螺旋藻(Spirulina)藻蓝蛋白具有独特的理化特性及生理功能,是药物、食品和化妆品的天然原料,具有较大的开发潜力。为探讨螺旋藻藻蓝蛋白的研究现状与发展前景,对中国知网和Web of Science数据库中1990—2023年发表的文献进行检索并筛选...螺旋藻(Spirulina)藻蓝蛋白具有独特的理化特性及生理功能,是药物、食品和化妆品的天然原料,具有较大的开发潜力。为探讨螺旋藻藻蓝蛋白的研究现状与发展前景,对中国知网和Web of Science数据库中1990—2023年发表的文献进行检索并筛选,使用Cite Space软件对文章发文量、研究团队及研究热点进行图谱分析。综合分析可知,国内年发文量偏少,呈平稳趋势;国外年发文量持续上升,尤其近几年发文量迅速增长,且发文量超过了100篇;国外研究热点集中于藻蓝蛋白在食品、医药行业的应用方面,而国内研究热点集中在提取纯化、稳定性、功能活性的研究与应用,下一步应结合研究现状开发适合规模化生产的提取纯化工艺,进一步加强藻蓝蛋白研究的广度与深度;国内外研究群体主要是高校的相关生物技术学院或研究机构等,总体来讲,学者间存在较为密切的合作,但研究机构间尚未形成紧密的合作关系,在地域上比较分散,各大高校和研究机构应突破地区或机构间的各种限制,促进该研究领域的深度融合和快速发展,深入挖掘藻蓝蛋白在各个领域的潜在应用。展开更多
The ε-subunit is the smallest subunit of chloroplast ATP synthase, and is known to inhibit ATPase activity in isolated CF1-ATPase. As a result ε is sometimes called an inhibitory subunit. In addition, and perhaps mo...The ε-subunit is the smallest subunit of chloroplast ATP synthase, and is known to inhibit ATPase activity in isolated CF1-ATPase. As a result ε is sometimes called an inhibitory subunit. In addition, and perhaps more importantly, the ε -subunit is essential for the coupling of proton translocation to ATP synthesis (as proton gate). The relation between the structure and function of ε -subunit of ATP synthase in higher plant chloroplast has been studied by molecular biological methods such as site-directed mu-tagenesis and truncations for C- or N-terminus of ε -subunit. The results showed that: (1) Thr42 of ε-subunit is important for its structure and function; (2) compared with the ε-subunit in E.coli, the ε-subunit of chloroplast ATP synthase is more sensitive to C- or N-terminus truncations.展开更多
Nine pigment-protein complexes were separated and characterized from intact Chroomonasplacoidea chloroplasts by IEF. The bands Ⅰ-Ⅵ with their isoelectric points (pI) values from 4 to 6 were phycocyanin components;...Nine pigment-protein complexes were separated and characterized from intact Chroomonasplacoidea chloroplasts by IEF. The bands Ⅰ-Ⅵ with their isoelectric points (pI) values from 4 to 6 were phycocyanin components; bands Ⅷ and Ⅸ (pI = 2.8-3.6) were chlorophyll-protein complexes. According to absorption and fluorescence spectra, band VII was designated as a novel phycocyanin-Chla/c2-protein complex (pI ≈ 3.4-3.7). These results indicated that phycocyanin is structurally and functionally coupled with chlorophyll-protein complex in C. placoidea, and probably interacted with electrostatic force in combination.展开更多
Phycocyanin (PC), a natural algal protein, is reported for having anti-oxidant and antiinfl ammatory properties. We investigated its ability to attenuate lung infl ammation in mice subjected to X-ray radiation. Male C...Phycocyanin (PC), a natural algal protein, is reported for having anti-oxidant and antiinfl ammatory properties. We investigated its ability to attenuate lung infl ammation in mice subjected to X-ray radiation. Male C57BL/6 mice were assigned to the control, total body irradiation, PC pretreatment, and PC treatment groups. Mice in the PC pretreatment group were gavaged with 200 mg/kg PC for 7 consecutive days before irradiation, and those in the PC treatment group were gavaged with 200 mg/kg PC for 7 consecutive days after irradiation. Lungs were collected on Day 7 after irradiation exposure. Hematoxylin and eosin staining of mouse lung sections showed considerable infl ammation damage 7 days after irradiation compared with the control lung but a reduction in pathological injury in the PC treatment group. Pretreatment or treatment with PC signifi cantly decreased levels of interleukin-6 and tumor necrosis factor-α in the lung, and also increased the relative mRNA expression of superoxide dismutase and glutathione. In vivo, PC signifi cantly reduced the expression of Toll-like receptor TLR2, myeloid diff erentiation primary response Myd88, and nuclear factor NF-κB, at both the transcriptional and translation level. Taken together, these data indicated that PC attenuated lung infl ammatory damage induced by radiation by blocking the TLR2- MyD88-NF-κB signaling pathway. Therefore, PC could be a protective agent against radiation-induced infl ammatory damage in normal tissues.展开更多
