The agp gene encoding the ADP-glucose pyrophosphorylase involved in cyanobacterial glycogen synthesis was amplified by PCR. The resulting agp fragment was cloned in plasmid pUC118 to generate plasmid pUCA. Part of the...The agp gene encoding the ADP-glucose pyrophosphorylase involved in cyanobacterial glycogen synthesis was amplified by PCR. The resulting agp fragment was cloned in plasmid pUC118 to generate plasmid pUCA. Part of the fragment within the agp DNA was deleted and replaced by an erythromycin resistance cassette to generate plasmid pUCAE, which was used to transform the Synechocystis sp. PCC 6803 wild-type strain and a mutant with resistance to erythromycin was obtained. PCR analysis of the genomic DNA from the resulting mutant indicated that the appropriate deletion and insertion indeed had occurred. The cell growth and Chl a, glycogen content in the mutant showed difference from those in the wild-type strain. The obtained biomass as well as the Chl a content in the mutant strain was higher than that of the wild-type strain, which suggested that the photosynthesis efficiency in the agp(-) strain was higher than that in the wild-type strain. No glycogen was found in the mutant, providing evidence for the correction of the mutant in physiological level.展开更多
A novel lipid occurred when cyanobacterium Synechocystis sp. PCC 6803 cells were grown in BG-11 medium with glucose applied. This lipid was determined to be a glycolipid, designated glycolipid-x (Glyco-x), by staining...A novel lipid occurred when cyanobacterium Synechocystis sp. PCC 6803 cells were grown in BG-11 medium with glucose applied. This lipid was determined to be a glycolipid, designated glycolipid-x (Glyco-x), by staining with alpha-naphthol and concentrated sulfuric acid. The occurrence of Glyco-x accompanies the disappearance of other lipids, especially DGDG. Glyco-x can also be observed in cells grown in BG-11 medium with the application of other carbon sources: fructose, maltose and lactose. Sodium thiosulfate, an effective scavenger of reactive oxygen intermediates, showed strong capability to inhibit glucose-induced occurrence of Glyco-x. In the presence of 0.3% sodium thiosulfate, Glyco-x could only be detected in cells grown in BG-11 medium with 100 mmol/L glucose applied in late-exponential phase. These results suggest that reactive oxygen species might be involved in the occurrence of Glyco-x in cyanobacterium Synechocystis sp. PCC 6803 cells grown in the presence of glucose.展开更多
Compositions of glycerolipids and fatty acid compositions of glycerolipids were compared among Synechocystis sp. PCC 6803 cells grown in the BG-11 medium containing different concentrations of glucose and Na2S2O3 in t...Compositions of glycerolipids and fatty acid compositions of glycerolipids were compared among Synechocystis sp. PCC 6803 cells grown in the BG-11 medium containing different concentrations of glucose and Na2S2O3 in this study. It was found that Na2S2O3 can effectively increase the percentage of sulphoquinovosyl diacylglycerol (SQDG) and phosphatidylglycerol (PG) to total membrane lipids and the simultaneous application of glucose with Na2S2O3 can counteract the effect of Na2S2O3. In addition, Na2S2O3 can significantly increase the percentage of palmitic acid (C, 16:0) in fatty acid composition of monogalactosyl diacylglycerol (MGDG) and digalactosyl diacylglycerol (DGDG) and decrease the fatty acid unsaturation degree accordingly, and these effects can also be eliminated by glucose. These results indicate that Na2S2O3 can take as a reductant to make membrane lipids in a low unsaturated state, and the simultaneous application of glucose can decrease the reducing power of Na2S2O3. In addition, Na2S2O3 can take as a sulfur donor for the synthesis of SQDG.展开更多
Calcium carbonate (CaCO3) crystals in their preferred orientation were obtained in BG11 culture media inoculated with Synechocystis sp. PCC6803 (inoculated BG11). In this study, the features of calcium carbonate d...Calcium carbonate (CaCO3) crystals in their preferred orientation were obtained in BG11 culture media inoculated with Synechocystis sp. PCC6803 (inoculated BG11). In this study, the features of calcium carbonate deposition were investigated. Inoculated BGll in different calcium ion concentrations was used for the experimental group, while the BGll culture medium was used for the control group. The surface morphologies of the calcium carbonate deposits in the experimental and control groups were determined by scanning and transmission electron microscopy. The deposits were analyzed by electronic probe micro-analysis, Fourier transform infrared spectrum, X-ray diffraction, thermal gravimetric analysis and differential scanning calorimetry. The results show that the surfaces of the crystals in the experimental group were hexahedral in a scaly pattern. The particle sizes were micrometer-sized and larger than those in the control group. The deposits of the control group contained calcium (Ca), carbon (C), oxygen (O), phosphorus (P), iron (Fe), copper (Cu), zinc (Zn), and other elements. The deposits in the experimental group contained Ca, C, and O only. The deposits of both groups contained calcite. The thermal decomposition temperature of the deposits in the control group was lower than those in the experimental group. It showed that the CaCO3 deposits of the experimental group had higher thermal stability than those of the control group. This may be due to the secondary metabolites produced by the algae cells, which affect the carbonate crystal structure and result in a close-packed structure. The algae cells that remained after thermal weight loss were heavier in higher calcium concentrations in BGll culture media. There may be more calcium- containing crystals inside and outside of these cells. These results shall be beneficial for understanding the formation mechanism of carbonate minerals.展开更多
