Harenicola maris gen. nov., sp. nov. isolated from the Sea of Japan shallow sediments.

A Gram-negative, non-motile bacterium КMM 3653T was isolated from a sediment sample from the Sea of Japan seashore, Russia. On the basis of the 16S rRNA gene sequence analysis the strain КMM 3653T was positioned within the family Rhodobacteraceae (class Alphaproteobacteria) forming a distinct lineage with the highest gene sequence similarities to the members of the genera Pacificibacter (95.2-94.7%) and Nioella (95.1-94.5%), respectively. According to the phylogenomic tree based on 400 conserved protein sequences, strain КMM 3653T was placed in the cluster comprising Vannielia litorea, Nioella nitratireducens, Litoreibacter albidus and Pseudoruegeria aquimaris as a separate lineage adjacent to V. litorea KCTC 32083T. The average nucleotide identity values between strain КMM 3653T and V. litorea KCTC 32083T, N. nitratireducens KCTC 32417T, L. albidus KMM 3851T, and P. aquimaris CECT 7680T were 71.1, 70.3, 69.6, and 71.0%, respectively. Strain КMM 3653T contained Q-10 as the predominant ubiquinone and C18:1ω7c as the major fatty acid followed by C16:0. The polar lipids were phosphatidylcholine, phosphatidylglycerol, diphosphatidylglycerol, an unidentified phospholipid, two unidentified aminolipids, and five unidentified lipids. The DNA G+C content of 61.8% was calculated from the genome sequence. Based on the phylogenetic evidence and distinctive phenotypic characteristics, we proposed strain KMM 3653T (= KCTC 82575T) to be classified as a novel genus and species Harenicola maris gen. nov., sp. nov.

Thalassobius aquimarinus sp. nov., isolated from the Sea of Japan seashore.

An aerobic, Gram-negative, non-pigmented non-motile bacterium designed КMM 8518T was isolated from a seawater sampled from the Sea of Japan seashore. Strain КMM 8518T grew at 7-42 °C and in the presence of 1-7% NaCl. The phylogenetic analyses based on 16S rRNA gene and whole-genome sequences placed the novel strain КMM 8518T into the genus Thalassobius as a separate lineage. Strain КMM 8518T shared the highest 16S rRNA gene sequence similarity of 98% to Thalassobius gelatinovorus KCTC 22092T and similarity values of ≤ 97% to other recognized Thalassobius species. The average nucleotide identity and digital DNA-DNA hybridization values between strain КMM 8518T and T. gelatinovorus KCTC 22092T were 79.6% and 23.5%, respectively. The major respiratory quinone was ubiquinone-10. The major fatty acid was C18:1ω7c followed by 11-methyl C18:1ω7c. Polar lipids comprised phosphatidylcholine, phosphatidylglycerol, diphosphatidylglycerol, an unidentified aminolipid, an unidentified phospholipid, and three unidentified lipids. The DNA G+C content of 62.7% was calculated from genome sequence analysis. Based on the phylogenetic analyses and distinctive phenotypic characteristics, the marine bacterium КMM 8518T is concluded to represent a novel species of the genus Thalassobius for which the name Thalassobius aquimarinus sp. nov. is proposed. The type strain of the species is strain KMM 8518T (= KCTC 82576T).

Characterization of Labrenzia polysiphoniae sp. nov. isolated from red alga Polysiphonia sp.

A group of five Gram-negative aerobic halophilic bacteria was isolated from the red alga Polysiphonia sp. specimen collected from the Sea of Japan seashore and subjected to a taxonomic study. On the basis of 16S rRNA gene sequence analysis, the novel isolates were affiliated to the genus Labrenzia sharing the highest gene sequence similarities of 98.1-98.4% with the type strain of Labrenzia suaedae KACC 13772T. The DNA-DNA hybridization values of 83-91% obtained between five novel strains, and 26 and 36% between two of the five novel strains and the closest neighbor Labrenzia suaedae KACC 13772T confirmed their assignment to the same separate species. Novel isolates were characterized by Q-10 as the major ubiquinone, by the predominance of C18:1ω7c followed by 11-methyl C18:1ω7c and C14:0 3-ОН in their fatty acid profiles. Polar lipids consisted of phosphatidylcholine, phosphatidylethanolamine, phosphatidylmonomethylethanolamine, phosphatidylglycerol, an unknown aminophospholipid, and an unknown phospholipid. Some of novel strains were found to inhibit growth of Gram-negative and Gram-positive test microorganisms. On the basis of phylogenetic analysis, DNA-DNA hybridization and phenotypic traits, a novel species with the name Labrenzia polysiphoniae sp. nov. (type strain KMM 9699T = rh46T = KACC 19711T), is proposed.

