Search Results (4)

In the recent past, various microalgae have been considered a renewable energy source for biofuel production, and their amount and extent can be enhanced by applying certain types of stress including salinity. Although microalgae growing under salinity stress result in a higher lipid content, they simultaneously reduce in growth and biomass output. To resolve this issue, the physiochemical changes in microalgae Scenedesmus sp. BHU1 have been assessed through two-stage cultivation. In stage-I, the maximum carbohydrate and lipid contents (39.55 and 34.10%) were found at a 0.4 M NaCl concentration, while in stage-II, the maximum carbohydrate and lipid contents (42.16 and 38.10%) were obtained in the 8-day-old culture. However, under increased salinity, Scenedesmus sp. BHU1 exhibited a decrease in photosynthetic attributes, including Chl-a, Chl-b, Fv/Fm, Y(II), Y(NPQ), NPQ, qP, qL, qN, and ETRmax but increased Y(NO) and carotenoids content. Apart from physiological attributes, osmoprotectants, stress biomarkers, and nonenzymatic antioxidants were also studied to elucidate the role of reactive oxygen species (ROS) facilitated lipid synthesis. Furthermore, elemental and mineral ion analysis of microalgal biomass was performed to evaluate the biomass quality for biofuel and cell homeostasis. Based on fluorometry analysis, we found the maximum neutral lipids in the 8-day-old grown culture at stage-II in Scenedesmus sp. BHU1. Furthermore, the use of Fourier-transform infrared (FT-IR) and nuclear magnetic resonance (NMR) spectroscopy analyses confirmed the presence of higher levels of hydrocarbons and triacylglycerides (TAGs) composed of saturated fatty acids (SFAs) and monounsaturated fatty acids (MUFAs) in the 8-day-old culture. Therefore, Scenedesmus sp. BHU1 can be a promising microalga for potential biodiesel feedstock. Full article

The mRubyFT is a monomeric genetically encoded fluorescent timer based on the mRuby2 fluorescent protein, which is characterized by the complete maturation of the blue form with the subsequent conversion to the red one. It has higher brightness in mammalian cells and higher photostability compared with other fluorescent timers. A high-resolution structure is a known characteristic of the mRubyFT with the red form chromophore, but structural details of its blue form remain obscure. In order to obtain insight into this, we obtained an S148I variant of the mRubyFT (mRubyFT^S148I) with the blocked over time blue form of the chromophore. X-ray data at a 1.8 Å resolution allowed us to propose a chromophore conformation and its interactions with the neighboring residues. The imidazolidinone moiety of the chromophore is completely matured, being a conjugated π-system. The methine bridge is not oxidized in the blue form bringing flexibility to the phenolic moiety that manifests itself in poor electron density. Integration of these data with the results of molecular dynamic simulation disclosed that the OH group of the phenolic moiety forms a hydrogen bond with the side chain of the T163 residue. A detailed comparison of mRubyFT^S148I with other available structures of the blue form of fluorescent proteins, Blue102 and mTagBFP, revealed a number of characteristic differences. Molecular dynamic simulations with the combined quantum mechanic/molecular mechanic potentials demonstrated that the blue form exists in two protonation states, anion and zwitterion, both sharing enolate tautomeric forms of the C=C–O⁻ fragment. These two forms have similar excitation energies, as evaluated by calculations. Finally, excited state molecular dynamic simulations showed that excitation of the chromophore in both protonation states leads to the same anionic fluorescent state. The data obtained shed light on the structural features and spectral properties of the blue form of the mRubyFT timer. Full article

True genetically encoded monomeric fluorescent timers (tFTs) change their fluorescent color as a result of the complete transition of the blue form into the red form over time. Tandem FTs (tdFTs) change their color as a consequence of the fast and slow independent maturation of two forms with different colors. However, tFTs are limited to derivatives of the mCherry and mRuby red fluorescent proteins and have low brightness and photostability. The number of tdFTs is also limited, and there are no blue-to-red or green-to-far-red tdFTs. tFTs and tdFTs have not previously been directly compared. Here, we engineered novel blue-to-red tFTs, called TagFT and mTagFT, which were derived from the TagRFP protein. The main spectral and timing characteristics of the TagFT and mTagFT timers were determined in vitro. The brightnesses and photoconversions of the TagFT and mTagFT tFTs were characterized in live mammalian cells. The engineered split version of the TagFT timer matured in mammalian cells at 37 °C and allowed the detection of interactions between two proteins. The TagFT timer under the control of the minimal arc promoter, successfully visualized immediate-early gene induction in neuronal cultures. We also developed and optimized green-to-far-red and blue-to-red tdFTs, named mNeptusFT and mTsFT, which were based on mNeptune-sfGFP and mTagBFP2-mScarlet fusion proteins, respectively. We developed the FucciFT2 system based on the TagFT-hCdt1-100/mNeptusFT2-hGeminin combination, which could visualize the transitions between the G1 and S/G2/M phases of the cell cycle with better resolution than the conventional Fucci system because of the fluorescent color changes of the timers over time in different phases of the cell cycle. Finally, we determined the X-ray crystal structure of the mTagFT timer and analyzed it using directed mutagenesis. Full article

The present study was carried out to determine the bioactivity of polysaccharides extracted from Euglena gracilis (EgPs). These were characterized by FT-IR and GC-MS. Cytotoxicity analyses (MTT) were performed on healthy human gingival fibroblast cell lines (HGF-1), obtaining an IC₅₀ of 228.66 µg mL⁻¹, and cell lines with anticancer activity for colon cancer (HCT-116), breast cancer (MCF-7), human leukemia (U-937, HL-60) and lung cancer (NCl-H460), showing that EgPs have anticancer activity, mainly in HTC-116 cells (IC₅₀ = 26.1 µg mL⁻¹). The immunological assay determined the immunomodulatory capacity of polysaccharides for the production of proinflammatory cytokines IL-6 and TNF-α in murine macrophages (RAW 264.7) and TNF-α in human monocytes (THP-1). It was observed that the EgPs had a stimulating capacity in the synthesis of these interleukins. The antioxidant capacity of polysaccharides and their biomass were analyzed using the ABTS method (18.30 ± 0.14% and (5.40 ± 0.56%, respectively, and the DPPH method for biomass (17.79 ± 0.57%). We quantitatively profiled HGF-1 proteins by liquid chromatography–tandem mass spectrometry analysis, coupled with 2-plex tandem mass tag labelling, in normal cells. In total, 1346 proteins were identified and quantified with high confidence, of which five were considered to be overexpressed. The data is available through ProteomeXchange, under identifier PXD029076. Full article