Exploring the physio-biochemical and metabolic shifts in a green alga, Caulerpa lentillifera (J.Agardh) in response to elevated pCO2 level

Objective:Algae contain many symbiotic bacteria, often considered pollutants in algal cultivation. Recent studies indicate that these connections enhance the longevity of both organisms. Researching the bioactive metabolites of marine bacteria has emerged as a promising strategy for drug discovery. Green algae, including Caulerpa lentillifera, have anticancer activity and possess antioxidant qualities. The research emphasizes the isolation and identification of beneficial proteins from symbiotic bacteria, particularly Caulerpa lentillifera. Methods:The stages of the research included isolation and identification of the endophytic bacteria of the green alga symbiont C. lentillifera, isolation of protein from the bacterial symbiont, fractionation, hydrolysis, ultrafiltration of protein into peptides, and testing for activity. Screening activities used the BSLT to obtain the value of LC50, and the mitotic test of the sea urchin zygote cell Tripneustes gratilla Linn to find the value of IC50. Results: The results of this study indicated that the bacterial symbiont of the algae C. lentillifera was a species of Cobetia marina strain CL2-2. The peptide with molecular weight < 3 kDa from Cobetia marina strain CL2-2 was active. The peptide was from protein deposited with 40-60% saturated ammonium sulfate and hydrolyzed using pepsin enzyme. BSLT toxicity tests indicate that peptides with a molecular weight of less than 3 kDa showed significant toxicity, indicated by an LC50 value of 4.061 ppm. In a mitotic cytotoxicity test involving sea urchin zygote cells, peptides with a molecular weight of less than 3 kDa indicated significant cytotoxic activity, resulting in an IC50 value of 7.236 μg/mL. Conclusion:The bioactive peptide with molecular weight of less than 3 kDa resulting from protein hydrolysis isolated from the green algae symbiont C. lentillifera has the potential as an anticancer agent.

Caulerpa lentillifera (CL), also called sea grape, is a type of edible green alga which was reported to have antioxidative and immunomodulatory potential. This study aimed to investigate the hepatoprotective effects of CL in a rat model of chronic ethanol exposure. Wistar rats were assigned to four groups and supplied with an isocaloric control liquid diet (group C), an ethanol liquid diet (group E), a control liquid diet supplemented with 5% CL (group CC), or an ethanol liquid diet supplemented with 5% CL (group EC) for a 12-week experimental period. Ethanol feeding induced steatosis, inflammation, and changes in the gut microbiota by the end of the study, whereas CL supplementation significantly improved liver injuries and decreased circulatory endotoxin levels. Moreover, we also found that CL reversed ethanol-induced elevation of hepatic toll-like receptor 4 (TLR4), MyD88 protein expression, the phosphorylated-nuclear factor (NF)-κB-to–NF–κB ratio, and proinflammatory cytokine concentrations. Additionally, CL also increased the abundance of Akkermansia and tight junction proteins and diminished the Firmicutes-to-Bacteroidetes ratio. Dietary CL inhibited the progression of alcoholic liver disease, and some of the possible mechanisms may be strengthening the intestinal barrier function, alleviating dysbiosis, and modulating the TLR4 pathway.

The edible seaweed Caulerpa lentillifera, a powerful natural food source that is rich in protein, minerals, dietary fibers, vitamins, saturated fatty acids and unsaturated fatty acids, has been mass cultured in some Asian countries and has been the focus of researchers in recent years. Here, the operational conditions of its culture, application in wastewater treatment, and bioactive components are summarized and comparatively analyzed. Based on previous studies, salinity, nutrient concentrations, irradiance and temperature are stress factors for algal growth. Moreover, dried Caulerpa lentillifera seaweed is efficient in the biosorption of heavy metals and cationic dyes in wastewater, and fresh seaweed can be introduced as a biofilter in aquaculture system treatment. In addition, among the rich bioactive compounds in Caulerpa lentillifera, the phenolic compounds show the potential ability for regulating glucose metabolism in vivo. Polysaccharides and oligosaccharides exhibit anticoagulant, immunomodulatory effects and cancer-preventing activity. Siphonaxanthin is a compound with attractive novel functions in cancer-preventing activity and lipogenesis-inhibiting effects. Furthermore, the antioxidant activity of siphonaxanthin extracted from Caulerpa lentillifera could be stronger than that of astaxanthin. This review offers an overview of studies of Caulerpa lentillifera addressing various aspects including cultivation, wastewater treatment and biological active components which may provide valuable information for the cultivation and utilization of this green alga.

