Denovo production of resveratrol by engineered Saccharomyces cerevisiae W303-1a using pretreated Gracilaria corticata extracts.

OBJECTIVE: Assembly and construction of resveratrol production pathway in Saccharomyces cerevisiae for denovo production of resveratrol using seaweed extract as fermentation medium. RESULTS: Genes involved in the production of resveratrol from tyrosine pathway, tyrosine ammonia lyase (FTAL) gene from Flavobacterium johnsoniae (FjTAL), the 4-coumarate:CoA ligase gene from Arabidopsis thaliana (4CL1) and the stilbene synthase gene from Vitis vinifera (VvSTS) were introduced into low copy, high copy and integrative vector and transformed into S. cerevisiae W303-1a. The resulting strains W303-1a/pARS-res5, W303-1a/2µ-res1 and W303-1a/IntUra-res9 produced a level of 2.39 ± 0.01, 3.33 ± 0.03 and 8.34 ± 0.03 mg resveratrol l-1 respectively. CRISPR mediated integration at the δ locus resulted in 17.13 ± 1.1 mg resveratrol l-1. Gracilaria corticata extract was tested as a substrate for the growth of transformant to produce resveratrol. The strain produced a comparable level, 13.6 ± 0.54 mg resveratrol l-1 when grown in seaweed extract medium. CONCLUSIONS: The strain W303-1a/IntδC-res1 utilized Gracillaria hydrolysate and produced 13.6 ± 0.54 mg resveratrol l-1 and further investigations are being carried out focusing on pathway engineering and optimization of process parameters to enhance resveratrol yield.

Edible-algae base composite film containing gelatin for food packaging from macroalgae, Gracilaroid (Gracilaria fisheri).

BACKGROUND: Conventional petroleum-based packaging films cause severe environmental problems. In the present study, bio-edible film was introduced as being safe to replace petroleum-based polymers. A food application for edible sachets and a composite edible film (EF) from marine algae, Gracilaria fisheri (GF) extract, were proposed. RESULTS: Carbohydrates were the most prevalent component in fresh GF fronds. Under neutral conditions comprising 90 °C for 40 min, the structure of the extract was determined by Fourier transform infrared to be a carrageenan-like polysaccharide. Glycerol was the best plasticizer for EF formation because it had the highest tensile strength (TS). The integration of gelatin into the algal composite film with gelatin (CFG) was validated to be significant. The best casting temperatures for 2 h were 70 and 100 °C among the four tested temperatures (25, 60, 70 and 100 °C). Temperatures did not result in any significant (P ≤ 0.05) differences in any character (color values, TS, water vapor permeability, oxygen transmission, thickness and water activity), except elongation at break. Visually, the CFG had a slightly yellow appearance. The best-to-worst order of film stability in the three tested solvents was oil, distilled water (DW) and ethanol. Its stability in ethanol (0-100%), temperature of DW (30-100 °C) and pH (3-7 in DW) demonstrated inverse relationships with the concentration or different conditions, except for pH 8-10 in DW. All treatments were significantly (P ≤ 0.05) different. CONCLUSION: The novel material made from polysaccharides from algae, G. fisheri, was used to improve EF. The edible sachet application is plausible from the EF. © 2024 Society of Chemical Industry.

Sexual selection in seaweed? Testing Bateman's principles in the red alga Gracilaria gracilis.

In anisogamous species, sexual selection is expected to be stronger in males. Bateman's principles state that the variance in (i) reproductive and (ii) mating success is greater for males, and (iii) the relationship between reproductive success and mating success (the Bateman gradient) is also stronger for males than for females. Sexual selection, based on Bateman's principles, has been demonstrated in animals and some angiosperms, but never in a seaweed. Here we focus on the oogamous haploid-diploid rhodophyte Gracilaria gracilis in which previous studies have shown evidence for non-random mating, suggesting the existence of male-male competition and female choice. We estimated mating and reproductive success using paternity analyses in a natural population where up to 92% of fertilizations occurred between partners of that population. The results show that the variance in mating success is significantly greater in males than in females and that the Bateman gradient is positive only in males. Distance to female partners also explains a minor part of the variance in male mating success. Although there is no evidence for sexual dimorphism, our study supports the hypothesis that sexual selection occurs in G. gracilis, probably on male traits, even if we cannot observe, characterize or quantify them yet.

