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A Gram-negative, aerobic, yellow pigmented, non-flagellated, non-gliding, rod-shaped bacterial strain, designated SM1355 T , was isolated from Antarctic intertidal sediment collected near the Chinese Antarctic Great Wall Station. The strain grew at 4-35 °C and with 0.5-7.0 % (w/v) NaCl. It hydrolysed aesculin but didn't reduce nitrate to nitrite. Phylogenetic analysis based on 16S rRNA gene sequences revealed that strain SM1355 T formed a distinct phylogenetic lineage within the family Flavobacteriaceae, sharing the highest 16S rRNA gene sequence similarity with Flaviramulus ichthyoenteri (96.3 %) and fairly high sequence similarities (95.0-96.0 %) with over 20 recognized species in eight genera of the family Flavobacteriaceae. The predominant fatty acids were anteiso-C15 : 0, iso-C15 : 0 and iso-C15 : 1 G. The major polar lipids were phosphatidylethanolamine and one unidentified lipid. The genomic DNA G+C content of strain SM1355 T was 36.2 mol%. Based on the results of the polyphasic characterization for strain SM1355 T , it is identified as the representative of a novel species in a new genus of the family Flavobacteriaceae, for which the name Changchengzhania lutea gen. nov., sp. nov. is proposed. The type strain of Changchengzhania lutea is SM1355 T (=JCM 30336 T =CCTCC AB 2014246 T ).

Unlabelled: Trimethylamine N-oxide (TMAO) is an important nitrogen source for marine bacteria. TMAO can also be metabolized by marine bacteria into volatile methylated amines, the precursors of the greenhouse gas nitrous oxide. However, it was not known how TMAO is recognized and imported by bacteria. Ruegeria pomeroyi DSS-3, a marine Roseobacter, has an ATP-binding cassette transporter, TmoXWV, specific for TMAO. TmoX is the substrate-binding protein of the TmoXWV transporter. In this study, the substrate specificity of TmoX of R. pomeroyi DSS-3 was characterized. We further determined the structure of the TmoX/TMAO complex and studied the TMAO-binding mechanism of TmoX by biochemical, structural, and mutational analyses. A Ca(2+) ion chelated by an extended loop in TmoX was shown to be important for maintaining the stability of TmoX. Molecular dynamics simulations indicate that TmoX can alternate between "open" and "closed" states for binding TMAO. In the substrate-binding pocket, four tryptophan residues interact with the quaternary amine of TMAO by cation-π interactions, and Glu131 forms a hydrogen bond with the polar oxygen atom of TMAO. The π-π stacking interactions between the side chains of Phe and Trp are also essential for TMAO binding. Sequence analysis suggests that the TMAO-binding mechanism of TmoX may have universal significance in marine bacteria, especially in the marine Roseobacter clade. This study sheds light on how marine microorganisms utilize TMAO., Importance: Trimethylamine N-oxide (TMAO) is an important nitrogen source for marine bacteria. The products of TMAO metabolized by bacteria are part of the precursors of the greenhouse gas nitrous oxide. It is unclear how TMAO is recognized and imported by bacteria. TmoX is the substrate-binding protein of a TMAO-specific transporter. Here, the substrate specificity of TmoX of Ruegeria pomeroyi DSS-3 was characterized. The TMAO-binding mechanism of TmoX was studied by biochemical, structural, and mutational analyses. Moreover, our results suggest that the TMAO-binding mechanism may have universal significance in marine bacteria. This study sheds light on how marine microorganisms utilize TMAO and should lead to a better understanding of marine nitrogen cycling., (Copyright © 2015, American Society for Microbiology. All Rights Reserved.)

A Gram-stain-negative, yellow-pigmented, aerobic, non-flagellated, non-gliding bacterial strain, designated SM1203(T), was isolated from surface seawater of Kongsfjorden, Svalbard. Phylogenetic analysis based on 16S rRNA gene sequences revealed that strain SM1203(T) was affiliated with the genus Bizionia in the family Flavobacteriaceae. The strain shared the highest 16S rRNA gene sequence similarity (>96%) with the type strains of Formosa spongicola (96.8%), Bizionia paragorgiae (96.3%), B. saleffrena (96.3%) and B. echini (96.1%) and 95.4-95.7% sequence similarity with the type strains of other known species of the genus Bizionia. The strain grew at 4-30 °C and in the presence of 1.0-5.0% (w/v) NaCl. The major fatty acids of strain SM1203(T) were iso-C15 : 0, iso-C15 : 1, anteiso-C15 : 0 and C15 : 0 and the main polar lipids were phosphatidylethanolamine and an unidentified lipid. The major respiratory quinone of strain SM1203(T) was menaquinone 6 (MK-6). The genomic DNA G+C content of strain SM1203(T) was 34.8 mol%. Based on the polyphasic characterization of strain SM1203(T) in this study, the strain represents a novel species in the genus Bizionia, for which the name Bizionia arctica sp. nov. is proposed. The type strain is SM1203(T) ( = CGMCC 1.12751(T) = JCM 30333(T)). An emended description of the genus Bizionia is also given.

