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Antonio Ventosa - One of the best experts on this subject based on the ideXlab platform.
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Diversity and distribution of Halomonas in Rambla Salada, a hypersaline environment in the southeast of Spain
FEMS microbiology ecology, 2013Co-Authors: Nahid Oueriaghli, Antonio Ventosa, Carmen M. González-domenech, Fernando Martínez-checa, Emilia Quesada, Gerard Muyzer, Victoria BéjarAbstract:We have studied the diversity and distribution of Halomonas populations in the hypersaline habitat Rambla Salada (Murcia, southeastern Spain) by using different molecular techniques. Denaturing gradient gel electrophoresis (DGGE) using specific primers for the 16S rRNA gene of Halomonas followed by a multivariate analysis of the results indicated that richness and evenness of the Halomonas populations were mainly influenced by the season. We found no significant differences between the types of samples studied, from either watery sediments or soil samples. The highest value of diversity was reached in June 2006, the season with the highest salinity. Furthermore, canonical correspondence analysis (CCA) demonstrated that both salinity and pH significantly affected the structure of the Halomonas community. Halomonas almeriensis and two denitrifiers, H. ilicicola and H. ventosae were the predominant species. CARD-FISH showed that the percentage of Halomonas cells with respect to the total number of microorganisms ranged from 4.4% to 5.7%. To study the functional role of denitrifying species, we designed new primer sets targeting denitrification nirS and nosZ genes. Using these primers, we analyzed sediments from the upwelling zone collected in June 2006, where we found the highest percentage of denitrifiers (74%). Halomonas ventosae was the predominant denitrifier in this site.
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Halomonas titanicae sp. nov., a halophilic bacterium isolated from the RMS Titanic.
International Journal of Systematic and Evolutionary Microbiology, 2010Co-Authors: Cristina Sánchez-porro, Bhavleen Kaur, Henrietta Mann, Antonio VentosaAbstract:A Gram-negative, heterotrophic, aerobic, non-endospore-forming, peritrichously flagellated and motile bacterial strain, designated BH1T, was isolated from samples of rusticles, which are formed in part by a consortium of micro-organisms, collected from the RMS Titanic wreck site. The strain grew optimally at 30–37 °C, pH 7.0–7.5 and in the presence of 2–8 % (w/v) NaCl. We carried out a polyphasic taxonomic study in order to characterize the strain in detail. Phylogenetic analyses based on 16S rRNA gene sequence comparison indicated that strain BH1T clustered within the branch consisting of species of Halomonas. The most closely related type strains were Halomonas neptunia (98.6 % 16S rRNA sequence similarity), Halomonas variabilis (98.4 %), Halomonas boliviensis (98.3 %) and Halomonas sulfidaeris (97.5 %). Other closely related species were Halomonas alkaliphila (96.5 % sequence similarity), Halomonas hydrothermalis (96.3 %), Halomonas gomseomensis (96.3 %), Halomonas venusta (96.3 %) and Halomonas meridiana (96.2 %). The major fatty acids of strain BH1T were C18 : 1 ω7c (36.3 %), C16 : 0 (18.4 %) and C19 : 0 cyclo ω8c (17.9 %). The DNA G+C content was 60.0 mol% (T m). Ubiquinone 9 (Q-9) was the major lipoquinone. The phenotypic features, fatty acid profile and DNA G+C content further supported the placement of strain BH1T in the genus Halomonas. DNA–DNA hybridization values between strain BH1T and H. neptunia CECT 5815T, H. variabilis DSM 3051T, H. boliviensis DSM 15516T and H. sulfidaeris CECT 5817T were 19, 17, 30 and 29 %, respectively, supporting the differential taxonomic status of BH1T. On the basis of the phenotypic, chemotaxonomic and phylogenetic data, strain BH1T is considered to represent a novel species, for which the name Halomonas titanicae sp. nov. is proposed. The type strain is BH1T (=ATCC BAA-1257T =CECT 7585T =JCM 16411T =LMG 25388T).
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Halomonas titanicae sp. nov., a halophilic bacterium isolated from the RMS Titanic.
