Antioxidative activities of a mycosporine-like amino acid,porphyra-334

Antioxidative activities of porphyra-334, a mycosporine-like amino acid extracted from laver were evaluated. Oxidation of linoleic acid induced by an alkyl-radical 2,2′-azobis (2-amidinopropane) dihydrochloride (AAPH) was successfully suppressed by porphyra-334 (0–200 μM). The simultaneous application of 0.02 μM α-tocopherol and 50 μM porphyra-334 effectively suppressed the AAPH induced oxidation level to approximately 40% of a single application of porphyra-334 after 10 min reaction. Porphyra-334 (0–200 μM) efficiently suppressed the lipid peroxidation induced by singlet oxygen although the antioxidative effect observed was relatively moderate at the initial stage of oxidation. These results suggested that porphyra-334 may function as an antioxidant which influences the storage stability of laver.     

Porphyra-334 Isolated from the Marine Algae Bangia atropurpurea: Conformational Performance for Energy Conversion

Prophyra-334 (p-334) may play a role of energy transfer under an uncertain mechanism, and we speculate the possible model. Via 1D and 2D NMR experiments, it was simulated the correlation between dissociation and conformation of p-334. Intramolecular interactions were observed based on a series of changes in the 1H and 13C chemical shifts. Nuclear Overhauser effect spectroscopy experiments and molecular models in various pD conditions indicated the p-334 molecular dissociation process status. In addition, we also used Chem3D software to find the most possible molecular conformation. The relationship between the structural status and energy conversion is explained. Those are the primary results. More researches on it are highly expected in the future.     

Anti-Inflammation Activities of Mycosporine-Like Amino Acids (MAAs) in Response to UV Radiation Suggest Potential Anti-Skin Aging Activity

Certain photosynthetic marine organisms have evolved mechanisms to counteract UV-radiation by synthesizing UV-absorbing compounds, such as mycosporine-like amino acids (MAAs). In this study, MAAs were separated from the extracts of marine green alga Chlamydomonas hedleyi using HPLC and were identified as porphyra-334, shinorine, and mycosporine-glycine (mycosporine-Gly), based on their retention times and maximum absorption wavelengths. Furthermore, their structures were confirmed by triple quadrupole MS/MS. Their roles as UV-absorbing compounds were investigated in the human fibroblast cell line HaCaT by analyzing the expression levels of genes associated with antioxidant activity, inflammation, and skin aging in response to UV irradiation. The mycosporine-Gly extract, but not the other MAAs, had strong antioxidant activity in the 2,2-diphenyl-1-picryhydrazyl (DPPH) assay. Furthermore, treatment with mycosporine-Gly resulted in a significant decrease in COX-2 mRNA levels, which are typically increased in response to inflammation in the skin, in a concentration-dependent manner. Additionally, in the presence of MAAs, the UV-suppressed genes, procollagen C proteinase enhancer (PCOLCE) and elastin, which are related to skin aging, had increased expression levels equal to those in UV-mock treated cells. Interestingly, the increased expression of involucrin after UV exposure was suppressed by treatment with the MAAs mycosporine-Gly and shinorine, but not porphyra-334. This is the first report investigating the biological activities of microalgae-derived MAAs in human cells.     

Nitrite production by Nitrosomonas europaea and Nitrosospira sp. AV in soils at different solution concentrations of ammonium

Although it remains unclear why NH₃-oxidizing bacteria (AOB) of the genus Nitrosospira dominate soil environments, and why Nitrosomonas spp. are less common, virtually no studies have compared their behavior in soil. In this study, the NH₃ oxidation rates of Nitrosomonas europaea (ATCC 19718) and Nitrosospira sp. AV were compared in three differently textured soils containing a range of extractable contents (2-11 μg soil). Soils were adjusted to pH 7.0-7.4 with CaCO₃ and sterilized with γ-radiation. Cell suspensions of each bacterium were inoculated into the soils to bring them to two-third of water-holding capacity and cell densities ∼2.5×10⁶ g⁻¹ soil. In virtually all cases, rates of production for both N. europaea and Nitrosospira sp. AV were linear over 48 h, and represented between 13 and 75%, respectively, of the maximum rates achieved in soil-free bacterial suspensions. Soil solution concentrations that supported these rates ranged between 0.2 and 1.5 mM. Addition of 21-36 μg soil raised soil solution levels to 1.8-2.5 mM and stimulated production to a greater extent in N. europaea (3.3-6.6-fold) than in Nitrosospira sp. AV (1-2.1-fold). Maximum rates of production were obtained by raising soil solution levels to 3-4 mM with a supplement of ∼80-90 μg soil. K_s values in soil for Nitrosospira sp. AV and N. europaea were estimated as 0.14 and 1.9 mM , respectively, and estimates of V_max were about 3.5-times higher for N. europaea (0.007 pmol h⁻¹ cell⁻¹) than for Nitrosospira sp. AV (0.002 pmol h⁻¹ cell⁻¹). The cell density of N. europaea increased in sterile Steiwer soil independent of supplemental . In the case of treatments receiving supplemental , growth yields of N. europaea calculated from either produced or consumed were similar to those reported in literature (3.5×10⁶-6×10⁶ cells μmol⁻¹). A higher growth yield was measured in the case of zero added (2.7×10⁷ cells μmol⁻¹), indicating that use of organic carbon compounds might have occurred and resulted in some energy sparing. Our results suggest that Nitrosospira spp. with a K_s similar to Nitrosospira sp. AV may have an advantage for survival in soil environments where soil solution levels are less than 1 mM. However, it is apparent that AOB like N. europaea are poised to take advantages of modest increases in extractable that raise soil solution levels to about 2.0-2.5 mM.     

