Optimization of Recombinant Protein Production in Synechococcus elongatus PCC 7942: Utilizing Native Promoters and Magnetic Fields.

The cyanobacterium Synechococcus elongatus PCC 7942 holds significant potential as a biofactory for recombinant protein (RP) production due to its capacity to harness light energy and utilize CO2. This study aimed to enhance RP production by integration of native promoters and magnetic field application (MF) in S. elongatus PCC 7942. The psbA2 promoter, which responds to stress conditions, was chosen for the integration of the ZsGreen1 gene. Results indicated successful gene integration, affirming prior studies that showed no growth alterations in transgenic strains. Interestingly, exposure to 30 mT (MF30) demonstrated a increase in ZsGreen1 transcription under the psbA2 promoter, revealing the influence of MF on cyanobacterial photosynthetic machinery. This enhancement is likely attributed to stress-induced shifts in gene expression and enzyme activity. MF30 positively impacted photosystem II (PSII) without disrupting the electron transport chain, aligning with the "quantum-mechanical mechanism" theory. Notably, fluorescence levels and gene expression with application of 30 mT were significantly different from control conditions. This study showcases the efficacy of utilizing native promoters and MF for enhancing RP production in S. elongatus PCC 7942. Native promoters eliminate the need for costly exogenous inducers and potential cell stress. Moreover, the study expands the scope of optimizing RP production in photoautotrophic microorganisms, providing valuable insights for biotechnological applications.

Magnetic Fields as Inducers of Phycobiliprotein Production by Synechococcus elongatus PCC 7942.

This study aimed to analyze the effect of magnetic field (MF) application on the metabolism of Synechococcus elongatus PCC 7942. Concentrations of biomass, carbohydrate, protein, lipid, and photosynthetic pigments (chlorophyll-a, C-phycocyanin, allophycocyanin and phycoerythrin) were determined. In cultures with MF application (30 mT for 24 h d-1), there were increases of 47.5% in total protein content, 87.4% in C-phycocyanin, and 332.8% in allophycocyanin contents, by comparison with the control. Allophycocyanin is the most affected pigment by MF application. Therefore, its biosynthetic route was investigated, and four genes related to its synthesis were found. However, the analysis of the gene expression showed no statistical differences from the control culture, which suggests that induction of such genes may occur soon after MF application with consequent stabilization over time. MF application may be a cost-effective alternative to increase production of compounds of commercial interest by cyanobacteria.

Influence of Cell Wall on Biomolecules Biosynthesis in Chlamydomonas reinhardtii Strains Exposed to Magnetic Fields.

The application of magnetic fields (MF) has attracted the attention of researchers due to their efficiency to change cell metabolism. Chlamydomonas reinhardtii is a biotechnologically useful microalga with versatile metabolism that may be a valuable organism to study the effects of the MF in biology. Therefore, two C. reinhardtii strains, one with cell wall (2137) and other which lacks the cell wall (Wt-S1-cc4694), were evaluated that a new sensitivity factor in the analysis could be included. Comparative studies were undertaken with the two C. reinhardtii strains under the MF intensities of 0.005 mT (terrestrial MF - control), 11  and 20 mT. Results indicated that the physical cell wall barrier protected cells against the MF applied during the assays. Only with the highest MF applied (20 mT) a slight increase in lipid concentration in the cell wall strain was detected. The lowest growth of the strain that lacks cell wall (Wt-S1) indicated that these cells are under a negative effect. To cope with the two MF stresses conditions, Wt-S1 cells produced more pigments (chlorophylls and carotenoids) and lipids and enhanced the antioxidant defense system. The raise of these compounds under MF could potentially have a positive biotechnological impact on algal biomass.

Simultaneous Application of Mixotrophic Culture and Magnetic Fields as a Strategy to Improve Spirulina sp. LEB 18 Phycocyanin Synthesis.

Spirulina is a filamentous microalga which is considered a promising alternative source of essential nutrients and active biomolecules. High production cost and the space required to install a photobioreactor are two of the greatest challenges in the industrial application of microalga-based products. Thus, this study aimed to improve Spirulina sp. LEB 18 biomass and phycocyanin content by combining the application of mixotrophic culture and magnetic fields (MF). Zarrouk medium was modified with 1 and 3 g/L liquid molasses and the application of 30 mT for 1·h/d was investigated. Mixotrophic culture with 1 g/L molasses showed the highest biomass concentration (1.62 g/L), carbohydrate content (25.6%), and lipid contents (8.7%) after 15 days. Although the combination of 30 mT and 1 g/L liquid molasses decreased biomass production (1.44 g/L), there was increase in protein yield (76.9%) and protein productivity (73.8 mg/L·d). The proposed method increased phycocyanin production by 145% and its purity from 0.584 in the control culture to 0.627. Data described by this study show that the combination of mixotrophic culture and MF application is a promising alternative to increase microalga protein and phycocyanin production.

Innovative development of membrane sparger for carbon dioxide supply in microalgae cultures.

The present study was aimed to develop a membrane sparger (MS) integrated into a tubular photobioreactor to promote the increase of the carbon dioxide (CO2 ) fixation by Spirulina sp. LEB 18 cultures. The use of MS for the CO2 supply in Spirulina cultures resulted not only in the increase of DIC concentrations but also in the highest accumulated DIC concentration in the liquid medium (127.4 mg L-1 d-1 ). The highest values of biomass concentration (1.98 g L-1 ), biomass productivity (131.8 mg L-1 d-1 ), carbon in biomass (47.9% w w-1 ), CO2 fixation rate (231.6 mg L-1 d-1 ), and CO2 use efficiency (80.5% w w-1 ) by Spirulina were verified with MS, compared to the culture with conventional sparger for CO2 supply. Spirulina biomass in both culture conditions had high protein contents varying from 64.9 to 69% (w w-1 ). MS can be considered an innovative system for the supply of carbon for the microalgae cultivation and biomass production. Moreover, the use of membrane system might contribute to increased process efficiency with a reduced cost of biomass production.

Bioprocess strategies for enhancing biomolecules productivity in Chlorella fusca LEB 111 using CO2 a carbon source.

The aim of this study was to evaluate the influence of different carbon dioxide (CO2 ) concentrations on the distribution of carbon forms in the culture medium and the biomass production and biomolecules productivity of the strain Chlorella fusca LEB 111. In this study, experiments were carried out in which C. fusca cultures were exposed to different CO2 concentrations, 0.03% (0.08 mlCO2 mlmedium-1 days-1 ), 5% (0.18 mlCO2 mlmedium-1 days-1 ), and 15% vol/vol CO2 (0.54 mlCO2 mlmedium-1 days-1 ). Among the carbon chemical species distributions in the culture medium, bicarbonate was predominant (94.2-98.9%), with the highest quantitative percentage in the experiment receiving a 15% CO2 injection. C. fusca LEB 111 cultivated with 15% CO2 showed the highest biomass productivity (194.3 mg L-1 days-1 ) and CO2 fixation rate (390.9 mg L-1 days-1 ). The carbohydrate productivity in the culture that received 15% CO2 was 46.2% higher than the value verified for the culture with the addition of CO2 from the air (0.03% CO2 ). In addition, CO2 concentration providing increases of 0.03-15% to C. fusca cultures resulted in a 31.6% increase in the lipid productivity. These results showed that C. fusca can be used for CO2 bioconversion and for producing biomass with potential applications for biofuels and bioproducts.