Winogradskyella profunda sp. nov. isolated from the Chukchi Sea bottom sediments.

An aerobic, Gram-negative, yellow-pigmented non-motile rod-shaped bacterium, designated Ch38T, was isolated from a sediment sample collected from the Chukchi Sea in the Arctic Ocean. Comparative 16S rRNA gene sequence analysis positioned strain Ch38T into the genus Winogradskyella as a distinct line adjacent to Winogradskyella multivorans KCTC 23891T, sharing the highest similarities of 97.5%, 97.2%, and 97.1% with Winogradskyella eximia KCTC 12219T, Winogradskyella damuponensis KCTC 23552T, and Winogradskyella multivorans KCTC 23891T, respectively. Strain Ch38T grew at 5-36 °C and in the presence of 1-6% (w/v) NaCl. It contained MK-6 as the predominant menaquinone and iso-C16:0 3-OH, anteiso-C15:0 followed by iso-C15:0 and iso-C16:1 as the major fatty acids. Polar lipids consisted of phosphatidylethanolamine, three unknown aminolipids, an unknown lipid and an unknown phospholipid. The DNA C + C content was 31.7 mol%. Based on the phylogenetic analysis and distinctive phenotypic characteristics, strain Ch38T is concluded to represent a novel species of the genus Winogradskyella, for which the name Winogradskyella profunda sp. nov. is proposed. The type strain of the species is strain Ch38T (= KMM 9725T = KACC 19710T).

Winogradskyella algae sp. nov., a marine bacterium isolated from the brown alga.

An aerobic, Gram-negative, yellow-pigmented non-motile rod-shaped bacterium Kr9-9T was isolated from a brown alga specimen collected near the Kuril Islands. Based on the 16S rRNA gene sequence analysis strain Kr9-9T was assigned to the genus Winogradskyella, and its close phylogenetic neighbors were found to be Winogradskyella damuponensis KCTC 23552T, Winogradskyella sediminis LMG 28075T, and Winogradskyella rapida CCUG 59098T showing high similarities of 98.1%, 97.5%, and 97.1%, respectively. It contained MK-6 as the predominant menaquinone and iso-C15:0, anteiso-C15:0, iso-C16:0 3-OH followed by iso-C15:1 as the major fatty acids. Polar lipids included phosphatidylethanolamine, three unidentified aminolipids and an unidentified lipid. The DNA C+C content was 32.3 mol%. Based on the phylogenetic analysis and distinctive phenotypic characteristics, strain Kr9-9T is concluded to represent a novel species of the genus Winogradskyella, for which the name Winogradskyella algae sp. nov. is proposed. The type strain of the species is strain Kr9-9T (= KMM 8180T = KACC 19709T).

Flavobacterium maris sp. nov. isolated from shallow sediments of the Sea of Japan.

