Chemical constituents from the heterotrophic marine microalgae Aurantiochytrium sp. SC145 and their antimicrobial activities.

One new indol, N-methoxymethyltryptophol (1), one new phenolic, (2 R)-2-(4-hydroxyphenyl)ethyl 2-hydroxy-3-phenylpropanoate (2) and fifteen known compounds (3-17) were isolated from the methanol extract of the fermentation of marine microalgae Aurantiochytrium sp. SC145. Their structures were elucidated by 1D-, 2D-NMR spectroscopic analysis, HR-ESI-MS, quantum chemical calculation methods and by comparing their NMR data with those reported in the literature. All compounds were evaluated for their antimicrobial activities against microorganisms. Compounds 2, 3 and 11 significantly exhibited antimicrobial activities on all tested Gram-(+), Gram-(-) bacteria and the yeast C. albicans with MIC values ranging from 32 to 256 µg/mL.

Isolation of Fucoxanthin from Sargassum oligocystum Montagne, 1845 Seaweed in Vietnam and Its Neuroprotective Activity.

Fucoxanthin extracted and purified from Vietnamese Sargassum oligocystum Montagne, 1845 exhibits various biological activities. In this study, the ability of fucoxanthin to inhibit acetylcholinesterase (AChE), the antioxidant activities, and the expression of antioxidant enzymes were investigated. Fucoxanthin isolated from Vietnamese S. oligocystum showed no cytotoxic effects; moreover, it exhibited AChE inhibitory activity (with an IC50 value of 130.12 ± 6.65 µg mL-1) and antioxidant activity (with an IC50 value of 3.42 ± 0.15 mg mL-1). At concentrations of 50 and 100 µg mL-1, fucoxanthin provided protection against amyloid ß-protein fragment 25-35-induced neurotoxicity in a C6 neuronal cell line, and the survival of C6 cells was higher than 81.01% and 80.98%, respectively, compared to the control group (59%). Moreover, antioxidant enzyme activity and quantitative PCR analysis suggested that the neuroprotective effect of fucoxanthin resulted from regulation of the gene expression of antioxidant enzymes (CAT and GPx) and ER pathways (caspase-3 and Bax), as well as the promotion of expression of genes involved in PI3K/Akt signaling (GSK-3ß), autophagy (p62 and ATG5), and the biosynthesis of ACh (VAChT and ChAT). Therefore, fucoxanthin extracted from the seaweed S. oligocystum in Vietnam is a potential feedstock source for the production of health foods that exert neuroprotective effects.

Characterization and Optimization of Culture Conditions for Aurantiochytrium sp. SC145 Isolated from Sand Cay (Son Ca) Island, Vietnam, and Antioxidative and Neuroprotective Activities of Its Polyunsaturated Fatty Acid Mixture.

Aurantiochytrium is a heterotrophic marine microalga that has potential industrial applications. The main objectives of this study were to isolate an Aurantiochytrium strain from Sand Cay (Son Ca) Island, Vietnam, optimize its culture conditions, determine its nutritional composition, extract polyunsaturated fatty acids (PUFAs) in the free (FFA) and the alkyl ester (FAAE) forms, and evaluate the antioxidation and neuroprotection properties of the PUFAs. Aurantiochytrium sp. SC145 can be grown stably under laboratory conditions. Its culture conditions were optimized for a dry cell weight (DCW) of 31.18 g/L, with total lipids comprising 25.29%, proteins 7.93%, carbohydrates 15.21%, and carotenoid at 143.67 µg/L of DCW. The FAAEs and FFAs extracted from Aurantiochytrium sp. SC145 were rich in omega 3-6-9 fatty acids (40.73% and 44.00% of total fatty acids, respectively). No acute or subchronic oral toxicity was determined in mice fed with the PUFAs in FFA or FAAE forms at different doses over 90 days. Furthermore, the PUFAs in the FFA or FAAE forms and their main constituents of EPA, DHA, and ALA showed antioxidant and AChE inhibitory properties and neuroprotective activities against damage caused by H2O2- and amyloid-ß protein fragment 25-35 (Aß25-35)-induced C6 cells. These data suggest that PUFAs extracted from Aurantiochytrium sp. SC145 may be a potential therapeutic target for the treatment of neurodegenerative disorders.

Genome analysis of the heterotrophic microalga Thraustochytrium sp. TN22 to identify genes involved in exopolysaccharide and carotenoid biosynthesis pathways.

In this study, the genome of Thraustochytrium sp. TN22, a heterotrophic marine microalga, was sequenced, assembled and annotated using the Illumina NovaSeq 6000, PacBio RS II and MinION platform to identify genes and EPS and carotenoid biosynthesis pathways. The genome of Thraustochytrium sp. TN22 consists of 38,842,079 bp with a G + C content of 66.52% and reached 90.7% gene completeness. In total, 15,208 genes were annotated, among which 14,236 genes were identified using the NCBI-nr, KEGG, GO, COG and InterProScan databases. Pathway analysis showed that EPS biosynthesis in Thraustochytrium sp. TN22 follows an ABC-dependent pathway. The carotenoids of Thraustochytrium TN22 are predicted to be synthesized via the mevalonate pathway, and the conversion of ß-carotene to xanthophyll proceeds through ß-cryptoxanthin and zeaxanthin intermediates. Our results will be useful for improving high-value coproducts for industrial application of thraustochytrids.