The management of cyanobacteria and potential exposure to associated biotoxins requires the allocation of scarce resources across a range of freshwater resources within various jurisdictions. Cost effective and reliab...The management of cyanobacteria and potential exposure to associated biotoxins requires the allocation of scarce resources across a range of freshwater resources within various jurisdictions. Cost effective and reliable methods for sample processing and analysis form the foundation of the protocol yielding reliable data from which to derive important decisions. In this study the utilization of new methods to collect, process and analyze samples enhanced our ability to evaluate cyanobacterial populations. Extraction of phycocyanin using the single freeze thaw method provided more accurate and precise measurements (CV 4.7% and 6.4%), offering a simple and cost-effective means to overcome the influence of morphological variability. In-vacuo concentration of samples prior to ELISA analysis provided a detection limit of 0.001 μg·L?1 MC. Fractionation of samples (?1) = ?0.279 + (1.368 ? Log PC (μg·L?1) while in an Aphanizomemon spp. dominant system Log MC (ng·L?1) = 0.385 + (0.449 ? Log PC (μg·L?1). These methods and sampling protocol could be used in other aquatic systems across a broader regional landscape to estimate the levels of microcystins.展开更多
Na^+/K^+-ATPases are membrane-associated enzymes responsible for the active transport of Na^+ and K^+ ions across cell membranes, generating chemical and electrical gradients. These enzymes' α-subunit provides c...Na^+/K^+-ATPases are membrane-associated enzymes responsible for the active transport of Na^+ and K^+ ions across cell membranes, generating chemical and electrical gradients. These enzymes' α-subunit provides catalytic function, binding and hydrolyzing ATP, and itself becoming phosphorylated during the transport cycle. In this study, Na^+/K^+-ATPase α-subunit eDNA was cloned from gill tissue of the swimming crab Portunus trituberculatus by reverse-transcription polymerase chain reaction (RT-PCR) and rapid amplification of eDNA end methods. Analysis of the nucleotide sequence revealed that the eDNA had a full-length of 3 833 base pairs (bp), with an open reading frame of 3 120 bp, 5' untranslated region (UTR) of 317 bp, and 3' UTR of 396 bp. The sequence encoded a 1 039 amino acid protein with a predicted molecular weight of 115.57 kDa and with estimated pI of 5.21. It was predicted here to possess all expected features of Na^+/K^+-ATPase members, including eight transmembrane domains, putative ATP-binding site, and phosphorylation site. Comparison of arnino acid sequences showed that the P. tritubereulatus α-subunit possessed an overall identity of 75%-99% to that of other organisms. Phylogenetic analysis revealed that this α-subunit was in the same category as those of crustaceans. Quantitative real-time RT-PCR analysis indicated that this α-subunit's transcript were most highly expressed in gill and lowest in muscle. RT-PCR analysis also revealed that α-subunit expression in crab gill decreased after 2 and 6 h, but increased after 12, 24, 48, and 72 h. In addition, α-subunit expression in hepatopancreas of crab decreased after 2-72 h. These facts indicated that the crab's Na^+/K^+-ATPase α-subunit was potentially involved in the observed acute response to low salinity stress.展开更多
Phycoerythrin and phycocyanin were purified from Porphyra yezoensis Ueda with their bioactivity determined in this study. Continuous precipitation with ammonium sulfate at different concentrations(10%, 20%, 40% and 50...Phycoerythrin and phycocyanin were purified from Porphyra yezoensis Ueda with their bioactivity determined in this study. Continuous precipitation with ammonium sulfate at different concentrations(10%, 20%, 40% and 50%) increased the purity(A564:A280) of phycoerythrin to 1.49, 3.92 fold of the raw extract(0.38) and the purity(A615:A280) of phycocyanin to 0.70, 3.33 fold of the raw extract(0.21). Two more times of chromatography with hydroxylapatites finally made the purity of phycoerythrin and phycocyanin reach 5.50, 14.47 fold of the raw extract, and 5.10, 24.29 fold of