Sea ice is one of the most frigid environments for marine microbes. In contrast to other ocean ecosystems, microbes in permanent sea ice are space confined and subject to many extreme conditions, which change on a seasonal basis. How these microbial communities are regulated to survive the extreme sea ice environment is largely unknown. Here, we show that filamentous phages regulate the host bacterial community to improve survival of the host in permanent Arctic sea ice. We isolated a filamentous phage, f327, from an Arctic sea ice Pseudoalteromonas strain, and we demonstrated that this type of phage is widely distributed in Arctic sea ice. Growth experiments and transcriptome analysis indicated that this phage decreases the host growth rate, cell density and tolerance to NaCl and H2O2, but enhances its motility and chemotaxis. Our results suggest that the presence of the filamentous phage may be beneficial for survival of the host community in sea ice in winter, which is characterized by polar night, nutrient deficiency and high salinity, and that the filamentous phage may help avoid over blooming of the host in sea ice in summer, which is characterized by polar day, rich nutrient availability, intense radiation and high concentration of H2O2. Thus, while they cannot kill the host cells by lysing them, filamentous phages confer properties advantageous to host survival in the Arctic sea ice environment. Our study provides a foremost insight into the ecological role of filamentous phages in the Arctic sea ice ecosystem.

A Gram-reaction-negative, aerobic, non-flagellated, rod-shaped bacterium, designated strain SM1211T, was isolated from Antarctic seawater. The isolate grew at 4-35 °C and with 0-10% (w/v) NaCl. It could produce bacteriochlorophyll a, but did not reduce nitrate to nitrite or hydrolyse DNA. Phylogenetic analysis of 16S rRNA gene sequences revealed that strain SM1211T constituted a distinct phylogenetic line within the family Rhodobacteraceae and was closely related to species in the genera Litorimicrobium, Leisingera, Seohaeicola and Phaeobacter with 95.1-96.0% similarities. The predominant cellular fatty acid was C18:1ω7c. The major polar lipids were phosphatidylethanolamine, phosphatidylglycerol, phosphatidylcholine, an unidentified aminolipid and two unidentified phospholipids. The genomic DNA G+C content of strain SM1211T was 60.7 mol%. Based on the phylogenetic, chemotaxonomic and phenotypic data obtained in this study, strain SM1211T is considered to represent a novel species in a new genus within the family Rhodobacteraceae, for which the name Puniceibacterium antarcticum gen. nov., sp. nov. is proposed. The type strain of Puniceibacterium antarcticum is SM1211T (=CCTCC AB 2013147T=KACC 16875T).

A Gram-stain-negative, aerobic, oxidase- and catalase-positive, flagellated, rod-shaped bacterial strain, designated SM1222(T), was isolated from the deep-sea sediment of the South China Sea. The strain grew at 4-35 °C and with 0.5-8 % NaCl (w/v). Phylogenetic analysis based on the 16S rRNA gene sequences revealed that strain SM1222(T) was affiliated with the genus Oceanisphaera in the class Gammaproteobacteria. It shared the highest sequence similarity with the type strain of Oceanisphaera ostreae (96.8 %) and 95.4-96.6 % sequence similarities with type strains of other species of the genus Oceanisphaera with validly published names. Strain SM1222(T) contained summed feature 3 (C16 : 1ω7c and/or iso-C15 : 0 2-OH), C18 : 1ω7c, C16 : 0, C12 : 0 and summed feature 2 (C14 : 0 3-OH and/or iso-C16 : 1 I) as the major fatty acids and ubiquinone Q-8 as the predominant respiratory quinone. The major polar lipids were phosphatidylethanolamine, phosphatidylglycerol and diphosphatidylglycerol. The genomic DNA G+C content of strain SM1222(T) was 51.5 mol%. On the basis of the evidence presented in this study, strain SM1222(T) represents a novel species of the genus Oceanisphaera, for which the name Oceanisphaera profunda sp. nov. is proposed. The type strain of Oceanisphaera profunda is SM1222(T) ( = CCTCC AB 2013241(T) = KCTC 32510(T)). An emended description of the genus Oceanisphaera Romanenko et al. 2003 emend. Choi et al. 2011 is also proposed.