Genome-scale signatures of adaptive gene expression changes in an invasive seaweed Gracilaria vermiculophylla.

Invasive species can successfully and rapidly colonize new niches and expand ranges via founder effects and enhanced tolerance towards environmental stresses. However, the underpinning molecular mechanisms (i.e., gene expression changes) facilitating rapid adaptation to harsh environments are still poorly understood. The red seaweed Gracilaria vermiculophylla, which is native to the northwest Pacific but invaded North American and European coastal habitats over the last 100 years, provides an excellent model to examine whether enhanced tolerance at the level of gene expression contributed to its invasion success. We collected G. vermiculophylla from its native range in Japan and from two non-native regions along the Delmarva Peninsula (Eastern United States) and in Germany. Thalli were reared in a common garden for 4 months at which time we performed comparative transcriptome (mRNA) and microRNA (miRNA) sequencing. MRNA-expression profiling identified 59 genes that were differently expressed between native and non-native thalli. Of these genes, most were involved in metabolic pathways, including photosynthesis, abiotic stress, and biosynthesis of products and hormones in all four non-native sites. MiRNA-based target-gene correlation analysis in native/non-native pairs revealed that some target genes are positively or negatively regulated via epigenetic mechanisms. Importantly, these genes are mostly associated with metabolism and defence capability (e.g., metal transporter Nramp5, senescence-associated protein, cell wall-associated hydrolase, ycf68 protein and cytochrome P450-like TBP). Thus, our gene expression results indicate that resource reallocation to metabolic processes is most likely a predominant mechanism contributing to the range-wide persistence and adaptation of G. vermiculophylla in the invaded range. This study, therefore, provides molecular insight into the speed and nature of invasion-mediated rapid adaption.

Gracilaria salicornia as potential substratum for green synthesis of Cerium Oxide Nanoparticles coupled hydrogel: An effective antimicrobial thin film.

Sodium alginate based (SA) hydrogel supplemented Cerium Oxide nanoparticles (CeO2NPs) was produced to fabricate an antimicrobial thin film using an aqueous extract of G. salicornia (Gs). The Gs-CeO2NPs were characterized via SEM, FT-IR, EDX, XRD and DLS, the particle size was 200 nm, agreed with XRD. Gs-SA powder was extracted and incorporated with CeO2NPs. The Gs-SA and its composite thin film (Gs-CeO2NPs-SATF) were characterized including viscosity, FT-IR, TGA, and SEM. The adhesion of Gs-SA coating around Gs-CeO2NPs confirmed via FTIR. The antimicrobial properties of Gs-CeO2NPs and CeO2NPs-SATF were proved in MICs for E. coli and Candida albicans at 62.5 and 250.0 µg/mL. The biofilm inhibition efficiency of CeO2NPs-SATF was 74.67 ± 0.98% and 65.45 ± 0.40% for E. coli and Candida albicans. The CeO2NPs-SATF was polydisperse in nature and film structure gets fluctuated with NPs concentration. Increased NPs into SATF enhances pore size of gel and corroborated with viscous behaviour. The cytotoxicity of Gs-CeO2NP-SA in Artemia salina at higher concentration 100 µg/mL provides less lethal effect into the adult. The antioxidant activity of Gs-CeO2NP-SA in DPPH assay was noticed at 0.6 mg ml-1 with radical scavenging activity at 65.85 ± 0.81%. Thus the Gs-CeO2NP-SATF would be suitable in antimicrobial applications.

Effects of dietary supplementation of Gracilaria lemaneiformis-derived sulfated polysaccharides on the growth, antioxidant capacity, and innate immunity of rabbitfish (Siganus canaliculatus).

