Polaribacter Irgensii Research Articles

An intrinsically disordered domain in Polaribacter irgensii KOPRI 22228 CspB confers extraordinary freeze-tolerance

Organisms living in extremely cold environments possess mechanisms to survive low temperatures. Among the known cold-induced genes, cold-shock proteins (Csps) are the most prominent. A csp-homologous gene, cspBPi, has been cloned from the Arctic bacterium Polaribacter irgensii KOPRI 22228, and overexpression of this gene greatly increased the freezing tolerance of its host. This protein consists of a unique N-terminal domain and a well conserved C-terminal cold shock domain. To elucidate the detailed mechanisms involved in the extraordinary freeze-tolerance conferred by CspBPi, we identified the responsible domain by mutational analysis. Changes of residues in the cold shock domain that are crucial for binding RNA or single-stranded DNA did not impair the ability of the host to survive freezing stress. All domain-shuffled CspBPi variants containing the N-terminal domain retained the ability to confer superior freeze-tolerance. Slow electrophoretic mobility and far-UV circular dichroism spectra of the N-terminal domain suggested an intrinsically disordered structure for this region. The N-terminal domain also bound to lipid vesicles in vitro. This lipid vesicle binding characteristic is shared with other intrinsically disordered proteins, such as α-synuclein and plant dehydrins, known to confer cold-tolerance when overexpressed, suggesting a mechanism for cold-survival through membrane binding.

CspB of an arctic bacterium, Polaribacter irgensii KOPRI 22228, confers extraordinary freeze-tolerance.

Freezing temperatures are a major challenge for life at the poles. Decreased membrane fluidity, uninvited secondary structure formation in nucleic acids, and protein cold-denaturation all occur at cold temperatures. Organisms adapted to polar regions possess distinct mechanisms that enable them to survive in extremely cold environments. Among the cold-induced proteins, cold shock protein (Csp) family proteins are the most prominent. A gene coding for a Csp-family protein, cspB, was cloned from an arctic bacterium, Polaribacter irgensii KOPRI 22228, and overexpression of cspB greatly increased the freeze-survival rates of Escherichia coli hosts, to a greater level than any previously reported Csp. It also suppressed the cold-sensitivity of an E. coli csp-quadruple deletion strain, BX04. Sequence analysis showed that this protein consists of a unique domain at its N-terminal end and a well conserved cold shock domain at its C-terminal end. The most common mechanism of Csp function in cold adaption is melting of the secondary structures in RNA and DNA molecules, thus facilitating transcription and translation at low temperatures. P. irgensii CspB bound to oligo(dT)-cellulose resins, suggesting single-stranded nucleic acid-binding activity. The unprecedented level of freeze-tolerance conferred by P. irgensii CspB suggests a crucial role for this protein in survival in polar environments.

Polaribacter marinaquae sp. nov., isolated from seawater.

A novel Gram-stain-negative, rod-shaped, non-spore-forming, non-flagellated, strictly aerobic strain, designated RZW3-2T, was isolated from seawater from near the Yellow Sea coast of PR China (35.475° N 119.613° E). The organism grew optimally between 24 and 28 °C, at pH 6.0-7.0 and in the presence of 2-3 % (w/v) NaCl. The strain requires seawater or artificial seawater for growth and NaCl alone does not support growth. RZW3-2T contained iso-C15 : 0, C16 : 1ω7c and/or C16 : 1ω6c and anteiso-C15 : 0 as the dominant fatty acids. The respiratory quinone detected in RZW3-2T was menaquinone 6 (MK6). The polar lipids of RZW3-2T comprised phosphatidylethanolamine (PE), four unidentified phospholipids (PL), two unidentified aminolipids (AL) and one unknown lipid (L). The DNA G+C content of RZW3-2T was 30.5 mol%. Phylogenetic analysis based on 16S rRNA gene sequences showed that the novel strain was related most closely to Polaribacter reichenbachii KCTC 23969T, Polaribacter dokdonensis KCTC 12392T and Polaribacter irgensii CIP 106478T with 98.0, 97.8 and 97.0 % sequence similarities, respectively. The DNA-DNA hybridization values between RZW3-2T and its closest phylogenetic relatives, P. reichenbachii KCTC 23969T and P. dokdonensis KCTC 12392T, were 52.0±0.6 % and 49.8±1.21 % respectively. On the basis of polyphasic analyses, RZW3-2T represents a novel species of the genus Polaribacter, for which the name Polaribacter marinaquae sp. nov. is proposed. The type strain is RZW3-2T (=JCM 30825T=KCTC 42664T=MCCC 1K00696T).

Mevalonosomes: specific vacuoles containing the mevalonate pathway in Plocamium brasiliense cortical cells (Rhodophyta).

