Identification and evaluation of spoilage potential of four Bacillus strains isolated from slimy rice noodles.

This study isolated four strains of Bacillus from slimy fresh rice noodles (FRN) and preliminary identified them as B. cereus, B. amyloliquefaciens, B. altitudinis, and B. subtilis, respectively, using morphological, physiological, and genetic analyses. The spoilage potential of each of these strains was then evaluated in FRN. The results indicated that both B. amyloliquefaciens and B. subtilis can cause FRN to become sticky and smelly, accompanied by an increase of acidity and high amylase activity. B. cereus and B. altitudinis mainly caused odor deterioration of FRN. There were 29, 20, 25, 25, and 27 volatile organic compounds (VOCs) identified from control FRN and FRN samples inoculated with B. cereus, B. amyloliquefaciens, B. altitudinis, and B. subtilis, respectively. The compositions of VOCs in the samples inoculated with B. amyloliquefaciens and B. subtilis were similar; esters, volatile organic acids, and acetoin were the main volatile compounds in the FRN. For B. cereus and B. altitudinis, the main adverse flavor substances were acetic acid and ammonia. This study provides a theoretical basis for quality control in the production, storage, and sale of FRN.

Novel Antioxidant Peptides Identified from Arthrospira platensis Hydrolysates Prepared by a Marine Bacterium Pseudoalteromonas sp. JS4-1 Extracellular Protease.

Crude enzymes produced by a marine bacterium Pseudoalteromonas sp. JS4-1 were used to hydrolyze phycobiliprotein. Enzymatic productions showed good performance on DPPH radical and hydroxyl radical scavenging activities (45.14 ± 0.43% and 65.11 ± 2.64%, respectively), especially small peptides with MWCO <3 kDa. Small peptides were fractioned to four fractions using size-exclusion chromatography and the second fraction (F2) had the highest activity in hydroxyl radical scavenging ability (62.61 ± 5.80%). The fraction F1 and F2 both exhibited good antioxidant activities in oxidative stress models in HUVECs and HaCaT cells. Among them, F2 could upregulate the activities of SOD and GSH-Px and reduce the lipid peroxidation degree to scavenge the ROS to protect Caenorhabditis elegans under adversity. Then, 25 peptides total were identified from F2 by LC-MS/MS, and the peptide with the new sequence of INSSDVQGKY as the most significant component was synthetized and the ORAC assay and cellular ROS scavenging assay both illustrated its excellent antioxidant property.

Characteristics and Application of a Novel Cold-Adapted and Salt-Tolerant Protease EK4-1 Produced by an Arctic Bacterium Mesonia algae K4-1.

Mesonia algae K4-1 from the Arctic secretes a novel cold-adapted and salt-tolerant protease EK4-1. It has the highest sequence similarity with Stearolysin, an M4 family protease from Geobacillus stearothermophilus, with only 45% sequence identity, and is a novel M4 family protease. Ek4-1 has a low optimal catalytic temperature (40 °C) and is stable at low temperatures. Moreover, EK4-1 is still active in 4 mol/L NaCl solution and is tolerant to surfactants, oxidizing agents and organic solvents; furthermore, it prefers the hydrolysis of peptide bonds at the P1' position as the hydrophobic residues, such as Leu, Phe and Val, and amino acids with a long side chain, such as Phe and Tyr. Mn2+and Mg2+ significantly promoted enzyme activity, while Fe3+, Co+, Zn2+ and Cu2+ significantly inhibited enzyme activity. Amino acid composition analysis showed that EK4-1 had more small-side-chain amino acids and fewer large-side-chain amino acids. Compared with a thermophilic protease Stearolysin, the cold-adapted protease EK4-1 contains more random coils (48.07%) and a larger active pocket (727.42 Å3). In addition, the acidic amino acid content of protease EK4-1 was higher than that of the basic amino acid, which might be related to the salt tolerance of protease. Compared with the homologous proteases EB62 and E423, the cold-adapted protease EK4-1 was more efficient in the proteolysis of grass carp skin, salmon skin and casein at a low temperature, and produced a large number of antioxidant peptides, with DPPH, ·OH and ROO· scavenging activities. Therefore, cold-adapted and salt-tolerant protease EK4-1 offers wide application prospects in the cosmetic and detergent industries.