The dietary supplementation of red seaweed-derived polysaccharides has been shown to be beneficial to fish and shellfish aquaculture. However, the function of red seaweed (Gracilaria lemaneiformis)-extracted polysaccharide (GLP) on the health status of rabbitfish (Siganus canaliculatus) is still unknown. This study explored the influences of GLP on growth performance, antioxidant activity, and immunity of rabbitfish. Herein, the fish were fed commercial pelleted feed incorporated with the diverse amount of GLP: 0 (control), 0.10 (GLP0.10), and 0.15 g kg-1 (GLP0.15) for 60 days. The results demonstrated that dietary GLP0.15 significantly elevated FBW and WG, while feed utilization efficiency improved (reduced feed conversion ratio and increased protein efficiency ratio) upon GLP0.10 treatment, regarding the control (P < 0.05). Also, dietary administration of GLP0.15 suggestively improved the serum acid phosphatase and lysozyme activity as well as hepatic total antioxidant capacity, catalase, and superoxide dismutase activity. In contrast, GLP0.15decreased the serum alkaline phosphatase, aspartate aminotransferase, alanine aminotransferase, and malonaldehyde activity when compared to the control (P<0.05). Moreover, the lipase (36.08 and 16.46 U/mgprot in GLP0.10 and GLP0.15, respectively) and amylase (0.43 and 0.23 U/mgprot in GLP0.10 and GLP0.15, respectively) activity recorded the maximum values than the control (8.61 and 0.13 U/mgprot, respectively).Further, the intestinal morphometry was developed (such as increased villus length, width, and area) in the fish fed with a GLP-supplemented diet compared to the control. The KEGG pathway analysis unveiled that several differentially expressed genes (DEGs) in control vs. GLP0.10 and control vs. GLP0.15 were associated with metabolic or immune-associated pathways like antigen processing and presentation, phagosome, complement and coagulation cascades, and platelet activation. The DEGs, namely C3, f5, fgb, MHC1, and cfb, were evaluated in control vs. GLP0.10 and C3 and MHC1 in control vs. GLP0.15, suggesting their possible contributions to GLP-regulated immunity. Additionally, the cumulative mortality of rabbitfish after the Vibrio parahaemolyticus challenge was lower in both GLP0.10 (8.88%) and GLP0.15 (11.11%) than in control (33.33%) (P<0.05). Thus, these findings direct the potential use of GLP as an immunostimulant and growth promoter in rabbitfish aquaculture.

In vitro investigation of silver nanoparticles synthesized using Gracilaria veruccosa - A seaweed against multidrug resistant Staphylococcusaureus.

In recent years, the biosynthesis of silver (Ag) nanoparticles has attracted a great deal of interest for applications in biomedicine and bioremediation. In the present study, Gracilaria veruccosa extract was used to synthesize Ag nanoparticles for investigating their antibacterial and antibiofilm potentials. The color shift from olive green to brown indicated the synthesis of AgNPs by plasma resonance at 411 nm. Physical and chemical characterization revealed that AgNPs of 20-25 nm sizes were synthesized. Detecting functional groups, such as carboxylic acids and alkenes, suggested that the bioactive molecules in the G. veruccosa extract assisted the synthesis of AgNPs. X-ray diffraction verified the s purity and crystallinity of the AgNPs with an average diameter of 25 nm, while DLS analysis showed a negative surface charge of -22.5 mV. Moreover, AgNPs were tested in vitro for antibacterial and antibiofilm efficacies against S. aureus. The minimum inhibitory concentration (MIC) of AgNPs against S. aureus was 3.8 µg/mL. Light and fluorescence microscopy proved the potential of AgNPs to disrupt the mature biofilm of S. aureus. Therefore, the present report has deciphered the potential of G. veruccosafor the synthesis of AgNPs and targeted the pathogenic bacteria S. aureus.

A Blend Consisting of Agaran from Seaweed Gracilaria birdiae and Chromium Picolinate Is a Better Antioxidant Agent than These Two Compounds Alone.