The edible seaweed Caulerpa lentillifera, a powerful natural food source that is rich in protein, minerals, dietary fibers, vitamins, saturated fatty acids and unsaturated fatty acids, has been mass cultured in some Asian countries and has been the focus of researchers in recent years. Here, the operational conditions of its culture, application in wastewater treatment, and bioactive components are summarized and comparatively analyzed. Based on previous studies, salinity, nutrient concentrations, irradiance and temperature are stress factors for algal growth. Moreover, dried Caulerpa lentillifera seaweed is efficient in the biosorption of heavy metals and cationic dyes in wastewater, and fresh seaweed can be introduced as a biofilter in aquaculture system treatment. In addition, among the rich bioactive compounds in Caulerpa lentillifera, the phenolic compounds show the potential ability for regulating glucose metabolism in vivo. Polysaccharides and oligosaccharides exhibit anticoagulant, immunomodulatory effects and cancer-preventing activity. Siphonaxanthin is a compound with attractive novel functions in cancer-preventing activity and lipogenesis-inhibiting effects. Furthermore, the antioxidant activity of siphonaxanthin extracted from Caulerpa lentillifera could be stronger than that of astaxanthin. This review offers an overview of studies of Caulerpa lentillifera addressing various aspects including cultivation, wastewater treatment and biological active components which may provide valuable information for the cultivation and utilization of this green alga.

Several species of Caulerpa have been investigated and bioactive principles, such as caulerpin and caulerpicine have been isolated from a number of them. Caulerpin produces mild anesthetic action, difficulty in breathing, sedation, and loss of balance. The toxic syndrome has been reported to be somewhat similar to that produced by ciguatera fish poisoning. The neurotrophic activity of caulerpicine is thought to be of clinical value. Tropical green algae and a few of their temperate relatives have yielded a number of bioactive metabolites and some of these are believed to be used by the algae as a chemical defense against herbivorous animals. This research was aimed to determine the secondary metabolites compound of some varieties of Caulerpa species. Chromatography and Spectrophotometric methods were used to conduct this research. The result shows that Caulerpin and caulerpicin presence on five species of Caulerpa species Caulerpa racemosa Siquijor, Caulerpa lentillifera wild and farm from Mactan, Caulerpa clavifera Mactan and Caulerpa lentillifera cultured in the common garden. Higher absorbance of both compounds caulerpin and caulerpicin found in wild Caulerpa lentillifera from Mactan while the lowest absorbance of caulerpin found in Caulerpa racemosa collected from Siquijor, whereas the lowest absorbance of caulerpicin found in the cultured Caulerpa lentillifera in the common garden.

β-1,3-Xylan was prepared from the green alga, Caulerpa lentillifera, and hydrolyzed to oligosaccharides by a mild acid treatment. The average degree of polymerization was about 5. The oligosaccharides reduced the number of viable human breast cancer MCF-7 cells in a dose-dependent manner, and induced chromatin condensation and degradation of poly ADP-ribose polymerase, indicating that they induced apoptosis in MCF-7 cells.

The green alga, Caulerpa lentillifera, is composed of a single cell with multiple nuclei, but it possesses structures analogous to leaves or fronds, stems or stolons, and roots or rhizoids. To understand molecular mechanisms involved in formation and function of these structures, we carried out RNA-seq analysis of fronds and stolons (including rhizoids). Taking advantage of the decoded genome of C. lentillifera, the present RNA-seq analysis addressed transcripts corresponding to 9,311 genes identified in the genome. RNA-seq data suggested that 8,734 genes are expressed in sporophytes. Despite the siphonous body of the alga, differential gene expression was evident in the two structures. 1,027 (11.8%) and 1,129 (12.9%) genes were preferentially expressed in fronds and stolons, respectively, while the remaining 6,578 (75.3%) genes were expressed at the same level in both. Most genes preferentially expressed in fronds are associated with photosynthesis and plant hormone pathways, including abscisic acid signaling. In contrast, those preferentially expressed in stolons are associated with translation and DNA replication. These results indicate that gene expression is regulated differently between fronds and stolons, which probably governs the function of each structure. Together with genomic information, the present transcriptomic data provide genic information about development and physiology of this unique, siphonous organism.

