Effects of temperature shifts on growth rate and lipid characteristics of Synechocystis sp. PCC6803 in a bench-top photobioreactor

Synechocystis sp. PCC6803 exhibited a high degree of variation in biomass and lipid production rates in response to temperature changes in a photobioreactor. Compared with an optimal temperature of 30-33°C, a higher temperature of 44°C and lower temperatures of 22°C and 18°C severely inhibited the specific growth rate (up to a 66% decrease), biomass production rate (up to a 71% decrease), nutrient utilization rates (up to a 77% decrease), and lipid production rate (up to a 80% decrease). Temperature stress triggered changes in the relative percentage of individual fatty acids (mainly for C16:0 and C18:3), and degree of unsaturation significantly changed: 0.87 at 30°C, 0.62 at 44°C, and 1.29 at 18°C. Although PCC6803 survived temperature stress and maintained its predominate position in the culture, it could not fully recover from long-term temperature stress. Thus, avoiding prolonged exposure to extreme temperature is crucial for using PCC6803 as feedstock for biofuel production.     

Cyanobacteria as a biosorbent for mercuric ion

The biosorption of Hg(2+) by two strains of cyanobacteria, Spirulina platensis and Aphanothece flocculosa, was studied under a batch stirred reaction system. Essential process parameters, including pH, biomass concentration, initial metal concentration, and presence of co-ions were shown to influence the Hg(2+) uptake. Hg(2+) uptake was optimal at pH 6.0 for both strains. The maximum loading capacities per gram of dry biomass were found to be 456 mg Hg(2+) for A. flocculosa and 428 mg Hg(2+) for S. platensis. At an initial concentration of 10 ppm Hg(2+), A. flocculosa was able to remove more than 98% of the mercury ion from solution. The biosorption kinetics of both strains showed that the metal uptake is bi-phasic, exhibiting a rapid initial uptake followed by a slower absorption process. The presence of dissolved Co(2+), Ni(2+), and Fe(3+) were found to play a synergistic role for Hg(2+) uptake by both strains. Regeneration of the biomass was examined by treating Hg(2+)-loaded samples with HCl and NH(4)Cl over four cycles of sorption and desorption.     

Photoautotrophic nutrient utilization and limitation during semi‐continuous growth of Synechocystis sp. PCC6803

Microbial photosynthesis presents a valuable opportunity to capture abundant light energy to produce renewable bioenergy and biomaterials. To understand the factors that control the productivity of photosynthetic microorganisms, we conducted a series of semi‐continuous experiments using bench‐scale photobioreactor (PBR) systems, the cyanobacterium Synechocystis PCC6803 (PCC6803), and light conditions imitating actual day–night light irradiance (LI). Our results demonstrate that using normal BG‐11 medium resulted in severe phosphate (P_i) limitation for continuous operation. Mitigation of P_i‐limitation, by augmenting the P_i content of BG‐11, allowed higher biomass productivity; however, once P_i‐limitation was alleviated, limitation by inorganic carbon (C_i) or LI occurred. C_i‐limitation was detected by a low total C_i concentration (<5 mg C/L) and high and fluctuating pH. C_i‐limitation was relieved by delivering more CO₂, which led to a stable pH in the range of 7–9 and at least 5 mg/L of C_i in HCO. LI limitation, evidenced by an average LI <14 W/m² for PCC6803, was induced by a high biomass concentration of 1,300 mg/L. Thus, this work provides quantitative tools of stoichiometry and kinetics to evaluate limitation on PBRs. Biotechnol. Bioeng. 2010;106: 553–563. © 2010 Wiley Periodicals, Inc.     

