Light and carbon dioxide-driven synthesis of high-density fuel in Synechococcus elongates UTEX 2973 / 生物工程学报

Development of "liquid sunshine" could be a key technology to deal with the issue of fossil fuel depletion. β-caryophyllene is a terpene compound with high energy density and has attracted attention for its potential application as a jet fuel. The high temperature and high light-tolerant photosynthetic cyanobacterium Synechococcus elongatus UTEX 2973 (hereafter Synechococcus 2973), whose doubling time is as short as 1.5 h, has great potential for synthesizing β-caryophyllene using sunlight and CO₂. In this study, a production of ~121.22 μg/L β-caryophyllene was achieved at 96 h via a combined strategy of pathway construction, key enzyme optimization and precursor supply enhancement. In addition, a final production of ~212.37 μg/L at 96 h was realized in a high-density cultivation. To our knowledge, this is the highest production reported for β-caryophyllene using cyanobacterial chassis and our study provide important basis for high-density fuel synthesis in cyanobacteria.

Pentoses Used in Cultures of Synechococcus nidulans and Spirulina paracas: Evaluation of Effects in Growth and in Content of Proteins and Carbohydrates

Abstract The biological assimilation of the sugars present in lignocellulosic residues has gained prominence since these residues are the most abundant and economic residues in nature. Thus, the objective of this work was to determine whether the use of D-xylose and L-arabinose as sources of carbon in Synechococcus nidulans and Spirulina paracas cultures affects the growth and production of proteins and carbohydrates. Kinetic growth parameters, pentose consumption, protein content and carbohydrates were evaluated. Synechococcus nidulans and Spirulina paracas consumed all concentrations of pentose used. The highest cellular concentration (1.37 g.L-1) and the highest protein productivity (54 mg.L-1.d-1) were obtained for Spirulina paracas, which was submitted to the addition of 38.33 mg.L-1 D-xylose and 1.79 mg.L-1 L-arabinose. The use of pentose promoted the accumulation of proteins for the studied microalgae. This is one of the first works to report protein bioaccumulation as a result of pentose addition.

Effects of light quality on cell growth and psbA promoter of engineered Synechococcus sp. PCC7002 / 生物工程学报

Light quality can regulate both psbA genes and vector promoter psbA of the engineered Synechococcus. Through light regulation, we tried to improve yield of the recombinant protein for vp28 gene-expressed Synechococcus sp. PCC7002. To drive photon-capturing efficiently, three limiting factors (irradiance, temperature and pH) were optimized by measuring net photosynthesis. High cell density cultures were performed with variant ratios of white, red and blue light in a 5-L photo-bioreactor. Yields of biomass, expressions of vp28 and transcription levels of psbA were compared. High ratio blue light-induced vp28 transcription had tripled and the relative accumulation of VP28 protein was doubled. The relative expressions of psbAII and psbAIII had positive correlations with higher ratio of blue light, not the red light. With high ratio red light inducing, dry biomass reached 1.5 g/L in three days. Therefore, we speculated that red light accelerated biomass accumulation of the transgenic strain and blue light promoted transcription for PpsbA and psbA. These results provided useful information for mass production of cyanobacteria and its secondary metabolites.

Biodiesel production from marine cyanobacteria cultured in plate and tubular photobioreactors.

Carbon (neutral) based renewable liquid biofuels are alternative to petroleum derived transport fuels that contribute to global warming and are of a limited availability. Microalgae based biofuels are considered as promising source of energy. Lyngbya sp. and Synechococcus sp. were studied for the possibility of biodiesel production in different media such as ASNIII, sea water enrichment medium and BG11. The sea water enrichment medium was found superior in enhancing the growth rate of these microalgae. Nitrogen depletion has less effect in total chlorophyll a content, at the same time the lipid content was increased in both Lyngbya sp. and Synechococcus sp. by 1.4 and 1.2 % respectively. Increase in salinity from 0.5-1.0 M also showed an increase in the lipid content to 2.0 and 0.8 % in these strains; but a salinity of 1.5 M has a total inhibitory effect in the growth. The total biomass yield was comparatively higher in tubular LED photobioreactor than the fluorescent flat plated photobioreactor. Lipid extraction was obtained maximum at 60 ºC in 1:10 sample: solvent ratio. GC-MS analysis of biodiesel showed high content of polyunsaturated fatty acids (PUFA; 4.86 %) than saturated fatty acid (SFA; 4.10 %). Biodiesel production was found maximum in Synechococcus sp. than Lyngbya sp. The viscosity of the biodiesel was closely related to conventional diesel. The results strongly suggest that marine microalgae could be used as a renewable energy source for biodiesel production.