This paper has identified, for the first time in a member of the Rhodophyta, a vacuolar organelle containing enzymes that are involved in the mevalonate pathway-an important step in red algal isoprenoid biosynthesis. These organelles were named mevalonosomes (Mev) and were found in the cortical cells (CC) of Plocamium brasiliense, a marine macroalgae that synthesizes several halogenated monoterpenes. P. brasiliense specimens were submitted to a cytochemical analysis of the activity of the 3-hydroxy-3-methylglutaryl-CoA synthase (HMGS). Using transmission electron microscopy (TEM), we confirmed the presence of HMGS activity within the Mev. Because HMGS is necessary for the biosynthesis of halogenated monoterpenes, we isolated a hexanic fraction (HF) rich in halogenated monoterpenes from P. brasiliense that contained a pentachlorinated monoterpene as a major metabolite. Because terpenes are often related to chemical defense, the antifouling (AF) activity of pentachlorinated monoterpene was tested. We found that the settlement of the mussel Perna perna was reduced by HF treatment (2.25 times less than control; 40% and 90% of fouled surface, respectively; P = 0.001; F9,9 = 1.13). The HF (at 10 μg · mL(-1) ) also inhibited three species of fouling microalgae (Chlorarachnion reptans, Cylindrotheca cloisterium, and Exanthemachrysis gayraliae), while at a higher concentration (50 μg · mL(-1) ), it inhibited the bacteria Halomonas marina, Polaribacter irgensii, Pseudoalteromonas elyakovii, Shewanella putrefaciens, and Vibrio aestuarianus. The AF activity of P. brasiliense halogenated monoterpenes and the localization of HMGS activity inside Mev suggest that this cellular structure found in CC may play a role in thallus protection against biofouling.

Antifouling activity of novel polyisoprene-based coatings made from photocurable natural rubber derived oligomers

Natural rubber is a renewable resource with a potential as precursor of a very wide range of novel polymers, including polyisoprene-based surfaces with antifouling (AF) activity. In this work, new ionic and non-ionic coatings were prepared by the photocrosslinking reaction of photosensitive cis-1,4-oligoisoprenes, bearing a variable number of ammonium groups. The photochemical crosslinking was achieved using radical (via acrylate groups) or cationic (via epoxy groups) processes. Surface properties of these coatings were studied by static contact angle measurements and AFM imaging. Assessment of bioactivity demonstrated that most of the resulting coatings showed AF potential against fouling organisms: growth inhibition of marine bacteria (Pseudoalteromonas elyakovii, Shewanella putrefaciens, Cobetia marina, Polaribacter irgensii, Vibrio aestuarianus) and fungi (Halosphaeriopsis mediosetigera, Asteromyces cruciatus, Lulworthia uniseptata, Zalerion sp., Monodictys pelagica); decreased adhesion of microalgae (Navicula jeffreyi, Cylindrotheca closterium, Chlorarachnion globosum, Pleurochrysis roscoffensis, Exanthemachrysis gayraliae, Amphora coffeaeformis); inhibition of attachment and/or germination of spores of Ulva intestinalis. The best AF activity was obtained with the ionic surfaces. These new coatings prepared from precursors obtained from natural rubber are in essence active by contact. As the biocidal functions are fixed covalently to the polymer chain, detectable release of biocidal products in the marine ecosystem is prevented so that a valuable environment-friendly alternative for new AF coatings is hereby proposed.

Greening effect on oysters and biological activities of the blue pigments produced by the diatom Haslea karadagensis (Naviculaceae)

Haslea karadagensis is a recently-described diatom, the second species of blue diatom to be identified, after Haslea ostrearia, the type species of the genus Haslea, which produces marennine, the water soluble pigment involved in the greening of oysters. Haslea karadagensis also produces a blue grey pigment, with different spectral characteristics from marennine itself. This study demonstrates that the pigment from H. karadagensis can colour the gills of two oysters of economic importance, Crassostrea gigas and Ostrea edulis, when they are fed with a suspension of algae, in the same way as the marennine produced by H. ostrearia. Like marennine, the purified pigment produced by H. karadagensis displays several different biological activities. Both the intracellular and extracellular forms of the pigment have been shown to inhibit the growth of marine bacteria (Polaribacter irgensii, Vibrio aestuarianus, Pseudoalteromonas elyakowii) and fungi (Corollospora maritima, Lulworthia sp., and Dendryphiella salina). The pigment also displays antiviral activity against Herpes simplex virus type 1 (HSV-1). In light of these preliminary results, the use of H. karadagensis and putative applications of its pigment in aquaculture, food chemistry and ecophysiological research, are discussed.

