Isolation and antibacterial activity of anabaena phycocyanin

The isolation and antibacterial activity of anabaena phycocyanin were investigated. The result indicates that three kinds of protein ingredients: PC-A, PC-B and PC-C were obtained using high performance liquid chromatography. The estimated molecular masses of PC-A and PC-B were 14 to 18 kD. PC-B and PC-C had certain antibacterial activity on Bibrio parahemolyticus, Bacillus mucilaginosus and Sarcina lutea. In addition, PC-C had certain antibacterial activity on Vibrio harveyi. PC-A did not possess antibacterial activity in the study.Keywords: Anabaena, phycocyanin, liquid chromatogram, antibacterialAfrican Journal of Biotechnology Vol. 12(15), pp. 1869-187

Characterization of Spermidine Synthase (SPDS) Gene and RNA−Seq Based Identification of Spermidine (SPD) and Spermine (SPM) Involvement in Improving High Temperature Stress Tolerance in Gracilariopsis lemaneiformis (Rhodophyta)

Gracilariopsis lemaneiformis, an important commercial red macroalga, is facing significant impacts from global warming, which limits algal growth and yield in China. Polyamines (PAs), spermidine (SPD) and spermine (SPM), are ubiquitous polycations important for growth and environmental stress responses including high temperature (HT) tolerance. Spermidine synthase (SPDS) gene is one of the important genes in higher PA biosynthesis, which plays critical roles in HT stress response. Here, we isolated an SPDS gene from G. lemaneiformis and further analyzed its phylogenetic tree, subcellular localization, and gene expression patterns under stress conditions. Meanwhile, supplemented with SPD and SPM were used to study the effects of PAs on HT tolerance in G. lemaneiformis. It showed exogenous 0.5 mM SPD and SPM, respectively, remarkably improved the algal relative growth rate (RGR) compared to those in the CK treatment groups under HT conditions. In addition, they both significantly elevated the activities of antioxidant enzymes and significantly upregulated the expression of genes encoding antioxidant enzymes, triggered transcription factors (TFs) signaling, and improved the expression of genes encoding small heat shock proteins (sHSP20s) during HT stress. Moreover, exogenous PA also enhanced the expression of genes involved in pyruvate metabolism, ascorbate and aldarate metabolism, and nucleotide excision repair in G. lemaneiformis, which helped to maintain better energy supply, redox homeostasis, and genome integrity under HT stress. Taken together, these data provided valuable information for functional characterization of specific gene in endogenous PA synthesis and uncovered the importance of exogenous PAs in promoting algae adaptation to HT stress

Genome-wide identification of the GT7 and GH16 genes and their correlations with agar metabolism in the agarophyte Gracilariopsis lemaneiformis

The economically important seaweed Gracilariopsis lemaneiformis (Gp. lemaneiformis) is the main resource for agar production and abalone bait. Few studies have focused on the functions of glycosyltransferases (GTs) and glycoside hydrolases (GHs) involved in saccharometabolism, including that of agar. To investigate the possible functions of glycosyltransferase family 7 (GT7) and glycoside hydrolase family 16 (GH16) members in agar metabolism, analyses of these gene families were conducted using bioinformatics-based and physiological methods. In total, five GlGT7 genes and four GlGH16 genes from the Gp. lemaneiformis genome were identified and analyzed. These GT7 and GH16 members are absent in higher plants, and the majority of GlGT7s and GlGH16s have no introns. Catalytic pocket residue analysis revealed that GlGT7s may function as β-1,4-galactosyltransferases and that GlGH16s play roles as agarases. Promoter prediction and qRT-PCR experiments verified that these genes can be regulated by light and phytohormones. GlGT7s were predicted to interact with carbohydrate sulfotransferases (STs), whereas yeast two-hybrid (Y2H) assays revealed no interactions between these proteins. Under heat stress, no significant difference was found in agar content; however, the expression of GlGT7s fluctuated and that of GlGH16 increased. Low nitrogen stress significantly increased the agar content, and the expression of GlGT7s increased, whereas that of GlGH16s decreased. In three cultivars of Gp. lemaneiformis, the expression of GlGT7 genes in cultivars 981 and Lulong No. 1 was upregulated relative to that in the wild type, whereas GlGH16 expression levels were significantly decreased. Fluorescence microscopy further showed that β-1,4-galactose accumulation was consistent with increases in agar content and GlGT7 expression. Pearson correlation analysis confirmed that the expression levels of GlGT7s and GlGH16s were positively and negatively correlated with agar accumulation, respectively. Taken together, these results demonstrated that GlGT7 and GlGH16 are intimately correlated with agar metabolism; in particular, GlGT7-2, GlGT7-5, and GlGH16-4 could act as molecular markers to indicate agar yield. This study will provide a valuable basis for breeding new cultivars with high agar content