The unicellular cyanobacterium Synechocystis sp. PCC 6803, a model organism known for its unique combination of highly desirable molecular genetic, physiological and morphological characteristics, was employed in the ...The unicellular cyanobacterium Synechocystis sp. PCC 6803, a model organism known for its unique combination of highly desirable molecular genetic, physiological and morphological characteristics, was employed in the present study. The species was cultured in BG11 liquid medium contained various initial concentrations of Pb^2+ and Cd〉 (0, 0.5, 1, 2, 4, 6 and 8 mg/L). The experiment was conducted for six days and the metal induced alterations in the ultrastructure, growth and pigment contents were assessed. Alterations in the ultrastructure of the Synechocystis sp. PCC 6803 cells became evident with the increased (〉4 mg/L Pb^2+) metal concentration. The photosynthetic apparatus (thylakoid membranes) were found to be the worst affected. Deteriorated or completely destroyed thylakoid membranes have made large empty spaces in the cell interior. In addition, at the highest concentration (8 mg/L pb^2+), the polyphosphate granules became more prominent both in size and number. Despite the initial slight stimulations (0.2, 3.8 and 6.5% respectively at 0.5, 1 and 2 mg/L pb^2+), both metals inhibited the growth in a dose-dependent manner as incubation progressed. Pigment contents (chlorophyll a, 13 carotene and phycocyanin) were also decreased with increasing metal concentration. Cells exposed to 6 mg/L Pb^2+, resulted in 36.56, 37.39 and 29.34% reductions of chlorophyll a, 13 carotene and phycocyanin respectively over the control. Corresponding reductions for the same CdZ+concentrations were 57.83, 48.94 and 56.90%. Lethal concentration (96 h LC50) values (3.47 mg/L Cd^2+ and 12.11 mg/L Pb^2+) indicated that Synechocystis sp. PCC 6803 is more vulnerable to Cd^2+ than Pb^2+.展开更多
Hemolysin produced by various bacteria,may destroy erythrocyte membranes via a pore-forming mechanism,a deter-gent action,or a lipase activity.Previous to this experiment,the mode of action used by cyanobacterial hemo...Hemolysin produced by various bacteria,may destroy erythrocyte membranes via a pore-forming mechanism,a deter-gent action,or a lipase activity.Previous to this experiment,the mode of action used by cyanobacterial hemolysin had not been re-ported.To characterize the action mode of hemolysin produced by the wild-type strain of Synechocystis sp.PCC6803,hemolysis of erythrocytes originating from human,mouse,sheep,rabbit and goldfish was studied.The erythrocytes of mouse,sheep and rabbit were sensitive,while those of human and fish were resistant,to this hemolysin.Using rabbit erythrocytes,it was shown that hemoly-sis occurred in two steps:a binding step within the first 10 min of treatment and a lytic step after 30 min.Both binding and lysis were highly temperature-dependent.Effects of erythrocyte density on hemolysis suggest that the hemolysin might target erythrocytes via a multiple-hit mechanism.In the osmotic protection experiment,all tested osmotic protectants,with molecular diameters ranging from 0.9 ?5.66 nm,failed to effectively inhibit hemolysis.Scanning electron micrographs showed that the hemolysin caused protuberances or echinocytes in rabbit erythrocytes,and then disrupted and ruptured the erythrocytes.Characteristics of hemolysis showed distinct differences from other pore-forming mechanisms,suggesting that this hemolysin might act through a detergent-like or lipase mecha-nism,rather than a pore-forming mechanism.展开更多
In this study, cDNA microarrays were developed from 3569 mRNA reads to analyze the expression profiles of the transcriptomes of Synechocystis sp. PCC6803 under low temperature (LT) stress. Among the genes on the cDN...In this study, cDNA microarrays were developed from 3569 mRNA reads to analyze the expression profiles of the transcriptomes of Synechocystis sp. PCC6803 under low temperature (LT) stress. Among the genes on the cDNA microarrays, 899 LT-affected genes exhibited a 1.5-fold (or greater) difference in expression compared with the genes from normal unstressed Synechocystis sp. PCC6803. Of the differentially expressed genes, 353 were up-regulated and 246 were down-regulated. The results showed that genes involved in photosynthesis were activated at LT (10℃), including genes for photosystem I, photosystem II, photosynthetic electron transport, and cytochrome b6/f complex. Moreover, desg, one of four genes that encode the fatty acid desaturases, was also induced by LT. However, the LT conditions to some degree enhanced the transcription of some genes. In addition, LT (10℃) may reduce cellular motility by regulating the transcription of spkA (sll1575), a serine/threonine protein kinase. The results reported in this study may contribute to a better understanding of the responses of the Synechocystis cell to LT, including pathways involved in photosynthesis and repair.展开更多