the raw extract, respectviely. The yield of high purity phycoerythrin and phycocyanin were 0.21% and 0.09% of dried P. yezoensis blade, respectively. The photodynamic cytotoxic experiment showed that both phycoerythrin and phycocyanin inhibited the growth of liver tumor cells significantly. It was found that 250 mg L-1 purified phycoerythrin and phycocyanin inhibited the growth of hepatocellular carcinoma cells 24 h after laser-irradiation by 80% and 59%, respectively, and 100 mg L-1 purified phycoerythrin and phycocyanin induced the apoptosis of 31.54% and 32.54% of the cells, respectively, 8 h after photodynamic therapy. Oue findings demonstrated that P. yezoensis can serve as photosensitizer(phycoerythrin and phycocyanin) producer.展开更多
BACKGROUND: Phycocyanin can relieve decrease of mitochondrial membrane potential through reducing production of active oxygen so as to protect neurons after hypoxia/reoxygenation. OBJECTIVE: To observe the effect of...BACKGROUND: Phycocyanin can relieve decrease of mitochondrial membrane potential through reducing production of active oxygen so as to protect neurons after hypoxia/reoxygenation. OBJECTIVE: To observe the effect of phycocyanin on activity of PC12 cells and mitochondrial membrane potential after hypoxia/reoxygenation. DESIGN: Randomized controlled study SETTING : Cerebrovascular Disease Institute of Affiliated Hospital, Medical College of Qingdao University MATERIALS: The experiment was carried out at the Key Laboratory of Prevention and Cure for cerebropathia in Shandong Province from October to December 2005. PC12 cells, rat chromaffin tumor cells, were provided by Storage Center of Wuhan University; phycocyanin was provided by Ocean Institute of Academia Sinica; Thiazoyl blue tetrazolium bromide (MTT) and rhodamine 123 were purchased from Sigma Company, USA; RPMI-1640 medium, fetal bovine serum and equine serum were purchased from Gibco Company, USA. METHODS: ① Culture of PC12 cells: PC12 cells were put into RPMI-1640 medium which contained 100 g/L heat inactivation equine serum and 0.05 volume fraction of fetal bovine serum and incubated in CO2 incubator at 37℃. Number of cells was regulated to 4 × 10^5 L 1, and cells were inoculated at 96-well culture plate. The final volume was 100μL. ② Model establishing and grouping: Cultured PC12 cells were randomly divided into three groups: phycocyanin group, model control group and non-hypoxia group. At 24 hours before hypoxia, culture solution in phycocyanin group was added with phycocyanin so as to make sure the final concentration of 3 g/L , but cells in model control group did not add with phycocyanin. Cells in non-hypoxia group were also randomly divided into adding phycocyanin group (the final concentration of 3 g/L) and non-adding phycocyanin group. Cells in model control group and phycocyanin group were cultured with hypoxia for 1 hour and reoxygenation for 1, 2 and 3 hours; meanwhile, cells in non-hypoxia group were cultured with oxygen and were measured at 1 hour after hypoxia/reoxygenation. ③ Detecting items: At 1, 2 and 3 hours after reoxygenation, absorbance (A value) of PC12 cells was measured with MTT technique so as to observe activity and quantity of cells. Fluorescence intensity of PC12 cells marked by rhodamine 123 was measured with confocal microscope in order to observe changes of mitochondrial membrane potential. MAEN OUTCOME MEASURES: Comparisons between quantity and activity of PC12 cells and mitochondria membrane potential at 1, 2 and 3 hours after reoxygenation. RESULTS: ① Effect of phycocyanin on quantity and activity of PC12 cells: A value was 0.924±0.027 in adding phycocyanin group and 0.924±0.033 in non-adding phycocyanin group. A value was lower in model control group and phycocyanin group than that in non-hypoxia group at 1, 2 and 3 hours after reoxygenation (0.817±0.053, 0.838±0.037, 0.875±0.029; 0.842±0.029, 0.872±0.025, 0.906±0.023, P 〈 0.05). A value was higher in phycocyanin group than that in model control group at 1, 2 and 3 after culture (P 〈 0.05). With culture time being longer, A value was increased gradually in phycocyanin group and model control group after reoxygenation (P 〈 