Collagen is an insoluble protein that widely distributes in the extracellular matrix of marine animals. Collagen degradation is an important step in the marine nitrogen cycle. However, the mechanism of marine collagen degradation is still largely unknown. Here, a novel subtilisin-like collagenolytic protease, myroicolsin, which is secreted by the deep sea bacterium Myroides profundi D25, was purified and characterized, and its collagenolytic mechanism was studied. Myroicolsin displays low identity (<30%) to previously characterized subtilisin-like proteases, and it contains a novel domain structure. Protein truncation indicated that the Pro secretion system C-terminal sorting domain in the precursor protein is involved in the cleavage of the N-propeptide, and the linker is required for protein folding during myroicolsin maturation. The C-terminal β-jelly roll domain did not bind insoluble collagen fiber, suggesting that myroicolsin may degrade collagen without the assistance of a collagen-binding domain. Myroicolsin had broad specificity for various collagens, especially fish-insoluble collagen. The favored residue at the P1 site was basic arginine. Scanning electron microscopy and atomic force microscopy, together with biochemical analyses, confirmed that collagen fiber degradation by myroicolsin begins with the hydrolysis of proteoglycans and telopeptides in collagen fibers and fibrils. Myroicolsin showed strikingly different cleavage patterns between native and denatured collagens. A collagen degradation model of myroicolsin was proposed based on our results. Our study provides molecular insight into the collagen degradation mechanism and structural characterization of a subtilisin-like collagenolytic protease secreted by a deep sea bacterium, shedding light on the degradation mechanism of deep sea sedimentary organic nitrogen.

A Gram-stain-negative, aerobic, catalase- and oxidase-positive, non-flagellated, rod-shaped bacterium, designated strain P-50-3(T), was isolated from seawater of the Pacific. The strain grew at 10-40 °C (optimum at 30 °C) and with 0-12 % (w/v, optimum 2 %) NaCl. It reduced nitrate to nitrite but did not hydrolyse gelatin, starch or Tween 80. Analysis of 16S rRNA gene sequences showed that strain P-50-3(T) clustered tightly with the genus Albimonas and shared the highest 16S rRNA gene sequence similarity (94.3 %) with the type strain of Albimonas donghaensis. The major respiratory quinone was Q-10 and the major cellular fatty acids were C18 : 1ω7c, C18 : 0, 11-methyl C18 : 1ω7c and C16 : 0. Polar lipids included phosphatidylglycerol (PG), phosphatidylcholine (PC), two unidentified aminolipids and an unidentified lipid. The genomic DNA G+C content of strain P-50-3(T) was 69.0 mol%. On the basis of the data obtained in this polyphasic study, strain P-50-3(T) represents a novel species within the genus Albimonas, for which the name Albimonas pacifica sp. nov. is proposed. The type strain of Albimonas pacifica is P-50-3(T) ( = KACC 16527(T) = CGMCC 1.11030(T)). An emended description of the genus Albimonas Lim et al. 2008 is also proposed.

A yellow, rod-shaped, Gram-negative, facultatively aerobic, gliding bacterium, designed strain P7-3-5(T), was isolated from intertidal sand of the Yellow Sea, China. Analysis of 16S rRNA gene sequences revealed that strain P7-3-5(T) formed a distinct lineage within the family Flavobacteriaceae, sharing 94.2-96.9 % sequence similarity with type strains of species of the most closely related genera, including Hyunsoonleella, Jejuia, Marinivirga and Algibacter. The strain grew at 4-40 °C and with 0.5-5.0 % (w/v) NaCl. It reduced nitrate to nitrite and hydrolysed gelatin and DNA. The major cellular fatty acids were iso-C15 : 0, iso-C15 : 1 G and anteiso-C15 : 0 and the major respiratory quinone was MK-6. Polar lipids included phosphatidylethanolamine (PE), three unidentified aminolipids (AL1-3) and four unidentified lipids (L1-4). The genomic DNA G+C content of strain P7-3-5(T) was 32.1 mol%. Data from this polyphasic study suggest that strain P7-3-5(T) represents a novel species in a new genus in the family Flavobacteriaceae, for which the name Arenitalea lutea gen. nov., sp. nov. is proposed. The type strain of Arenitalea lutea is P7-3-5(T) ( = CGMCC 1.12213(T) = KACC 16457(T)).