Co2+, Cu2+, and Zn2+ accumulation by cyanobacterium Spirulina platensis

The Spirulina platensis biomass was characterized for its metal accumulation as a function of pH, external metal concentration, equilibrium isotherms, kinetics, effect of co-ions under free (living cells, lyophilized, and oven-dried) and immobilized (Ca-alginate and polyacrylamide gel) conditions. The maximum metal biosorption by S. platensis biomass was observed at pH 6.0 with free and immobilized biomass. The studies on equilibrium isotherm experiments showed highest maximum metal loading by living cells (181.0 +/- 13.1 mg Co(2+)/g, 272.1 +/- 29.4 mg Cu(2+)/g and 250.3 +/- 26.4 mg Zn(2+)/g) followed by lyophilized (79.7 +/- 9.6 mg Co(2+)/g, 250.0 +/- 22.4 mg Cu(2+)/g and 111.2 +/- 9.8 mg Zn(2+)/g) and oven-dried (25.9 +/- 1.9 mg Co(2+)/g, 160.0 +/- 14.2 mg Cu(2+)/g and 35.1 +/- 2.7 mg Zn(2+)/g) biomass of S. platensis on a dry weight basis. The polyacrylamide gel (PAG) immobilization of lyophilized biomass found to be superior over Ca-alginate (Ca-Alg) and did not interfere with the S. platensis biomass biosorption capacity, yielding 25% of metal loading after PAG entrapment. The time-dependent metal biosorption in both the free and immobilized form revealed existence of two phases involving an initial rapid phase (which lasted for 1-2 min) contributing 63-77% of total biosorption, followed by a slower phase that continued for 2 h. The metal elution studies conducted using various reagents showed more than 90% elution with mineral acids, calcium salts, and Na(2)EDTA with free (lyophilized or oven-dried) as well as immobilized biomass. The experiments conducted to examine the suitability of PAG-immobilized S. platensis biomass over multiple cycles of Co(2+), Cu(2+), and Zn(2+) sorption and elution showed that the same PAG cubes can be reused for at least seven cycles with high efficiency.     

Cu2+ Removal and recovery by Spi SORB: batch stirred and up-flow packed bed columnar reactor systems

The biosorption of Cu²⁺ by free and poly acrylamide gel (PAG) immobilized Spirulina platensis (SpiSORB) was characterized under batch and continuous packed bed columnar reaction systems. The biosorption of Cu²⁺ was shown to be highest at pH of 6.0 for both types of biomass. The PAG immobilization process did not interfere with the Cu²⁺ binding sites present on biomass leading to cent percent (ca. 250 mg g⁻¹ of dry biomass) retention of biosorption as compared to free cells. Transmission electron microscopy on Cu²⁺ localization revealed that majority of metal is being sequestered by the cell wall only. The infrared spectrum of metal treated S. platensis biomass indicated the possible involvement of amide, amino, and carboxyl groups in metal binding. Up-flow packed bed columnar reactor containing 2.0 g of PAG immobilized S. platensis shown a maximum of 143-fold volume reduction factor at the residence time of 4.6 min for Cu²⁺ alone and found to decrease dramatically when Zn²⁺ is present in a bimetallic solution.     

Nutrient acquisition and limitation for the photoautotrophic growth of Synechocystis sp. PCC6803 as a renewable biomass source

Photoautotrophic microorganisms (cyanobacteria and algae) offer high promise as a source of biomass for renewable energy due to their rapid growth rates and high biomass yields. To provide a framework for evaluating the feasibility of growing phototrophic microorganisms with high biomass production rates, we operated a bench‐scale photobioreactor using Synechocystis sp. PCC6803 and with light conditions imitating actual day–night light irradiance (LI). During the time of peak LI, PCC6803's specific growth rate (1.7 day⁻¹) and the nitrate uptake rate (0.46 g N/g DW day) were high compared to past reports. Analysis employing the stoichiometry of photosynthesis of PCC6803 and ionic speciation showed that bicarbonate and phosphate were driven to very low concentrations for the high‐LI conditions. In particular, the systematic evaluation of rate‐limiting factors identified when the CO₂–C_i supply rate needed to be increased to mitigate HCO depletion and a large pH increase. It also showed that the traditional BG‐11 medium needs to be augmented with phosphate to avoid severe P depletion. This work exploits quantitative understanding the stoichiometry and kinetics of cyanobacteria for the high‐rate production of a renewable biomass. Biotechnol. Bioeng. 2011;108: 277–285. © 2010 Wiley Periodicals, Inc.