A PadR family transcriptional repressor regulates the transcription of chromate efflux transporter in Enterobacter sp. Z1.

Chromium is a prevalent toxic heavy metal, and chromate [Cr(VI)] exhibits high mutagenicity and carcinogenicity. The presence of the Cr(VI) efflux protein ChrA has been identified in strains exhibiting resistance to Cr(VI). Nevertheless, certain strains of bacteria that are resistant to Cr(VI) lack the presence of ChrB, a known regulatory factor. Here, a PadR family transcriptional repressor, ChrN, has been identified as a regulator in the response of Enterobacter sp. Z1(CCTCC NO: M 2019147) to Cr(VI). The chrN gene is cotranscribed with the chrA gene, and the transcriptional expression of this operon is induced by Cr(VI). The binding capacity of the ChrN protein to Cr(VI) was demonstrated by both the tryptophan fluorescence assay and Ni-NTA purification assay. The interaction between ChrN and the chrAN operon promoter was validated by reporter gene assay and electrophoretic mobility shift assay. Mutation of the conserved histidine residues His14 and His50 resulted in loss of ChrN binding with the promoter of the chrAN operon. This observation implies that these residues are crucial for establishing a DNA-binding site. These findings demonstrate that ChrN functions as a transcriptional repressor, modulating the cellular response of strain Z1 to Cr(VI) exposure.

Chromate-induced methylglyoxal detoxification system drives cadmium and chromate immobilization by Cupriavidus sp. MP-37.

The detoxification of cadmium (Cd) or chromium (Cr) by microorganisms plays a vital role in bacterial survival and restoration of the polluted environment, but how microorganisms detoxify Cd and Cr simultaneously is largely unknown. Here, we isolated a bacterium, Cupriavidus sp. MP-37, which immobilized Cd(II) and reduced Cr(VI) simultaneously. Notably, strain MP-37 exhibited variable Cd(II) immobilization phenotypes, namely, cell adsorption and extracellular immobilization in the co-presence of Cd(II) and Cr(VI), while cell adsorption in the presence of Cd(II) alone. To unravel Cr(VI)-induced extracellular Cd(II) immobilization, proteomic analysis was performed, and methylglyoxal-scavenging protein (glyoxalase I, GlyI) and a regulator (YafY) showed the highest upregulation in the co-presence of Cd(II) and Cr(VI). GlyI overexpression reduced the intracellular methylglyoxal content and increased the immobilized Cd(II) content in extracellular secreta. The addition of lactate produced by GlyI protein with methylglyoxal as substrate increased the Cd(II) content in extracellular secreta. Reporter gene assay, electrophoretic mobility shift assay, and fluorescence quenching assay demonstrated that glyI expression was induced by Cr(VI) but not by Cd(II), and that YafY positively regulated glyI expression by binding Cr(VI). In the pot experiment, inoculation with the MP-37 strain reduced the Cd content of Oryza sativa L., and their secreted lactate reduced the Cr accumulation in Oryza sativa L. This study reveals that Cr(VI)-induced detoxification system drives methylglyoxal scavenging and Cd(II) extracellular detoxification in Cd(II) and Cr(VI) co-existence environment.

A Coculture of Enterobacter and Comamonas Species Reduces Cadmium Accumulation in Rice.

The accumulation of cadmium (Cd) in plants is strongly impacted by soil microbes, but its mechanism remains poorly understood. Here, we report the mechanism of reduced Cd accumulation in rice by coculture of Enterobacter and Comamonas species. In pot experiments, inoculation with the coculture decreased Cd content in rice grain and increased the amount of nonbioavailable Cd in Cd-spiked soils. Fluorescence in situ hybridization and scanning electron microscopy detection showed that the coculture colonized in the rhizosphere and rice root vascular tissue and intercellular space. Soil metagenomics data showed that the coculture increased the abundance of sulfate reduction and biofilm formation genes and related bacterial species. Moreover, the coculture increased the content of organic matter, available nitrogen, and potassium and increased the activities of arylsulfatase, ß-galactosidase, phenoloxidase, arylamidase, urease, dehydrogenase, and peroxidase in soils. In subsequent rice transcriptomics assays, we found that the inoculation with coculture activated a hypersensitive response, defense-related induction, and mitogen-activated protein kinase signaling pathway in rice. Heterologous protein expression in yeast confirmed the function of four Cd-binding proteins (HIP28-1, HIP28-4, BCP2, and CID8), a Cd efflux protein (BCP1), and three Cd uptake proteins (COPT4, NRAM5, and HKT6) in rice. Succinic acid and phenylalanine were subsequently proved to inhibit rice divalent Cd [Cd(II)] uptake and activate Cd(II) efflux in rice roots. Thus, we propose a model that the coculture protects rice against Cd stress via Cd immobilization in soils and reducing Cd uptake in rice. [Formula: see text] Copyright © 2023 The Author(s). This is an open access article distributed under the CC BY-NC-ND 4.0 International license.

