Phycoerythrin （PE） is one of the most important proteins involved in light capturing during photosynthesis in red algae. Its potential biological activities had gained wide concerns. In the present study, tumor cyto...Phycoerythrin （PE） is one of the most important proteins involved in light capturing during photosynthesis in red algae. Its potential biological activities had gained wide concerns. In the present study, tumor cytotoxic and hydroxyl radical assay were preformed to detect the bioactivity of recombinant PE. Recombinant plasmids pGEX-PE and pBGL were transformed into E.coli BL21 to make two recombinant strains BEX （pGEX-PE） and BGL （pBGL）. PE expressing in BEX （pGEX-PE） was validated by SDS-PAGE and Western blotting analysis. SDS-PAGE analysis indicated that the PE-GST fusion protein was mostly inclusion bodies. Specific expression of PE was confirmed by Western blotting analysis. The recombinant E. coli BEX （pGEX-PE） cells were collected and sonicated. The supernatants were reserved for the tumor cytotoxic experiments. The result of tumor cytotoxic assay indicated that the supernatants containing PE had the activity of inhibiting the growth of Hela cells and with the increase of protein concentration, the inhibiting rate increased from 37.31% to 63.26%, which showed significant difference from the control. Hydroxyl radical scavenging effect was tested with supernatants of BEX （pGEX-PE） and BGL （pBGL） cell lysates treated with sonication and heating. For the sonication samples, the scavenging rates of the supernatants of BEX （pGEX-PE） and BGL （pBGL） cell lysates were significantly higher than the negative control BL2 I（pGEX-4T） （P〈0.02）, and the scavenging rates increased slowly following the increase of the protein content. For the heating samples, except for the 0.2 mgmL^-1 BGL （pBGL） products, the scavenging effects of the supernatants of BEX （pGEX-PE） and BGL （pBGL） cell lysates were stronger than that of negative control BL21（pGEX-4T）. However, the effect intensity was not positively correlated with the increase of the protein concentration. Though a partially decreased hydroxyl radical scavenging activity was led by heating, the biological activity was still retained and conspicuous. This research showed that phycoerythrin protein expressing in E. coli has the potential medical and sanitarian value.展开更多
Three surveys were carried out in Pearl River Estuary and adjacent coastal area in May, August, and November, 2013, to investigate the temporal and spatial variations of abundance of phycoerythrin-rich Synechococcus(P...Three surveys were carried out in Pearl River Estuary and adjacent coastal area in May, August, and November, 2013, to investigate the temporal and spatial variations of abundance of phycoerythrin-rich Synechococcus(PE-rich SYN) and phycocyanin-rich Synechococcus(PC-rich SYN). The effects of environmental factors on the alternation of the different Synechococcus groups were also elucidated. PE-rich SYN was detected in three surveys, whereas PC-rich SYN was detected in May and August, but not in November. The highest abundances of PE-rich SYN and PC-rich SYN were recorded in August and May, with mean values of 74.17×103 and 189.92×103 cells m L-1, respectively. From May to November, the relative abundance of PE-rich SYN increased, whereas that of PC-rich SYN declined. PE-rich and PC-rich SYN presented similar horizontal distributions with high abundance in the southern estuary in May, and in the western estuary in August. The abundances of PE-rich and PC-rich SYN were high at 27–32℃and salinity of 10–20. PC-rich SYN was not detected at < 24℃, and PC:PE-rich SYN decreased in abundance with salinity increase. When less than 20 mg L-1, suspended particulate matter(SPM) was helpful for Synechococcus growth. PE-rich SYN decreased in abundance when the concentration of dissolved inorganic nitrogen increased in May and November, and the concentration of phosphate increased in November. However, PC-rich SYN abundance and nutrients showed no correlation. Principal component analysis and regression analysis indicated that PE-rich SYN significantly correlated with the principal components that were affected by environmental factors.展开更多
文摘Phycoerythrin （PE） is one of the most important proteins involved in light capturing during photosynthesis in red algae. Its potential biological activities had gained wide concerns. In the present study, tumor cytotoxic and hydroxyl radical assay were preformed to detect the bioactivity of recombinant PE. Recombinant plasmids pGEX-PE and pBGL were transformed into E.coli BL21 to make two recombinant strains BEX （pGEX-PE） and BGL （pBGL）. PE expressing in BEX （pGEX-PE） was validated by SDS-PAGE and Western blotting analysis. SDS-PAGE analysis indicated that the PE-GST fusion protein was mostly inclusion bodies. Specific expression of PE was confirmed by Western blotting analysis. The recombinant E. coli BEX （pGEX-PE） cells were collected and sonicated. The supernatants were reserved for the tumor cytotoxic experiments. The result of tumor cytotoxic assay indicated that the supernatants containing PE had the activity of inhibiting the growth of Hela cells and with the increase of protein concentration, the inhibiting rate increased from 37.31% to 63.26%, which showed significant difference from the control. Hydroxyl radical scavenging effect was tested with supernatants of BEX （pGEX-PE） and BGL （pBGL） cell lysates treated with sonication and heating. For the sonication samples, the scavenging rates of the supernatants of BEX （pGEX-PE） and BGL （pBGL） cell lysates were significantly higher than the negative control BL2 I（pGEX-4T） （P〈0.02）, and the scavenging rates increased slowly following the increase of the protein content. For the heating samples, except for the 0.2 mgmL^-1 BGL （pBGL） products, the scavenging effects of the supernatants of BEX （pGEX-PE） and BGL （pBGL） cell lysates were stronger than that of negative control BL21（pGEX-4T）. However, the effect intensity was not positively correlated with the increase of the protein concentration. Though a partially decreased hydroxyl radical scavenging activity was led by heating, the biological activity was still retained and conspicuous. This research showed that phycoerythrin protein expressing in E. coli has the potential medical and sanitarian value.
基金supported by the National Natural Science Foundation of China (Nos. 41476098, 41276154)Special Scientific Research Funds for Central Non-profit Institutes of Yellow Sea Fisheries Research Institutes (2060302201502)
文摘Three surveys were carried out in Pearl River Estuary and adjacent coastal area in May, August, and November, 2013, to investigate the temporal and spatial variations of abundance of phycoerythrin-rich Synechococcus(PE-rich SYN) and phycocyanin-rich Synechococcus(PC-rich SYN). The effects of environmental factors on the alternation of the different Synechococcus groups were also elucidated. PE-rich SYN was detected in three surveys, whereas PC-rich SYN was detected in May and August, but not in November. The highest abundances of PE-rich SYN and PC-rich SYN were recorded in August and May, with mean values of 74.17×103 and 189.92×103 cells m L-1, respectively. From May to November, the relative abundance of PE-rich SYN increased, whereas that of PC-rich SYN declined. PE-rich and PC-rich SYN presented similar horizontal distributions with high abundance in the southern estuary in May, and in the western estuary in August. The abundances of PE-rich and PC-rich SYN were high at 27–32℃and salinity of 10–20. PC-rich SYN was not detected at < 24℃, and PC:PE-rich SYN decreased in abundance with salinity increase. When less than 20 mg L-1, suspended particulate matter(SPM) was helpful for Synechococcus growth. PE-rich SYN decreased in abundance when the concentration of dissolved inorganic nitrogen increased in May and November, and the concentration of phosphate increased in November. However, PC-rich SYN abundance and nutrients showed no correlation. Principal component analysis and regression analysis indicated that PE-rich SYN significantly correlated with the principal components that were affected by environmental factors.