Objective:To explore the ability of Synechocystis sp.PCC6803 in transforming 6-deoxypseudoanisatin.Methods:The experiment was performed by incubating 6-deoxypseudoanisatin with the freshwater cyanobacterium Synechocys...Objective:To explore the ability of Synechocystis sp.PCC6803 in transforming 6-deoxypseudoanisatin.Methods:The experiment was performed by incubating 6-deoxypseudoanisatin with the freshwater cyanobacterium Synechocystis sp.PCC6803 under continuous white light at 30C for 5 days.The crude converted product was detected using thin-layer chromatography(TLC)and further analyzed using high-performance liquid chromatography(HPLC)as well as HPLC with electron spray ionization mass spectrometry(HPLC-ESI-MS).Results:TLC results showed that 6-deoxypseudoanisatin was converted into a less polar product.HPLC and MS data indicated that the retention time of the converted product increased in comparison with the standard of 6-deoxypseudoanisatin.Conclusion:Thus,the study appears to demonstrate that Synechocystis sp.PCC6803 can transform 6-deoxypseudoanisatin.The polarity of the converted product is less than that of 6-deoxypseudoanisatin.展开更多
Iron stress-induced protein A (IsiA), a major chlorophyll-binding protein in the thylakoid membrane, is significantly induced under iron deficiency conditions. Using immunoblot analysis and 77 K fluorescence spectro...Iron stress-induced protein A (IsiA), a major chlorophyll-binding protein in the thylakoid membrane, is significantly induced under iron deficiency conditions. Using immunoblot analysis and 77 K fluorescence spectroscopy combined with sucrose gradient fractionation, we monitored dynamic changes of IsiA- containing complexes in Synechocystis sp. PCC 6803 during exposure to long-term iron deficiency. Within 3 days of exposure to iron deficiency conditions, the initially induced free IsiA proteins preferentially con- jugated to PSI trimer to form IsiA18-PS I trimers, which serve as light energy collectors for efficiently trans- mitting energy to PS h With prolonged iron deficiency, IsiA proteins assembled either into IsiA aggregates or into two other types of IsiA-PS I supercomplexes, namely IsiA-PS I high fluorescence supercomplex (IHFS) and IsiA-PS I low fluorescence supercomplex (ILFS). Further analysis revealed a role for IsiA as an energy dissipater in the IHFS and as an energy collector in the ILFS. The trimeric structure of PS I mediated by PsaL was found to be indispensable for the formation of IHFS/ILFS. Dynamic changes in IsiA-containing complexes in cyanobacteria during long-term iron deficiency may represent an adaptation to iron limitation stress for flexible light energy distribution, which balances electron transfer between PS I and PS II, thus minimizing photooxidative damage.展开更多
The chlL gene encoding one component of light-independent (dark) protochlorophyllide oxido reductase (DPOR) was deleted in cyanobacterium Synechocystis sp. PCC 6803 (S.6803). The resulting chlL- mutant lost DPOR activ...The chlL gene encoding one component of light-independent (dark) protochlorophyllide oxido reductase (DPOR) was deleted in cyanobacterium Synechocystis sp. PCC 6803 (S.6803). The resulting chlL- mutant lost DPOR activity. No significant differences of chlorophyll (Chl) content and growth rate were observed between the wild and the mutant strains grown at 50 mE·m2·s1 light intensity for photomixtrophic and photoautotrophic growth. However, differences were observed at 1 mE·m2·s1 light intensity. For photomixtrophic growth, the mutant Chl content was 50% of the wild content with continuous light and 35.7% of the wild content with a 10 h light/ 14 h dark cycle. For photoautotriphic growth, the mutant Chl level was 76.3% of the wild content with continuous light and 63.2% with a 10 h light/ 14 h dark cycle. The results indicate that DPOR contributes to Chl synthesis and increases the growth rate in cyanobacteria phototrophically cultured at 1mE·m2·s1 light intensity. In contrast, the photosynthetic capacity on a per-cell basis of the mutant is 5% higher than that of the wild strain with continuous light and 27% higher than that of the wild strain with a 10 h light/14 h dark cycle at 1 mE·m2·s1 light intensity for photoautotrophic growth. With the low Chl content, the cyanobacteria have the ability to improve their photosynthetic capacity by decreasing the ratio of PSI to PSII by unknown morphological or physiological means.展开更多