A blend refers to the combination of two or more components to achieve properties that are superior to those found in the individual products used for their production. Gracilaria birdiae agaran (SPGb) and chromium picolinate (ChrPic) are both antioxidant agents. However, there is no documentation of blends that incorporate agarans and ChrPic. Hence, the objective of this study was to generate blends containing SPGb and ChrPic that exhibit enhanced antioxidant activity compared to SPGb or ChrPic alone. ChrPic was commercially acquired, while SPGb was extracted from the seaweed. Five blends (B1; B2; B3; B4; B5) were produced, and tests indicated B5 as the best antioxidant blend. B5 was not cytotoxic or genotoxic. H2O2 (0.6 mM) induced toxicity in fibroblasts (3T3), and this effect was abolished by B5 (0.05 mg·mL-1); neither ChrPic nor SPGb showed this effect. The cells also showed no signs of toxicity when exposed to H2O2 after being incubated with B5 and ChrPic for 24 h. In another experiment, cells were incubated with H2O2 and later exposed to SPGb, ChrPic, or B5. Again, SPGb was not effective, while cells exposed to ChrPic and B5 reduced MTT by 100%. The data demonstrated that B5 has activity superior to SPGb and ChrPic and points to B5 as a product to be used in future in vivo tests to confirm its antioxidant action. It may also be indicated as a possible nutraceutical agent.

Study of the Phytochemical Composition, Antioxidant Properties, and In Vitro Anti-Diabetic Efficacy of Gracilaria bursa-pastoris Extracts.

In this study, a comparison was made of the chemical makeup of different extracts obtained from Gracilaria bursa-pastoris, a type of red seaweed that was gathered from the Nador lagoon situated in the northern part of Morocco. Additionally, their anti-diabetic and antioxidant properties were investigated. The application of GC-MS technology to analyze the fatty acid content of the samples revealed that linoleic acid and eicosenoic acid were the most abundant unsaturated fatty acids across all samples, with palmitic acid and oleic acid following in frequency. The HPLC analysis indicated that ascorbic and kojic acids were the most prevalent phenolic compounds, while apigenin was the most common flavonoid molecule. The aqueous extract exhibited significant levels of polyphenols and flavonoids, registering values of 381.31 ± 0.33 mg GAE/g and 201.80 ± 0.21 mg QE/g, respectively. Furthermore, this particular extract demonstrated a remarkable ability to scavenge DPPH radicals, as evidenced by its IC50 value of 0.17 ± 0.67 mg/mL. In addition, the methanolic extract was found to possess antioxidant properties, as evidenced by its ability to prevent ß-carotene discoloration, with an IC50 ranging from 0.062 ± 0.02 mg/mL to 0.070 ± 0.06 mg/mL. In vitro study showed that all extracts significantly inhibited the enzymatic activity of α-amylase and α-glucosidase. Finally, molecular docking models were applied to assess the interaction between the primary phytochemicals identified in G. bursa-pastoris extracts and the human pancreatic α-amylase and α-glucosidase enzymes. The findings suggest that these extracts contain bioactive substances capable of reducing enzyme activity more effectively than the commercially available drug acarbose.

A Phytoprostane from Gracilaria longissima Increases Platelet Activation, Platelet Adhesion to Leukocytes and Endothelial Cell Migration by Potential Binding to EP3 Prostaglandin Receptor.

Plant phytoprostanes (PhytoPs) are lipid oxidative stress mediators that share structural similarities with mammal prostaglandins (PGs). They have been demonstrated to modulate inflammatory processes mediated by prostaglandins. The present study aims to test the effects of the most abundant oxylipin from Gracilaria longissima, ent-9-D1t-Phytoprostane (9-D1t-PhytoP), on platelet activation and vascular cells as well as clarify possible interactions with platelets and the endothelial EP3 receptor Platelet and monocyte activation was assessed by flow cytometry in the presence of purified 9-D1t-PhytoP. Cell migration was studied using the human Ea.hy926 cell line by performing a scratch wound healing assay. The RNA expression of inflammatory markers was evaluated by RT-PCR under inflammatory conditions. Blind docking consensus was applied to the study of the interactions of selected ligands against the EP3 receptor protein. The 9D1t-PhytoP exerts several pharmacological effects; these include prothrombotic and wound-healing properties. In endothelial cells, 9D1t-PhytP mimics the migration stimulus of PGE2. Computational analysis revealed that 9D1t-PhytP forms a stable complex with the hydrophobic pocket of the EP3 receptor by interaction with the same residues as misoprostol and prostaglandin E2 (PGE2), thus supporting its potential as an EP3 agonist. The potential to form procoagulant platelets and the higher endothelial migration rate of the 9-D1t-PhytoP, together with its capability to interact with PGE2 main target receptor in platelets suggest herein that this oxylipin could be a strong candidate for pharmaceutical research from a multitarget perspective.