Changes in membrane lipids and carotenoids during light acclimation in a marine cyanobacterium Synechococcus sp.

Time course of carotenoid and membrane lipid variation during high light (HL) acclimation (about 85 μmol m−2 s−1), after transfer from low light (LL) (5–10 μmol m−2 s−1), was determined in a marine Synechococcus strain. Highperformance liquid chromatography (HPLC) coupled to diode array detector (DAD) or electrospray ionization mass spectrometry (ESI-MS) was used for compound separation and detection. Myxoxanthophyll rose within a time interval of 8 h to 24 h after the onset of exposure to HL. β-carotene content started to decrease after 4 h of the onset of exposure to HL. Zeaxanthin content rose with exposure to HL, but it was only significant after 24 h of exposure. Carotenoid changes are in agreement with a coordinated activity of the enzymes of the myxoxanthophyll biosynthetic pathway, with no rate-limiting intermediate steps. Lipid analysis showed all species with a C18:3/C16:0 composition increased their content, the changes of PG(18:3/16:0) and MGDG(18:3/16:0) being primarily significant. Major lipid changes were also found to occur within 24 h. These changes might suggest reduction and reorganization of the thylakoid membrane structure. Hypotheses are also drawn on the role played by lipid molecule shape and their possible effect in membrane fluidity and protein accommodation.

Crecimiento y producción de metabolitos de la cianobacteria marina Synechococcus sp. (Chroococcales) en función de la irradiancia

Growth and metabolite production of the marine cyanobacterium Synechococcus sp. (Chroococcales) in function to irradiance. Changes in salinity, temperature and irradiance during wet and dry seasons have induced metabolic versatility in cyanobacteria from saline environments. Cyanobacteria from these environments have biotechnological potential for the production of metabolites with pharmaceutical and industrial interest. We studied the growth, dry mass and metabolite production of the cyanobacterium Synechococcus sp. MOF-03 in function of irradiance (78, 156 and 234 µmol q m-2 s-1). All batch cultures were maintained by triplicate in constant aeration, 12:12 h photoperiod, 30 ±2ºC and 35‰. Maximum values of protein, carbohydrates and lipids, of 530.19 ±11.16, 408.94 ±4.27 and 56.20 ±1.17 µg ml-1, respectively, were achieved at 78 µmol q m-2 s-1. Pigments, analyzed by HPLC, showed maximum values at 78 µmol q m-2 s-1 for chlorophyll a with 7.72 ±0.16 µg ml-1, and at 234 µmol q m-2 s-1 for ß-carotene and zeaxanthin with 0.70 ±0.01 and 0.67 ±0.05 µg ml-1. Chlorophyll a:ß-carotene ratio decreased from 17.15 to 6.91 at 78 and 234 µmol q m-2 s-1; whereas ß-carotene:zeaxanthin ratio showed no changes between 78 and 156 µmol q m-2 s-1, around 1.21, and decreased at 234 µmol q m-2 s-1, to 1.04. Also, this cyanobacterium produced the greatest cell density and dry mass at 156 µmol q m-2 s-1, with 406.13 ±21.74 x106 cell ml-1 and 1.49 ±0.11 mg ml-1, respectively. Exopolysaccharide production was stable between 156 y 234 µmol q m-2 s-1, around 110 µg ml-1. This Synechococcus strain shows a great potential for the production of enriched biomass with high commercial value metabolites. Rev. Biol. Trop. 56 (2): 421-429. Epub 2008 June 30.

