Yang Boyu, Li Shubin, Mu Wei, Wang Zhao, Han Xiaojun
State Key Laboratory of Urban Water Resource and Environment, School of Chemistry and Chemical Engineering, Harbin Institute of Technology, 92 West Da-Zhi Street, Harbin, 150001, China.
Small. 2023 Mar;19(13):e2201305. doi: 10.1002/smll.202201305. Epub 2022 Jul 29.
The bottom-up constructed artificial cells help to understand the cell working mechanism and provide the evolution clues for organisms. The energy supply and metabolism mimicry are the key issues in the field of artificial cells. Herein, an artificial cell containing cyanobacteria capable of light harvesting and carbon dioxide fixation is demonstrated to produce glucose molecules by converting light energy into chemical energy. Two downstream "metabolic" pathways starting from glucose molecules are investigated. One involves enzyme cascade reaction to produce H O (assisted by glucose oxidase) first, followed by converting Amplex red to resorufin (assisted by horseradish peroxidase). The other pathway is more biologically relevant. Glucose molecules are dehydrogenated to transfer hydrogens to nicotinamide adenine dinucleotide (NAD ) for the production of nicotinamide adenine dinucleotide hydride (NADH) molecules in the presence of glucose dehydrogenase. Further, NADH molecules are oxidized into NAD by pyruvate catalyzed by lactate dehydrogenase, meanwhile, lactate is obtained. Therefore, the cascade cycling of NADH/NAD is built. The artificial cells built here pave the way for investigating more complicated energy-supplied metabolism inside artificial cells.
自下而上构建的人工细胞有助于理解细胞的工作机制,并为生物体提供进化线索。能量供应和代谢模拟是人工细胞领域的关键问题。在此,展示了一种含有能够进行光捕获和二氧化碳固定的蓝细菌的人工细胞,其通过将光能转化为化学能来产生葡萄糖分子。研究了两条从葡萄糖分子开始的下游“代谢”途径。一条途径涉及酶级联反应,首先产生H₂O(由葡萄糖氧化酶辅助),随后将Amplex red转化为试卤灵(由辣根过氧化物酶辅助)。另一条途径与生物学相关性更强。在葡萄糖脱氢酶存在的情况下,葡萄糖分子脱氢将氢转移至烟酰胺腺嘌呤二核苷酸(NAD⁺)以产生还原型烟酰胺腺嘌呤二核苷酸(NADH)分子。此外,NADH分子在乳酸脱氢酶催化下被丙酮酸氧化为NAD⁺,同时得到乳酸。因此,构建了NADH/NAD⁺的级联循环。这里构建的人工细胞为研究人工细胞内更复杂的能量供应代谢铺平了道路。