An aerobic, gram-negative, yellow-pigmented, and non-motile bacterium designated KMM 9535(T) was isolated from a marine sediment sample obtained from the Sea of Japan seashore and subjected to a phylogenetic and phenotypic study. On the basis of 16S rRNA gene sequence analysis, strain KMM 9535(T) was placed to the genus Flavobacterium sharing the highest sequence similarities to Flavobacterium ahnfeltiae KCTC 32467(T) (99.3%), Flavobacterium jumunjinense KCTC 23618(T) (96.5%), Flavobacterium ponti KCTC 22802(T) (96.3%), Flavobacterium urocaniciphilum JCM 19142(T) (96.1%), and Flavobacterium gelidilacus LMG 21477(T) (95.8%). The DNA-DNA hybridization value between strain KMM 9535(T) and the closest related F. ahnfeltiae KCTC 32467(T) was 33%. Strain KMM 9535(T) grew at 5-36 °C and in the presence of 0-3% (w/v) NaCl. It contained MK-6 as the predominant menaquinone, and the major fatty acids were iso-C15:0, iso-C17:1, iso-C15:1, and iso-C17:0 3-OH. The DNA G+C content was 28.8 mol%. On the basis of the results obtained, it is proposed strain KMM 9535(T) to be classified as a novel species of the genus Flavobacterium, Flavobacterium maris sp. nov., with the type strain of the species KMM 9535(T) (=NRIC 0920(T) = KCTC 42093(T)).

Tamlana sedimentorum sp. nov., isolated from shallow sand sediments of the Sea of Japan.

An aerobic, Gram-stain-negative, saffron-pigmented, non-motile bacterial strain KMM 9545(T), was isolated from a marine sand sediment sample obtained from the Sea of Japan seashore and characterized in a taxonomic study using a polyphasic approach. The 16S rRNA gene sequence of strain KMM 9545(T) showed a high level of similarity to species of the genus Gaetbulibacter (95.1-96.2%), the type strains of species of the genus Tamlana (94.9-96.1%) and members of the genus Algibacter (94.8-96.1%). Phylogenetic analysis based on the 16S rRNA gene sequences positioned strain KMM 9545(T) as a distinct lineage in the cluster comprising species of the genus Tamlana. Strain KMM 9545(T) grew at temperatures between 5-36 °C and in the presence of 2-4% (w/v) NaCl. It contained MK-6 as the predominant menaquinone and iso-C(15 : 0), iso-C(15 : 1), iso-C(17 : 1) 3-OH, iso-C(15 : 0) 3-OH and iso-C(15 : 0) 2-OH as the major fatty acids. The genomic DNA G+C content was 31.3 mol%. On the basis of the phenotypic characteristics and phylogenetic distance, it can be concluded that strain KMM 9545(T) represents a novel species of the genus Tamlana, for which the name Tamlana sedimentorum sp. nov. is proposed. The type strain is KMM 9545(T) ( = NRIC 0921(T) = JCM 19808(T)).

Loktanella maritima sp. nov. isolated from shallow marine sediments.

An aerobic, Gram-stain-negative, non-motile bacterium, KMM 9530(T), was isolated from a sediment sample collected from the Sea of Japan seashore. Comparative 16S rRNA gene sequence analysis positioned novel strain KMM 9530(T) in the genus Loktanella as a separate line adjacent to Loktanella sediminilitoris KCTC 32383(T), Loktanella tamlensis JCM 14020(T) and Loktanella maricola JCM 14564(T) with 98.5-98.2% sequence similarity. Strain KMM 9530(T) was characterized by its weak hydrolytic capacity and inability to assimilate most organic substrates. The major isoprenoid quinone was Q-10, polar lipids consisted of phosphatidylcholine, phosphatidylglycerol, diphosphatidylglycerol, an unknown phospholipid, an unknown aminolipid and unknown lipids, and the major fatty acid was C18 : 1ω7c. On the basis of phylogenetic analysis, DNA-DNA hybridization and phenotypic characterization, it can be concluded that the novel strain KMM 9530(T) represents a novel species in the genus Loktanella, for which the name Loktanella maritima sp. nov. is proposed. The type strain of the species is KMM 9530(T) ( = NRIC 0919(T) = JCM 19807(T)).

Luteimonas vadosa sp. nov., isolated from seashore sediment.