Genome-Wide Identification, Characterization and Experimental Expression Analysis of CNGC Gene Family in Gossypium.

Cyclic nucleotide-gated ion channels (CNGCs) are channel proteins for calcium ions, and have been reported to play important roles in regulating survival and environmental response of various plants. However, little is known about how the CNGC family works in Gossypium. In this study, 173 CNGC genes, which were identified from two diploid and five tetraploid Gossypium species, were classified into four groups by phylogenetic analysis. The collinearity results demonstrated that CNGC genes are integrally conservative among Gossypium species, but four gene losses and three simple translocations were detected, which is beneficial to analyzing the evolution of CNGCs in Gossypium. The various cis-acting regulatory elements in the CNGCs' upstream sequences revealed their possible functions in responding to multiple stimuli such as hormonal changes and abiotic stresses. In addition, expression levels of 14 CNGC genes changed significantly after being treated with various hormones. The findings in this study will contribute to understanding the function of the CNGC family in cotton, and lay a foundation for unraveling the molecular mechanism of cotton plants' response to hormonal changes.

Probing the Internal pH and Permeability of a Carboxysome Shell.

The carboxysome is a protein-based nanoscale organelle in cyanobacteria and many proteobacteria, which encapsulates the key CO2-fixing enzymes ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) and carbonic anhydrase (CA) within a polyhedral protein shell. The intrinsic self-assembly and architectural features of carboxysomes and the semipermeability of the protein shell provide the foundation for the accumulation of CO2 within carboxysomes and enhanced carboxylation. Here, we develop an approach to determine the interior pH conditions and inorganic carbon accumulation within an α-carboxysome shell derived from a chemoautotrophic proteobacterium Halothiobacillus neapolitanus and evaluate the shell permeability. By incorporating a pH reporter, pHluorin2, within empty α-carboxysome shells produced in Escherichia coli, we probe the interior pH of the protein shells with and without CA. Our in vivo and in vitro results demonstrate a lower interior pH of α-carboxysome shells than the cytoplasmic pH and buffer pH, as well as the modulation of the interior pH in response to changes in external environments, indicating the shell permeability to bicarbonate ions and protons. We further determine the saturated HCO3- concentration of 15 mM within α-carboxysome shells and show the CA-mediated increase in the interior CO2 level. Uncovering the interior physiochemical microenvironment of carboxysomes is crucial for understanding the mechanisms underlying carboxysomal shell permeability and enhancement of Rubisco carboxylation within carboxysomes. Such fundamental knowledge may inform reprogramming carboxysomes to improve metabolism and recruit foreign enzymes for enhanced catalytical performance.

Assessment of features in facial hyperpigmentation: Comparison study between VISIA and CSKIN.

BACKGROUND: Hyperpigmentary disorder is one of the commonest skin concerns in dermatology clinics. The availability of noninvasive instruments provided a convenient, objective, and reproducible methodology for the evaluation of pigmentation and skin color. The aim of this study is to compare CSKIN and VISIA in measuring facial hyperpigmentation, as well as to assess the correlation between the instrumental analyzing and clinical evaluation. METHODS: Eighty Chinese patients were enrolled. Images were taken and analyzed by VISIA from Canfield and CSKIN from Yanyun Technology, and the facial hyperpigmentation was graded by three dermatologists. RESULTS: Feature counts within the facial pigmented areas analyzed by VISIA showed positive correlations with brown pixels (r = 0.331, p < 0.05) and brown percent (r = 0.395, p < 0.0001) measured by CSKIN. The parameters measured by CSKIN and VISIA were significantly correlated with visual scores graded by the dermatologists, with VISIA presenting a moderate correlation (r = 0.509, p < 0.001) and CSKIN a slightly stronger correlation with the visual scores (r = 0.653, p < 0.001). CONCLUSION: CSKIN could serve as an alternative in the assessment and follow-up of skin disease featuring with facial hyperpigmentation.