Chemical Composition and Antioxidant Potential of Five Algae Cultivated in Fully Controlled Closed Systems.

In this study, the chemical composition and antioxidant profile of five edible macroalgae, Fucus vesiculosus, Palmaria palmata, Porphyra dioica, Ulva rigida, and Gracilaria gracilis, cultivated in fully controlled closed systems, were determined. Protein, carbohydrates, and fat contents ranged between 12.4% and 41.8%, 27.6% and 42.0%, and 0.1% and 3.4%, respectively. The tested seaweeds presented considerable amounts of Ca, Mg, K, Mn, and Fe, which reinforce their favorable nutritional profile. Regarding their polysaccharide composition, Gracilaria gracilis and Porphyra dioica were rich in sugars common to agar-producing red algae, and Fucus vesiculosus was composed mainly of uronic acids, mannose, and fucose, characteristic of alginate and fucoidans, whereas rhamnose and uronic acid, characteristic of ulvans, predominated in Ulva rigida. Comparatively, the brown F. vesiculosus clearly stood out, presenting a high polysaccharide content rich in fucoidans, and higher total phenolic content and antioxidant scavenging activity, determined by DPPH and ABTS. The remarkable potential of these marine macroalgae makes them excellent ingredients for a wide range of health, food, and industrial applications.

Pre-domestication bottlenecks of the cultivated seaweed Gracilaria chilensis.

Gracilaria chilensis is the main cultivated seaweed in Chile. The low genetic diversity observed in the Chilean populations has been associated with the over-exploitation of natural beds and/or the founder effect that occurred during post-glacial colonization from New Zealand. How these processes have affected its evolutionary trajectory before farming and incipient domestication is poorly understood. In this study, we used 2232 single nucleotide polymorphisms (SNPs) to assess how the species' evolutionary history in New Zealand (its region of origin), the founder effect linked to transoceanic dispersion and colonization of South America, and the recent over-exploitation of natural populations have influenced the genetic architecture of G. chilensis in Chile. The contrasting patterns of genetic diversity and structure observed between the two main islands in New Zealand attest to the important effects of Quaternary glacial cycles on G. chilensis. Approximate Bayesian Computation (ABC) analyses indicated that Chatham Island and South America were colonized independently near the end of the Last Glacial Maximum and emphasized the importance of coastal and oceanic currents during that period. Furthermore, ABC analyses inferred the existence of a recent and strong genetic bottleneck in Chile, matching the period of over-exploitation of the natural beds during the 1970s, followed by rapid demographic expansion linked to active clonal propagation used in farming. Recurrent genetic bottlenecks strongly eroded the genetic diversity of G. chilensis prior to its cultivation, raising important challenges for the management of genetic resources in this incipiently domesticated species.

Dietary supplementation with Gracilaria gracilis by-products modulates the immune status and oxidative stress response of gilthead seabream (Sparus aurata) stimulated with Photobacterium damselae subsp. piscicida.