Synechocystis sp. PCC 6803 cell growth and poly β hydroxybutyrate (PHB) biosynthesis were studied in the presence of sodium acetate (NaAc). For nitrogen sufficient conditions, 15 mmol/L NaAc improved the PHB co...Synechocystis sp. PCC 6803 cell growth and poly β hydroxybutyrate (PHB) biosynthesis were studied in the presence of sodium acetate (NaAc). For nitrogen sufficient conditions, 15 mmol/L NaAc improved the PHB content up to 9.9% (w/w) while for nitrogen starved conditions, the PHB content was up to 15.2% (w/w). NaAc at levels below 20 mmol/L promoted cell growth in the first six days, but the growth slowed on the seventh day when the NaAc concentration exceeded 15 mmol/L. The PHB content in the final biomass reached 11.0% of the dry cellular weight in the presence of 20 mmol/L NaAc. Two adjacent open reading frames (ORFs) in the genome of Synechocystis sp. PCC 6803, slr1993 and slr1994, were assigned to phbA and phbB, respectively, while the phbC gene was found to be far from these genes. This may account for the low expression of PHB in cyanobacteria.展开更多
Two-component systems are signal transduction systems which enable bacteria to regulate cellular functions in response to changing environmental conditions. The unicellular Synechocystis sp. PCC 6803 has become a mode...Two-component systems are signal transduction systems which enable bacteria to regulate cellular functions in response to changing environmental conditions. The unicellular Synechocystis sp. PCC 6803 has become a model organism for a range of biochemical and molecular biology studies aiming at investigating environmental stress response. The publication of the complete genome sequence of the cyanobacterium Synechocystis sp. PCC 6803 provided a tremendous stimulus for research in this field, and at least 80 open reading frames were identified as members of the two-component signal transduction systems in this single species of cyanobacteria. To date, functional roles have been determined for only a limited number of such proteins. This review summarizes our current knowledge about the two-component signal transduction systems in Synechocystis sp. PCC 6803 and describes recent achievements in elucidating the functional roles of these systems.展开更多
Exploiting light to drive redox reactions is currently a hot topic since light is considered as an environmentally friendly source of energy.Consequently,cyanobacteria,which can use light e.g.,for generating NADPH,are...Exploiting light to drive redox reactions is currently a hot topic since light is considered as an environmentally friendly source of energy.Consequently,cyanobacteria,which can use light e.g.,for generating NADPH,are in the focus of research.Previously,it has been shown that various heterologous redox enzymes could be expressed in these microorganisms.Here we demonstrated the successful inducer-free expression of𝛼-keto-acid dehydroge-nases(L-HicDH and D-HicDH)from Lactobacillus confusus DSM 20196 and Lactobacillus paracasei DSM 20008 in Synechocystis sp.PCC 6803ΔhoxYH mutant using replicative plasmids.While the L-HicDH showed poor activity limited by the amount of expressed enzyme,the D-HicDH was applied both in vivo and in vitro,transforming the selected𝛼-keto acids to the corresponding optically pure(R)-𝛼-hydroxy acids(ee>99%)in up to 53%and 90%conversion,respectively.展开更多
Genetic modifi cation is useful for improving the nutritional qualities of cyanobacteria. To increase the total unsaturated fatty acid content, along with the ratio of ω-3/ω-6 fatty acids, genetic engineering can be...Genetic modifi cation is useful for improving the nutritional qualities of cyanobacteria. To increase the total unsaturated fatty acid content, along with the ratio of ω-3/ω-6 fatty acids, genetic engineering can be used to modify fatty acid metabolism. S ynechococcus sp. PCC7002, a fast-growing cyanobacterium, does not contain a Δ6 desaturase gene and is therefore unable to synthesize γ-linolenic acid(GLA) and stearidonic acid(SDA), which are important in human health. In this work, we constructed recombinant vectors Syd6 D, Syd15 D and Syd6Dd15 D to express the Δ15 desaturase and Δ6 desaturase genes from Synechocystis PCC6803 in Synechococcus sp. PCC7002, with the aim of expressing polyunsaturated fatty acids. Overexpression of the Δ15 desaturase gene in S ynechococcus resulted in 5.4 times greater accumulation of α-linolenic acid compared with the wild-type while Δ6 desaturase gene expression produced both GLA and SDA. Co-expression of the two genes resulted in low-level accumulation of GLA but much larger amounts of SDA, accounting for as much to 11.64% of the total fatty acid content.展开更多
文摘The agp gene encoding the ADP-glucose pyrophosphorylase involved in cyanobacterial glycogen synthesis was amplified by PCR. The resulting agp fragment was cloned in plasmid pUC118 to generate plasmid pUCA. Part of the fragment within the agp DNA was deleted and replaced by an erythromycin resistance cassette to generate plasmid pUCAE, which was used to transform the Synechocystis sp. PCC 6803 wild-type strain and a mutant with resistance to erythromycin was obtained. PCR analysis of the genomic DNA from the resulting mutant indicated that the appropriate deletion and insertion indeed had occurred. The cell growth and Chl a, glycogen content in the mutant showed difference from those in the wild-type strain. The obtained biomass as well as the Chl a content in the mutant strain was higher than that of the wild-type strain, which suggested that the photosynthesis efficiency in the agp(-) strain was higher than that in the wild-type strain. No glycogen was found in the mutant, providing evidence for the correction of the mutant in physiological level.