Effective and stable adsorptive removal of Cadmium(II) and Lead(II) using selenium nanoparticles modified by microbial SmtA metallothionein.

Metallothionein SmtA-modified selenium nanoparticles (SmtA-SeNPs), efficient adsorbents for Cd(II) and Pb(II), were synthesized in the present work. The ligand, microbial SmtA protein, was synthesized using an engineered strain Escherichia coli, posing the benefits of simplicity, safety, and high production. SmtA-SeNPs were spheres with diameters between 68.1 and 122.4 nm, containing amino, hydroxyl, and sulfhydryl functional groups with negatively charged (pH > 5). SmtA-SeNPs displayed better adsorption performance than dissociative SmtA and SeNPs. The adsorption of Cd(II) and Pb(II) mainly depends on the electrostatic attractions and the metal chelation of abundant functional groups. The maximum adsorption capacity was 506.3 mg/g for Cd(II) and 346.7 mg/g for Pb(II), which were higher than the values of most nanoparticles. In addition, SmtA-SeNPs were immobilized with a membrane filter to produce a SmtA-SeNPs filter, and the percentage removal of Cd(II) and Pb(II) increased from 26.75% to 98.13% for Cd(II) and from 9.95% to 99.20% compared with the blank filter. Moreover, the SmtA-SeNPs filter was regenerated using subacid deionized water, and the filter exhibited a stable removal ratio of Cd(II) and Pb(II) in ten continuous cycles of Cd(II)- or Pb(II)-containing wastewater treatment. The residual amounts of Cd and Pb met national standard levels of wastewater discharge.

Reducing cadmium in rice using metallothionein surface-engineered bacteria WH16-1-MT.

Cadmium (Cd) accumulation in rice grains poses a health risk for humans. In this study, a bacterium, Alishewanella sp. WH16-1-MT, was engineered to express metallothionein on the cell surface. Compared with the parental WH16-1 strain, Cd2+ adsorption efficiency of WH16-1-MT in medium was increased from 1.2 to 2.6 mg/kg dry weight. The WH16-1-MT strain was then incubated with rice in moderately Cd-contaminated paddy soil. Compared with WH16-1, inoculation with WH16-1-MT increased plant height, panicle length and thousand-kernel weight, and decreased the levels of ascorbic acid and glutathione and the activity of peroxidase. Compared with WH16-1, WH16-1-MT inoculation significantly reduced the concentrations of Cd in brown rice, husks, roots and shoots by 44.0 %, 45.5 %, 36.1 % and 47.2 %, respectively. Moreover, inoculation with WH16-1-MT reduced the bioavailability of Cd in soil, with the total Cd proportion in oxidizable and residual states increased from 29 % to 32 %. Microbiome analysis demonstrated that the addition of WH16-1-MT did not significantly alter the original bacterial abundance and community structure in soil. These results indicate that WH16-1-MT can be used as a novel microbial treatment approach to reduce Cd in rice grown in moderately Cd-contaminated paddy soil.

Hyphomicrobium album sp. nov., isolated from mountain soil and emended description of genus Hyphomicrobium.