0.05). ~ Effect of phycocyanin on mitochondrial membrane potential of PC12 cells: Fluorescence intensity was 2.967±0.253 in adding phycocyanin group and 2.962±0.294 in non-adding phycocyanin group. Fluorescence intensity was lower in model control group and phycocyanin group than that in non-hypoxia group at 1, 2 and 3 hours after hypoxia/reoxygenation (1.899±0.397, 2.119±0.414, 2.287±0.402; 2.191±0.377, 2.264±0.359, 2.436±0.471, P 〈 0.05); but it was higher in phycocyanin group than that in model control group at 1, 2 and 3 after reoxygenation (P 〈 0.05). With culture time being longer, fluorescence intensity was increased gradually in phycocyanin group and model control group after reoxygenation (P 〈 0.05). CONCLUSION: Phycocyanin and reoxygenation can protect PC12 cells after hypoxia injury through increasing mitochondrial membrane potential and cellular activity, and the effect is improved gradually with prolonging time of reoxygenation.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
The synthesized C-phycocyanins (C-PCs) doped silica biomaterials were characterized by the SEM and BET surface area analysis measurement. The morphology of C-PCs doped silica biomaterials indicates that the surface ...The synthesized C-phycocyanins (C-PCs) doped silica biomaterials were characterized by the SEM and BET surface area analysis measurement. The morphology of C-PCs doped silica biomaterials indicates that the surface of the silica cluster is formed by a great number of silica particles with an average size of between 30 and 40 nm. Silica itself is a porous structure with the average pore diameter of 2.95 nm. Pores with their diameter less than 5 nm account for 84.07%. In addition, the C-PCs can be utilized as a fluorescent protein probe to monitor influence of the protein encapsulation and to study matrix and protein interaction and stability of protein in silica matrix. Application of protein encapsulation silica materials requires biomolecules to keep bioactivity and stability on potentially unfavorable industrial conditions. The C-PCs in solution or in silicate matrix irradiated by ultraviolet ray can result in photobleaching, whereas the protein in the silica is less affected. The measured photodamage rate constant of C-PCs in buffer solution is 25 times faster than that of C-PCs in silica matrix. However, the lifetime of C-PCs in silica matrix or phosphate buffer is unaffected. These studies suggest that entrapment of C-PCs into silica matrixes not only can maintain their biological activity but also noticeably improve their photostability.展开更多
This study investigated the abnormal expression of ATP synthase β-subunit(ATPsyn-β) in pancreas islets of rat model of polycystic ovary syndrome(PCOS) with type 2 diabetes mellitus(T2DM),and the secretion func...This study investigated the abnormal expression of ATP synthase β-subunit(ATPsyn-β) in pancreas islets of rat model of polycystic ovary syndrome(PCOS) with type 2 diabetes mellitus(T2DM),and the secretion function changes after up-regulation of ATP5 b.Sixty female SD rats were divided into three groups randomly and equally.The rat model of PCOS with T2 DM was established by free access to the high-carbohydrate/high-fat diet,subcutaneous injections of DHEA,and a single injection of streptozotocin.The pancreas was removed for the detection of the ATPsyn-β expression by immunohistochemical staining,Western blotting and reverse transcription-PCR(RT-PCR).The pancreas islets of the rats were cultured,isolated with collagenase Ⅴ and purified by gradient centrifugation,and the insulin secretion after treatment with different glucose concentrations was tested.Lentivirus ATP5 b was successfully constructed with the vector of GV208 and transfected into the pancreas islets for the over-expression of ATPsyn-β.The insulin secretion and intracellular ATP content were determined after transfection of the PCOS-T2 DM pancreas islets with Lenti-ATP5 b.The results showed that the expression of ATPsyn-β protein and m RNA was significantly decreased in the pancreas of PCOS-T2 DM rats.The ATP content in the pancreas islets was greatly increased and the insulin secretion was improved after the up-regulation of ATPsyn-β in the pancreas islets transfected with lenti-ATP5 b.These results indicated that for PCOS,the ATPsyn-β might be one of the key factors for the attack of T2 DM.展开更多