The response regularity of biohydrogen production by anthracite H2-producing bacteria consortium to six conventional veterinary antibiotics.

The impact of antibiotics on H2-producing bacteria must be considered in the industrialization of biological H2 production using livestock manure as raw resources. However, whether antibiotics that may be contained in excreta will threaten the safety of biohydrogen production needs to be researched. This study explored the impact characteristics and mechanism of six single antibiotics and three groups of compound antibiotics on H2 production. Experiments confirmed that most antibiotics have different degrees of H2 production inhibition, while some antibiotics, which like Penicillin G, Streptomycin Sulfate, and their compound antibiotics, could promote the growth of Ethanoligenens sp. and improve H2 yield on the contrary. Comprehensive analysis shows that the main inhibitory mechanisms were: (1) board-spectrum inhibition, (2) partial inhibition, (3) H2 consumption enhancement; and the enhancement mechanisms were: (1) enhance the growth of H2-producing bacteria, (2) enhanced starch hydrolysis, (3) inhibitory H2 consumption or release of acid inhibition. Meanwhile, experiment found that the effect of antibiotics on H2 producing was not only related to type, but also to dosage. Even one kind of antibiotic may have completely opposite effects on H2-producing bacteria under different dosage conditions. Inhibition of H2 yield was highest with Levofloxacin at 6.15 mg/L, gas production was reduced by 88.77%; and enhancement of H2 yield was highest with Penicillin G at 7.20 mg/L, the gas production increased by 72.90%. In the selection of raw material, the type and content of antibiotics demand a detailed investigation and analysis to ensure that the sustainability of H2 yield.

Recent Advances in Detection for Breast-Cancer-Derived Exosomes.

Breast cancer is the most common malignant tumor in women, its incidence is secret, and more than half of the patients are diagnosed in the middle and advanced stages, so it is necessary to develop simple and efficient detection methods for breast cancer diagnosis to improve the survival rate and quality of life of breast cancer patients. Exosomes are extracellular vesicles secreted by all kinds of living cells, and play an important role in the occurrence and development of breast cancer and the formation of the tumor microenvironment. Exosomes, as biomarkers, are an important part of breast cancer fluid biopsy and have become ideal targets for the early diagnosis, curative effect evaluation, and clinical treatment of breast cancer. In this paper, several traditional exosome detection methods, including differential centrifugation and immunoaffinity capture, were summarized, focusing on the latest research progress in breast cancer exosome detection. It was summarized from the aspects of optics, electrochemistry, electrochemiluminescence and other aspects. This review is expected to provide valuable guidance for exosome detection of clinical breast cancer and the establishment of more reliable, efficient, simple and innovative methods for exosome detection of breast cancer in the future.

Diversity of protease-producing bacteria in the soils of the South Shetland Islands, Antarctica.

Proteases secreted from bacteria into soil play a key role in the degradation of organic nitrogen, which is the first and, usually, the rate-limiting step of nitrogen cycling. However, the diversity of protease-producing bacteria and their excreted proteases in Antarctic soil have not yet been fully explored. Here we studied 20 soil samples from the South Shetland Islands, Antarctica and isolated 253 strains with protease activity. These protease-producing bacteria belonged to the phyla Bacteroidetes, Proteobacteria, Actinobacteria, Firmicutes, and Deinococcus-Thermus. Thhe predominant genera were Arthrobacter (14.9%), Chryseobacterium (14.5%), Flavobacterium (14.5%), and Pseudomonas (14.5%). Most of these bacteria secreted serine proteases and metalloproteases. There was quite a large distribution in activity as quantified by protease and inhibition assays. Only a few strains secreted aspartic and/or cysteine proteases. Together these data provided novel insight into the diversity and mechanism of organic nitrogen degradation in Antarctic soils by various proteases, which may have potential in new biotechnological applications.