A soil bacterium, designated XQ2T, was isolated from Lang Mountain in Hunan province, P. R. China. The strain is Gram stain negative, facultative anaerobic, and the cells are motile and rod-shaped. The 16S rRNA gene sequence of strain XQ2T shared the highest similarities with Hyphomicrobium sulfonivorans S1T (97.1%), Pedomicrobium manganicum ACM 3038T (95.9%) and Hyphomicrobium aestuarii DSM 1564T (95.4%) and grouped with H. sulfonivorans S1T. The average nucleotide identity (ANI) values and the DNA-DNA hybridization (dDDH) values between strain XQ2T and H. sulfonivorans S1T were 86.6% and 55.4% respectively. Strain XQ2T had a genome size of 3.91 Mb and the average G+C content was 65.1%. The major fatty acids (> 5%) were C18:1ω6c, C18:1ω7c, C19:0 cyclo ω8c, C16:0 and C18:0. The major respiratory quinone was Q-9 (82.8%) and the minor one was Q-8 (17.2%). The polar lipids were diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine, phosphatidylcholine, unidentified phospholipid and two unidentified lipids. On the basis of phenotypic, chemotaxonomic and phylogenetic characteristics, strain XQ2T represents a novel species of the genus Hyphomicrobium, for which the name Hyphomicrobium album sp. nov. is proposed. The type strain is XQ2T (= KCTC 82378T = CCTCC AB 2020178T). The genus description is also emended.

NemA Catalyzes Trivalent Organoarsenical Oxidation and Is Regulated by the Trivalent Organoarsenical-Selective Transcriptional Repressor NemR.

Synthetic aromatic arsenicals such as roxarsone (Rox(V)) and nitarsone (Nit(V)) have been used as animal growth enhancers and herbicides. Microbes contribute to redox cycling between the relatively less toxic pentavalent and highly toxic trivalent arsenicals. In this study, we report the identification of nemRA operon from Enterobacter sp. Z1 and show that it is involved in trivalent organoarsenical oxidation. Expression of nemA is induced by chromate (Cr(VI)), Rox(III), and Nit(III). Heterologous expression of NemA in Escherichia coli confers resistance to Cr(VI), methylarsenite (MAs(III)), Rox(III), and Nit(III). Purified NemA catalyzes simultaneous Cr(VI) reduction and MAs(III)/Rox(III)/Nit(III) oxidation, and oxidation was enhanced in the presence of Cr(VI). The results of electrophoretic mobility shift assays and fluorescence assays demonstrate that the transcriptional repressor, NemR, binds to either Rox(III) or Nit(III). NemR has three conserved cysteine residues, Cys21, Cys106, and Cys116. Mutation of any of the three resulted in loss of response to Rox(III)/Nit(III), indicating that they form an Rox(III)/Nit(III) binding site. These results show that NemA is a novel trivalent organoarsenical oxidase that is regulated by the trivalent organoarsenical-selective repressor NemR. This discovery expands our knowledge of the molecular mechanisms of organoarsenical oxidation and provides a basis for studying the redox coupling of environmental toxic compounds.

Comamonas testosteroni antA encodes an antimonite-translocating P-type ATPase.

Antimony, like arsenic, is a toxic metalloid widely distributed in the environment. Microbial detoxification of antimony has recently been identified. Here we describe a novel bacterial P1B-type antimonite (Sb(III))-translocating ATPase from the antimony-mining bacterium Comamonas testosterone JL40 that confers resistance to Sb(III). In a comparative proteomics analysis of strain JL40, an operon (ant operon) was up-regulated by Sb(III). The ant operon includes three genes, antR, antC and antA. AntR belongs to the ArsR/SmtB family of metalloregulatory proteins that regulates expression of the ant operon. AntA belongs to the P1B family of the P-type cation-translocating ATPases. It has both similarities to and differences from other members of the P1B-1 subfamily and appears to be the first identified member of a distinct subfamily that we designate P1B-8. Expression AntA in E. coli AW3110 (Δars) conferred resistance to Sb(III) and reduced the intracellular concentration of Sb(III) but not As(III) or other metals. Everted membrane vesicles from cells expressing antA accumulated Sb(III) but not As(III), where uptake in everted vesicles reflects efflux from cells. AntC is a small protein with a potential Sb(III) binding site, and co-expression of AntC with AntA increased resistance to Sb(III). We propose that AntC functions as an Sb(III) chaperone to AntA, augmenting Sb(III) efflux. The identification of a novel Sb(III)-translocating ATPase enhances our understanding of the biogeochemical cycling of environmental antimony by bacteria.