An aerobic, Gram-stain-negative, yellow-pigmented, non-motile, rod-shaped bacterium designated strain KMM 9005(T) was isolated from a sediment sample collected from the Sea of Japan seashore. Comparative 16S rRNA gene sequence analysis showed that strain KMM 9005(T) belonged to the genus Luteimonas and was most closely related to Luteimonas cucumeris KCTC 23627(T) (96.5 % sequence similarity) and Luteimonas aquatica LMG 24212(T) (96.1 % sequence similarity). Strain KMM 9005(T) was characterized by the presence of thin fimbriae, the major ubiquinone Q-8, by the predominance of iso-C17 : 1 followed by iso-C16 : 0, iso-C15 : 0 and iso-C17 : 0 in its fatty acid profile, weak hydrolytic capacity and the inability to assimilate most organic substrates. Based on these distinctive phenotypic characteristics and phylogenetic analysis, strain KMM 9005(T) represents a novel species of the genus Luteimonas, for which the name Luteimonas vadosa sp. nov. is proposed. The type strain is KMM 9005(T) ( = NRIC 0881(T) = JCM 18392(T)).

Alteromonas australica sp. nov., isolated from the Tasman Sea.

A non-pigmented, motile, Gram-negative bacterium designated H 17(T) was isolated from a seawater sample collected in Port Phillip Bay (the Tasman Sea, Pacific Ocean). The new organism displayed optimal growth between 4 and 37 °C, was found to be neutrophilic and slightly halophilic, tolerating salt water environments up to 10 % NaCl. Strain H 17(T) was found to be able to degrade starch and Tween 80 but unable to degrade gelatin or agar. Phosphatidylglycerol (27.7 %) and phosphatidylethanolamine (72.3 %) were found to be the only associated phospholipids. The major fatty acids identified are typical for the genus Alteromonas and include C16:0, C16:1ω7, C17:1ω8 and C18:1ω7. The G+C content of the DNA was found to be 43.4 mol%. A phylogenetic study, based on the 16S rRNA gene sequence analysis and Multilocus Phylogenetic Analysis, clearly indicated that strain H 17(T) belongs to the genus Alteromonas. The DNA-DNA relatedness between strain H 17(T) and the validly named Alteromonas species was between 30.7 and 46.4 mol%. Based on these results, a new species, Alteromonas australica, is proposed. The type strain is H 17(T) (= KMM 6016(T) = CIP 109921(T)).

Hydrolysis of fucoidan by fucoidanase isolated from the marine bacterium, Formosa algae.

Intracellular fucoidanase was isolated from the marine bacterium, Formosa algae strain KMM 3553. The first appearance of fucoidan enzymatic hydrolysis products in a cell-free extract was detected after 4 h of bacterial growth, and maximal fucoidanase activity was observed after 12 h of growth. The fucoidanase displayed maximal activity in a wide range of pH values, from 6.5 to 9.1. The presence of Mg2+, Ca2+ and Ba2+ cations strongly activated the enzyme; however, Cu2+ and Zn2+ cations had inhibitory effects on the enzymatic activity. The enzymatic activity of fucoidanase was considerably reduced after prolonged (about 60 min) incubation of the enzyme solution at 45 °C. The fucoidanase catalyzed the hydrolysis of fucoidans from Fucus evanescens and Fucus vesiculosus, but not from Saccharina cichorioides. The fucoidanase also did not hydrolyze carrageenan. Desulfated fucoidan from F. evanescens was hydrolysed very weakly in contrast to deacetylated fucoidan, which was hydrolysed more actively compared to the native fucoidan from F. evanescens. Analysis of the structure of the enzymatic products showed that the marine bacteria, F. algae, synthesized an α-l-fucanase with an endo-type action that is specific for 1→4-bonds in a polysaccharide molecule built up of alternating three- and four-linked α-l-fucopyranose residues sulfated mainly at position 2.

Fucoidan Sulfatases from Marine Bacterium Wenyingzhuangia fucanilytica CZ1127T.

