College of Biology and the Environment, Nanjing Forestry University, Nanjing 210037, China.
Molecules. 2018 Jun 6;23(6):1369. doi: 10.3390/molecules23061369.
Phycocyanin, which covalently binds phycocyanobilin chromophores, is not only a candidate fluorescent probe for biological imaging, but also a potential antioxidative agent for healthcare. Herein, a plasmid harboring two cassettes was constructed, with from in one cassette and the fusion gene in the other, and then expressed in . PCB-CpcB(C-82), a fluorescent phycocyanin β subunit, was biosynthesized in , exhibiting an absorption maximum at 620 nm and fluorescence emission maximum at 640 nm. When was replaced by , PCB-CpcB(C-153), another fluorescent phycocyanin β subunit, was produced, exhibiting an absorption maximum at 590 nm and fluorescence emission maximum at 620 nm. These two fluorescent biliproteins showed stronger scavenging activity toward hydroxyl and DPPH free radicals than apo-CpcB. The IC values for hydroxyl radical scavenging by PCB-CpcB(C-82), PCB-CpcB(C-153), and apo-CpcB were 38.72 ± 2.48 µg/mL, 51.06 ± 6.74 µg/mL, and 81.82 ± 0.67 µg/mL, respectively, and the values for DPPH radical scavenging were 201.00 ± 5.86 µg/mL, 240.34 ± 4.03 µg/mL, and 352.93 ± 26.30 µg/mL, respectively. The comparative antioxidant capacities of the proteins were PCB-CpcB(C-82) > PCB-CpcB(C-153) > apo-CpcB, due to bilin binding. The two fluorescent biliproteins exhibited a significant effect on relieving the growth of cells injured by H₂O₂. The results of this study suggest that the fluorescent phycocyanin β subunits of were reconstructed by one expression vector in , and could be developed as potential antioxidants.
藻蓝蛋白与藻蓝胆素发色团共价结合,不仅是生物成像的候选荧光探针,也是医疗保健的潜在抗氧化剂。本文构建了一个含有两个盒的质粒,一个盒中含有 ,另一个盒中含有融合基因 ,并在 中表达。在 中生物合成了荧光藻蓝蛋白β亚基 PCB-CpcB(C-82),其吸收最大值为 620nm,荧光发射最大值为 640nm。当 被 取代时,产生了另一个荧光藻蓝蛋白β亚基 PCB-CpcB(C-153),其吸收最大值为 590nm,荧光发射最大值为 620nm。这两种荧光双蛋白对羟基和 DPPH 自由基的清除活性均强于脱辅基藻蓝蛋白。PCB-CpcB(C-82)、PCB-CpcB(C-153)和脱辅基藻蓝蛋白清除羟基自由基的 IC 值分别为 38.72±2.48µg/mL、51.06±6.74µg/mL 和 81.82±0.67µg/mL,清除 DPPH 自由基的 IC 值分别为 201.00±5.86µg/mL、240.34±4.03µg/mL 和 352.93±26.30µg/mL。由于结合了胆红素,这三种蛋白的抗氧化能力比较为:PCB-CpcB(C-82)>PCB-CpcB(C-153)>脱辅基藻蓝蛋白。两种荧光双蛋白对 H₂O₂损伤 的细胞生长有明显的缓解作用。本研究表明,通过一个表达载体在 中重建了 的荧光藻蓝蛋白β亚基,可作为潜在的抗氧化剂进行开发。