Sediminibacterium soli sp. nov., isolated from soil.

A Gram-stain-negative, facultative anaerobic strain, designated WSJ-3T, was isolated from soil. Phylogenetic analyses based on 16S rRNA gene sequences indicated that strain WSJ-3T belongs to genus Sediminibacterium and exhibits the highest sequence similarities to Sediminibacterium roseum SYL130T (97.0%), Sediminibacterium goheungense DSM 28323T (96.9%), Sediminibacterium aquarii AA5T (96.7%), and Sediminibacterium salmoneum NBRC 103935T (95.2%). The average nucleotide identity values of strain WSJ-3T/S. roseum SYL130T and strain WSJ-3T/S. goheungense DSM 28323T are 72.2% and 70.4%, respectively, and digital DNA-DNA hybridization values for these are 19.2% and 19.1%, respectively. Strain WSJ-3T has a genome size of 3.88 Mb, with a DNA G + C content of 50.1 mol% and comprises of 3263 predicted genes. A phylogenetic tree constructed using the genomic core protein coding sequences revealed that strain WSJ-3T clusters with S. roseum SYL130T. Strain WSJ-3T has menaquinone-7 as the only respiratory quinone and phosphatidylethanolamine, three unidentified phospholipids, four unidentified aminophospholipids, two unidentified aminolipids, and three unidentified lipids as the polar lipids. The major fatty acids of strain WSJ-3T are iso-C15:0, iso-C17:0 3-OH, and iso-C15:1 G. On the basis of the polyphasic results, the isolate represents a novel species of the genus Sediminibacterium, for which the name Sediminibacterium soli sp. nov. is proposed. The type strain is WSJ-3T (= KCTC 72839T = CCTCC AB 2019408T).

Simultaneous degradation of triazophos, methamidophos and carbofuran pesticides in wastewater using an Enterobacter bacterial bioreactor and analysis of toxicity and biosafety.

Triazophos (TAP), methamidophos (MAP) and carbofuran (CF) pesticides are highly toxic, soluble and absorbable. Efficient co-degradation of multi-pesticides is rare reported. The objectives of this study were to investigate TAP, MAP and CF co-degradative ability of Enterobacter sp. Z1 and study the degradation mechanisms. Strain Z1 was shown to efficiently co-degrade TAP, MAP and CF when they were used as primary carbon sources. The degradation occurred over a wide range of temperatures, pH values and pesticide concentrations and followed first-order kinetics. Under the optimum conditions (37 °C, pH 7 and 100 mg/L of each pesticide), the degradation efficiencies were 100%, 100%, and 95.3% for TAP, MAP and CF, respectively. In addition, strain Z1 could simultaneously degrade TAP, MAP, CF and total nitrogen in wastewater in a batch bioreactor, with high removal efficiencies of 98.3%, 100%, 98.7% and 100%, respectively. Genomics, proteomics, qRT-PCR and gene overexpression analyses revealed that the degradation mechanisms involved the activities of multiple proteins, among which, organophosphorus hydrolase (Oph) and 3-hydroxyacyl-CoA dehydrogenase (PaaC) are primarily responsible for TAP and MAP degradation, while carbofuran hydrolase (Mcd) and amidohydrolase (RamA) primarily degrade CF. Among these enzymes, PaaC and RamA are newly identified pesticide-degrading enzymes. Toxicity assays of strain Z1 using reporter recombinase gene (recA) and zebrafish showed that there was no accumulation of toxic metabolites during the degradation process. Biosafety test using zebrafish showed that the strain was nontoxic toward zebrafish. Strain Z1 provides a good purification effect for pesticides-containing wastewater and novel microbial pesticide-degrading mechanisms were discovered.

Sphingomonas montanisoli sp. nov., isolated from mountain soil.