Biocharacteristics and draft genome sequence of Halomonas hydrothermalis C22, a pyruvate-producing halophilic bacterium isolated from a commercial Spirulina culture in Vietnam.

Halophilic bacteria are receiving increasing attention for industrial chemical production processes due to their unique properties. Herein, an alkaliphilic and halophilic bacterium was isolated from a commercial Spirulina culture at Nghe An province in Vietnam and found to secrete pyruvate. Pyruvate is widely used as a starting material in the industrial biosynthesis of pharmaceuticals, and is employed for production of crop protection agents, polymers, cosmetics, and food additives. Phenotypic and chemotaxonomic characterization, and the 16S rRNA gene sequence homology with Halomonas hydrothermalis strain DSM 15,725 (99.2%) predicted that the strain belongs to the Halomonas genus, thus we named this strain as H. hydrothermalis strain C22. We investigated the biocharacteristics and capacity of strain C22 and determined the draft genome sequence comprising 3,934,166 bp with a G + C content of 60.2% encoding 3,668 proteins, 58 tRNAs, 9 rRNAs, and 1 tmRNA. Maximal pyruvate secretion reached 51.1 g/l after 84 h of cultivation. The results will facilitate future studies on the genetic and metabolic diversity of halophilic bacteria and expand our understanding of important bioprocesses in this microorganism.

Squalene promotes cholesterol homeostasis in macrophage and hepatocyte cells via activation of liver X receptor (LXR) α and ß.

OBJECTIVE: To examine the effect of squalene on liver X receptors (LXRs) that regulate target genes associated with reverse cholesterol transport and thus control whole-body cholesterol homeostasis. RESULTS: To examine the effect of squalene on liver X receptors (LXRs) that regulate target genes associated with reverse cholesterol transport and thus control whole-body cholesterol homeostasis. Squalene significantly stimulated the transactivation of liver X receptor modulator LXRα and LXRß. The mRNA expression of LXRs and their target genes, including ABCA1, ABCG1 and ApoE, was significantly induced in macrophages stimulated with squalene, resulting in removal of cholesterol from the cells. Notably, squalene did not induce higher hepatic triacylglycerol levels nor did it alter expression of sterol regulatory element-binding protein 1c (SREBP-1c) and FAS in hepatocyte cells, primarily because of its upregulation of Insig-2a, which delays nuclear translocation of SREBP-1c, a key hepatic lipogenic transcription factor. CONCLUSION: Squalene has hypocholesterolemic effect through the activation of LXRα and ß without inducing hepatic lipogenesis.

Transcriptome sequencing and comparative analysis of Schizochytrium mangrovei PQ6 at different cultivation times.

OBJECTIVE: The heterotrophic marine microalga, Schizochytrium mangrovei PQ6, synthesizes large amounts of polyunsaturated fatty acids (PUFAs) with possible nutritional applications. We characterized the transcriptome of S. mangrovei PQ6, focusing on lipid metabolism pathways throughout growth. RESULT: Cell growth, total lipid, and docosahexaenoic acid (DHA, 22:6n-3) contents of S. mangrovei PQ6 in 500 ml batch cultures rapidly increased on day 1 in cultivation and reached their maximum levels on day 5. Maximum lipid accumulation in 500 ml batch cultures occurred on day 5, with total lipid and DHA contents reaching 33.2 ± 1.25% of dry cell weight (DCW) and 136 mg/g DCW, respectively. 11,025 unigenes, 28,617 unigenes and 18,480 unigenes from the transcriptomes of samples collected on day 1, 3, and 5 in cultivation were identified, respectively. These unigenes of the three samples were further assembled into 30,782 unigenes with an average size of 673 bp and N50 of 950 bp, and a total of 9,980 unigenes were annotated in public protein databases. 93 unigenes involved in lipid metabolism in which expression patterns corresponded with total lipid and DHA accumulation patterns were identified. CONCLUSION: The possible roles of PUFAs pathways, such as those mediated by fatty acid synthase, polyketide synthase, and desaturase/elongase, co-exist in S. mangrovei PQ6.

Extraction of squalene as value-added product from the residual biomass of Schizochytrium mangrovei PQ6 during biodiesel producing process.

