Comprehensive understanding of the mutant 'giant' Arthrospira platensis developed via ultraviolet mutagenesis.

Introduction: Cyanobacteria are typically of a size that can be observed under a microscope. Here, we present cyanobacteria of a size that can be observed with the naked eye. Arthrospira platensis NCB002 strain showed differentiated morphological characteristics compared to previously reported Arthrospira spp. Methods: Arthrospira platensis NCB002 was obtained by the UV irradiation of Arthrospira sp. NCB001, which was isolated from freshwater and owned by NCell Co., Ltd. A. platensis NIES-39 was obtained from the National Institute for Environmental Studies (Tsukuba, Japan). We used various analytical techniques to determine its overall characteristics. Results and discussion: The draft genome of strain NCB002 consists of five contigs comprising 6,864,973 bp with a G+C content of 44.3 mol%. The strain NCB002 had an average length of 11.69 ± 1.35 mm and a maximum of 15.15 mm, which is 23.4-50.5 times longer than the length (0.3-0.5 mm) of previously known Arthrospira spp., allowing it to be harvested using a thin sieve. Transcriptome analysis revealed that these morphological differences resulted from changes in cell wall formation mechanisms and increased cell division. Our results show that NCB002 has outstanding industrial value and provides a comprehensive understanding of it.

Green and efficient method to acquire high-value phycobiliprotein from microalgal biomass involving deep eutectic solvent-based ultrasound-assisted extraction.

Phycoerythrin (PE) is a phycobiliprotein holding great potential as a high-value food colorant and medicine. Deep eutectic solvent (DES)-based ultrasound-assisted extraction (UAE) was applied to extract B-PE by disrupting the resistant polysaccharide cell wall of Porphyridium purpureum. The solubility of cell wall monomers in 31 DESs was predicted using COSMO-RS. Five glycerol-based DESs were tested for extraction, all of which showed significantly higher B-PE yields by up to 13.5 folds than water. The DES-dependent B-PE extraction efficiencies were proposedly associated with different cell disrupting capabilities and protein stabilizing effects of DESs. The DES-based UAE method could be considered green according to a metric assessment tool, AGREEprep. The crude extract containing DES was further subjected to aqueous two-phase system, two-step ammonium sulfate precipitation, and ultrafiltration processes. The final purified B-PE had a PE purity ratio of 3.60 and a PC purity ratio of 0.08, comparable to the purity of commercial products.

Sparse ordinal discriminant analysis.

Ordinal class labels are frequently observed in classification studies across various fields. In medical science, patients' responses to a drug can be arranged in the natural order, reflecting their recovery postdrug administration. The severity of the disease is often recorded using an ordinal scale, such as cancer grades or tumor stages. We propose a method based on the linear discriminant analysis (LDA) that generates a sparse, low-dimensional discriminant subspace reflecting the class orders. Unlike existing approaches that focus on predictors marginally associated with ordinal labels, our proposed method selects variables that collectively contribute to the ordinal labels. We employ the optimal scoring approach for LDA as a regularization framework, applying an ordinality penalty to the optimal scores and a sparsity penalty to the coefficients for the predictors. We demonstrate the effectiveness of our approach using a glioma dataset, where we predict cancer grades based on gene expression. A simulation study with various settings validates the competitiveness of our classification performance and demonstrates the advantages of our approach in terms of the interpretability of the estimated classifier with respect to the ordinal class labels.

Endophyte Bacillus tequilensis improves the growth of microalgae Haematococcus lacustris by regulating host cell metabolism.

This study identified an endosymbiotic bacterium, Bacillus tequilensis, residing within the cells of the microalga Haematococcus lacustris through 16S rRNA analysis. To confirm the optimal interactive conditions between H. lacustris and B. tequilensis, the effects of different ratios of cells using H. lacustris of different growth stages were examined. Under optimized conditions, the cell density, dry weight, chlorophyll content, and astaxanthin content of H. lacustris increased significantly, and the fatty acid content improved 1.99-fold. Microscopy demonstrated the presence of bacteria within the H. lacustris cells. The interaction upregulated amino acid and nucleotide metabolism in H. lacustris. Interestingly, muramic and phenylacetic acids were found exclusively in H. lacustris cells in the presence of B. tequilensis. Furthermore, B. tequilensis delayed pigment degradation in H. lacustris. This study reveals the impact of the endosymbiont B. tequilensis on the metabolism of H. lacustris and offers new perspectives on the symbiotic relationship between them.

Hydrotrifluoroacetylation of Alkenes via Designer Masked Acyl Reagents.