International journal of systematic and evolutionary microbiology, 2010Co-Authors: Cristina Sánchez-porro, Bhavleen Kaur, Henrietta Mann, Antonio VentosaAbstract:A Gram-negative, heterotrophic, aerobic, non-endospore-forming, peritrichously flagellated and motile bacterial strain, designated BH1(T), was isolated from samples of rusticles, which are formed in part by a consortium of micro-organisms, collected from the RMS Titanic wreck site. The strain grew optimally at 30-37°C, pH 7.0-7.5 and in the presence of 2-8 % (w/v) NaCl. We carried out a polyphasic taxonomic study in order to characterize the strain in detail. Phylogenetic analyses based on 16S rRNA gene sequence comparison indicated that strain BH1(T) clustered within the branch consisting of species of Halomonas. The most closely related type strains were Halomonas neptunia (98.6 % 16S rRNA sequence similarity), Halomonas variabilis (98.4 %), Halomonas boliviensis (98.3 %) and Halomonas sulfidaeris (97.5 %). Other closely related species were Halomonas alkaliphila (96.5 % sequence similarity), Halomonas hydrothermalis (96.3 %), Halomonas gomseomensis (96.3 %), Halomonas venusta (96.3 %) and Halomonas meridiana (96.2 %). The major fatty acids of strain BH1(T) were C(18 : 1)ω7c (36.3 %), C(16 : 0) (18.4 %) and C(19 : 0) cyclo ω8c (17.9 %). The DNA G+C content was 60.0 mol% (T(m)). Ubiquinone 9 (Q-9) was the major lipoquinone. The phenotypic features, fatty acid profile and DNA G+C content further supported the placement of strain BH1(T) in the genus Halomonas. DNA-DNA hybridization values between strain BH1(T) and H. neptunia CECT 5815(T), H. variabilis DSM 3051(T), H. boliviensis DSM 15516(T) and H. sulfidaeris CECT 5817(T) were 19, 17, 30 and 29 %, respectively, supporting the differential taxonomic status of BH1(T). On the basis of the phenotypic, chemotaxonomic and phylogenetic data, strain BH1(T) is considered to represent a novel species, for which the name Halomonas titanicae sp. nov. is proposed. The type strain is BH1(T) (=ATCC BAA-1257(T) =CECT 7585(T) =JCM 16411(T) =LMG 25388(T)).
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Halomonas ilicicola sp. nov., a moderately halophilic bacterium isolated from a saltern.
INTERNATIONAL JOURNAL OF SYSTEMATIC AND EVOLUTIONARY MICROBIOLOGY, 2009Co-Authors: María I. Arenas, Antonio Ventosa, Cristina Sánchez-porro, Patricia I. Bañón, José Luis Copa-patiño, Juan SoliveriAbstract:A Gram-negative, heterotrophic, aerobic, pale orange-pigmented, non-endospore-forming and motile bacterial strain, designated strain SP8T, was isolated from a salty water sample from the solar salterns of Santa Pola, located on the Mediterranean coast of Spain. The strain grew optimally at 37 °C, pH 6.5 and in the presence of 10 % NaCl. A polyphasic taxonomic study was conducted in order to characterize the strain in detail. Phylogenetic analyses based on 16S rRNA gene sequence comparisons indicated that strain SP8T clustered within the branch constituted by species of the genus Halomonas. The closest phylogenetic neighbours of strain SP8T were Halomonas muralis LMG 20969T (96.0 % sequence similarity), Halomonas pantelleriensis AAPT (95.9 %) and Halomonas campaniensis 5AGT (95.8 %). Phenotypic features, the fatty acid profile and the DNA G+C content of the novel strain further supported its placement in the genus Halomonas. On the basis of phenotypic, chemotaxonomic and phylogenetic distinctiveness, it is suggested that strain SP8T represents a novel species for which the name Halomonas ilicicola sp. nov. is proposed. The type strain is SP8T (=CECT 7331T=CCM 7522T=DSM 19980T).
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Description of Kushneria aurantia gen. nov., sp. nov., a novel member of the family Halomonadaceae, and a proposal for reclassification of Halomonas marisflavi as Kushneria marisflavi comb. nov., of Halomonas indalinina as Kushneria indalinina comb.