This study evaluated the effects of agar waste (AW) dietary supplementation, obtained from the seaweed Gracilaria gracilis cultivated under two different spectral lights, neutral (NT) and blue (BL), on haematological parameters, inflammatory response, and antioxidant biomarkers of gilthead seabream (Sparus aurata). Three diets were prepared: i) a basal diet (CTR), ii) a diet supplemented with 2.5% NT, and iii) a diet supplemented with 2.5% BL. After 15 days of feeding, fish were injected with PBS (placebo) or inactivated Photobacterium damselae subsp. piscicida (stimulated) and sampled at 4 h and 24 h post-stimulus. Results indicated that fish fed NT and BL supplemented diets had lower Ht value and mean corpuscular volume (MCV) than fish fed the CTR diet, regardless of the stimulus and the sampling time. No differences in mean corpuscular haemoglobin (MCH) were found between fish fed the different diets, while the mean corpuscular haemoglobin concentration (MCHC) increased in fish fed AW supplemented diets compared to fish fed the CTR diet, regardless of the stimulus and the sampling time. In response to inflammation, fish fed the NT diet displayed higher neutrophils count in blood when compared to the CTR group, regardless of the stimulus and sampling time. Thrombocyte count was higher in fish fed NT and BL diets than in the CTR group, especially in the stimulated fish (Diet*injection (D*I), P = 0.004). An increase in plasma protease activity was detected in fish fed NT or BL diets in both placebo and stimulated fish regardless of the sampling time. Hepatic catalase activity was higher in fish fed the NT and BL than in the CTR group, particularly in the stimulated fish (D*I, P < 0.001). In addition, both stimulated and placebo fish that received the BL diet showed an increase in hepatic GR activity compared to the CTR group, regardless of the sampling time. Dietary supplementation with AW by-products obtained from G. gracilis cultured under NT and BL conditions showed to improve the inflammatory and antioxidant mechanisms in gilthead seabream in response to a UV-killed bacterial stimulus, having valuable applications for the sustainable use of seaweed toward improving the health and welfare of cultured fish.

Insights into agar and secondary metabolite pathways from the genome of the red alga Gracilaria domingensis (Rhodophyta, Gracilariales).

Gracilariales is a clade of florideophycean red macroalgae known for being the main source of agar. We present a de novo genome assembly and annotation of Gracilaria domingensis, an agarophyte alga with flattened thallus widely distributed along Central and South American Atlantic intertidal zones. In addition to structural analysis, an organizational comparison was done with other Rhodophyta genomes. The nuclear genome has 78 Mbp, with 11,437 predicted coding genes, 4,075 of which did not have hits in sequence databases. We also predicted 1,567 noncoding RNAs, distributed in 14 classes. The plastid and mitochondrion genome structures were also obtained. Genes related to agar synthesis were identified. Genes for type II galactose sulfurylases could not be found. Genes related to ascorbate synthesis were found. These results suggest an intricate connection of cell wall polysaccharide synthesis and the redox systems through the use of L-galactose in Rhodophyta. The genome of G. domingensis should be valuable to phycological and aquacultural research, as it is the first tropical and Western Atlantic red macroalgal genome to be sequenced.

Green synthesis of gold nanoparticles using Gracilaria crassa leaf extract and their ecotoxicological potential: Issues to be considered.

The use of vegetal species for gold nanoparticles (AuNPs) biosynthesis can constitute an alternative to replacing the extensive use of several hazardous chemicals commonly used during NPs synthesis and, therefore, can reduce biological impacts induced by the release of these products into the natural environment. However, the "green nanoparticles" and/or "eco-friendly nanoparticles" label does not ensure that biosynthesized NPs are harmless to non-target organisms. Thus, we aimed to synthesize AuNPs from seaweed Gracilaria crassa aqueous extract through an eco-friendly, fast, one-pot synthetic route. The formation of spherical, stable, polycrystalline NPs with a diameter of 32.0 nm ± 4.0 nm (mean ±SEM) was demonstrated by UV-vis spectroscopy, field emission scanning electron microscopy, and high-resolution transmission electron microscopy, energy-dispersive X-ray and X-ray diffraction measurement, and Fourier-transform infrared spectroscopy analysis. In addition, different phytocomponents were identified in the biosynthesized AuNPs, using Gas Chromatography-Mass Spectrometry (GC-MS). However, both G. crassa aqueous extract and the biosynthesized AuNPs showed high ecotoxicity in Anopheles stephensi larvae exposed to different concentrations. Therefore, our study supports the potential of seaweed G. crassa as a raw material source for AuNPs biosynthesis while also shedding light on its ecotoxicological potential, which necessitates consideration of its risk to aquatic biota.

Depolymerized Fractions of Sulfated Galactans Extracted from Gracilaria fisheri and Their Antibacterial Activity against Vibrio parahaemolyticus and Vibrio harveyi.