Because of its impressive ability to promote pharmaceutical activity, the introduction of trifluoromethylacyl (CF3CO) functionality into organic compounds has become an important and growing research area. Although various protocols have been developed to access trifluoroketones, the use of trifluoroacetyl radicals remains virtually undeveloped. Herein, we disclose a novel method for trifluoroacetylation through an umpolung reagent, thereby transforming an electrophilic radical into a nucleophilic radical. The applicability of this transformation is highlighted by large-scale, late-stage reactions of complex bioactive molecules sclareolide and loratadine. Furthermore, the direct transformation of trifluoromethyl ketones into various fluorinated analogues illustrates the potential synthetic application of our developed method.

C-H amidation of 2-aryl azlactones under iridium(III) catalysis: access to chiral amino acids.

In this study, we examine the site-selctive iridium(III)-catalyzed C-H amidation between 2-aryl azlactones and acyl azides. This transformation produces a range of ortho-amidated azlactones, which act as precursors for the synthesis of chiral amino acids via organocatalyzed ring-opening reactions. To test its effectiveness, the method developed is applied to the late-stage C-H amidation of complex drug molecules. The isolation of an iridacycle species supports a proposed reaction pathway.

Optimal Composition of Li Argyrodite with Harmonious Conductivity and Chemical/Electrochemical Stability: Fine-Tuned Via Tandem Particle Swarm Optimization.

A tandem (two-step) particle swarm optimization (PSO) algorithm is implemented in the argyrodite-based multidimensional composition space for the discovery of an optimal argyrodite composition, i.e., with the highest ionic conductivity (7.78 mS cm-1 ). To enhance the industrial adaptability, an elaborate pellet preparation procedure is not used. The optimal composition (Li5.5 PS4.5 Cl0.89 Br0.61 ) is fine-tuned to enhance its practical viability by incorporating oxygen in a stepwise manner. The final composition (Li5.5 PS4.23 O0.27 Cl0.89 Br0.61 ), which exhibits an ionic conductivity (σion ) of 6.70 mS cm-1 and an activation barrier of 0.27 eV, is further characterized by analyzing both its moisture and electrochemical stability. Relative to the other compositions, the exposure of Li5.5 PS4.23 O0.27 Cl0.89 Br0.61 to a humid atmosphere results in the least amount of H2 S released and a negligible change in structure. The improvement in the interfacial stability between the Li(Ni0.9 Co0.05 Mn0.05 )O2 cathode and Li5.5 PS4.23 O0.27 Cl0.89 Br0.61 also results in greater specific capacity during fast charge/discharge. The structural and chemical features of Li5.5 PS4.5 Cl0.89 Br0.61 and Li5.5 PS4.23 O0.27 Cl0.89 Br0.61 argyrodites are characterized using synchrotron X-ray diffraction, Raman spectroscopy, and X-ray photoelectron spectroscopy. This work presents a novel argyrodite composition with favorably balanced properties while providing broad insights into material discovery methodologies with applications for battery development.

Establishment of ultrasonic stimulation to enhance growth of Haematococcus lacustris.

In this study, ultrasonication at a frequency of 40 kHz was used to shorten the sonication period and enhance the growth of Haematococcus lacustris. To confirm the optimal conditions, the effects of ultrasound output and treatment interval were examined. Under optimal conditions (20 W and 15-day cycle), the maximum cell density and chlorophyll content were 66.75 × 104 cells mL-1 and 36.54 mg g-1, respectively, which were increased by 50.00% and 39.01%, respectively, compared to the control. Transmission electron microscopy analysis showed that ultrasonication caused tiny cracks in the W4 and W6 strata but did not disrupt the inner W2 layer. Additionally, RT-qPCR analysis showed that ultrasonication upregulated both cell division and nitrogen uptake. No difference were detected in the composition or quantity of fatty acids. This study demonstrates a novel ultrasonic approach for enhancing the growth of H. lacustris.

Microalgal secondary metabolite productions as a component of biorefinery: A review.

The interest in developing microalgae for industrial use has been increasing because of concerns about the depletion of petroleum resources and securing sustainable energy sources. Microalgae have high biomass productivity and short culture periods. However, despite these advantages, various barriers need to be overcome for industrial applications. Microalgal cultivation has a high unit price, thus rendering industrial application difficult. It is indispensably necessary to co-produce their primary and secondary metabolites to compensate for these shortcomings. In this regard, this article reviews the following aspects, (1) co-production of primary and secondary metabolites in microalgae, (2) induction methods for the promotion of the biosynthesis of secondary metabolites, and (3) perspectives on the co-production and co-extraction of primary and secondary metabolites. This paper presents various approaches for producing useful metabolites from microalgae and suggests strategies that can be utilized for the co-production of primary and secondary metabolites.

Complete chloroplast genome of Gypsophila oldhamiana Miq. (Caryophyllales: Caryophyllaceae).