A soil bacterium, designated ZX9611T, was isolated from Taihang Mountain in Henan province, PR China. The strain was Gram-stain-negative and strictly aerobic. The cells were motile, rod-shaped and formed light pink-colored colonies. The 16S rRNA gene sequence of ZX9611T shared the highest similarities with those of Sphingomonas crocodyli CCP-7T (97.0%), Sphingomonas jatrophae S5-249T (96.6%) and Sphingomonas starnbergensis 382T (95.9%). Phylogenetic analyses based on 16S rRNA gene sequences demonstrated that ZX9611T clustered with S. crocodyli CCP-7T, S. jatrophae S5-249T and S. starnbergensis 382T. The average nucleotide identity (ANI) values between ZX9611T and two type strains (S. crocodyli BCRC 81096T and S. jatrophae DSM 27345T) were 88.3 and 68.6% respectively. ZX9611T exhibited genome-sequence-based digital DNA-DNA hybridization (dDDH) values of 53.3 % and 15.3 %, compared with S. crocodyli BCRC 81096T and S. jatrophae DSM 27345T, respectively. ZX9611T had a genome size of 4.12 Mb and an average DNA G+C content of 64.8 %. ZX9611T had major fatty acids (>5 %) including summed feature 8 (C18 : 1 ω7c and/or C18 : 1 ω6c), C14 : 0 2-OH, C16 : 0 and summed feature 3 (C16 : 1 ω7c and/or C16 : 1 ω6c), and the major polyamine was sym-homospermidine. The only respiratory quinone was ubiquinone-10. The polar lipids were diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylglycerol, phosphatidylcholine and sphingoglycolipid. On the basis of phenotypic, chemotaxonomic and phylogenetic characteristics, strain ZX9611T represents a novel species of genus Sphingomonas, for which the name Sphingomonas montanisoli sp. nov. is proposed. The type strain is ZX9611T (=KCTC 72622T=CCTCC AB 2019350T).

Luteimonas gilva sp. nov., isolated from farmland soil.

A Gram-stain-negative, rod-shaped bacterium, strain H23T, was isolated from farmland soil sampled in Enshi City, Hubei Province, PR China. The isolate grew optimally at 28-32 °C, pH 8.0 and with 0.5 % (w/v) NaCl. Based on the results of 16S rRNA gene sequence and phylogenetic analyses, strain H23T belonged to the genus Luteimonas with the highest degree of 16S rRNA gene sequence similarity to Luteimonas cucumeris Y4T (97.41 %). The DNA G+C content was 65.88 mol%. The average nucleotide identity and the Genome-to-Genome Distance Calculator results also showed low relatedness (below 95 and 70 %, respectively) between strain H23T and type strains in the genus Luteimonas. Ubiquinone-8 was the predominant quinone. The major fatty acids were iso-C15 : 0, iso-C16 : 0, iso-C17 : 0 and iso-C17 : 1 ω9c. Polar lipids were dominated by diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylglycerol and unidentified phospholipids. Low digital DNA-DNA hybridization values, as well as physiological and biochemical differences, such as no casein hydrolysis, being catalase-negative, and tesing positive for cystine arylamidase, α-chymotrypsin and N-acetyl-ß-glucosaminidase, could distinguish strain H23T from its closely related species. Strain H23T is considered to represent a novel species in the genus Luteimonas, for which the name Luteimonas gilva sp. nov. is proposed, with strain H23T (=CCTCC AB 2019255T=KCTC 72593T) as the type strain.

Simultaneous removal of chromate and arsenite by the immobilized Enterobacter bacterium in combination with chemical reagents.