A Label-Free Fluorometric Glutathione Assay Based on a Conformational Switch of G-quadruplex.

In this paper, a label-free fluorescent method for glutathione (GSH) detection based on a thioflavin T/G-quadruplex conformational switch is developed. The sensing assay is fabricated depending on the virtue of mercury ions to form a thymine-thymine mismatch, which collapses the distance between two ssDNA and directs the guanine-rich part to form an intra-strand asymmetric split G-quadruplex. The newly formed G-quadruplex efficiently reacts with thioflavin T and enhances the fluorescent intensity. In the presence of GSH, Hg2+ is absorbed, destroying the G-quadruplex formation with a significant decrease in fluorescence emission. The proposed fluorescent assay exhibits a linear range between 0.03-5 µM of GSH with a detection limit of 9.8 nM. Furthermore, the efficacy of this method is examined using human serum samples to detect GSH. Besides GSH, other amino acids are also investigated in standard samples, which display satisfactory sensitivity and selectivity. Above all, we develop a method with features including potentiality, facility, sensitivity, and selectivity for analyzing GSH for clinical diagnostics.

Analysis of facial redness by comparing VISIA® from Canfield and CSKIN® from Yanyun Technology.

BACKGROUND: Redness is the most common symptom among many facial dermatoses. With the rapid development of optical instruments, spectral imaging, and image processing technology, there appear varieties of skin color analysis methods and instruments. The aim of this study is to reveal the differences and correlations in measuring the facial redness between CSKIN® and VISIA® , as well as the relevance between the instrument parameters and clinical evaluation. MATERIALS & METHODS: Forty-three Chinese patients were enrolled. Images were taken and analyzed by VISIA® from Canfield and CSKIN® from Yanyun Technology, and the facial erythema was graded by the dermatologists. RESULTS: Feature counts within the red areas measured by VISIA® were found to have significantly positive correlations with red pixels and percent which were measured by CSKIN® on both sides of the face (r = .45 ~ .566, P < .01). The parameters analyzed by CSKIN® and VISIA® feature counts were correlated with visual scores graded by the dermatologists, VISIA® presented with a weak correlation (r = .213, P < .05), while CSKIN® had a moderate correlation with the visual scores (r = .472 ~ .492, P < .001). CONCLUSION: CSKIN® may be another alternative option when encountering with measurement and follow-up of facial erythema.

Novel PMM2 missense mutation in a Chinese family with non-syndromic premature ovarian insufficiency.

PURPOSE: This study sought to identify a disease-related gene in a consanguineous Chinese family in which there were two premature ovarian insufficiency (POI) sisters. METHOD: We used whole-exome sequencing and Sanger sequencing to identify the disease-causing gene. Results were verified using an assay of mutant protein and in silico analyses. RESULT: We identified a novel missense mutation (NM_000303: c.556G>A, p.Gly186Arg) in the PMM2 gene. The two sisters suffer from premature ovarian insufficiency (POI) only and have no other symptoms of congenital disorder of glycosylation type-1a (CDG-Ia). We found that the enzymic activity of the mutant PMM2 protein was reduced by 55.21% (p < 0.05) when compared with wild type, and many in silico tools suggested the mutation is disease-related. CONCLUSION: This particular gene modification results in changes in activity of phosphomannomutase modification, which could lead to PMM2-CDG-Ia with an uncommon phenotype.

Mechanistic Insight into the Binding and Swelling Functions of Prepeptidase C-Terminal (PPC) Domains from Various Bacterial Proteases.