Transcriptome Profiling of the Microalga \u3ci\u3eChlorella pyrenoidosa\u3c/i\u3e In Response to Different Carbon Dioxide Concentrations

To enrich our knowledge of carbon dioxide (CO2)-concentrating mechanism (CCM) in eukaryotic algae, we used high-throughput sequencing to investigate the transcriptome profiling of the microalga Chlorella pyrenoidosa (Chlorophyta) response to different CO2 levels. Altogether, 53.86 million (M) and 62.10 M clean short reads of 100 nucleotides (nt) were generated from this microalga cultured at 4-fold air CO2 (control) and air CO2 concentrations by Illumina sequencing. A total of 32,662 unigenes were assembled from the two pooled samples. With an E-value cut-off of 1e-5, 9590, 6782, 5954, and 9092 unigenes were annotated in NR, Gene Ontology (GO), Eukaryotic Cluster of Orthologous Groups of proteins (KOG), and Kyoto Encyclopedia of Genes and Genomes (KEGG) databases, respectively. After screening, 51 differentially expressed unigenes were up-regulated and 8 were down-regulated in the air CO2 group, relative to the control. The transcript levels of eight differentially expressed unigenes were validated by real-time quantitative PCR, which manifested that thioredoxin-like protein, laminin subunit beta-1, and chlorophyll a/b binding protein might be associated with the utilization of inorganic carbon at low CO2 levels

Preparation and Identification of ACE Inhibitory Peptides from the Marine Macroalga Ulva intestinalis

Angiotensin I-converting enzyme (ACE) inhibitory peptides derived from seaweed represent a potential source of new antihypertensive. The aim of this study was to isolate and purify ACE inhibitory peptides (ACEIPs) from the protein hydrolysate of the marine macroalga Ulva intestinalis. U. intestinalis protein was hydrolyzed by five different proteases (trypsin, pepsin, papain, α-chymotrypsin, alcalase) to prepare peptides; compared with other hydrolysates, the trypsin hydrolysates exhibited the highest ACE inhibitory activity. The hydrolysis conditions were further optimized by response surface methodology (RSM), and the optimum conditions were as follows: pH 8.4, temperature 28.5 °C, enzyme/protein ratio (E/S) 4.0%, substrate concentration 15 mg/mL, and enzymolysis time 5.0 h. After fractionation and purification by ultrafiltration, gel exclusion chromatography and reverse-phase high-performance liquid chromatography, two novel purified ACE inhibitors with IC50 values of 219.35 μM (0.183 mg/mL) and 236.85 μM (0.179 mg/mL) were obtained. The molecular mass and amino acid sequence of the ACE inhibitory peptides were identified as Phe-Gly-Met-Pro-Leu-Asp-Arg (FGMPLDR; MW 834.41 Da) and Met-Glu-Leu-Val-Leu-Arg (MELVLR; MW 759.43 Da) by ultra-performance liquid chromatography-tandem mass spectrometry. A molecular docking study revealed that the ACE inhibitory activities of the peptides were mainly attributable to the hydrogen bond and Zn(II) interactions between the peptides and ACE. The results of this study provide a theoretical basis for the high-valued application of U. intestinalis and the development of food-derived ACE inhibitory peptides

Transcriptome Profiling Reveals the Antitumor Mechanism of Polysaccharide from Marine Algae Gracilariopsis lemaneiformis.

Seaweed is one of the important biomass producers and possesses active metabolites with potential therapeutic effects against tumors. The red alga Gracilariopsis lemaneiformis (Gp. lemaneiformis) possesses antitumor activity, and the polysaccharide of Gp. lemaneiformis (PGL) has been demonstrated to be an ingredient with marked anticancer activity. However, the anticancer mechanism of PGL remains to be elucidated. In this study, we analyzed the inhibitory effect of PGL on the cell growth of 3 human cancer cell lines and found that PGL inhibited cell proliferation, reduced cell viability, and altered cell morphology in a time- and concentration-dependent manner. Our transcriptome analysis indicates that PGL can regulate the expression of 758 genes, which are involved in apoptosis, the cell cycle, nuclear division, and cell death. Furthermore, we demonstrated that PGL induced apoptosis and cell cycle arrest and modulated the expression of related genes in the A549 cell line. Our work provides a framework to understand the effects of PGL on cancer cells, and can serve as a resource for delineating the antitumor mechanisms of Gp. lemaneiformis

Comparative proteomic analysis of Ulva prolifera response to high temperature stress

