Induction of apoptosis in cancer cell lines by the Red Sea brine pool bacterial extracts.

BACKGROUND: Marine microorganisms are considered to be an important source of bioactive molecules against various diseases and have great potential to increase the number of lead molecules in clinical trials. Progress in novel microbial culturing techniques as well as greater accessibility to unique oceanic habitats has placed the marine environment as a new frontier in the field of natural product drug discovery. METHODS: A total of 24 microbial extracts from deep-sea brine pools in the Red Sea have been evaluated for their anticancer potential against three human cancer cell lines. Downstream analysis of these six most potent extracts was done using various biological assays, such as Caspase-3/7 activity, mitochondrial membrane potential (MMP), PARP-1 cleavage and expression of γH2Ax, Caspase-8 and -9 using western blotting. RESULTS: In general, most of the microbial extracts were found to be cytotoxic against one or more cancer cell lines with cell line specific activities. Out of the 13 most active microbial extracts, six extracts were able to induce significantly higher apoptosis (>70%) in cancer cells. Mechanism level studies revealed that extracts from Chromohalobacter salexigens (P3-86A and P3-86B(2)) followed the sequence of events of apoptotic pathway involving MMP disruption, caspase-3/7 activity, caspase-8 cleavage, PARP-1 cleavage and Phosphatidylserine (PS) exposure, whereas another Chromohalobacter salexigens extract (K30) induced caspase-9 mediated apoptosis. The extracts from Halomonas meridiana (P3-37B), Chromohalobacter israelensis (K18) and Idiomarina loihiensis (P3-37C) were unable to induce any change in MMP in HeLa cancer cells, and thus suggested mitochondria-independent apoptosis induction. However, further detection of a PARP-1 cleavage product, and the observed changes in caspase-8 and -9 suggested the involvement of caspase-mediated apoptotic pathways. CONCLUSION: Altogether, the study offers novel findings regarding the anticancer potential of several halophilic bacterial species inhabiting the Red Sea (at the depth of 1500-2500 m), which constitute valuable candidates for further isolation and characterization of bioactive molecules.

The extremely halophilic bacterium Salicola marasensis IC10 accumulates the compatible solute betaine.

The extremely halophilic bacterium strain IC10 was isolated from a solar saltern on Isla Cristina (southern Spain). Phylogenetic, genotypic and phenotypic data supported the inclusion of this strain in the species Salicola marasensis. An analysis of intracellular organic osmotic solutes showed glycine betaine to be present, contributing to the overall osmotic balance, and this was the only compatible solute accumulated when S. marasensis IC10 was grown over a wide range of external NaCl concentrations (10-25%, w/v).

Description of Kushneria aurantia gen. nov., sp. nov., a novel member of the family Halomonadaceae, and a proposal for reclassification of Halomonas marisflavi as Kushneria marisflavi comb. nov., of Halomonas indalinina as Kushneria indalinina comb. nov. and of Halomonas avicenniae as Kushneria avicenniae comb. nov.

An aerobic, moderately halophilic, Gram-negative, motile, non-sporulating rod-shaped bacterium, designated strain A10(T), was isolated from the surface of leaves of the black mangrove Avicennia germinans and was subjected to a polyphasic taxonomic study. Strain A10(T) was able to grow at NaCl concentrations in the range 5-17.5 % (w/v) with optimum growth at 10 % (w/v) NaCl. Growth occurred at temperatures of 20-40 degrees C (optimal growth at 37 degrees C) and pH 5.5-8.5 (optimal growth at pH 7.0-8.0). The major respiratory quinone was ubiquinone 9. The major fatty acids were C(16 : 0), C(18 : 1)omega7c, C(19 : 0) cyclo omega8c and C(12 : 0) 3-OH. The polar lipids were phosphatidylglycerol, diphosphatidylglycerol, phosphatidylethanolamine, and unidentified phospholipids, glycolipids and an aminoglycolipid. Phylogenetic analysis based on 16S rRNA gene sequence comparisons revealed that strain A10(T) is closely related to Halomonas avicenniae MW2a(T) (95.7 % sequence similarity), Halomonas marisflavi SW32(T) (95.2 %) and Halomonas indalinina GC2.1(T) (95.0 %). Strain A10(T) formed a coherent phylogenetic branch with these three species, separated from other species of Halomonas and closely related genera (with 16S rRNA gene sequence similarities below 94.0 %). A complete 23S rRNA gene sequence comparison of strain A10(T) with closely related species confirmed the phylogenetic position of the novel isolate, forming a branch with the species Halomonas avicenniae, Halomonas indalinina and Halomonas marisflavi, separated from other species of the genera belonging to the family Halomonadaceae (showing sequence similarities below 91.7 %). DNA-DNA hybridization studies between strain A10(T) and Halomonas avicenniae MW2a(T), Halomonas marisflavi DSM 15357(T) and Halomonas indalinina CG2.1(T) were 21, 17 and 10 %, respectively. These levels of DNA-DNA relatedness were low enough to classify strain A10(T) as representing a genotypically distinct species. Overall, the phenotypic, genotypic, chemotaxonomic and phylogenetic results demonstrated that strain A10(T) represents a new genus and species. The name Kushneria aurantia gen. nov., sp. nov. is proposed, with strain A10(T) (=CCM 7415(T)=CECT 7220(T)) as the type strain. This is the type species of the new proposed genus, which belongs to the family Halomonadaceae. In addition, our data support the placement of the species Halomonas marisflavi, Halomonas indalinina and Halomonas avicenniae within this new genus, as Kushneria marisflavi comb. nov., Kushneria indalinina comb. nov. and Kushneria avicenniae comb. nov., respectively.

Chromohalobacter japonicus sp. nov., a moderately halophilic bacterium isolated from a Japanese salty food.

A Gram-negative, non-spore-forming, rod-shaped, motile bacterium, designated strain 43(T), was isolated from a Japanese salty food and then subjected to a polyphasic taxonomic study. Strain 43(T) is moderately halophilic, growing at NaCl concentrations in the range 5-25 % (w/v), with optimum growth between 7.5 and 12.5 % (w/v) NaCl. Growth occurs at temperatures from 15 to 42 degrees C (optimally at 28-37 degrees C) and at pH 5.5-9.0 (optimally at pH 7.0-8.0). A phylogenetic analysis based on 16S rRNA gene sequence comparisons revealed that strain 43(T) belongs to the genus Chromohalobacter. The closest relatives were Chromohalobacter canadensis ATCC 43984(T) (99.3 % 16S rRNA gene sequence similarity), Chromohalobacter beijerinckii ATCC 19372(T) (99.1 %), Chromohalobacter sarecensis LV4(T) (98.3 %), Chromohalobacter nigrandesensis LTS-4N(T) (97.9 %) and Chromohalobacter marismortui ATCC 17056(T) (97.9 %). The DNA G+C content was 62.9 mol%, which is within the range described for the genus Chromohalobacter. DNA-DNA hybridization studies between strain 43(T) and C. canadensis CECT 5385(T) and C. beijerinckii DSM 7218(T) showed 38 and 49 % relatedness, respectively; lower DNA-DNA hybridization percentages were obtained with respect to other related Chromohalobacter species. The major fatty acids of strain 43(T) were C(16 : 0), C(19 : 0) cyclo omega8c and C(12 : 0) 3-OH. Overall, the phenotypic, genotypic and phylogenetic results demonstrated that strain 43(T) represents a novel species within the genus Chromohalobacter. The name Chromohalobacter japonicus sp. nov. is proposed, with strain 43(T) (=CECT 7219(T) =CCM 7416(T)) as the type strain.