Evolution of Antifreeze Protein Genes in the Diatom Genus Fragilariopsis: Evidence for Horizontal Gene Transfer, Gene Duplication and Episodic Diversifying Selection

Hypotheses about horizontal transfer of antifreeze protein genes to ice-living diatoms were addressed using two different statistical methods available in the program Prunier. The role of diversifying selection in driving the differentiation of a set of antifreeze protein genes in the diatom genus Fragilariopsis was also investigated. Four horizontal gene transfer events were identified. Two of these took place between two major eukaryote lineages, that is from the diatom Chaetoceros neogracile to the copepod Stephos longipes and from a basidiomycete clade to a monophyletic group, consisting of the diatom species Fragilariopsis curta and Fragilariopsis cylindrus. The remaining two events included transfers from an ascomycete lineage to the proteobacterium Stigmatella aurantiaca and from the proteobacterium Polaribacter irgensii to a group composed of 4 proteobacterium species. After the Fragilariopsis lineage acquired the antifreeze protein gene from the basidiomycetes, it duplicated and went through episodic evolution, characterized by strong positive selection acting on short segments of the branches in the tree. This selection pattern suggests that the paralogs differentiated functionally over relatively short time periods. Taken together, the results obtained here indicate that the group of antifreeze protein genes considered here have a complex evolutionary history.

Rescue of a cold-sensitive mutant at low temperatures by cold shock proteins from Polaribacter irgensii KOPRI 22228

Exposure to low temperatures induces the biosynthesis of specific sets of proteins, including cold shock proteins (Csps). Since many of the specific functions of pychrophilic Csps are unknown, the roles of Csps from an Arctic bacterium, Polaribacter irgensii KOPRI 22228, were examined. The genes encoding CspA and CspC of P. irgensii were cloned in this study. Sequence analysis showed that these proteins have cold shock domains containing two RNA-binding motifs, RNP1 and RNP2. Both proteins bound oligo(dT)-cellulose resins, suggesting single-stranded nucleic acid-binding activity. When the P. irgensii Csps were overexpressed in Escherichia coli, the cold-resistance of the host was increased by more than five-fold. The P. irgensii Csps also rescued a cold-sensitive E. coli csp-quadruple deletion strain, BX04, at low temperatures. These results suggest that Csps from P. irgensii play a role in survival in polar environments.

Anti-microfouling activity of lipidic metabolites from the invasive brown alga Sargassum muticum (Yendo) Fensholt.

The purification of the chloroform extract from the brown invasive macroalga Sargassum muticum, through a series of chromatographic separations, yielded 12 fractions that were tested against strains of bacteria, microalgae, and fungi involved in marine biofilm formation. The chemical composition of four (a, c, g, and k) out of the six fractions that exhibited anti-microfouling activity was investigated. Fraction a contained saturated and unsaturated linear hydrocarbons (C12-C27). Arachidonic acid was identified as the major metabolite in fraction c whereas fraction g contained mainly palmitic, linolenic, and palmitoleic acids. Fraction k was submitted to further purification yielding the fraction kAcaF1e that was composed of galactoglycerolipids, active against the growth of two of the four bacterial strains (Shewanella putrefaciens and Polaribacter irgensii) and all tested fungi. These promising results, in particular the isolation and the activity of galactoglycerolipids, attest the potential of the huge biomass of S. muticum as a source of new environmentally friendly antifouling compounds.

Polaribacter gen. nov., with three new species, P. irgensii sp. nov., P. franzmannii sp. nov. and P. filamentus sp. nov., gas vacuolate polar marine bacteria of the Cytophaga-Flavobacterium-Bacteroides group and reclassification of 'Flectobacillus glomeratus' as Polaribacter glomeratus comb. nov.

Several psychrophilic, gas vacuolate strains of the Cytophage-Flavobacterium-Bacteroides (CFB) phylogenetic group were isolated from sea ice and water from the Arctic and the Antarctic. The closest taxonomically defined species by 16S rRNA sequence analysis is 'Flectobacillus glomeratus'. However, 'Flc. glomeratus' is phylogenetically distant from the Flectobacillus type species, Flc. major. On the basis of phenotypic, genotypic and 16S rRNA sequence analyses we propose a new genus, Polaribacter, with three new species, Polaribacter irgensii strain 23-P (ATCC 700398), Polaribacter franzmannii strain 301 (ATCC 700399) and Polaribacter filamentus strain 215 (ATCC 700397). P. filamentus is the type species of the genus. None of these species exhibits a cosmopolitan or bipolar distribution. This is the first taxonomic description of gas vacuolate bacteria in the CFB group. Additionally, we propose that 'Flc. glomeratus' be reclassified to the genus Polaribacter as P. glomeratus, comb. nov.