Simultaneous chromate [Cr(VI)] reduction and arsenite [As(III)] oxidation is a promising pretreatment process for Cr and As removal. Here, a facultative anaerobic bacterium, Enterobacter sp. Z1, presented capacities of simultaneous Cr(VI) reduction and As(III) oxidation during anoxic cultivation in a wild range of temperature (20-45 °C) and pH (Cerkez et al., 2015; Chen et al., 2015; China Environmental Prote, 1996; Fan et al., 2008, 2019) conditions. Strikingly, strain Z1 could simultaneously contribute up to 92.8% of the reduction of Cr(VI) and 45.8% of the oxidation of As(III) in wastewater. The cells of strain Z1 were embedded with sodium alginate to produce biobeads, and the biobeads exhibited stable ratio of Cr(VI) reduction (91.8%) and As(III) oxidation (29.6%) even in the 5 continuous cycles of wastewater treatment. Moreover, in a process pretreated with the Z1 biobeads followed a precipitation with Ca(OH)2 and FeCl3, the removal efficiencies in wastewater were 98.9% and 98.3% for total Cr and As, respectively, which were 44.1% and 9.8% higher than those of using Ca(OH)2 and FeCl3, only. The residual amounts of Cr and As met the national standard levels of wastewater discharge. Proteomics analysis showed that cysteine, sulfur and methionine metabolisms, As resistance and oxidoreductase (CysH, CysI, CysJ, NemA and HemF) were induced by Cr(VI) and As(III). Moreover, the addition of cysteine to the medium also significantly improved bacterial Cr(VI) reduction rate. Our results provide a novel microbial pretreatment approach for enhancing remediation of Cr(VI) and As(III) pollution in wastewater, and reveal the evident that cysteine, sulfur and methionine metabolisms, As resistance and oxidoreductases are associated with the redox conversion of Cr(VI) and As(III).

Mucilaginibacter hurinus sp. nov., isolated from briquette warehouse soil.

A novel bacterial strain, designated ZR32T, was isolated from briquette warehouse soil in Ulsan (Korea). The strain was aerobic, showing pink-colored colonies on R2A agar. Phylogenetic analyses based on 16S rRNA gene sequences showed that strain ZR32T was closely related to Mucilaginibacter soli R9-65T (97.0%), Mucilaginibacter gynuensis YC7003T (96.9%), and Mucilaginibacter lutimaris BR-3T (96.8%). The values of DNA-DNA relatedness related two highest strains M. soli R9-65T and M. gynuensis YC7003T were 31.2 ± 6.9% and 19.7 ± 0.3%, respectively. Its genome size was 3.9 Mb, comprising 3402 predicted genes. The DNA G+C content of strain ZR32T was 43.0 mol%. The major cellular fatty acids (> 5% of total) were summed feature 3 (C16:1ω6c and/or C16:1ω7c), C16:0, C16:1ω5c, iso-C15:0, iso-C17:0 3-OH, and C17:1ω9c. The major respiratory quinine was menaquinone-7 (MK-7). The major polar lipids were phosphatidylethanolamine, two unidentified phospholipids, one unidentified sphingolipid, and one unidentified polar lipid. Strain ZR32T showed distinctive characteristics such as the temperature and pH for growth ranges, being positive for ß-glucosidase, salicin production, negative for N-acetyl-glucosamine assimilation, being resistant to carbenicillin and piperacillin to related species. On the basis of phenotypic, chemotaxonomic, and phylogenetic data, strain ZR32T represents a novel species of the genus Mucilaginibacter, for which the name Mucilaginibacter hurinus sp. nov. is proposed. The type strain is ZR32T (= KCTC 62193 = CCTCC AB 2017285).

Fibrisoma montanum sp. nov., isolated from soil of Mountain Danxia, China.

A Gram-stain-negative, non-motile, rod-shaped, aerobic bacterium, designated HYT19T, was isolated from soil of Mountain Danxia in southern China. It showed the highest similarity of 16S rRNA gene sequence (97.0%) and formed a monophyletic clade with Fibrisoma limi BUZ 3T. Strain HYT19T grew at 16-37 °C (optimum 28-30 °C) and at pH 6-7. The draft genome size of strain HYT19T was 7.8 Mb with a DNA G+C content of 54.0 mol%. The digital DDH and average nucleotide identity values between strain HYT19T and F. limi BUZ 3T were 28.8% and 85.1%, respectively. MK-7 was the sole respiratory quinone. The major polar lipids were phosphatidylethanolamine, unidentified aminophospholipid, two unidentified aminolipids, unidentified phospholipid and unidentified lipid. The strain contained C16:1ω5c, iso-C15:0, summed feature 3 (C16:1ω6c and/or C16:1ω7c), C16:0, iso-C17:0 3-OH and anteiso-C15:0 as the major fatty acids. On the basis of phylogenetic, genomic, phenotypic and chemotaxonomic analysis, we propose a new species Fibrisoma montanum sp. nov. of genus Fibrisoma. The type strain is HYT19T (= CCTCC AB 2018342T = JCM 33105T).

