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首页> 外文期刊>World Journal of Microbiology & Biotechnology >Type 2 IDI performs better than type 1 for improving lycopene production in metabolically engineered E. coli strains
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Type 2 IDI performs better than type 1 for improving lycopene production in metabolically engineered E. coli strains

机译:在代谢工程化的大肠杆菌菌株中,类型2 IDI在改善番茄红素产生方面比类型1更好。

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摘要

In this study a comparison was made between type 1 and type 2 isopentenyl diphosphate isomerases (IDI) in improving lycopene production in Escherichia coli. The corresponding genes of Bacillus licheniformis and the host (i (Bl) and i (Ec) , respectively) were expressed in lycopene producing E. coli strains by pTlyci(Bl) and pTlyci(Ec) plasmids, under the control of tac promoter. The results showed that the overexpression of i (Ec) improved the lycopene production from 33 +/- A 1 in E. coli Tlyc to 68 +/- A 3 mg/gDCW in E. coli Tlyci(Ec). In contrast, the expression of i (Bl) increased the lycopene production more efficiently up to 80 +/- A 9 mg/gDCW in E. coli Tlyci(Bl). The introduction of a heterologous mevalonate pathway to elevate the IPP abundance resulted in a lycopene production up to 132 +/- A 5 mg/gDCW with i (Ec) in E. coli Tlyci(Ec)-mev and 181 +/- A 9 mg/gDCW with i (Bl) in E. coli Tlyci(Bl)-mev, that is, 4 and 5.6 times respectively. When fructose, mannose, arabinose, and acetate were each used as an auxiliary substrate with glycerol, lycopene production was inhibited by different extents. Among auxiliary substrates tested, only citrate was an improving one for lycopene production in all strains with a maximum of 198 +/- A 3 mg/gDCW in E. coli Tlyci(Bl)-mev. It may be concluded that the type 2 IDI performs better than the type 1 in metabolic engineering attempts for isoprenoid production in E. coli. In addition, the metabolic engineering of citrate pathway seems a promising approach to have more isoprenoid accumulation in E. coli.
机译:在这项研究中,比较了1型和2型异戊烯基二磷酸异构酶(IDI)在改善大肠杆菌中番茄红素生产中的作用。在tac启动子的控制下,pTlyci(B1)和pTlyci(Ec)质粒在产番茄红素的大肠杆菌中表达了地衣芽孢杆菌和宿主的相应基因(分别为i(B1)和i(Ec))。结果表明,i(Ec)的过表达将番茄红素的产量从大肠杆菌Tlyc中的33 +/- A 1提高到大肠杆菌Tlyci(Ec)中的68 +/- A 3 mg / gDCW。相反,i(B1)的表达在大肠杆菌Tlyci(B1)中更有效地提高了番茄红素的产量,最高可达80 +/- A 9 mg / gDCW。引入异源甲羟戊酸途径来提高IPP的丰度导致在番茄Tlyci(Ec)-mev和181 +/- A 9中含i(Ec)的番茄红素产量高达132 +/- A 5 mg / gDCW mg / gDCW在大肠杆菌Tlyci(B1)-mev中具有i(B1)的倍数,分别是4倍和5.6倍。当果糖,甘露糖,阿拉伯糖和乙酸盐分别用作甘油的辅助底物时,番茄红素的产生受到不同程度的抑制。在测试的辅助底物中,只有柠檬酸盐是所有菌株中番茄红素生产的改良剂,在大肠杆菌Tlyci(B1)-mev中最大为198 +/- A 3 mg / gDCW。可以得出结论,在大肠杆菌中生产类异戊二烯的代谢工程学尝试中,类型2的IDI比类型1的性能更好。此外,柠檬酸途径的代谢工程似乎是在大肠杆菌中具有更多类异戊二烯积累的有前途的方法。

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