineering, University of California, Los Angeles, CA 90034 * Author (). Received: , Accepted: Abstract Summary of metabolic engineering in plex and diverse host of exogenous metabolites has made gratifying achievements. But the metabolic engineering approach mainly to expand the necessary enzymes and the discharged cell control and therefore is not controlled metabolic pathway metabolic imbalance and undesirable product. We have described the metabolic engineering of another process, the status of the regulator of metabolism in the cells to regulate gene expression through the design of regulatory circuits. Recovery and especially in E. coli Ntr regulator to control the biosynthesis pathway of the lycopene change in the adjustment loop system. Excess sugar deposition artificial stimulation regulators, the ACP control lycopene two key enzymes expression, which are in response to the fluid dynamics. Thus, when reducing the negative impact caused by metabolic imbalance, intracellular control system will significantly improve the yield of lycopene. Although we demonstrate that this method can improve the metabolic yield, but it can also be extended to other areas of gene expression in these areas must be subject to the stringent constraints of cell physiology, such as gene therapy. Keywords: carotenoids, isoprenoid, metabolic engineering, metabolic control, nitrogen regulator Metabolic engineering in the last century thanks to a full understanding of the biosynthetic capacity to bioinformatics and functional genomics disciplines to enter as a result of advances. The direct advantage of this knowledge is to allow reasonable novel path design and exclude a priori the reaction, in order to achieve the desired results of this innate reaction is unnecessary or harmful. However, with increasingly effective metabolic pathways, metabolic engineering will be faced with a new challenge: to adjust the level of process reengineering control of gene expression in these paths. To construct the geic ponents of metabolic pathways, we have plans to make him bee like the gene expression path of dynamics as important. Many scholars believe that the high level of induction of rebinant proteins or cell path will lead to growth delay and reduction of metabolic activity. The fact these surface features: continuous product demand is higher than changing the demand for cell growth. We hope that this S amp。包含 glnAP2啟動(dòng)子的 PCR片段被克隆到質(zhì)粒 pAROG、 pPS706和 pTacIDI的 EcoRV EcoRI位點(diǎn)以分別產(chǎn)生質(zhì)粒 p2AROG pPSG706和 p2IDI。 實(shí)驗(yàn)方案 材料 所有的化學(xué)藥品來(lái)自 Sigma(St. Louis, MO).修飾和限制酶來(lái)自生命工藝。（ C） 丙酮酸鹽 的分泌物。自從丙酮酸鹽成為 類(lèi)異戊二烯路徑的先驅(qū)之一來(lái)自細(xì)胞內(nèi)代謝物的排泄指出碳仍然不能有效地轉(zhuǎn)移到番茄紅素。另一方面包含 Ptacidi (pTacIDI)的菌株，在相同的條 圖 4. 番茄紅素產(chǎn)品的代謝控制工程 （ A） 用 基因 dxs 、 gps 、 idi和 crtBI、 G3P、 丙酮酸鹽 、 3磷酸鹽、 IPP、 Pyr、 DMAPP、二甲基二磷酸、 FPP、 GPP、 GGPP等 重建番茄紅素路徑 （ B）利用 IDI 和 PPS的動(dòng)力學(xué)控制來(lái)改變代謝流量到番茄紅素路徑策略。 大腸桿菌中的 類(lèi)異戊二烯 利用丙酮酸鹽和 G3P作為先驅(qū)，如圖 4 A所示。宿主菌株是 BW18302，它被單獨(dú)轉(zhuǎn)化為 pAROG、 p2AROG3或者沒(méi)有 質(zhì)粒（▽?zhuān)? 多種復(fù)制基因表達(dá)的動(dòng)力控制 為了測(cè)定由 glnAp2提供的動(dòng)力控制是否能減輕因高水平的蛋白質(zhì)表達(dá)導(dǎo)致的代謝失衡和生長(zhǎng)延遲，而代謝失衡和生長(zhǎng)延遲通常在代謝工程中它是不可避免的。這種菌株包含 glnL2020等位基因。 glnAp2核 , glnAp2 核序列 。在 NRII缺失時(shí)， NRI才能響應(yīng) ACP水平。 我們最初的任務(wù)是鑒別一種可以放映細(xì)胞內(nèi)代謝狀態(tài)的信號(hào)分子和一種能夠監(jiān)控它的傳感器。如圖