Abstract Background Ulva prolifera belongs to green macroalgae and is the dominant species of green tide. It is distributed worldwide and is therefore subject to high-temperature stress during the growth process. However, the adaptation mechanisms of the response of U. prolifera to high temperatures have not been clearly investigated yet. Methods In this study, isobaric tags for relative and absolute quantitation (iTRAQ) labelling was applied in combination with the liquid chromatography-tandem mass spectrometry (LC-MS/MS) to conduct comparative proteomic analysis of the response of U. prolifera to high-temperature stress and to elucidate the involvement of this response in adaptation mechanisms. Differentially expressed proteins (DEPs) of U. prolifera under high temperature (denote UpHT) compared with the control (UpC) were identified. Bioinformatic analyses including GO analysis, pathway analysis, and pathway enrichment analysis was performed to analyse the key metabolic pathways that underlie the thermal tolerance mechanism through protein networks. Quantitative real-time PCR and western blot were performed to validate selected proteins. Results In the present study, 1223 DEPs were identified under high temperature compared with the control, which included 790 up-regulated and 433 down-regulated proteins. The high-temperature stimulus mainly induced the expression of glutathione S-transferase, heat shock protein, ascorbate peroxidase, manganese superoxide dismutase, ubiquitin-related protein, lhcSR, rubisco activase, serine/threonine protein kinase 2, adenylate kinase, Ca2+-dependent protein kinase (CDPK), disease resistance protein EDS1, metacaspase type II, NDPK2a, 26S proteasome regulatory subunit, ubiquinone oxidoreductase, ATP synthase subunit, SnRK2s, and cytochrome P450. The down-regulated proteins were photosynthesis-related proteins, glutathione reductase, catalase-peroxidase, thioredoxin, thioredoxin peroxidase, PP2C, and carbon fixation-related proteins. Furthermore, biological index analysis indicated that protein content and SOD activity decreased; the value of Fv/Fm dropped to the lowest point after culture for 96 h. However, APX activity and MDA content increased under high temperature. Conclusion The present study implied an increase in proteins that were associated with the stress response, oxidative phosphorylation, the cytokinin signal transduction pathway, the abscisic acid signal transduction pathway, and the glutathione metabolism pathway. Proteins that were associated with photosynthesis, carbon fixation in photosynthesis organisms, and the photosynthesis antenna protein pathway were decreased. These pathways played a pivotal role in high temperature regulation. These novel proteins provide a good starting point for further research into their functions using genetic or other approaches. These findings significantly improve the understanding of the molecular mechanisms involved in the tolerance of algae to high-temperature stress

MicroRNA and Putative Target Discoveries in Chrysanthemum Polyploidy Breeding

MicroRNAs (miRNAs), around 22 nucleotides (nt) in length, are a class of endogenous and noncoding RNA molecule that play an essential role in plant development, either by suppressing the transcription of target genes at a transcriptional level or inhibiting translation at a posttranscriptional level. To understand the roles of miRNAs and their target genes in chrysanthemum polyploidy breeding, three sRNA libraries of normal and abnormal embryos after hybridization were performed by RNA-Seq. As a result, a total of 170 miRNAs were identified and there are 41 special miRNAs in cross of paternal chromosome doubling, such as miR169b, miR440, and miR528-5p. miR164c and miR159a were highly expressed in a normal embryo at 18 days after pollination, suggesting the regulatory role at the late stage of embryonic development. miR172c was only detected in the normal embryo at 18 days after pollination, which means that miR172c mainly mediates gene expression in postembryonic development and these genes may promote embryo maturation. Other miRNAs, including miR414, miR2661, and miR5021, may regulate the genes participated in pathways of auxin response and energy metabolism; then they regulate the complex embryonic development together

Enzymatic Degradation of Gracilariopsis lemaneiformis Polysaccharide and the Antioxidant Activity of Its Degradation Products

Gracilariopsis lemaneiformis polysaccharides (GLP) were degraded using pectinase, glucoamylase, cellulase, xylanase, and β-dextranase into low-molecular-weight polysaccharides, namely, GPP, GGP, GCP, GXP, and GDP, respectively, and their antioxidant capacities were investigated. The degraded GLP showed higher antioxidant activities than natural GLP, and GDP exhibited the highest antioxidant activity. After the optimization of degradation conditions through single-factor and orthogonal optimization experiments, four polysaccharide fractions (GDP1, GDP2, GDP3, and GDP4) with high antioxidant abilities (hydroxyl radical scavenging activity, DPPH radical scavenging activity, reduction capacity, and total antioxidant capacity) were obtained. Their cytoprotective activities against H2O2-induced oxidative damage in human fetal lung fibroblast 1 (HFL1) cells were examined. Results suggested that GDP pretreatment can significantly improve cell viability, reduce reactive oxygen species and malonaldehyde levels, improve antioxidant enzyme activity and mitochondria membrane potential, and alleviate oxidative damage in HFL1 cells. Thus, the enzyme degradation of GLP with β-dextranase can significantly improve its antioxidant activity, and GDP might be a suitable source of natural antioxidants