可以推算无法被测量到的反应其物质的通量及判断碳源流向!.PPTVIP

System Biology Term ProjectTryptophan Production Group 5 Teammates: 林瑋耿, 吳東耘, 許博凱, 彭嘉冠 在現存途徑中提高目標產物代謝流(五個方法) 增加代謝途徑中限速步驟酶編碼基因的複製數 強化啟動子為主的關鍵基因的表現系統 在現存途徑中提高目標產物代謝流(五個方法) 提高目標途徑活化因子的合成速率 去活化目標途徑抑制因子的編碼基因 阻斷與目標途徑相競爭的代謝途徑 Tryptophan synthesis pathway Strategy of enhancing the tryptophan production Increase the glucose uptake without depleting the available PEP. Recycle PEP Reduce PEP usage in PTS. Increase the E4P supply, especially the late stage of fermentation. Non-oxidatively increase E4P. Decrease the feedback-inhibition. Feedback-insensitive isozyme Trp operon repressor mutant. Optimize the glucose feed rate. PEP:carbohydratephosho-transferase system (PTS) PEP supply Recycle PEP: PEP synthase (ppsA) recycles pyruvate generated by PTS back to PEP. (Yi et al., 2002. Biotechnol. Prog., 18, 1141-1148) Reduce PEP usage in PTS: replace PTS with glucose facilitator protein (Zymomonas mobilis’ glf) or galactose permease (E.coli gal-P ). (Yi et al., 2003. Biotechnol. Prog., 19, 1450-1459) E4P supply At the late stage of fermentation, carbon flow will shift to TCA cycle, and the available E4P is limited. modify the pentose phosphate pathway (PPP) by overexpressing transketolase in order to change the carbon flow with a non-oxidative pathway. (Ikeda and Katsumata, Applied and Environmental Microbiol., 2497–2502) Decrease feedback inhibition DAHP synthase isoenzymes: Encoding by aroF– feedback inhibited by Tyr. Encoding by aroG– feedback inhibited by Phe. Encoding by aroH– feedback inhibited by Trp. Trp operon repressor mutant: TrpR2 has a frame-shift mutation in trpR that eliminates the production of functional trp repressor. Anthranilate synthase mutant (trpE19): trpE19 has a mutation in trpE that eliminates feedback inhibition of anthranilate synthase by tryptophan. Optimize the glucose feed rate The glucose feed rate profile have substantial effects on both tryptophan and glutamate (by-product) production. A feed rate profile wa