舍瓦内拉
联合囊肿
电子传输链
化学
生物物理学
细胞色素
异源表达
恶臭假单胞菌
细胞生物学
光电流
生物化学
生物
重组DNA
细菌
材料科学
光电子学
突变体
基因
酶
遗传学
作者
Melania Reggente,Nils Schürgers,Mohammed Mouhib,Sara Politi,Alessandra Antonucci,Ardemis A. Boghossian
标识
DOI:10.1101/2023.02.28.530417
摘要
Abstract At the center of microbial bioelectricity applications lies the critical need to express foreign heme proteins that are capable of redirecting the electron flux of the cell’s metabolism. This study presents bioengineered Synechocystis sp. PCC 6803 cells capable of increased electrogenicity through the introduction of a genetic construct for cytochrome expression. We could demonstrate the functional expression of the periplasmic MtrA decaheme c-type cytochrome from Shewanella oneidensis , a dissimilatory metal-reducing exoelectrogen, inside Synechocystis . Protein expression was verified through western-blotting and immunostaining, and oxygen evolution, optical density, and absorption measurements confirm sustained cell activity and viability under the tested expression conditions. Furthermore, the bioengineered cells show enhanced mediated exoelectrogenicity, as confirmed through a colorimetric iron assay and electrochemical measurements. Compared to wildtype cells on graphite electrodes, the bioengineered cells show a 2-fold increase in light-dependent, extracellular electron transfer, achieving photocurrent densities of 4 μA/cm 2 under white light illumination of ∼500 μmol m -2 s -1 . The increased capacitance obtained under illumination and suppressed photocurrents in the presence of the photosynthetic inhibitor, 3-(3,4-dichlorophenyl)-1,1-dimethylurea (DCMU) suggest increased extraction of photosynthetically derived electrons from the recombinant cells. The improved bioelectricity transport across the outer membranes, as achieved through the heterologous heme expression inside cyanobacteria, presents new opportunities for re-wiring the metabolisms of light-harvesting microbes.
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