文摘A novel lipid occurred when cyanobacterium Synechocystis sp. PCC 6803 cells were grown in BG-11 medium with glucose applied. This lipid was determined to be a glycolipid, designated glycolipid-x (Glyco-x), by staining with alpha-naphthol and concentrated sulfuric acid. The occurrence of Glyco-x accompanies the disappearance of other lipids, especially DGDG. Glyco-x can also be observed in cells grown in BG-11 medium with the application of other carbon sources: fructose, maltose and lactose. Sodium thiosulfate, an effective scavenger of reactive oxygen intermediates, showed strong capability to inhibit glucose-induced occurrence of Glyco-x. In the presence of 0.3% sodium thiosulfate, Glyco-x could only be detected in cells grown in BG-11 medium with 100 mmol/L glucose applied in late-exponential phase. These results suggest that reactive oxygen species might be involved in the occurrence of Glyco-x in cyanobacterium Synechocystis sp. PCC 6803 cells grown in the presence of glucose.
文摘Compositions of glycerolipids and fatty acid compositions of glycerolipids were compared among Synechocystis sp. PCC 6803 cells grown in the BG-11 medium containing different concentrations of glucose and Na2S2O3 in this study. It was found that Na2S2O3 can effectively increase the percentage of sulphoquinovosyl diacylglycerol (SQDG) and phosphatidylglycerol (PG) to total membrane lipids and the simultaneous application of glucose with Na2S2O3 can counteract the effect of Na2S2O3. In addition, Na2S2O3 can significantly increase the percentage of palmitic acid (C, 16:0) in fatty acid composition of monogalactosyl diacylglycerol (MGDG) and digalactosyl diacylglycerol (DGDG) and decrease the fatty acid unsaturation degree accordingly, and these effects can also be eliminated by glucose. These results indicate that Na2S2O3 can take as a reductant to make membrane lipids in a low unsaturated state, and the simultaneous application of glucose can decrease the reducing power of Na2S2O3. In addition, Na2S2O3 can take as a sulfur donor for the synthesis of SQDG.
基金Supported by the National Natural Science Foundation of China(Nos.40972043,41040018,41210104058,21176145,41372108,41302079)the Higher Educational Science and Technology Program of Shandong Province(No.J10LC15)+4 种基金the China Postdoctoral Science Foundation(No.2013M540560)the Program for Scientific Research Innovation Team in Colleges and Universities of Shandong Province,and SDUST Research Fund(No.2010KYTD103)the Open Project of Key Lab of Marine Bioactive Substance and Modern Analytical Technique,State Oceanic Administration,China(No.MBSMAT-2012-03)the Scientific and Technological Program of Qingdao(No.13-1-4-232-jch)the Domestic Visiting Scholar Program for Young Core Teachers in Shandong Universities,Shandong Province,China
文摘Calcium carbonate (CaCO3) crystals in their preferred orientation were obtained in BG11 culture media inoculated with Synechocystis sp. PCC6803 (inoculated BG11). In this study, the features of calcium carbonate deposition were investigated. Inoculated BGll in different calcium ion concentrations was used for the experimental group, while the BGll culture medium was used for the control group. The surface morphologies of the calcium carbonate deposits in the experimental and control groups were determined by scanning and transmission electron microscopy. The deposits were analyzed by electronic probe micro-analysis, Fourier transform infrared spectrum, X-ray diffraction, thermal gravimetric analysis and differential scanning calorimetry. The results show that the surfaces of the crystals in the experimental group were hexahedral in a scaly pattern. The particle sizes were micrometer-sized and larger than those in the control group. The deposits of the control group contained calcium (Ca), carbon (C), oxygen (O), phosphorus (P), iron (Fe), copper (Cu), zinc (Zn), and other elements. The deposits in the experimental group contained Ca, C, and O only. The deposits of both groups contained calcite. The thermal decomposition temperature of the deposits in the control group was lower than those in the experimental group. It showed that the CaCO3 deposits of the experimental group had higher thermal stability than those of the control group. This may be due to the secondary metabolites produced by the algae cells, which affect the carbonate crystal structure and result in a close-packed structure. The algae cells that remained after thermal weight loss were heavier in higher calcium concentrations in BGll culture media. There may be more calcium- containing crystals inside and outside of these cells. These results shall be beneficial for understanding the formation mechanism of carbonate minerals.