Fucoidans belong to a structurally heterogeneous class of sulfated polysaccharides isolated from brown algae. They have a wide spectrum of biological activities. The complex structures of these polysaccharides hinder structure-activity relationships determination. Fucoidan sulfatases can make useful tools for the determination of the fine chemical structure of fucoidans. In this study, identification and preparation of two recombinant sulfatases able to catalyze the cleavage of sulfate groups from fragments of fucoidan molecules is described for the first time. Two genes of sulfatases swf1 and swf4 of the marine bacterium Wenyingzhuangia fucanilytica CZ1127T were cloned and the proteins were produced in Escherichia coli cells. Sulfatases SWF1 and SWF4 are assigned to S1_17 and S1_25 subfamilies of formylglycine-dependent enzymes of S1 family (SulfAtlas). Some molecular and biochemical characteristics of recombinant fucoidan sulfatases have been studied. Detailed specificity and catalytic features of sulfatases were determined using various sulfated fucooligosaccharides. Structures of products produced by SWF1 and SWF4 were established by nuclear magnetic resonance (NMR) spectroscopy. Based on the obtained data, the enzymes are classified as fucoidan exo-2O-sulfatase (SWF1) and fucoidan exo-3O-sulfatase (SWF4). In addition, we demonstrated the sequential action of sulfatases on 2,3-di-O-sulfated fucooligosacchrides, which indicates an exolitic degradation pathway of fucoidan by a marine bacterium W. fucanilytica CZ1127T.

Celeribacter neptunius gen. nov., sp. nov., a new member of the class Alphaproteobacteria.

A whitish Gram-negative, motile, aerobic bacterium, designated strain H 14(T), was isolated from seawater collected at St Kilda beach in Port Phillip Bay, Melbourne, Australia. Analysis of 16S rRNA gene sequences revealed that the organism belonged to the Roseobacter lineage of the class Alphaproteobacteria, forming a distinct evolutionary lineage at the genus level. Strain H 14(T) was distantly related to the genera Nautella, Ruegeria and Pseudoruegeria (family Rhodobacteraceae). Strain H 14(T) was unable to degrade gelatin, casein, chitin, agar and starch, did not produce any carotenoids, did not possess bacteriochlorophyll a and had a limited ability to utilize carbon sources. Strain H 14(T) grew with concentrations of 1-8 % (w/v) NaCl and over a temperature range of 5-35 degrees C. Phosphatidylglycerol was the major phospholipid (90 %); phosphatidylcholine (7.9 %) and phosphatidylethanolamine (2.0 %) were present in minor quantities. The predominant fatty acids were C(18 : 1)omega7c (82.4 %), C(18 : 1)omega9c (5.1 %) and C(18 : 0) (3.8 %). The DNA G+C composition for strain H 14(T) was 59.1 mol%. Based on the results of physiological, biochemical, chemotaxonomic and phylogenetic investigations, a new genus, Celeribacter gen. nov., with the type species Celeribacter neptunius sp. nov. is proposed. The type strain of the type species is H 14(T) (=KMM 6012(T)=CIP 109922(T)).

Winogradskyella exilis sp. nov., isolated from the starfish Stellaster equestris, and emended description of the genus Winogradskyella.

A pale-yellowish-pigmented strain, 022-2-26(T), was isolated from a starfish, Stellaster equestris. Cells of strain 022-2-26(T) were Gram-negative short rods that were chemo-organotrophic, alkalitolerant and mesophilic. The predominant menaquinone was MK-6. The major cellular fatty acids were iso-C(15 : 0), iso-C(15 : 1), C(15 : 0), iso-C(15 : 0) 2-OH and iso-C(17 : 0) 3-OH (together representing 87 % of the total fatty acids). The DNA G+C content was 30.1 mol%. A 16S rRNA gene sequence of the isolate was determined and phylogenetic analyses revealed that strain 022-2-26(T) formed a robust clade (neighbour-joining algorithm with a bootstrap value of 95 % and parsimony and maximum-likelihood algorithms) with type strains of species in the genus Winogradskyella. The closest phylogenetic neighbour of strain 022-2-26(T) was Winogradskyella poriferorum UST030701-295(T) (96 % 16S rRNA gene sequence similarity; 59 differences between sequences). On the basis of the phenotypic and chemotaxonomic characteristics and the phylogenetic evidence, it is proposed that strain 022-2-26(T) represents a novel species, Winogradskyella exilis sp. nov. The type strain is 022-2-26(T) (=KMM 6013(T) =CIP 109976(T)).