Sphingomonas gilva sp. nov., isolated from mountain soil.

A Gram-stain-negative, strictly aerobic, motile, yellow, rod-shaped bacterium, designated ZDH117T, was isolated from soilsampled atthe Danxialandformin Guangdong Province, PR China. The 16S rRNA gene sequence of strain ZDH117T had highest similarityvalues to Sphingomonas adhaesivaDSM 7418T (97.5 %), SphingomonasdesiccabilisCP1DT (97.3 %) and Sphingomonas ginsenosidimutans KACC 14949T (97.2 %). However, phylogenetic analyses based on 16S rRNA gene sequences demonstrated that strain ZDH117T clustered with Sphingomonas zeicaulis 541T (96.17 %) and Sphingomonas sanxanigenens DSM 19645T (95.95 %). The genomic average nucleotide identity values of ZDH117T with S. adhaesiva DSM 7418T, S. desiccabilis CP1DTand S. ginsenosidimutans KACC TT were 75.1, 75.2 and 75.0 %, respectively. The G+C content of the genomic DNA was 67.6 mol%. Strain ZDH117T was characterized to have ubiquinone-10 as the predominant respiratory quinone, sym-homospermidine as the major polyamine and summed feature 8 (C18 : 1ω6c and/or C18 : 1ω7c), C14 : 0-2OH, C16 : 0 and summed feature 3 (C16 : 1ω6c and/or C16 : 1ω7c) as the major cellular fatty acids (>5 % of total). The predominant polar lipids were diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylglycerol, phosphatidylcholine, sphingoglycolipid, an unidentified phospholipid and three unidentified lipids. On the basis of its phenotypic, chemotaxonomic and phylogenetic characteristics, strain ZDH117T represents a novel species of the genus Sphingomonas, for which the name Sphingomonas gilva sp. nov. is proposed. The type strain is ZDH117T (=KCTC 62894T=CCTCCAB 2018262T).

Hymenobacter edaphi sp. nov., isolated from abandoned arsenic-contaminated farmland soil.

A Gram-stain-negative, aerobic, rod-shaped, non-motile, pink-pigmented bacterium, designated NLT, was isolated from arsenic-contaminated farmland soil. Strain NLT showed the highest 16S rRNA gene sequence similarities with those of Hymenobacter jeollabukensis 1-3-3-8T (98.9 %), Hymenobacter gummosus ANT-18T (97.5 %), Hymenobacter paludis KBP-30T (97.4 %), Hymenobacter ocellatus Myx2105T (97.1 %) and Hymenobacter coalescens WW84T (96.4 %). The values of genomic orthoANI and dDDH between strain NLT and Hymenobacter jeollabukensis KCTC 52741T was 90.5 and 41.2 %, respectively, and those between strain NLT and Hymenobacter gummosus KCTC 52166T was 84.4 and 28.4 %, respectively. Strain NLT exhibited DNA-DNA hybridisation values of 41.3 and 44.1 % with Hymenobacter paludis KCTC 32237T and Hymenobacter ocellatus DSM 11117T, respectively. Strain NLT had major fatty acids (>10 %) of summed feature 4 (iso-C17 : 1 I and/or anteiso-C17 : 1 B), iso-C15 : 0 and anteiso-C15 : 0 and the predominant polyamine of homospermidine. The only respiratory quinone was menaquinone-7. The polar lipids were phosphatidylethanolamine, phospholipid, three unidentified lipids and two amino lipids. Strain NLT had a genome size of 6.04 Mb and the average G+C content of 65.6 %. Compared to the other Hymenobacter spp., strain NLT is different in polar lipid profile (without aminophospholipid) and leucine arylamidase activity. Based on the data of the polyphasic analysis, it is considered that strain NLT represented a novel species of genus Hymenobacter, for which the name Hymenobacter edaphisp. nov. is proposed. The type strain is NLT (=KCTC 62521T=CCTCC AB 2018028T).

Cooperation between two strains of Enterobacter and Klebsiella in the simultaneous nitrogen removal and phosphate accumulation processes.