文摘The unicellular cyanobacterium Synechocystis sp. PCC 6803, a model organism known for its unique combination of highly desirable molecular genetic, physiological and morphological characteristics, was employed in the present study. The species was cultured in BG11 liquid medium contained various initial concentrations of Pb^2+ and Cd〉 (0, 0.5, 1, 2, 4, 6 and 8 mg/L). The experiment was conducted for six days and the metal induced alterations in the ultrastructure, growth and pigment contents were assessed. Alterations in the ultrastructure of the Synechocystis sp. PCC 6803 cells became evident with the increased (〉4 mg/L Pb^2+) metal concentration. The photosynthetic apparatus (thylakoid membranes) were found to be the worst affected. Deteriorated or completely destroyed thylakoid membranes have made large empty spaces in the cell interior. In addition, at the highest concentration (8 mg/L pb^2+), the polyphosphate granules became more prominent both in size and number. Despite the initial slight stimulations (0.2, 3.8 and 6.5% respectively at 0.5, 1 and 2 mg/L pb^2+), both metals inhibited the growth in a dose-dependent manner as incubation progressed. Pigment contents (chlorophyll a, 13 carotene and phycocyanin) were also decreased with increasing metal concentration. Cells exposed to 6 mg/L Pb^2+, resulted in 36.56, 37.39 and 29.34% reductions of chlorophyll a, 13 carotene and phycocyanin respectively over the control. Corresponding reductions for the same CdZ+concentrations were 57.83, 48.94 and 56.90%. Lethal concentration (96 h LC50) values (3.47 mg/L Cd^2+ and 12.11 mg/L Pb^2+) indicated that Synechocystis sp. PCC 6803 is more vulnerable to Cd^2+ than Pb^2+.
基金the National Natural Science Fund of China (No. 30870250)Shandong Provincial NaturalScience Fund (No. Q2006D09)
文摘Hemolysin produced by various bacteria,may destroy erythrocyte membranes via a pore-forming mechanism,a deter-gent action,or a lipase activity.Previous to this experiment,the mode of action used by cyanobacterial hemolysin had not been re-ported.To characterize the action mode of hemolysin produced by the wild-type strain of Synechocystis sp.PCC6803,hemolysis of erythrocytes originating from human,mouse,sheep,rabbit and goldfish was studied.The erythrocytes of mouse,sheep and rabbit were sensitive,while those of human and fish were resistant,to this hemolysin.Using rabbit erythrocytes,it was shown that hemoly-sis occurred in two steps:a binding step within the first 10 min of treatment and a lytic step after 30 min.Both binding and lysis were highly temperature-dependent.Effects of erythrocyte density on hemolysis suggest that the hemolysin might target erythrocytes via a multiple-hit mechanism.In the osmotic protection experiment,all tested osmotic protectants,with molecular diameters ranging from 0.9 ?5.66 nm,failed to effectively inhibit hemolysis.Scanning electron micrographs showed that the hemolysin caused protuberances or echinocytes in rabbit erythrocytes,and then disrupted and ruptured the erythrocytes.Characteristics of hemolysis showed distinct differences from other pore-forming mechanisms,suggesting that this hemolysin might act through a detergent-like or lipase mecha-nism,rather than a pore-forming mechanism.