The isolation of biliproteins from the Spirulina platensis cultured in southern China was accomplished with gel filtration on Sephacryl S 200 and chromatography on hydroxylapatite. The spectrophotometry, isoelect...The isolation of biliproteins from the Spirulina platensis cultured in southern China was accomplished with gel filtration on Sephacryl S 200 and chromatography on hydroxylapatite. The spectrophotometry, isoelectric point, and amino acid composition of C phycocyanin were determined, respectively.展开更多
文摘螺旋藻(Spirulina)藻蓝蛋白具有独特的理化特性及生理功能,是药物、食品和化妆品的天然原料,具有较大的开发潜力。为探讨螺旋藻藻蓝蛋白的研究现状与发展前景,对中国知网和Web of Science数据库中1990—2023年发表的文献进行检索并筛选,使用Cite Space软件对文章发文量、研究团队及研究热点进行图谱分析。综合分析可知,国内年发文量偏少,呈平稳趋势;国外年发文量持续上升,尤其近几年发文量迅速增长,且发文量超过了100篇;国外研究热点集中于藻蓝蛋白在食品、医药行业的应用方面,而国内研究热点集中在提取纯化、稳定性、功能活性的研究与应用,下一步应结合研究现状开发适合规模化生产的提取纯化工艺,进一步加强藻蓝蛋白研究的广度与深度;国内外研究群体主要是高校的相关生物技术学院或研究机构等,总体来讲,学者间存在较为密切的合作,但研究机构间尚未形成紧密的合作关系,在地域上比较分散,各大高校和研究机构应突破地区或机构间的各种限制,促进该研究领域的深度融合和快速发展,深入挖掘藻蓝蛋白在各个领域的潜在应用。
文摘The ε-subunit is the smallest subunit of chloroplast ATP synthase, and is known to inhibit ATPase activity in isolated CF1-ATPase. As a result ε is sometimes called an inhibitory subunit. In addition, and perhaps more importantly, the ε -subunit is essential for the coupling of proton translocation to ATP synthesis (as proton gate). The relation between the structure and function of ε -subunit of ATP synthase in higher plant chloroplast has been studied by molecular biological methods such as site-directed mu-tagenesis and truncations for C- or N-terminus of ε -subunit. The results showed that: (1) Thr42 of ε-subunit is important for its structure and function; (2) compared with the ε-subunit in E.coli, the ε-subunit of chloroplast ATP synthase is more sensitive to C- or N-terminus truncations.
文摘Nine pigment-protein complexes were separated and characterized from intact Chroomonasplacoidea chloroplasts by IEF. The bands Ⅰ-Ⅵ with their isoelectric points (pI) values from 4 to 6 were phycocyanin components; bands Ⅷ and Ⅸ (pI = 2.8-3.6) were chlorophyll-protein complexes. According to absorption and fluorescence spectra, band VII was designated as a novel phycocyanin-Chla/c2-protein complex (pI ≈ 3.4-3.7). These results indicated that phycocyanin is structurally and functionally coupled with chlorophyll-protein complex in C. placoidea, and probably interacted with electrostatic force in combination.
基金Supported by the National Key Research and Development Program of China(No.2018YFD0901102)
文摘Phycocyanin (PC), a natural algal protein, is reported for having anti-oxidant and antiinfl ammatory properties. We investigated its ability to attenuate lung infl ammation in mice subjected to X-ray radiation. Male C57BL/6 mice were assigned to the control, total body irradiation, PC pretreatment, and PC treatment groups. Mice in the PC pretreatment group were gavaged with 200 mg/kg PC for 7 consecutive days before irradiation, and those in the PC treatment group were gavaged with 200 mg/kg PC for 7 consecutive days after irradiation. Lungs were collected on Day 7 after irradiation exposure. Hematoxylin and eosin staining of mouse lung sections showed considerable infl ammation damage 7 days after irradiation compared with the control lung but a reduction in pathological injury in the PC treatment group. Pretreatment or treatment with PC signifi cantly decreased levels of interleukin-6 and tumor necrosis factor-α in the lung, and also increased the relative mRNA expression of superoxide dismutase and glutathione. In vivo, PC signifi cantly reduced the expression of Toll-like receptor TLR2, myeloid diff erentiation primary response Myd88, and nuclear factor NF-κB, at both the transcriptional and translation level. Taken together, these data indicated that PC attenuated lung infl ammatory damage induced by radiation by blocking the TLR2- MyD88-NF-κB signaling pathway. Therefore, PC could be a protective agent against radiation-induced infl ammatory damage in normal tissues.