The bacterial prepeptidase C-terminal (PPC) domain can be found in the C termini of a wide variety of proteases that are secreted by marine bacteria. However, the functions of these PPC domains remain unknown due to a lack of systematic research. Here, the binding and swelling abilities of eight PPC domains from six different proteases were compared systematically via scanning electron microscopy (SEM), enzyme assays, and fluorescence spectroscopy. These PPC domains all possess the ability to bind and swell insoluble collagen. PPC domains can expose collagen monomers but cannot disrupt the pyridinoline cross-links or unwind the collagen triple helix. This ability can play a synergistic role alongside collagenase in collagen hydrolysis. Site-directed mutagenesis of the PPC domain from Vibrio anguillarum showed that the conserved polar and aromatic residues Y6, D26, D28, Y30, W42, E53, C55, and Y65 and the hydrophobic residues V10, V18, and I57 played key roles in substrate binding. Molecular dynamic simulations were conducted to investigate the interactions between PPC domains and collagen. Most PPC domains have a similar mechanism for binding collagen, and the hydrophobic binding pocket of PPC domains may play an important role in collagen binding. This study sheds light on the substrate binding mechanisms of PPC domains and reveals a new function for the PPC domains of bacterial proteases in substrate degradation.IMPORTANCE Prepeptidase C-terminal (PPC) domains commonly exist in the C termini of marine bacterial proteases. Reports examining PPC have been limited, and its functions remain unclear. In this study, eight PPCs from six different bacteria were examined. Most of the PPCs possessed the ability to bind collagen, feathers, and chitin, and all PPCs could significantly swell insoluble collagen. PPCs can expose collagen monomers but cannot disrupt pyridinoline cross-links or unwind the collagen triple helix. This swelling ability may also play synergistic roles in collagen hydrolysis. Comparative structural analyses and the examination of PPC mutants revealed that the hydrophobic binding pockets of PPCs may play important roles in collagen binding. This study provides new insights into the functions and ecological significance of PPCs, and the molecular mechanism of the collagen binding of PPCs was clarified, which is beneficial for the protein engineering of highly active PPCs and collagenase in the pharmaceutical industry and of artificial biological materials.

Comprehensive analysis of the phylogeny and extracellular proteases in genus Vibrio strain.

As one of the dominant bacteria in the ocean, Vibrio play important roles in maintaining the aquatic ecosystem. In this study, we studied the phylogenetic relationships of 32 Vibrio based on the 16S rRNA genes sequences and utilized substrate immersing zymography method to detect the trend of protease production and components of multiprotease system of Vibrio extracellular proteases. The result showed that different extracellular proteolytic profiles among various Vibrio strains demonstrated a large interspecific variation, and for strains from the same environments, the closer the evolutionary relationship of them, the more similar their zymograms were. In addition, these proteases displayed very different hydrolysis abilities to casein and gelatin. Moreover, the results of the inhibitor-substrate immersing zymography indicated that the proteases secreted by marine Vibrio mostly belonged to serine proteases or metalloproteases. These results implied that combined taxonomic information of the Vibrios with their extracellular protease zymograms maybe contributed to the study of the classification, phylogeny and pathogenic mechanism of Vibrio, and can serve as a theoretical basis for controlling the pathogenic Vibrio disease as well as exploiting proteases. More importantly, we can also eliminate many similar strains by this way, thus can greatly reduce the workload of the experiments for us.

Detection of coralyne and heparin by polymerase extension reaction using SYBR Green I.

Polydeoxyadenosine (poly (dA)) has been extensively applied for detecting many drug molecules. Herein, we developed a sensitive method for detecting coralyne and heparin using a modified DNA probe with poly (dA) at one end. In the absence of coralyne, the DNA probe was digested by the Exonuclease I (Exo I), and therefore the SYBR Green I (SG I) emitted an extremely low fluorescent signal. While coralyne specifically binding to poly (dA) with strong propensity could remarkably restrain the disintegration of the DNA probe, through which as a template the second strand of DNA sequence was formed with the introduction of DNA polymerase. Therefore, the fluorescent signal of SG I was intensified to quantify coralyne. Based on this method, heparin can be determined due to its strong affinity towards coralyne. This method showed a linear range from 2 to 500 nM for coralyne with a low detection limit of 0.98 nM, and the linear range of heparin was from 1 to 100 nM when 1.25 nm was the detection limit. The proposed method was also implemented successfully in biological samples and showed a potential application for screening potential therapeutic molecules.