基金Supported by the National Natural Science Foundation of China(No.40876082)the International Innovation Partnership Program:Typical Environmental Process and Effects on Resources in Coastal Zone Area+3 种基金the Public Science and Technology Research Funds Projects of the Ocean(Nos.200905021,201205027)the Outstanding Young Scholars Fellowship of Shandong Province(Molecular Phycology,No.JQ200914)the Natural Science Foundation of Shandong Province(No.ZR2012DQ015)the Guangdong Province Comprehensive Strategic Cooperation Project of the Chinese Academy of Sciences(No.2011A090100040)
文摘In this study, cDNA microarrays were developed from 3569 mRNA reads to analyze the expression profiles of the transcriptomes of Synechocystis sp. PCC6803 under low temperature (LT) stress. Among the genes on the cDNA microarrays, 899 LT-affected genes exhibited a 1.5-fold (or greater) difference in expression compared with the genes from normal unstressed Synechocystis sp. PCC6803. Of the differentially expressed genes, 353 were up-regulated and 246 were down-regulated. The results showed that genes involved in photosynthesis were activated at LT (10℃), including genes for photosystem I, photosystem II, photosynthetic electron transport, and cytochrome b6/f complex. Moreover, desg, one of four genes that encode the fatty acid desaturases, was also induced by LT. However, the LT conditions to some degree enhanced the transcription of some genes. In addition, LT (10℃) may reduce cellular motility by regulating the transcription of spkA (sll1575), a serine/threonine protein kinase. The results reported in this study may contribute to a better understanding of the responses of the Synechocystis cell to LT, including pathways involved in photosynthesis and repair.
基金Beijing University of Chinese Medicine(2013-JYBZZ-JS-139)Independent Project Topics Foundation.
文摘Objective:To explore the ability of Synechocystis sp.PCC6803 in transforming 6-deoxypseudoanisatin.Methods:The experiment was performed by incubating 6-deoxypseudoanisatin with the freshwater cyanobacterium Synechocystis sp.PCC6803 under continuous white light at 30C for 5 days.The crude converted product was detected using thin-layer chromatography(TLC)and further analyzed using high-performance liquid chromatography(HPLC)as well as HPLC with electron spray ionization mass spectrometry(HPLC-ESI-MS).Results:TLC results showed that 6-deoxypseudoanisatin was converted into a less polar product.HPLC and MS data indicated that the retention time of the converted product increased in comparison with the standard of 6-deoxypseudoanisatin.Conclusion:Thus,the study appears to demonstrate that Synechocystis sp.PCC6803 can transform 6-deoxypseudoanisatin.The polarity of the converted product is less than that of 6-deoxypseudoanisatin.
文摘Iron stress-induced protein A (IsiA), a major chlorophyll-binding protein in the thylakoid membrane, is significantly induced under iron deficiency conditions. Using immunoblot analysis and 77 K fluorescence spectroscopy combined with sucrose gradient fractionation, we monitored dynamic changes of IsiA- containing complexes in Synechocystis sp. PCC 6803 during exposure to long-term iron deficiency. Within 3 days of exposure to iron deficiency conditions, the initially induced free IsiA proteins preferentially con- jugated to PSI trimer to form IsiA18-PS I trimers, which serve as light energy collectors for efficiently trans- mitting energy to PS h With prolonged iron deficiency, IsiA proteins assembled either into IsiA aggregates or into two other types of IsiA-PS I supercomplexes, namely IsiA-PS I high fluorescence supercomplex (IHFS) and IsiA-PS I low fluorescence supercomplex (ILFS). Further analysis revealed a role for IsiA as an energy dissipater in the IHFS and as an energy collector in the ILFS. The trimeric structure of PS I mediated by PsaL was found to be indispensable for the formation of IHFS/ILFS. Dynamic changes in IsiA-containing complexes in cyanobacteria during long-term iron deficiency may represent an adaptation to iron limitation stress for flexible light energy distribution, which balances electron transfer between PS I and PS II, thus minimizing photooxidative damage.
基金the National Natural Science Foundation of China (No. 39870064)
文摘The chlL gene encoding one component of light-independent (dark) protochlorophyllide oxido reductase (DPOR) was deleted in cyanobacterium Synechocystis sp. PCC 6803 (S.6803). The resulting chlL- mutant lost DPOR activity. No significant differences of chlorophyll (Chl) content and growth rate were observed between the wild and the mutant strains grown at 50 mE·m2·s1 light intensity for photomixtrophic and photoautotrophic growth. However, differences were observed at 1 mE·m2·s1 light intensity. For photomixtrophic growth, the mutant Chl content was 50% of the wild content with continuous light and 35.7% of the wild content with a 10 h light/ 14 h dark cycle. For photoautotriphic growth, the mutant Chl level was 76.3% of the wild content with continuous light and 63.2% with a 10 h light/ 14 h dark cycle. The results indicate that DPOR contributes to Chl synthesis and increases the growth rate in cyanobacteria phototrophically cultured at 1mE·m2·s1 light intensity. In contrast, the photosynthetic capacity on a per-cell basis of the mutant is 5% higher than that of the wild strain with continuous light and 27% higher than that of the wild strain with a 10 h light/14 h dark cycle at 1 mE·m2·s1 light intensity for photoautotrophic growth. With the low Chl content, the cyanobacteria have the ability to improve their photosynthetic capacity by decreasing the ratio of PSI to PSII by unknown morphological or physiological means.