Today microalgae represent a viable alternative source of squalene for commercial application. The species Schizochytrium mangrovei, a heterotrophic microalga, has been widely studied and provides a high amount of squalene, polyunsaturated fatty acids and has good profiles for biodiesel production. Our work was aimed at examining the squalene contents in Vietnam's heterotrophic marine microalga S. mangrovei PQ6 biomass and residues of the biodiesel process from this strain. Thin-layer chromatography and high-performance liquid chromatography (HPLC) methods were successfully applied to the determination of squalene in S. mangrovei PQ6. The squalene content and production of S. mangrovei PQ6 reached 33.00 ± 0.02 and 33.04 ± 0.03 mg g(-1) of dry cell weight; and 0.992 g L(-1) and 1.019 g L(-1) in 30 and 150 L bioreactors, respectively after 96 h of fermentation. In addition, squalene was also detected in spent biomass (approximately 80.10 ± 0.03 mg g(-1) of spent biomass) from the S. mangrovei PQ6 biodiesel production process. The structure of squalene in residues of the biodiesel process was confirmed from its nuclear magnetic resonance spectra. The results obtained from our work suggest that there is tremendous potential in the exploitation of squalene as a value-added by-product besides biodiesel from S. mangrovei PQ6 to reduce biodiesel price.

Biodiesel production from Vietnam heterotrophic marine microalga Schizochytrium mangrovei PQ6.

This work is to explore the potential of producing biodiesel and valuable co-products from the biomass of Schizochytrium mangrovei PQ6 which was isolated from Phu Quoc Island, Kien Giang province, Vietnam. This microalga contains high lipid content (up to 70% of dry cell weight) and high level of total fatty acids, which is ideal for making biodiesel. The production of fatty acid methyl esters (FAME) from this marine microalga resulted in a yield of 88% based on algal oil and 44% based on algal biomass. The process of separating the obtained FAME into a first fraction enriched with saturated FAME (SFAME) and a second fraction enriched with unsaturated FAME (UFAME) was then investigated to exploit the valuable co-products. The obtained results shown that the mass fraction of SFAME and UFAME were 70% and 30%, respectively. The UFAME fraction contains a high content of DHA (accounting for 69.000% of TFAs). The test results of the SFAME fraction indicated that specific gravity at 15°C, flash point, water and sediment, kinematic viscosity at 40°C, sulfated ash, sulfur, copper strip corrosion at 50°C, cetane number, carbon residue, iodine number, workmanship meet Vietnam Biodiesel B100 Standard. Moreover, the utilization of waste glycerol from biodiesel process as carbon source for the cultivation of the microalgae S. mangrovei PQ6 and Spirulina platensis was also investigated.

Residual congener pattern of dioxins in human breast milk in southern Vietnam.

This study evaluated residual congener patterns of dioxin/furan (=PCDD/DF) related to tactical herbicides aerially sprayed over the regions of southern Vietnam through Operation Ranch Hand. The study focused on Cam Chinh (CC) commune, Quang Tri province (an area sprayed with tactical herbicides), and the Cam Phuc (CP) commune, Ha Tinh province (a non-sprayed area). Breast milk samples for analysis were collected in September 2002 and July 2003 from lactating primiparous and multiparous mothers born after the war (<31 years old). We found the levels of each congener in the CC commune were higher than in the CP commune, and determined specificity in the PCDD/DF congener pattern in CC commune samples by cluster analysis. The congener pattern is characterized by higher (the hexa-, the hepta-, and the octa-) chlorinated PCDD/DFs; this appears to be the same profile as that presented by pentachlorophenol (PCP), rather than 2,4,5-trichlorophenoxy acid (2,4,5-T) contaminated with 2,3,7,8-TCDD. A GC/MS study in the 1970s detected the chlorophenols 2,4-di and 2,4,6-trichlorophenol in some Agent Orange samples, which contained, like PCP, a wide variety of PCDD/DF congeners. In this context, it may be expected that certain tactical herbicides contaminated with various chlorophenol impurities, have a unique congener pattern when compared with pure 2,4,5-T formulations.

A GIS study of dioxin contamination in a Vietnamese region sprayed with herbicide.

OBJECTIVES: The American-Vietnam War ended over 35 years ago, but dioxins still remain in human tissue and Vietnam's environment. The main objective of this study was to assess the correlation between dioxin concentration in soil, sediment and breast milk in one of the areas sprayed with herbicide, namely Cam Chinh commune, Quang Tri province, using a geographic information system (GIS). METHODS: The concentration of polychlorinated dibenzo-p-dioxins (PCDDs) and polychlorinated dibenzofurans (PCDFs) in soil, sediment and breast milk samples was determined and the data analyzed using the geostatistical log-normal Kriging algorithm. RESULTS: The mean dioxin levels in soil and breast milk in the sprayed area were significantly higher than those in the non-sprayed area. The distribution pattern of PCDDs and PCDFs in soil was different, and there were no significant correlations between the estimated dioxin levels in soil obtained by the Kriging method and those in breast milk. CONCLUSIONS: The possibility remains that another exposure route, such as exposure to herbicides used during the Vietnam War, might affect dioxin levels in breast milk, although more soil data are needed to make more reliable geographical estimations.