Vadali Ravishankar V, Fu Yuchun, Bennett George N, San Ka-Yiu
Department of Bioengineering, Rice University, Houston, Texas 77005, USA.
Biotechnol Prog. 2005 Sep-Oct;21(5):1558-61. doi: 10.1021/bp050124l.
Lycopene is a useful phytochemical that holds great commercial value. In our study the lycopene production pathway in E. coli originating from the precursor isopentenyl diphosphate (IPP) of the non-mevalonate pathway was reconstructed. This engineered strain of E. coli accumulated lycopene intracellularly under aerobic conditions. As a next step, the production of lycopene was enhanced through metabolic engineering methodologies. Various competing pathways at the pyruvate and acetyl-CoA nodes were inactivated to divert more carbon flux to IPP and subsequently to lycopene. It was found that the ackA-pta, nuo mutant produced a higher amount of lycopene compared to the parent strain. To further enhance lycopene production, a novel mevalonate pathway, in addition to the already existing non-mevalonate pathway, was engineered. This pathway utilizes acetyl-CoA as precursor, condensing it to form acetoacetyl-CoA and subsequently leading to formation of IPP. Upon the introduction of this new pathway, lycopene production increased by over 2-fold compared to the ackA-pta, nuo mutant strain.
番茄红素是一种具有重要商业价值的有用植物化学物质。在我们的研究中,源自非甲羟戊酸途径的前体异戊烯基二磷酸(IPP)的大肠杆菌中番茄红素生产途径得以重建。这种工程化的大肠杆菌菌株在有氧条件下在细胞内积累番茄红素。下一步,通过代谢工程方法提高了番茄红素的产量。丙酮酸和乙酰辅酶A节点处的各种竞争途径被灭活,以将更多的碳通量转移到IPP,进而转移到番茄红素。结果发现,与亲本菌株相比,ackA-pta、nuo突变体产生的番茄红素量更高。为了进一步提高番茄红素产量,除了现有的非甲羟戊酸途径外,还设计了一种新的甲羟戊酸途径。该途径利用乙酰辅酶A作为前体,将其缩合形成乙酰乙酰辅酶A,随后导致IPP的形成。引入这条新途径后,与ackA-pta、nuo突变体菌株相比,番茄红素产量增加了两倍多。