基金Supported by the National Natural Science F oundation of China (No.2 0 0 760 2 4)
文摘Synechocystis sp. PCC 6803 cell growth and poly β hydroxybutyrate (PHB) biosynthesis were studied in the presence of sodium acetate (NaAc). For nitrogen sufficient conditions, 15 mmol/L NaAc improved the PHB content up to 9.9% (w/w) while for nitrogen starved conditions, the PHB content was up to 15.2% (w/w). NaAc at levels below 20 mmol/L promoted cell growth in the first six days, but the growth slowed on the seventh day when the NaAc concentration exceeded 15 mmol/L. The PHB content in the final biomass reached 11.0% of the dry cellular weight in the presence of 20 mmol/L NaAc. Two adjacent open reading frames (ORFs) in the genome of Synechocystis sp. PCC 6803, slr1993 and slr1994, were assigned to phbA and phbB, respectively, while the phbC gene was found to be far from these genes. This may account for the low expression of PHB in cyanobacteria.
基金Supported by the National Natural Science Foundation of China (Nos. 40272054 and 40332022) and Research Grant of Doctoral Program for High Institue to Q. Wu
文摘Two-component systems are signal transduction systems which enable bacteria to regulate cellular functions in response to changing environmental conditions. The unicellular Synechocystis sp. PCC 6803 has become a model organism for a range of biochemical and molecular biology studies aiming at investigating environmental stress response. The publication of the complete genome sequence of the cyanobacterium Synechocystis sp. PCC 6803 provided a tremendous stimulus for research in this field, and at least 80 open reading frames were identified as members of the two-component signal transduction systems in this single species of cyanobacteria. To date, functional roles have been determined for only a limited number of such proteins. This review summarizes our current knowledge about the two-component signal transduction systems in Synechocystis sp. PCC 6803 and describes recent achievements in elucidating the functional roles of these systems.
文摘Exploiting light to drive redox reactions is currently a hot topic since light is considered as an environmentally friendly source of energy.Consequently,cyanobacteria,which can use light e.g.,for generating NADPH,are in the focus of research.Previously,it has been shown that various heterologous redox enzymes could be expressed in these microorganisms.Here we demonstrated the successful inducer-free expression of𝛼-keto-acid dehydroge-nases(L-HicDH and D-HicDH)from Lactobacillus confusus DSM 20196 and Lactobacillus paracasei DSM 20008 in Synechocystis sp.PCC 6803ΔhoxYH mutant using replicative plasmids.While the L-HicDH showed poor activity limited by the amount of expressed enzyme,the D-HicDH was applied both in vivo and in vitro,transforming the selected𝛼-keto acids to the corresponding optically pure(R)-𝛼-hydroxy acids(ee>99%)in up to 53%and 90%conversion,respectively.
基金Supported by the International S&T Cooperation Program of China(No.2012DFA30450)the National Natural Science Foundation of China(No.30871541)+1 种基金the Taishan Scholar Foundation of Shandong Province(No.tshw20091014)the Innovation Program of the University Institutes of Jinan,Shandong Province(No.201004044)
文摘Genetic modifi cation is useful for improving the nutritional qualities of cyanobacteria. To increase the total unsaturated fatty acid content, along with the ratio of ω-3/ω-6 fatty acids, genetic engineering can be used to modify fatty acid metabolism. S ynechococcus sp. PCC7002, a fast-growing cyanobacterium, does not contain a Δ6 desaturase gene and is therefore unable to synthesize γ-linolenic acid(GLA) and stearidonic acid(SDA), which are important in human health. In this work, we constructed recombinant vectors Syd6 D, Syd15 D and Syd6Dd15 D to express the Δ15 desaturase and Δ6 desaturase genes from Synechocystis PCC6803 in Synechococcus sp. PCC7002, with the aim of expressing polyunsaturated fatty acids. Overexpression of the Δ15 desaturase gene in S ynechococcus resulted in 5.4 times greater accumulation of α-linolenic acid compared with the wild-type while Δ6 desaturase gene expression produced both GLA and SDA. Co-expression of the two genes resulted in low-level accumulation of GLA but much larger amounts of SDA, accounting for as much to 11.64% of the total fatty acid content.