Caulerpa lentillifera is a green algae that distributes worldwide and is cultivated for food. We assessed vegetative propagation of C. lentillifera by measuring the specific growth rate (SGR) and chlorophyll fluorescence of the green algae cultured at different salinities and nutrient levels. The results indicated that C. lentillifera can survive in salinities ranging from 20 to 50, and can develop at salinities of 30 to 40. The maximum SGR for C. lentillifera occurred at a salinity of 35. Both chlorophyll content and the ratio of variable to maximum fluorescence (Fv/Fm) were also at a maximum at a salinity of 35. Photosynthesis was inhibited in salinities greater than 45 and less than 25. Both the maximum SGR and maximum chlorophyll content were found in algae treated with a concentration of 0.5 mmol/L of NO3-N and 0.1 mmol/L of PO4-P. The photosynthetic capacity of photosystem II (PSII) was inhibited in cultures of C. lentillifera at high nutrient levels. This occurred when NO3-N concentrations were greater than 1.0 mmol/L and when PO4-P concentrations were at 0.4 mmol/L. As there is strong need for large-scale cultivation of C. lentillifera, these data contribute important information to ensure optimal results.

Caulerpa lentillifera is a green alga that contains essential nutrients. This study investigated the effects of autoclave- and microwave-assisted C. lentillifera extracts (A-CLE and M-CLE) on the growth, reproduction, immunity, and oxidative stress in Daphnia magna. Both extracts were supplemented to D. magna at 1 to 5 g/L. The results showed that D. magna treated with 2 to 5 g/L of A-CLE produced juveniles faster than the control. Juveniles at 1 to 4 g/L A-CLE treatments increased. Moreover, juveniles in 1 g/L M-CLE treatment significantly increased, but the hatching time was not affected. A-CLE also stimulated the immunity of D. magna by increasing hemocytes in the 3 g/L treatment. Furthermore, both extracts did not affect the growth rate, phenoloxidase activity, malondialdehyde level, and catalase activity in D. magna. Therefore, our results indicated that both extracts could enhance the reproduction and immunity of D. magna. Our study is useful for the health improvement of D. magna.

Genome evolution and development of unicellular, multinucleate macroalgae (siphonous algae) are poorly known, although various multicellular organisms have been studied extensively. To understand macroalgal developmental evolution, we assembled the ∼26 Mb genome of a siphonous green alga, Caulerpa lentillifera, with high contiguity, containing 9,311 protein-coding genes. Molecular phylogeny using 107 nuclear genes indicates that the diversification of the class Ulvophyceae, including C. lentillifera, occurred before the split of the Chlorophyceae and Trebouxiophyceae. Compared with other green algae, the TALE superclass of homeobox genes, which expanded in land plants, shows a series of lineage-specific duplications in this siphonous macroalga. Plant hormone signalling components were also expanded in a lineage-specific manner. Expanded transport regulators, which show spatially different expression, suggest that the structural patterning strategy of a multinucleate cell depends on diversification of nuclear pore proteins. These results not only imply functional convergence of duplicated genes among green plants, but also provide insight into evolutionary roots of green plants. Based on the present results, we propose cellular and molecular mechanisms involved in the structural differentiation in the siphonous alga.

The increasing prevalence of obesity and its related diseases, including diabetes mellitus and metabolic dysfunction-associated fatty liver disease, has become a significant social problem. These diseases are believed to be preventable through healthy diet and exercise habits, and the investigation of food ingredients that are useful for prevention of these diseases is actively ongoing. Carotenoids are the major lipophilic pigments responsible for yellow-to-red colors in our diet, and the ingestion of certain carotenoids has been reported to prevent obesity. For example, β-carotene suppresses adipogenic differentiation of mouse preadipocyte line 3T3-L1 through its provitamin A activity. Fucoxanthin, a carotenoid found in brown algae, also has the similar effect via a different mechanism and is used as an active ingredient in foods with functional claims in Japan. In contrast, siphonaxanthin, a carotenoid found in some green algae such as Caulerpa lentillifera (commonly known as sea grape), exhibited stronger biological activities than other carotenoids in cell-based studies; it significantly suppressed adipogenic differentiation of 3T3-L1 cells even at low concentrations where β-carotene and fucoxanthin did not show inhibitory effects. However, its practical applications have not yet been realized. This review summarizes the studies on the anti-obesity effects of carotenoids and discusses the potential of siphonaxanthin as a novel functional food ingredient.Graphical