Two strains, Enterobacter sp. Z1 and Klebsiella sp. Z2, were exhibited great capacities for heterotrophic nitrification-aerobic denitrification (HNAD) and intracellular phosphate accumulation. Strikingly, the co-cultured strains enhanced the removal efficiency of total nitrogen and phosphate, with removal efficiencies of ammonia, nitrate, nitrite and soluble phosphate of 99.64%, 99.85%, 96.94% and 66.7% respectively. Furthermore, high removal efficiencies from wastewaters with high concentrations of ammonia (over 1000 mg/L) were achieved by inoculation with the co-strains, which left residual ammonia of less than 1 mg/L within 10 h. To elucidate the mechanism of HNAD in co-strains, quantitative PCR was carried out to examine the expression levels of hydroxylamine oxidase (Hao), nitrate reductase (NapA and NarG), nitrite reductase (NirS) and polyphosphate kinase (Ppk), and the results showed that the napA2, narG and ppk genes in the strains were significantly upregulated under the co-cultured conditions and provided an explanation for the nitrogen and phosphate removal.

Phenylobacterium soli sp. nov., isolated from arsenic and cadmium contaminated farmland soil.

A Gram-stain-negative, facultative anaerobic, non-motile and rod-shaped bacterial strain, designated LX32T, was isolated from arsenic and cadmium contaminated farmland soil. Phylogenetic analysis based on 16S rRNA gene sequence indicated that strain LX32T was closely related to Phenylobacterium hankyongense HKS-05T (97.7 % sequence similarity), Phenylobacterium kunshanense CCTCC AB 2013085T (97.4 %) and Phenylobacterium deserti CCTCC AB 2016297T (97.1 %). The average nucleotide identity values of the whole genome sequences of LX32T/P. hankyongense HKS-05T, LX32T/P. kunshanense CCTCC AB 2013085T and LX32T/P. deserti CCTCC AB 2016297T were 79.8, 77.9 and 77.5 %, respectively. Its genome size was 4.02 Mb, comprising 3998 predicted genes with a DNA G+C content of 70.1 mol%. The major fatty acids were C15 : 0, C16 : 0 and summed feature 8 (comprising C18 : 1ω7c and/or C18 : 1ω6c). The polar lipid profiles consisted of phosphatidylglycerol, aminophospholipid, seven glycolipids and two unidentified polar lipids. The predominantly respiratory quinone was ubiquinone-10. Based on polyphasic analyses, the isolate is considered to represent a novel species of the genus Phenylobacterium, for which the name Phenylobacterium soli sp. nov. is proposed. The type strain is LX32T (=KCTC 62522=CCTCC AB 2018055).

Pedobacter paludis sp. nov., isolated from wetland soil.

A Gram-stain-negative, aerobic, non-motile, rod-shaped and non-spore-forming bacterium, designated YXT, was isolated from wetland soil. Compared to strain YXT, Pedobacter yonginense HMD1002T had the highest 16S rRNA gene sequence identity (97.8%), and the remaining strains had the identities below 97%. Phylogenetic analysis of the 16S rRNA gene sequences showed that strain YXT grouped with P. yonginense HMD1002T. The values of DNA-DNA hybridization and genomic orthologous average nucleotide identity (orthoANI) between strain YXT and Pedobacter yonginense KCTC 22721T were 58.5% and 82.0%, respectively. The genome size of strain YXT was 5.0 Mb, comprising 4369 predicted genes with a DNA G+C content of 37.3 mol %. Strain YXT had menaquinone-7 as the only respiratory quinone and polar lipids of phosphatidylethanolamine, sphingolipid, aminolipid and three unidentified lipids. The predominant cellular fatty acids (> 10%) of strain YXT were summed feature 3 (iso-C15:0 2-OH and/or C16:1ω7c), iso-C15:0 and iso-C17:0 3-OH. Strain YXT could be distinguished from the other Pedobacter members based on the data of phylogenetic distance, DNA-DNA hybridization, genomic orthoANI, RecA MLSA, core-protein comparison, and hydrolyses of L-arginine, utilization of D/L-lactate, L-alanine, 5-ketonic gluconate and glycogen. Therefore, strain YXT represents a novel species of the genus Pedobacter, for which the name Pedobacter paludis sp. nov. is proposed. The type strain is YXT (= KCTC 62520T = CCTCC AB 2018029T).