文摘The management of cyanobacteria and potential exposure to associated biotoxins requires the allocation of scarce resources across a range of freshwater resources within various jurisdictions. Cost effective and reliable methods for sample processing and analysis form the foundation of the protocol yielding reliable data from which to derive important decisions. In this study the utilization of new methods to collect, process and analyze samples enhanced our ability to evaluate cyanobacterial populations. Extraction of phycocyanin using the single freeze thaw method provided more accurate and precise measurements (CV 4.7% and 6.4%), offering a simple and cost-effective means to overcome the influence of morphological variability. In-vacuo concentration of samples prior to ELISA analysis provided a detection limit of 0.001 μg·L?1 MC. Fractionation of samples (?1) = ?0.279 + (1.368 ? Log PC (μg·L?1) while in an Aphanizomemon spp. dominant system Log MC (ng·L?1) = 0.385 + (0.449 ? Log PC (μg·L?1). These methods and sampling protocol could be used in other aquatic systems across a broader regional landscape to estimate the levels of microcystins.
基金Supported by the National High Technology Research and Development Program of China(863 Program)(No.2012AA10A409)the National Natural Science Foundation of China(No.41306177)the Special Scientific Research Funds for Central Non-Profit Institutes,Yellow Sea Fisheries Research Institutes(No.20603022013027)
文摘Na^+/K^+-ATPases are membrane-associated enzymes responsible for the active transport of Na^+ and K^+ ions across cell membranes, generating chemical and electrical gradients. These enzymes' α-subunit provides catalytic function, binding and hydrolyzing ATP, and itself becoming phosphorylated during the transport cycle. In this study, Na^+/K^+-ATPase α-subunit eDNA was cloned from gill tissue of the swimming crab Portunus trituberculatus by reverse-transcription polymerase chain reaction (RT-PCR) and rapid amplification of eDNA end methods. Analysis of the nucleotide sequence revealed that the eDNA had a full-length of 3 833 base pairs (bp), with an open reading frame of 3 120 bp, 5' untranslated region (UTR) of 317 bp, and 3' UTR of 396 bp. The sequence encoded a 1 039 amino acid protein with a predicted molecular weight of 115.57 kDa and with estimated pI of 5.21. It was predicted here to possess all expected features of Na^+/K^+-ATPase members, including eight transmembrane domains, putative ATP-binding site, and phosphorylation site. Comparison of arnino acid sequences showed that the P. tritubereulatus α-subunit possessed an overall identity of 75%-99% to that of other organisms. Phylogenetic analysis revealed that this α-subunit was in the same category as those of crustaceans. Quantitative real-time RT-PCR analysis indicated that this α-subunit's transcript were most highly expressed in gill and lowest in muscle. RT-PCR analysis also revealed that α-subunit expression in crab gill decreased after 2 and 6 h, but increased after 12, 24, 48, and 72 h. In addition, α-subunit expression in hepatopancreas of crab decreased after 2-72 h. These facts indicated that the crab's Na^+/K^+-ATPase α-subunit was potentially involved in the observed acute response to low salinity stress.
