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
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
Inspired by iron fertilization experiments in HNLC（high-nitrate, low-chlorophyll） sea areas,we proposed the use of iron-rich engineered microalgae for microbial contaminant control in iron-free culture media. Based ...Inspired by iron fertilization experiments in HNLC（high-nitrate, low-chlorophyll） sea areas,we proposed the use of iron-rich engineered microalgae for microbial contaminant control in iron-free culture media. Based on the genome sequence and natural transformation system of Synechocystis sp. PCC6803, ftn A（encoding ferritin） was selected as our target gene and was cloned into wild-type Synechocystis sp. PCC6803. Tests at the molecular level confirmed the successful construction of the engineered Synechocystis sp. PCC6803-ftn A. After Fe3＋-EDTA pulsing, the intracellular iron content of Synechocystis sp. PCC6803-ftn A was significantly enhanced, and the algae was used in the microbial contamination control system. In the coupled Synechocystis sp. PCC6803-ftn A production and municipal wastewater（MW, including Scenedesmus obliquus and Bacillus） treatment, Synechocystis sp. PCC6803-ftn A accounted for all of the microbial activity and significantly increased from 70% of the microbial community to 95%.These results revealed that while the stored iron in the Synechocystis sp. PCC6803-ftn A cells was used for growth and reproduction of this microalga in the MW, the growth of other microbes was inhibited because of the iron limitation, and these results provide a new method for microbial contamination control during a coupling process.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
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.
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 Key Technologies R&D Program of China(No.2012BAJ25B02)
文摘Inspired by iron fertilization experiments in HNLC（high-nitrate, low-chlorophyll） sea areas,we proposed the use of iron-rich engineered microalgae for microbial contaminant control in iron-free culture media. Based on the genome sequence and natural transformation system of Synechocystis sp. PCC6803, ftn A（encoding ferritin） was selected as our target gene and was cloned into wild-type Synechocystis sp. PCC6803. Tests at the molecular level confirmed the successful construction of the engineered Synechocystis sp. PCC6803-ftn A. After Fe3＋-EDTA pulsing, the intracellular iron content of Synechocystis sp. PCC6803-ftn A was significantly enhanced, and the algae was used in the microbial contamination control system. In the coupled Synechocystis sp. PCC6803-ftn A production and municipal wastewater（MW, including Scenedesmus obliquus and Bacillus） treatment, Synechocystis sp. PCC6803-ftn A accounted for all of the microbial activity and significantly increased from 70% of the microbial community to 95%.These results revealed that while the stored iron in the Synechocystis sp. PCC6803-ftn A cells was used for growth and reproduction of this microalga in the MW, the growth of other microbes was inhibited because of the iron limitation, and these results provide a new method for microbial contamination control during a coupling process.
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.