The seaweed Caulerpa lentillifera is commonly utilized as food for human in many Asian countries. It is an efficient bio-filter, despite its ability to absorb pollutants from the habitat of sources.This study determine the influence of habitat on the quality characteristics of the green macro alga, Caulerpa lentillifera, taken from the different sources. Purposive sampling was used with samples taken from Bohol and Cebu islands, throughout the three (3) trials. Samples from Bohol islands were obtained from the culture ponds; while samples from Cebu islands were taken from both the culture ponds and from the wild. The sea-grapes were analyzed for the presence of heavy metals, lethality assay, bacterial count, direct pollution indicators and proximate composition. Data obtained were treated statistically using ANOVA at 5percent level of significance. Results of the study showed significant difference on heavy metals (Hg and Pb) from various seaweed sources. High concentrations of heavy metals were found in all the seaweed samples from Cebu compared with that of the Bohol samples. In terms of lethality assay (LC 50), the wild samples from Cebu significantly showed higher lethal values compared with that obtained from Bohol samples. As to the bacterial count (cfu/g), the wild samples from Cebu showed the lowest Aerobic Plate Count of 9.3 x 103 cfu/g. However, higher bacterial counts were obtained from Cebu samples that were taken from the two culture ponds co-cultured with milkfish. Across sites, the proximate composition of all the sea-grape samples from Cebu and Bohol also showed significant difference. In terms of E. coli, all the samples have shown no significant difference, with &lt;1.0 x 10 cfu.g-1. Regarding the indicators of direct pollution, all the samples from both islands showed low levels of direct pollutants, such as PO4 , NH3, and NO3 . Based on the results of this study, sea-grapes showed varying quality characteristics as influenced by the habitat or sources they were obtained. As a bio-filter, the quality characteristics of the seaweed are indicators of the various pollutants found from the habitat or sorces.

Disruption of macrophyte-induced algal defence by a cyanobacterial competitor through metabolic shifts and microbiome dysbiosis.

Marine algae have excellent bioresource properties with potential nutritional and bioactive therapeutic benefits, but studies regarding Caulerpa lentillifera are limited. This study aims to explore the metabolites profile and the antioxidant, anticancer, anti-obesity, and in vitro cytotoxicity properties of fractionated ethanolic extract of C. lentillifera using two maceration and soxhlet extraction methods. Dried simplicia of C. lentillifera was mashed and extracted in ethanol solvent, concentrated and evaporated, then sequentially partitioned with equal volumes of ethyl acetate and n-Hexane. Six samples were used in this study, consisting of ME (Maceration—Ethanol), MEA (Maceration—Ethyl Acetate), MH (Maceration—n-Hexane), SE (Soxhletation—Ethanol), SEA (Soxhletation—Ethyl Acetate), and SH (Soxhletation—n-Hexane). Non-targeted metabolomic profiling was determined using LC-HRMS, while antioxidant, anti-obesity, and anticancer cytotoxicity were determined using DPPH and ABTS, lipase inhibition, and MTT assay, respectively. This study demonstrates that C. lentillifera has several functional metabolites, antioxidant capacity (EC50 MH is very close to EC50 of Trolox), as well as anti-obesity properties (EC50 MH < EC50 orlistat, an inhibitor of lipid hydrolyzing enzymes), which are useful as precursors for new therapeutic approaches in improving obesity-related diseases. More interestingly, ME, MH, and SE are novel bioresource agents for anticancer drugs, especially for hepatoma, breast, colorectal, and leukemia cancers. Finally, C. lentillifera can be a nutraceutical with great therapeutic benefits.

Caulerpa lentillifera is a marine nutrient-rich edible green algae, with its external shape similar to ‘sea grape’, it has functions of purifying blood, anti-oxidation, anti-cancer, and anti-tumor. The mitogenome sequence of C. lentillifera is 209,894 bp long. A total of 67 genes were determined, including 17 protein-encoding genes, 3 rRNA genes, 27 tRNA genes, and 20 unidentified open reading frame (ORF). Phylogenetic analysis showed that C. lentillifera clustered together into a single branch. The mitogenome analysis will help the understanding of Ulvophyceae evolution.