基金financially supported by the National Key Technology R&D Program (2012BAC07B03)Shanghai Universities First-class Disciplines Project,Discipline name: Marine Science and Shanghai Municipal Education Commission (Preponderant Subject Program #S30701)Key Laboratory of Freshwater Fishery Germplasm Resources, Ministry of Agriculture, P. R. China, Shanghai Engineering Research Center of Aquaculture, Shanghai University Knowledge Service Platform, Shanghai Ocean University Aquatic Animal Breeding Center (ZF1206)
文摘Phycoerythrin and phycocyanin were purified from Porphyra yezoensis Ueda with their bioactivity determined in this study. Continuous precipitation with ammonium sulfate at different concentrations(10%, 20%, 40% and 50%) increased the purity(A564:A280) of phycoerythrin to 1.49, 3.92 fold of the raw extract(0.38) and the purity(A615:A280) of phycocyanin to 0.70, 3.33 fold of the raw extract(0.21). Two more times of chromatography with hydroxylapatites finally made the purity of phycoerythrin and phycocyanin reach 5.50, 14.47 fold of the raw extract, and 5.10, 24.29 fold of the raw extract, respectviely. The yield of high purity phycoerythrin and phycocyanin were 0.21% and 0.09% of dried P. yezoensis blade, respectively. The photodynamic cytotoxic experiment showed that both phycoerythrin and phycocyanin inhibited the growth of liver tumor cells significantly. It was found that 250 mg L-1 purified phycoerythrin and phycocyanin inhibited the growth of hepatocellular carcinoma cells 24 h after laser-irradiation by 80% and 59%, respectively, and 100 mg L-1 purified phycoerythrin and phycocyanin induced the apoptosis of 31.54% and 32.54% of the cells, respectively, 8 h after photodynamic therapy. Oue findings demonstrated that P. yezoensis can serve as photosensitizer(phycoerythrin and phycocyanin) producer.
基金the Natural Science Foundation of Shandong Province, No. Y2004C04
文摘BACKGROUND: Phycocyanin can relieve decrease of mitochondrial membrane potential through reducing production of active oxygen so as to protect neurons after hypoxia/reoxygenation. OBJECTIVE: To observe the effect of phycocyanin on activity of PC12 cells and mitochondrial membrane potential after hypoxia/reoxygenation. DESIGN: Randomized controlled study SETTING : Cerebrovascular Disease Institute of Affiliated Hospital, Medical College of Qingdao University MATERIALS: The experiment was carried out at the Key Laboratory of Prevention and Cure for cerebropathia in Shandong Province from October to December 2005. PC12 cells, rat chromaffin tumor cells, were provided by Storage Center of Wuhan University; phycocyanin was provided by Ocean Institute of Academia Sinica; Thiazoyl blue tetrazolium bromide (MTT) and rhodamine 123 were purchased from Sigma Company, USA; RPMI-1640 medium, fetal bovine serum and equine serum were purchased from Gibco Company, USA. METHODS: ① Culture of PC12 cells: PC12 cells were put into RPMI-1640 medium which contained 100 g/L heat inactivation equine serum and 0.05 volume fraction of fetal bovine serum and incubated in CO2 incubator at 37℃. Number of cells was regulated to 4 × 10^5 L 1, and cells were inoculated at 96-well culture plate. The final volume was 100μL. ② Model establishing and grouping: Cultured PC12 cells were randomly divided into three groups: phycocyanin group, model control group and non-hypoxia group. At 24 hours before hypoxia, culture solution in phycocyanin group was added with phycocyanin so as to make sure the final concentration of 3 g/L , but cells in model control group did not add with phycocyanin. Cells in non-hypoxia group were also randomly divided into adding phycocyanin group (the final concentration of 3 g/L) and non-adding phycocyanin group. Cells in model control group and phycocyanin group were cultured with hypoxia for 1 hour and reoxygenation for 1, 2 and 3 hours; meanwhile, cells in non-hypoxia group were cultured with oxygen and were measured at 1 hour after hypoxia/reoxygenation. ③ Detecting items: At 1, 2 and 3 hours after reoxygenation, absorbance (A value) of PC12 cells was measured with MTT technique so as to observe activity and quantity of cells. Fluorescence intensity of PC12 cells marked by rhodamine 123 was measured with confocal microscope in order to observe changes of mitochondrial membrane potential. MAEN OUTCOME MEASURES: Comparisons between quantity and activity of PC12 cells and mitochondria membrane potential at 1, 2 and 3 hours after reoxygenation. RESULTS: ① Effect of phycocyanin on quantity and activity of PC12 cells: A value was 0.924±0.027 in adding phycocyanin group and 0.924±0.033 in non-adding phycocyanin group. A value was lower in model control group and phycocyanin group than that in non-hypoxia group at 1, 2 and 3 hours after reoxygenation (0.817±0.053, 0.838±0.037, 0.875±0.029; 0.842±0.029, 0.872±0.025, 0.906±0.023, P 〈 0.05). A value was higher in phycocyanin group than that in model control group at 1, 2 and 3 after culture (P 〈 0.05). With culture time being longer, A value was increased gradually in phycocyanin group and model control group after reoxygenation (P 〈 0.05). ~ Effect of phycocyanin on mitochondrial membrane potential of PC12 cells: Fluorescence intensity was 2.967±0.253 in adding phycocyanin group and 2.962±0.294 in non-adding phycocyanin group. Fluorescence intensity was lower in model control group and phycocyanin group than that in non-hypoxia group at 1, 2 and 3 hours after hypoxia/reoxygenation (1.899±0.397, 2.119±0.414, 2.287±0.402; 2.191±0.377, 2.264±0.359, 2.436±0.471, P 〈 0.05); but it was higher in phycocyanin group than that in model control group at 1, 2 and 3 after reoxygenation (P 〈 0.05). With culture time being longer, fluorescence intensity was increased gradually in phycocyanin group and model control group after reoxygenation (P 〈 0.05). CONCLUSION: Phycocyanin and reoxygenation can protect PC12 cells after hypoxia injury through increasing mitochondrial membrane potential and cellular activity, and the effect is improved gradually with prolonging time of reoxygenation.