INTERNATIONAL JOURNAL OF SYSTEMATIC AND EVOLUTIONARY MICROBIOLOGY, 2009Co-Authors: Cristina Sánchez-porro, Rafael R. De La Haba, Nelís Soto-ramírez, M. Carmen Márquez, Rafael Montalvo-rodríguez, Antonio VentosaAbstract:An aerobic, moderately halophilic, Gram-negative, motile, non-sporulating rod-shaped bacterium, designated strain A10T, was isolated from the surface of leaves of the black mangrove Avicennia germinans and was subjected to a polyphasic taxonomic study. Strain A10T was able to grow at NaCl concentrations in the range 5–17.5 % (w/v) with optimum growth at 10 % (w/v) NaCl. Growth occurred at temperatures of 20–40 °C (optimal growth at 37 °C) and pH 5.5–8.5 (optimal growth at pH 7.0–8.0). The major respiratory quinone was ubiquinone 9. The major fatty acids were C16 : 0, C18 : 1 ω7c, C19 : 0 cyclo ω8c and C12 : 0 3-OH. The polar lipids were phosphatidylglycerol, diphosphatidylglycerol, phosphatidylethanolamine, and unidentified phospholipids, glycolipids and an aminoglycolipid. Phylogenetic analysis based on 16S rRNA gene sequence comparisons revealed that strain A10T is closely related to Halomonas avicenniae MW2aT (95.7 % sequence similarity), Halomonas marisflavi SW32T (95.2 %) and Halomonas indalinina GC2.1T (95.0 %). Strain A10T formed a coherent phylogenetic branch with these three species, separated from other species of Halomonas and closely related genera (with 16S rRNA gene sequence similarities below 94.0 %). A complete 23S rRNA gene sequence comparison of strain A10T with closely related species confirmed the phylogenetic position of the novel isolate, forming a branch with the species Halomonas avicenniae, Halomonas indalinina and Halomonas marisflavi, separated from other species of the genera belonging to the family Halomonadaceae (showing sequence similarities below 91.7 %). DNA–DNA hybridization studies between strain A10T and Halomonas avicenniae MW2aT, Halomonas marisflavi DSM 15357T and Halomonas indalinina CG2.1T were 21, 17 and 10 %, respectively. These levels of DNA–DNA relatedness were low enough to classify strain A10T as representing a genotypically distinct species. Overall, the phenotypic, genotypic, chemotaxonomic and phylogenetic results demonstrated that strain A10T represents a new genus and species. The name Kushneria aurantia gen. nov., sp. nov. is proposed, with strain A10T (=CCM 7415T=CECT 7220T) as the type strain. This is the type species of the new proposed genus, which belongs to the family Halomonadaceae. In addition, our data support the placement of the species Halomonas marisflavi, Halomonas indalinina and Halomonas avicenniae within this new genus, as Kushneria marisflavi comb. nov., Kushneria indalinina comb. nov. and Kushneria avicenniae comb. nov., respectively.
John A Baross - One of the best experts on this subject based on the ideXlab platform.
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Halomonas and marinobacter ecotypes from hydrothermal vent subseafloor and deep sea environments
FEMS Microbiology Ecology, 2011Co-Authors: Jonathan Z Kaye, Katrina J Edwards, Jason B Sylvan, John A BarossAbstract:Moderately halophilic and euryhaline bacteria are routinely found in cool to warm hydrothermal vent and nearby cold, deep-sea environments. To elucidate the diversity of these microorganisms – with the goal of determining which among them constitute ecotypes specifically associated with hydrothermal vent and subseafloor habitats – PCR primers were designed to detect natural populations of euryhaline Gammaproteobacteria belonging to the cosmopolitan genera Halomonas and Marinobacter. The distribution patterns of 16S rRNA gene sequence data revealed that Halomonas group 2A comprised a subseafloor population at Axial Seamount on the Juan de Fuca Ridge. Complementary biogeographic and physiological data suggested that other Halomonas clades include members that are cold adapted (Halomonas group 2B) or associated with massive sulfide deposits (Halomonas group 2C). Similarly, a monophyletic Marinobacter clade may represent Fe2+-oxidizing facultative chemoautotrophs based on the phylogenetic data presented here and previously reported phenotypic characterizations. The biogeographic distributions of Halomonas and Marinobacter isolates and clones reveal that these are cosmopolitan genera, commonly found in the deep sea and in hydrothermal vent settings. As such, they are good candidates for further laboratory investigations into the biogeochemical processes in these environments.
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Halomonas neptunia sp. nov., Halomonas sulfidaeris sp. nov., Halomonas axialensis sp. nov. and Halomonas hydrothermalis sp. nov.: halophilic bacteria isolated from deep-sea hydrothermal-vent environments.
International Journal of Systematic and Evolutionary Microbiology, 2004Co-Authors: Jonathan Z Kaye, Antonio Ventosa, M. Carmen Márquez, John A BarossAbstract:To assess the physiological and phylogenetic diversity of culturable halophilic bacteria in deep-sea hydrothermal-vent environments, six isolates obtained from low-temperature hydrothermal fluids, sulfide rock and hydrothermal plumes in North and South Pacific Ocean vent fields located at 1530–2580 m depth were fully characterized. Three strains were isolated on media that contained oligotrophic concentrations of organic carbon (0·002 % yeast extract). Sequencing of the 16S rRNA gene indicated that all strains belonged to the genus Halomonas in the γ-subclass of the Proteobacteria. Consistent with previously described species, the novel strains were slightly to moderately halophilic and grew in media containing up to 22–27 % total salts. The isolates grew at temperatures as low as −1 to 2 °C and had temperature optima of 30 or 20–35 °C. Both the minimum and optimum temperatures for growth were similar to those of Antarctic and sea-ice Halomonas species and lower than typically observed for the genus as a whole. Phenotypic tests revealed that the isolates were physiologically versatile and tended to have more traits in common with each other than with closely related Halomonas species, presumably a reflection of their common deep-sea, hydrothermal-vent habitat of origin. The G+C content of the DNA for all strains was 56·0–57·6 mol%, and DNA–DNA hybridization experiments revealed that four strains (Eplume1T, Esulfide1T, Althf1T and Slthf2T) represented novel species and that two strains (Eplume2 and Slthf1) were related to Halomonas meridiana. The proposed new species names are Halomonas neptunia (type strain Eplume1T=ATCC BAA-805T=CECT 5815T=DSM 15720T), Halomonas sulfidaeris (type strain Esulfide1T=ATCC BAA-803T=CECT 5817T=DSM 15722T), Halomonas axialensis (type strain Althf1T=ATCC BAA-802T=CECT 5812T=DSM 15723T) and Halomonas hydrothermalis (type strain Slthf2T=ATCC BAA-800T=CECT 5814T=DSM 15725T).