The complete chloroplast genome sequence of Gypsophila oldhamiana Miq., a species of the Caryophyllaceae family, was assembled and analyzed from the de novo assembly of Illumina paired-end sequencing data. The total length of the chloroplast genome of G. oldhamiana was 152,675 bp including a large single-copy (LSC) region of 83,552 bp, a small single-copy (SSC) region of 17,349 bp, and a pair of identical inverted repeat regions (IRs) of 25,887 bp. The genome possessed a total of 130 genes, including 85 protein-coding genes, 37 transfer RNA (tRNA) genes, and 8 ribosomal RNA (rRNA) genes. The phylogenetic analysis of G. oldhamiana with 14 related species discovered the closest taxonomical relationship with Gypsophila vaccaria voucher in the Caryophyllaceae family.

Semi-continuous immobilized cultivation of Porphyridium cruentum for sulfated polysaccharides production.

In this study, semi-continuous immobilized cultivation of Porphyridium cruentum through calcium alginate beads was performed for sulfated polysaccharides (SPs) production. The cell biomass and daily SPs productivity in the calcium alginate bead immobilized culture were increased by up to 79 ± 3.4% and 45.6 ± 3.2%, compared to those in the control, respectively. Furthermore, simultaneous application of immobilization and blue wavelength illumination further increased the phycobiliproteins content by 260 ± 9%, compared to those in the control. Similarly, nutrient deficiencies in combination with immobilization increased daily SPs productivity by about twice that of the control. The chemical composition and biological activity of the extracellular polymeric substances produced through immobilization were similar to those of the control. This study suggests the potential application of calcium alginate beads-based immobilization for continuous and high-efficiency SPs production using P. cruentum.

Enhancement of phycoerythrin productivity in Porphyridium purpureum using the clustered regularly interspaced short palindromic repeats/CRISPR-associated protein 9 ribonucleoprotein system.

Genetic manipulation of the Porphyridium sp. may increase the production of phycoerythrin. Since phycobiliproteins capture and transfer energy to both photosystems (PS I and PS II), it was hypothesized that the gene mutation involved increases phycoerythrin synthesis. The gene encoding chlorophyll synthase (CHS1) was selected as chlorophyll synthase plays an important role in photosynthesis, mediating the final process of chlorophyll synthesis. Clustered regularly interspaced short palindromic repeats/CRISPR-associated protein 9 ribonucleoprotein (CRISPR/Cas9 RNP) delivery system was used to generate the chlorophyll synthase loss-of-function mutants (Δchs1). Independent Δchs1 showed no differences in the growth and production of sulfated polysaccharide compared to control. Phycoerythrin contents of the two independent mutants substantially increased regardless of light source. This study provides a novel applicability for the CRISPR/Cas9 RNP method in red microalgae toward a bio-product of interest. The obtained mutants could serve as potential producers of phycoerythrin if Porphyridium is selected as a natural source.

C-H Methylation of Iminoamido Heterocycles with Sulfur Ylides*.

The direct methylation of N-heterocycles is an important transformation for the advancement of pharmaceuticals, agrochemicals, functional materials, and other chemical entities. Herein, the unprecedented C(sp2 )-H methylation of iminoamido heterocycles as nucleoside base analogues is described. Notably, trimethylsulfoxonium salt was employed as a methylating agent under aqueous conditions. A wide substrate scope and excellent level of functional-group tolerance were attained. Moreover, this method can be readily applied to the site-selective methylation of azauracil nucleosides. The feasibility of gram-scale reactions and various transformations of the products highlight the synthetic potential of the developed method. Combined deuterium-labeling experiments aided the elucidation of a plausible reaction mechanism.

C2-Selective C-H Methylation of Heterocyclic N-Oxides with Sulfonium Ylides.

A redox-neutral C2-selective methylation of heterocyclic N-oxides with sulfonium ylides is described herein. This report presents unprecedented findings for the utility of sulfonium ylides as the methylation source of N-heterocycles beyond the Corey-Chaykovsky reaction. Intriguingly, pyrrolidine plays a significant role in minimizing the reductive C2-methylation process. This method is characterized by its mild conditions, simplicity, and excellent site selectivity. The applicability of the developed protocol is showcased by the late-stage methylation and sequential transformations of complex drug molecules.

Ru(ii)-Catalyzed C-H addition and oxidative cyclization of 2-aryl quinazolinones with activated aldehydes.

The ruthenium(ii)-catalyzed cross-coupling reaction between 2-aryl quinazolinones and activated aldehydes is described. This method enables the site-selective hydroxyalkylation under redox-neutral conditions. Moreover, this protocol provides a facile access to various tetracyclic isoindoloquinazolinones by using Cu(OAc)2 as an external oxidant via C-H addition and subsequent intramolecular cyclization. A wide substrate scope and a high level of chemoselectivity as well as broad functional group tolerance are observed.

Astaxanthin biosynthesis promotion with pH shock in the green microalga, Haematococcus lacustris.