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.
基金Funded by the National Natural Science Foundation of China (No. 20873005)Beijing Natural Science Fundation (No.2083028)+1 种基金the Scientific Research Foundation for the Returned Overseas Chinese Scholars, State Education Ministry422 Funds from University of Science and Technology Beijing
文摘The synthesized C-phycocyanins (C-PCs) doped silica biomaterials were characterized by the SEM and BET surface area analysis measurement. The morphology of C-PCs doped silica biomaterials indicates that the surface of the silica cluster is formed by a great number of silica particles with an average size of between 30 and 40 nm. Silica itself is a porous structure with the average pore diameter of 2.95 nm. Pores with their diameter less than 5 nm account for 84.07%. In addition, the C-PCs can be utilized as a fluorescent protein probe to monitor influence of the protein encapsulation and to study matrix and protein interaction and stability of protein in silica matrix. Application of protein encapsulation silica materials requires biomolecules to keep bioactivity and stability on potentially unfavorable industrial conditions. The C-PCs in solution or in silicate matrix irradiated by ultraviolet ray can result in photobleaching, whereas the protein in the silica is less affected. The measured photodamage rate constant of C-PCs in buffer solution is 25 times faster than that of C-PCs in silica matrix. However, the lifetime of C-PCs in silica matrix or phosphate buffer is unaffected. These studies suggest that entrapment of C-PCs into silica matrixes not only can maintain their biological activity but also noticeably improve their photostability.
文摘This study investigated the abnormal expression of ATP synthase β-subunit(ATPsyn-β) in pancreas islets of rat model of polycystic ovary syndrome(PCOS) with type 2 diabetes mellitus(T2DM),and the secretion function changes after up-regulation of ATP5 b.Sixty female SD rats were divided into three groups randomly and equally.The rat model of PCOS with T2 DM was established by free access to the high-carbohydrate/high-fat diet,subcutaneous injections of DHEA,and a single injection of streptozotocin.The pancreas was removed for the detection of the ATPsyn-β expression by immunohistochemical staining,Western blotting and reverse transcription-PCR(RT-PCR).The pancreas islets of the rats were cultured,isolated with collagenase Ⅴ and purified by gradient centrifugation,and the insulin secretion after treatment with different glucose concentrations was tested.Lentivirus ATP5 b was successfully constructed with the vector of GV208 and transfected into the pancreas islets for the over-expression of ATPsyn-β.The insulin secretion and intracellular ATP content were determined after transfection of the PCOS-T2 DM pancreas islets with Lenti-ATP5 b.The results showed that the expression of ATPsyn-β protein and m RNA was significantly decreased in the pancreas of PCOS-T2 DM rats.The ATP content in the pancreas islets was greatly increased and the insulin secretion was improved after the up-regulation of ATPsyn-β in the pancreas islets transfected with lenti-ATP5 b.These results indicated that for PCOS,the ATPsyn-β might be one of the key factors for the attack of T2 DM.
文摘The isolation of biliproteins from the Spirulina platensis cultured in southern China was accomplished with gel filtration on Sephacryl S 200 and chromatography on hydroxylapatite. The spectrophotometry, isoelectric point, and amino acid composition of C phycocyanin were determined, respectively.