Efficient Isolation of Mycosporine-Like Amino Acids from Marine Red Algae by Fast Centrifugal Partition Chromatography

Marine rhodophyta are known to synthesize specific secondary metabolites, mycosporine-like amino acids (MAAs), to protect themselves from harmful UV-radiation. Shinorine and porphyra-334 are among the most abundant representatives of this compound class. In the present work, a novel approach for their isolation is described. As a first step, a fast centrifugal partition chromatography method, with an aqueous two-phase system comprising water, ethanol, ammonium sulfate and methanol in ascending mode, was developed to isolate the two MAAs from crude aqueous-methanolic extracts of three algal species within 90 min. The compounds could be isolated when just one of them was present in a sample or also both at the same time. By employing solid phase extraction as a second purification step, the individual MAAs were obtained in high purity and good quantity within a much shorter time frame than the established purification protocols, e.g., semi-preparative HPLC. For example, from 4 g Porphyra sp. (Nori) crude extract, 15.7 mg shinorine and 36.2 mg porphyra-334 were isolated. Both were highly pure, as confirmed by TLC, HPLC-MS and NMR analyses.     

Relationships between quality parameters and content of glycerol galactoside and porphyra-334 in dried laver nori <Emphasis Type="Italic">Porphyra yezoensis</Emphasis>

ABSTRACT:   Glycerol galactoside (GG; floridoside + isofloridoside) and porphyra-334 (P-334) are contained in nori (Susabinori Porphyra yezoensis and Asakusanori Porphyra tenera ). Glycerol galactoside has been found to have bifidogenic growth stimulator activity and P-334 is known to have ultraviolet-absorbing activity in the UVA region of sunlight. These substances have, respectively, potential for application to pre-biotic foods and in cosmetics as a sunscreen. In the present study, to investigate the relationships between GG and P-334 contents and the quality of nori, we measured the GG and P-334 contents with other components (total protein, chlorophyll-a, β-carotene and phycobillins) that are related to the quality of nori samples produced from different production areas and with different qualities. We found that the GG content was closely negatively correlated with the contents of other components, whereas P-334 was positively correlated with the other components. From these results, it is suggested that low-quality nori is a potential source of GG, and as a source for P-334, scraps of nori produced during nori processing should be suitable.     

Nrf2 and NF-κB Signaling Pathways Contribute to Porphyra-334-Mediated Inhibition of UVA-Induced Inflammation in Skin Fibroblasts

In this study, we examined the protective effects of porphyra-334 against UVA-irradiated cellular damage and elucidated the underlying mechanisms. Porphyra-334 prevented UVA-induced cell death and exhibited scavenging activities against intracellular oxidative stress induced by UVA irradiation in skin fibroblasts. We found that porphyra-334 significantly reduced the secretion and expression of IL-6 and TNF-α, reduced nuclear expression of Nuclear factor-κB (NF-κB), and sustained NF-E2-related factor 2 (Nrf2) activation. Further mechanism research revealed that porphyra-334 promoted the Nrf2 signaling pathway in UVA-irradiated skin fibroblasts. Our results show that the antioxidant effect of porphyra-334 is due to the direct scavenging of oxidative stress and its inhibitory effects on NF-κB-dependent inflammatory genes, such as IL-6 and TNF-κ. Therefore, we hypothesize that boosting the Nrf2- NF-κB-dependent response to counteract environmental stress is a promising strategy for the prevention of UVA-related damage.