A marine prosthecate bacterium, designated strain P-1 km-3(T), was isolated from deep seawater from the Pacific. Cells of strain P-1 km-3(T) were Gram-stain-negative, aerobic, catalase- and oxidase-positive, dimorphic rods with a single polar prostheca or flagellum. The strain hydrolysed gelatin and grew at 6-40 °C (optimum, 30 °C) and with 0.5-12% (w/v) NaCl (optimum, 2%). Phylogenetic analysis of the 16S rRNA gene sequences revealed that strain P-1 km-3(T) belonged to the family Hyphomonadaceae in the class Alphaproteobacteria and represented a separate lineage, located between the genera Oceanicaulis and Woodsholea. Sequence similarities of strain P-1 km-3(T) with type strains of species of the genera Oceanicaulis and Woodsholea were 93.2-93.9%. The predominant cellular fatty acids in strain P-1 km-3(T) were C18:1ω7c, C18:0, 11-methyl C18:1ω7c, C17:0 and C19:0 cyclo ω8c. The major respiratory quinone of strain P-1 km-3(T) was Q-10. The polar lipids of strain P-1 km-3(T) comprised glucuronopyranosyldiglyceride (GUDG), monoglycosyldiglyceride (MGDG), sulfo-quinovosyl diacylglycerol (SQDG), phosphatidylglycerol (PG), an unidentified phospholipid (PL) and an unidentified lipid (L). The genomic DNA G+C content of strain P-1 km-3(T) was 66.0 mol%. On the basis of the polyphasic data presented in this study, strain P-1 km-3(T) is proposed to represent a novel species in a new genus, Marinicauda pacifica gen. nov., sp. nov., within the family Hyphomonadaceae. The type strain of the type species is P-1 km-3(T) (=KACC 16526(T)=CGMCC 1.11031(T)).

A Gram-negative, aerobic, non-motile, pink-pigmented and rod-shaped strain, designated ZS3-33(T), was isolated from Antarctic intertidal sandy sediment. The strain grew optimally at 15 °C and with 1.0 % (w/v) NaCl. It reduced nitrate to nitrite and hydrolysed Tween 20. It could not produce bacteriochlorophyll a. The predominant cellular fatty acid was C18 : 1ω7c and the predominant respiratory quinone was Q-10. The major polar lipids were phosphatidylglycerol, phosphatidylcholine, two unidentified aminophospholipids and an unidentified aminolipid. Analyses of 16S rRNA gene sequences revealed that strain ZS3-33(T) belonged to the genus Pseudorhodobacter, showing 97.4 % similarity to the type strain of Pseudorhodobacter ferrugineus and 95.3 % similarity to the type strain of Pseudorhodobacter aquimaris. Levels of gyrB gene sequence similarity between strain ZS3-33(T) and the type strains of P. ferrugineus and P. aquimaris were 87.6 and 81.7 %, respectively. DNA-DNA relatedness between strain ZS3-33(T) and P. ferrugineus DSM 5888(T) was 56.6 %. The genomic DNA G+C content of strain ZS3-33(T) was 57.1 mol%. Based on data from this polyphasic study, strain ZS3-33(T) represents a novel species of the genus Pseudorhodobacter, for which the name Pseudorhodobacter antarcticus sp. nov. is proposed. The type strain is ZS3-33(T) ( = CGMCC 1.10836(T) = KCTC 23700(T)). An emended description of the genus Pseudorhodobacter Uchino et al. 2002 emend. Jung et al. 2012 is also proposed.

Elastin is a common insoluble protein that is abundant in marine vertebrates, and for this reason its degradation is important for the recycling of marine nitrogen. It is still unclear how marine elastin is degraded because of the limited study of marine elastases. Here, a novel protease belonging to the M23A subfamily, secreted by Pseudoalteromonas sp. CF6-2 from deep-sea sediment, was purified and characterized, and its elastolytic mechanism was studied. This protease, named pseudoalterin, has low identities (<40%) to the known M23 proteases. Pseudoalterin has a narrow specificity but high activity toward elastin. Analysis of the cleavage sites of pseudoalterin on elastin showed that pseudoalterin cleaves the glycyl bonds in hydrophobic regions and the peptide bonds Ala-Ala, Ala-Lys, and Lys-Ala involved in cross-linking. Two peptic derivatives of desmosine, desmosine-Ala-Ala and desmosine-Ala-Ala-Ala, were detected in the elastin hydrolysate, indicating that pseudoalterin can dissociate cross-linked elastin. These results reveal a new elastolytic mechanism of the M23 protease pseudoalterin, which is different from the reported mechanism where the M23 proteases only cleave glycyl bonds in elastin. Genome analysis suggests that M23 proteases may be popular in deep-sea sediments, implying their important role in elastin degradation. An elastin degradation model of pseudoalterin was proposed, based on these results and scanning electron microscopic analysis of the degradation by pseudoalterin of bovine elastin and cross-linked recombinant tropoelastin. Our results shed light on the mechanism of elastin degradation in deep-sea sediment.