In this study, the use of pH shock to improve astaxanthin synthesis in Haematococcus lacustris was investigated. It has been found that pH shock (pH = 4.5, 60 s) imposes stress in the cells and induces physiological changes, which result in astaxanthin accumulation. The optimal acid-base combination of pH shock was H2SO4-KOH, which increased the astaxanthin content per cell to 39 ± 6.92% than those of the control. In addition, pH shock can be applied simultaneously with the other inductive strategies such as high irradiance and carbon source supply. When high irradiance was applied simultaneously with pH shock, astaxanthin yield was increased 65 ± 0.541% than control. In addition, astaxanthin content per cell was increased 105 ± 6.66% than those of the control, with the concomitant application of carbon source addition with pH shock. Herein, these novel findings provide a useful technique for producing astaxanthin using H. lacustris.

Deoxygenative Amination of Azine-N-oxides with Acyl Azides via [3 + 2] Cycloaddition.

A transition-metal-free deoxygenative C-H amination reaction of azine-N-oxides with acyl azides is described. The initial formation of an isocyanate from the starting acyl azide via a Curtius rearrangement can trigger a [3 + 2] dipolar cycloaddition of polar N-oxide fragments to generate the aminated azine derivative. The applicability of this method is highlighted by the late-stage and sequential amination reactions of complex bioactive compounds, including quinidine and fasudil. Moreover, the direct transformation of aminated azines into various bioactive N-heterocycles illustrates the significance of this newly developed protocol.

Effect of Pseudoalteromonas sp. MEBiC 03485 on biomass production and sulfated polysaccharide biosynthesis in Porphyridium cruentum UTEX 161.

The effect of co-cultivation of Porphyridium cruentum UTEX 161 with Pseudoalteromonas sp. MEBiC 03485 on P. cruentum growth and its sulfated polysaccharide (EPS) production were examined. The strain MEBiC 03485 had beneficial effects on P. cruentum growth, EPS production, and EPS quality. These effects were due to a compound secreted by the strain MEBiC 03485. Notably, secretory compound treatment also increased intracellular phycoerythrin and phycocyanin content by 89.4% and 161%, respectively. In addition, the biological activities of EPS extracted from MEBiC 03485 treatment tended to be higher than the control without treatment. Our results suggest a novel approach for potentially enhancing the growth of P. cruentum and its EPS production and quality by co-culturing with the symbiotic strain MEBiC 03485.

ZnO Microfiltration Membranes for Desalination by a Vacuum Flow-Through Evaporation Method.

ZnO was deposited on macroporous α-alumina membranes via atomic layer deposition (ALD) to improve water flux by increasing their hydrophilicity and reducing mass transfer resistance through membrane pore channels. The deposition of ZnO was systemically performed for 4-128 cycles of ALD at 170 °C. Analysis of membrane surface by contact angles (CA) measurements revealed that the hydrophilicity of the ZnO ALD membrane was enhanced with increasing the number of ALD cycles. It was observed that a vacuum-assisted 'flow-through' evaporation method had significantly higher efficacy in comparison to conventional desalination methods. By using the vacuum-assisted 'flow-through' technique, the water flux of the ZnO ALD membrane (~170 L m-2 h-1) was obtained, which is higher than uncoated pristine membranes (92 L m-2 h-1). It was also found that ZnO ALD membranes substantially improved water flux while keeping excellent salt rejection rate (>99.9%). Ultrasonic membrane cleaning had considerable effect on reducing the membrane fouling.

Control of a toxic cyanobacterial bloom species, Microcystis aeruginosa, using the peptide HPA3NT3-A2.

Microcystis aeruginosa, a species of freshwater cyanobacteria, is known to be one of the dominant species causing cyanobacterial harmful algal blooms (CyanoHABs). M. aeruginosa blooms have the potential to produce neurotoxins and peptide hepatotoxins, such as microcystins and lipopolysaccharides (LPSs). Currently, technologies for CyanoHAB control do not provide any ultimate solution because of the secondary pollution associated with the control measures. In this study, we attempted to use the peptide HPA3NT3-A2, which has been reported to be nontoxic and has antimicrobial properties, for the development of an eco-friendly control against CyanoHABs. HPA3NT3-A2 displayed significant algicidal effects against M. aeruginosa cells. HPA3NT3-A2 induced cell aggregation and flotation (thereby facilitating harvest), inhibited cell growth through sedimentation, and eventually destroyed the cells. HPA3NT3-A2 had no algicidal effect on other microalgal species such as Haematococcus pluvialis and Chlorella vulgaris. Additionally, HPA3NT3-A2 was not toxic to Daphnia magna. The algicidal mechanism of HPA3NT3-A2 was intracellular penetration. The results of this study suggest the novel possibility of controlling CyanoHABs using HPA3NT3-A2.