Las cianobacterias de ambientes salinos presentan una versatilidad metabólica inducida por los cambios de salinidad, temperatura e irradiancia, durante los períodos de sequía y lluvias. Por ello es importante la búsqueda en estos ambientes de cianobacterias con potencial biotecnológico para la producción de metabolitos de interés farmacéutico e industrial. Se reporta el crecimiento, masa seca y producción de metabolitos de la cianobacteria Synechococcus sp. MOF-03 en función de la irradiancia (78, 156 y 234 µmol q m-2 s-1). Los cultivos discontinuos por triplicado, fueron mantenidos con aireación constante, fotoperiodo 12:12 h, 30 ±2ºC y a 35‰. Los máximos valores de proteínas, carbohidratos y lípidos de 530.19 ±11.16, 408.94 ±4.27 y 56.20 ±1.17 µg ml-1 respectivamente, fueron obtenidos a 78 µmol q m-2 s-1. Los pigmentos, analizados por HPLC, mostraron los máximos a 78 µmol q m-2 s-1 para clorofila a con 7.72 ±0.16 µg ml-1; y a 234 µmol q m-2 s-1 para ß-caroteno y zeaxantina con 0.70 ±0.01 and 0.67 ±0.05 µg ml-1. La relación clorofila a:ß-caroteno disminuyó de 17.15 hasta 6.91 a 78 y 234 µmol q m-2 s-1; mientras que la relación ß-caroteno:zeaxantina se mantuvo sin cambios entre 78 y 156 µmol q m-2 s-1, con cerca de 1.21 y disminuyó a 234 µmol q m-2 s-1 a 1.04. La cianobacteria produjo la mayor densidad celular y masa seca a 156 µmol q m-2 s-1, con 406.13 ±21.74 x106 cel ml-1 y 1.49 ±0.11 mg ml-1 respectivamente. La producción de exopolisacáridos se mantuvo alrededor de 110 µg ml-1 entre 156 y 234 µmol q m-2 s-1. Así, esta cepa de Synechococcus muestra un gran potencial para la producción de biomasa enriquecida con metabolitos de alto valor comercial.

Effect of Synechococcus sp.PCC7942 with trans-mutated hCu,Zn-SOD-gene on antioxidation activities in mice / 中国海洋药物

Objective To study the biological activities of Synechococcus sp.PCC7942 with trans-mutated hCu,Zn-SOD-gene.Methods Synechococcus sp.PCC7942 with trans-mutated hCu,Zn-SOD were administered orally for 20d to mice,then the activities of glutathione peroxidase(GSH-Px),catalase(CAT),superoxide dismutase(SOD) and the content of malondialdehyde(MDA) were determined.Results The activities of GSHPx in serum and the activities of CAT in blood increased obviously;the activity of SOD in serum and liver increased markedly;the content of MDA in serum and liver decreased obviously.Conclusion Synechococcus sp.PCC7942 with trans-mutated hCu,Zn-SOD-gene had obvious antioxidant effect in vivo.

Mercury ions inhibit photosynthetic electron transport at multiple sites in the cyanobacterium Synechococcus 6301.

Inhibition of electron transport activities in the spheroplasts of Synechococcus 6301 by HgCl2 is dependent on the concentration of mercury ions. The inhibition of whole chain electron transport activity occurs at low concentration of Hg2+ (6 μΜ). This inhibition occurs mostly due to interaction of Hg2+ on plastocyanin. At an elevated concentration (24 μΜ), mercury induces inhibition chiefly in photosystem II catalyzed electron transport. At this concentration it also alters both the absorption and emission characteristics of the phycocyanin. The photosystem I catalyzed electron transport was inhibited by 50% only at high concentrations (36 μΜ) of HgCl2. However, electron transport catalyzed by photosystems I and II from reduced duroquinone to methylviologen which involves intersystem electron transport is extremely sensitive to mercury (low concentration 6–9 μΜ) like that of whole chain assay indicating that the observed inhibition in whole chain electron transport at low concentrations is mostly contributed by the damage involving other intersystem electron transport carrier(s) like plastocyanin. Thus mercury ions depending on the concentration affects the electron transport at multiple sites in the spheroplasts of Synechococcus.

Preliminary Studies on the Growth Potential of Transgenic Synechococcus sp.PCC 7002 with Mouse Metallothionein-ⅠGene / 微生物学通报

The growth predominance and culturing potential of transgenic Synechococcus sp.PCC 7002 with mouse metallothionein-Igene are characterized and expatiated by room temperature absorption spectra,photosynthetic oxygen evolution rate,growth kinetic parameters.The results show that the spectral absorption,maximum net photosynthetic rate and saturated light intensity of transgenic Synechococcus sp.PCC 7002 are higher than wild strain,whereas its the respiration rate and compensating light intensity are lower than wild strain.The transgenic cells exhibit a higher growth rate,and the maximum cell concentration is 1.74 times higher than wild ones when cultured in shaking flask.The air lift photobioreactor is suitable for bringing its growth potential into play.