Fluorometric determination of the activity of uracil-DNA glycosylase by using graphene oxide and exonuclease I assisted signal amplification.

The base-excision repair enzyme uracil-DNA glycosylase (UDG) plays a crucial role in the maintenance of genome integrity. The authors describe a fluorometric method for the detection of the activity of UDG. It is making use of (a) a 3'-FAM-labeled hairpin DNA probe with two uracil deoxyribonucleotides in the self-complementary duplex region of its hairpin structure, (b) exonuclease I (Exo I) that catalyzes the release of FAM from the UDG-induced stretched ssDNA probe, and (c) graphene oxide that quenches the green FAM fluorescence of the intact hairpin DNA probe in the absence of UDG. If Exo I causes the release of FAM from the hairpin DNA probe, the fluorescence peaking at 517 nm is turned off in the absence of UDG but turned on in its presence. The resulting assay has a wide linear range (0.008 to 1 U·mL-1) and a detection limit as low as 0.005 U·mL-1. It has good specificity for UDG over potentially interfering enzymes and gave satisfactory results when applied to biological samples. Conceivably, the method may be used in a wide range of applications such as in diagnosis, drug screening, and in studying the repair of DNA lesions. Graphical abstract Schematic presentation of a fluorometric strategy for detection of the activity of uracil-DNA glycosylase by using on graphene oxide and exonuclease I assisted signal amplification.

Purification, Characterization, and Application for Preparation of Antioxidant Peptides of Extracellular Protease from Pseudoalteromonas sp. H2.

The study reported on the isolation of a metalloprotease named EH2 from Pseudoalteromonas sp. H2. EH2 maintained more than 80% activity over a wide pH range of 5-10, and the stability was also nearly independent of pH. Over 65% activity was detected at a wide temperature range of 20-70 °C. The high stability of the protease in the presence of different surfactants and oxidizing agents was also observed. Moreover, we also investigated the antioxidant activities of the hydrolysates generated from porcine and salmon skin collagen by EH2. The results showed that salmon skin collagen hydrolysates demonstrated higher DPPH (1,1-diphenyl-2-picrylhydrazyl) (42.88% ± 1.85) and hydroxyl radical (61.83% ± 3.05) scavenging activity than porcine skin collagen. For oxygen radical absorbance capacity, the hydrolysates from porcine skin collagen had higher efficiency (7.72 ± 0.13 µmol·TE/µmol). Even 1 nM mixed peptides could effectively reduce the levels of intracellular reactive oxygen species. The two types of substrates exerted the best antioxidant activity when hydrolyzed for 3 h. The hydrolysis time and type of substrate exerted important effects on the antioxidant properties of hydrolysates. The hydrolyzed peptides from meat collagens by proteases have good antioxidant activity, which may have implications for the potential application of marine proteases in the biocatalysis industry.

Classification and structural insight into vibriolysin-like proteases of Vibrio pathogenicity.

Vibriolysin-like proteases (VLPs) are important virulence agents in the arsenal of Vibrio causing instant cytotoxic effects during infection. Most of Vibrio secreted VLPs show serious pathogenicity, while some species of Vibrio with VLPs are non-pathogenic, like Vibrio tasmaniensis and Vibrio pacinii. To investigate the relation between VLPs and Vibrio pathogenicity, one phylogenetic tree of VLPs was constructed and compared consensus sequences at the N-terminus of VLPs. Based on these results, VLPs were defined into nine phylogenetic clades. Pathogenicity analysis of Vibrio showed that Vibrio species with VLPs III, VI, VII or VIII are serious pathogenic bacteria, while species with VLPs I, II, IV or IX are opportunistic pathogens. Multiple sequence alignment showed that the N-terminal 5-16 nucleotides of each clade are highly conservative. Topological analysis of VLPs exhibited the structural differences in N-terminal regions of each VLP clade. These results suggest that structure of N-terminus might play a key role in the pathogenicity of VLPs. Our findings give new insights into the classification of VLPs and the relationship between VLPs and Vibrio pathogenicity.