Various seaweed sulfated polysaccharides have been explored for antimicrobial application. This study aimed to evaluate the antibacterial activity of the native Gracilaria fisheri sulfated galactans (NSG) and depolymerized fractions against the marine pathogenic bacteria Vibrio parahaemolyticus and Vibrio harveyi. NSG was hydrolyzed in different concentrations of H2O2 to generate sulfated galactans degraded fractions (SGF). The molecular weight, structural characteristics, and physicochemical parameters of both NSG and SGF were determined. The results revealed that the high molecular weight NSG (228.33 kDa) was significantly degraded to SGFs of 115.76, 3.79, and 3.19 kDa by hydrolysis with 0.4, 2, and 10% H2O2, respectively. The Fourier transformed spectroscopy (FTIR) and 1H- and 13C-Nuclear magnetic resonance (NMR) analyses demonstrated that the polysaccharide chain structure of SGFs was not affected by H2O2 degradation, but alterations were detected at the peak positions of some functional groups. In vitro study showed that SGFs significantly exerted a stronger antibacterial activity against V. parahaemolyticus and V. harveyi than NSG, which might be due to the low molecular weight and higher sulfation properties of SGF. SGF disrupted the bacterial cell membrane, resulting in leakage of intracellular biological components, and subsequently, cell death. Taken together, this study provides a basis for the exploitation and utilization of low-molecular-weight sulfated galactans from G. fisheri to prevent and control the shrimp pathogens.

Increased Sulfation in Gracilaria fisheri Sulfated Galactans Enhances Antioxidant and Antiurolithiatic Activities and Protects HK-2 Cell Death Induced by Sodium Oxalate.

Urolithiasis is a common urological disease characterized by the presence of a stone anywhere along the urinary tract. The major component of such stones is calcium oxalate, and reactive oxygen species act as an essential mediator of calcium oxalate crystallization. Previous studies have demonstrated the antioxidant and antiurolithiatic activities of sulfated polysaccharides. In this study, native sulfated galactans (N-SGs) with a molecular weight of 217.4 kDa from Gracilaria fisheri were modified to obtain lower molecular weight SG (L-SG) and also subjected to sulfation SG (S-SG). The in vitro antioxidant and antiurolithiatic activities of the modified substances and their ability to protect against sodium oxalate-induced renal tubular (HK-2) cell death were investigated. The results revealed that S-SG showed more pronounced antioxidant activities (DPPH and O2- scavenging activities) than those of other compounds. S-SG exhibited the highest antiurolithiatic activity in terms of nucleation and aggregation, as well as crystal morphology and size. Moreover, S-SG showed improved cell survival and increased anti-apoptotic BCL-2 protein in HK-2 cells treated with sodium oxalate. Our findings highlight the potential application of S-SG in the functional food and pharmaceutical industries.

Antioxidant Activity of Gracilaria lemaneiformis Polysaccharide Degradation Based on Nrf-2/Keap-1 Signaling Pathway in HepG2 Cells with Oxidative Stress Induced by H2O2.

The objective of this research was to investigate the antioxidant activity of Gracilarialemaneiformis polysaccharide degradation and its underlying mechanism involved in the Nrf-2/Keap-1 signaling pathway in HepG2 cells with oxidative stress induced by H2O2. The result of the scavenging ability of free radicals showed that GLP-HV (polysaccharide degraded by H2O2-vitamin C (Vc)) performed a better scavenging ability than GLP (G.lemaneiformis polysaccharide). Moreover, the scavenging ability of polysaccharide to these free radicals from strong to weak was as follows: superoxide radical, ferric ion, ABTS+, and DPPH radical, and their IC50 values were 3.56 ± 0.0028, 4.97 ± 0.18, 9.62 ± 0.35, and 23.85 ± 1.78 mg/mL, respectively. Furthermore, GLP-HV obviously relieved oxidative stress in HepG2 cells, which strengthened the activity of T-AOC, CAT, GSH-PX, and SOD, and diminished the intensity of MDA, intracellular ROS, and calcium ion based on the Nrf-2/Keap-1 signaling pathway. The PCR result revealed that polysaccharide upregulated the expression of the genes Nrf-2, HO-1, NQO-1, and ZO-1 and downregulated Keap-1. The correlation between chemical properties and antioxidant mechanism of GLP-HV was evaluated via a heat map. The results illustrated that reducing sugar and active groups presented a positive correlation, and molecular weight and viscosity exhibited a negative relation with antioxidant activity.