Glaciecola nitratireducens strain FR1064(T) was isolated from seawater and described as a new species by Baik et al. in 2006. The genome size is about 1.01 to 1.26 Mb smaller than two reported Glaciecola genomes, indicating the gain or loss of large genome segments in the evolution of Glaciecola strains.

The peptide transporter from a cold-adapted bacterium has never been reported. In the present study, the dpp operon from the psychrophilic bacterium Pseudoalteromonas sp. strain SM9913 was cloned and analyzed. The dipeptide binding protein DppA of SM9913 was overexpressed in Escherichia coli, and its cold adaptation characteristics were studied. The recombinant DppA of SM9913 (PsDppA) displayed the highest ligand-binding affinity at 15 degrees C, whereas the recombinant DppA of E. coli (EcDppA) displayed the highest ligand-binding affinity at 35 degrees C. Thermal and guanidium hydrochloride unfolding analyses indicated that PsDppA has more structural instability than EcDppA. Six domain-exchanged mutants of PsDppA were expressed and purified. Analyses of these mutants indicated that domains III, I-2, and I-3 of PsDppA were less stable than those from EcDppA and that domains III and I-2 made a significant contribution to the high binding affinity of PsDppA at low temperatures. Structural and sequence analyses suggested that the state transition-involved regions in domain III and the alpha part of domain I-2 are the hot spots of optimization during cold adaptation and that decreasing the side-chain size in these regions is an important strategy for the cold adaptation of PsDppA.

Background: Zunongwangia profunda SM-A87, which was isolated from deep-sea sediment, is an aerobic, gram-negative bacterium that represents a new genus of Flavobacteriaceae. This is the first sequenced genome of a deep-sea bacterium from the phylum Bacteroidetes., Results: The Z. profunda SM-A87 genome has a single 5 128 187-bp circular chromosome with no extrachromosomal elements and harbors 4 653 predicted protein-coding genes. SM-A87 produces a large amount of capsular polysaccharides and possesses two polysaccharide biosynthesis gene clusters. It has a total of 130 peptidases, 61 of which have signal peptides. In addition to extracellular peptidases, SM-A87 also has various extracellular enzymes for carbohydrate, lipid and DNA degradation. These extracellular enzymes suggest that the bacterium is able to hydrolyze organic materials in the sediment, especially carbohydrates and proteinaceous organic nitrogen. There are two clustered regularly interspaced short palindromic repeats in the genome, but their spacers do not match any sequences in the public sequence databases. SM-A87 is a moderate halophile. Our protein isoelectric point analysis indicates that extracellular proteins have lower predicted isoelectric points than intracellular proteins. SM-A87 accumulates organic osmolytes in the cell, so its extracelluar proteins are more halophilic than its intracellular proteins., Conclusion: Here, we present the first complete genome of a deep-sea sedimentary bacterium from the phylum Bacteroidetes. The genome analysis shows that SM-A87 has some common features of deep-sea bacteria, as well as an important capacity to hydrolyze sedimentary organic nitrogen.

Nearly all high-molecular-weight (HMW) dissolved organic nitrogen and part of the particulate organic nitrogen in the deep sea are present in hydrolysis-resistant amides, and so far the mechanisms of biodegradation of these types of nitrogen have not been resolved. The M12 family is the second largest family in subclan MA(M) of Zn-containing metalloproteases and includes most enzymes from animals and only one enzyme (flavastacin) from a human-pathogenic bacterium (Flavobacterium meningosepticum). Here, we characterized the novel M12 protease myroilysin with elastinolytic activity and collagen-swelling ability from the newly described deep-sea bacterium Myroides profundi D25. Myroilysin is a monomer enzyme with 205 amino acid residues and a molecular mass of 22,936 Da. It has the same conserved residues at the four zinc ligands as astacin and very low levels of identity (