Flavobacterium phocarum sp. nov., isolated from soils of a seal habitat in Antarctica.

A Gram-stain-negative, yellow-pigmented, non-flagellated, gliding, rod-shaped, oxidase-negative and catalase-positive bacterium, designated SE14T, was isolated from soil on King George Island, South Shetland Islands, Antarctica. Strain SE14T grew at 4-25 °C (optimum, 20 °C), at pH 6.0-9.0 (optimum, pH 7.0-7.5) and with 0-3.0 % NaCl (optimum, 1.0-1.5 %), and could not produce flexirubin-type pigments. 16S rRNA gene sequence analysis showed the the isolate belonged to the genus Flavobacterium. Strain SE14T had the highest 16S rRNA gene sequence similarity to Flavobacterium antarcticum, F. tegetincola and F. degerlachei with 95.8, 95.5 and 95.2 %, respectively. The strain SE14T consisted of a clade with Flavobacteriumnoncentrifugens (16S rRNA gene sequence similarity 94.9 %) and F. qiangtangense (16S rRNA gene sequence similarity 94.2 %) and simultaneously formed a distinct phyletic lineage in the neighbour-joining phylogenetic tree. Polar lipids of the strain included phosphatidylethanolamine and four unidentified aminolipids. Strain SE14T contained anteiso-C15 : 0, iso-C15 : 0 and a mixture of iso-C15 : 0 2-OH and/or C16 : 1ω7c as the main fatty acids, and the only respiratory quinone was menaquinone-6. The genomic DNA G+C content was 42.3 mol%. The polyphasic taxonomic study revealed that strain SE14T belongs to a novel species within the genus Flavobacterium , and the name Flavobacterium phocarum sp. nov. is proposed. The type strain is SE14T (=CCTCC AB 2017225T=KCTC 52612T).

Flavobacterium ardleyense sp. nov., isolated from Antarctic soil.

A Gram-stain-negative, aerobic, yellow-pigmented, non-flagellated, non-gliding, rod-like, oxidase- and catalase-positive bacterium, designated A2-1T, was isolated from soil on Ardley Island, South Shetland Islands, Antarctica. Strain A2-1T grew at 4-22 °C (optimum, 10 °C), at pH 6.0-8.0 (optimum, pH 6.5) and with 0-1.5 % NaCl (optimum, 0.5 %), but could not produce flexirubin-type pigments. 16S rRNA gene sequence analysis showed that the isolates belonged to the genus Flavobacterium. Strain A2-1T had the highest 16S rRNA gene sequence similarity to Flavobacterium cucumis, F. ahnfeltiae and F. cheniae with 95.7, 95.6 and 95.4 %, respectively. The strain A2-1T consisted of a clade with F. cucumis and F. cheniae and simultaneously formed a distinct phyletic lineage in the neighbour-joining phylogenetic tree. Polar lipids of the strain included phosphatidylethanolamine (PE), four unidentified aminolipids and one unidentified lipid. The strain A2-1T contained anteiso-C15 : 0 (20.2 %), iso-C15 : 0 (16.2 %) and C15 : 1 G (11.0 %) as the main fatty acids and the only respiratory quinone was menaquinone MK-6. The genomic DNA G+C content was 34.0 mol%. The polyphasic taxonomic study revealed that the strain A2-1T belongs to a novel species within the genus Flavobacterium and the name Flavobacterium ardleyense sp. nov. is proposed. The type strain is A2-1T (=CCTCC AB 2017157T=KCTC 52644T).