Mechanism of the allelopathic effect of macroalgae Gracilaria bailiniae on Nitzschia closterium.

With the rapid development of the seaweed industry in China, the scale and production of its commercial seaweed are ranked among the most significant worldwide. Consequently, the control of algal blooms, especially fouling diatoms, during macroalgae industrialisation is an important issue. Many diatom bloom studies have focused on physical and chemical controls, with limited economic and eco-friendly biological controls reported. In our study, Gracilaria bailiniae fresh thalli and aqueous extract profoundly suppressed Nitzschia closterium growth (50% inhibition concentration of the fourth day (IC50-4 day) was 0.667 × 10-3 g·mL-1 and 3.889 × 10-3 g·mL-1, respectively). The cellular morphology changes of N. closterium exposed to the G. bailiniae aqueous extract were severe atrophies and plasmolysis and dissolution of endocellular structures. To explore more potential allelochemicals to control N. closterium, the intracellular compounds of G. bailiniae were detected and screened. Three organic acids (citrate, hydroxyethanesulfonic acid (HA) and taurine) had allelopathic potential against N. closterium. Our results showed that citrate and HA markedly suppressed N. closterium (IC50-4 day: 1.035 mM and 1.151 mM, respectively); however, taurine poorly suppressed N. closterium (IC50-4 day: 2.500 mM). Therefore, HA is one of the main allelopathic compounds in G. bailiniae. Further, the allelopathic mechanism of HA against the N. closterium photosynthetic system broke its photosynthetic apparatus (oxygen-evolving complex, reaction centres, the effective antenna size and the donor side of photosystem II) and hindered electron transport. The experimental results provide a new and eco-friendly strategy to control diatom blooms.

Biomass decomposition and heavy metal release from seaweed litter, Gracilaria lemaneiformis, and secondary pollution evaluation.

The seaweed Gracilaria lemaneiformis can bioremediate heavy metals and improve the environmental quality of mariculture zones. However, the seaweed litter that is produced in the growth and harvest processes becomes one of the important bottlenecks and causes secondary pollution that restricts the development of sustainable seaweed cultivation. Seaweeds exist widely in the coastal areas of the world and are cultivated on a large scale in Asia, but their decomposition process is rarely studied. Experiments that compared decaying dry (dead) and fresh (falling and dying) Gracilaria were conducted to quantify the differences in decomposition rates and heavy metal release in different physiological states. The heavy metals in the seawater and sediment were investigated. The litterbag technique under controlled laboratory conditions was used. The results indicated that the decomposition rates (k) and decay times in 50% (t50%) and 95% (t95%) values varied between dry and fresh Gracilaria. Fresh Gracilaria exhibited a weight loss rate of 15%, and the dry weight loss was 44%. The variations in MAIs (accumulation index of metals) and MR (release rate of metals) between the dry and fresh Gracilaria litters differed significantly, which provides evidence that metals are released back into the environment from Gracilaria litters. The contacted sediments could accelerate the heavy metal release from Gracilaria. Based on our estimates obtained from a 45 d experiment, at least 27.5％ of Cd, 16％ of Cu, 60.1％ of Pb, 72.3％ of Zn, 49.4％ of Fe, 38.6％ of Mn, 68.1％ of Cr, and 67.5％ of Ni present in the fresh Gracilaria and 37.4％ of Cd, 46.2％ of Cu, 77.7％ of Pb, 53.7％ of Zn, 42.7％ of Fe, 67.2％ of Mn, 75.1％ of Cr, and 73.5％ of Ni present in the dried Gracilaria were released back into the water when the biomass was left to decay. This study simulates and underscores that Gracilaria has an strong effect on the heavy metal cycles in marine environments and offers a theoretical basis for the development of sustainable seaweed industries in mariculture zones.