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Halomonas neptunia sp. nov., Halomonas sulfidaeris sp. nov., Halomonas axialensis sp. nov. and Halomonas hydrothermalis sp. nov.: halophilic bacteria isolated from deep-sea hydrothermal-vent environments.
International journal of systematic and evolutionary microbiology, 2004Co-Authors: Jonathan Z Kaye, Antonio Ventosa, M. Carmen Márquez, John A BarossAbstract:To assess the physiological and phylogenetic diversity of culturable halophilic bacteria in deep-sea hydrothermal-vent environments, six isolates obtained from low-temperature hydrothermal fluids, sulfide rock and hydrothermal plumes in North and South Pacific Ocean vent fields located at 1530-2580 m depth were fully characterized. Three strains were isolated on media that contained oligotrophic concentrations of organic carbon (0.002 % yeast extract). Sequencing of the 16S rRNA gene indicated that all strains belonged to the genus Halomonas in the gamma-subclass of the Proteobacteria. Consistent with previously described species, the novel strains were slightly to moderately halophilic and grew in media containing up to 22-27 % total salts. The isolates grew at temperatures as low as -1 to 2 degrees C and had temperature optima of 30 or 20-35 degrees C. Both the minimum and optimum temperatures for growth were similar to those of Antarctic and sea-ice Halomonas species and lower than typically observed for the genus as a whole. Phenotypic tests revealed that the isolates were physiologically versatile and tended to have more traits in common with each other than with closely related Halomonas species, presumably a reflection of their common deep-sea, hydrothermal-vent habitat of origin. The G+C content of the DNA for all strains was 56.0-57.6 mol%, and DNA-DNA hybridization experiments revealed that four strains (Eplume1(T), Esulfide1(T), Althf1(T) and Slthf2(T)) represented novel species and that two strains (Eplume2 and Slthf1) were related to Halomonas meridiana. The proposed new species names are Halomonas neptunia (type strain Eplume1(T)=ATCC BAA-805(T)=CECT 5815(T)=DSM 15720(T)), Halomonas sulfidaeris (type strain Esulfide1(T)=ATCC BAA-803(T)=CECT 5817(T)=DSM 15722(T)), Halomonas axialensis (type strain Althf1(T)=ATCC BAA-802(T)=CECT 5812(T)=DSM 15723(T)) and Halomonas hydrothermalis (type strain Slthf2(T)=ATCC BAA-800(T)=CECT 5814(T)=DSM 15725(T)).
Jonathan Z Kaye - One of the best experts on this subject based on the ideXlab platform.
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Halomonas and marinobacter ecotypes from hydrothermal vent subseafloor and deep sea environments
FEMS Microbiology Ecology, 2011Co-Authors: Jonathan Z Kaye, Katrina J Edwards, Jason B Sylvan, John A BarossAbstract:Moderately halophilic and euryhaline bacteria are routinely found in cool to warm hydrothermal vent and nearby cold, deep-sea environments. To elucidate the diversity of these microorganisms – with the goal of determining which among them constitute ecotypes specifically associated with hydrothermal vent and subseafloor habitats – PCR primers were designed to detect natural populations of euryhaline Gammaproteobacteria belonging to the cosmopolitan genera Halomonas and Marinobacter. The distribution patterns of 16S rRNA gene sequence data revealed that Halomonas group 2A comprised a subseafloor population at Axial Seamount on the Juan de Fuca Ridge. Complementary biogeographic and physiological data suggested that other Halomonas clades include members that are cold adapted (Halomonas group 2B) or associated with massive sulfide deposits (Halomonas group 2C). Similarly, a monophyletic Marinobacter clade may represent Fe2+-oxidizing facultative chemoautotrophs based on the phylogenetic data presented here and previously reported phenotypic characterizations. The biogeographic distributions of Halomonas and Marinobacter isolates and clones reveal that these are cosmopolitan genera, commonly found in the deep sea and in hydrothermal vent settings. As such, they are good candidates for further laboratory investigations into the biogeochemical processes in these environments.
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Halomonas neptunia sp. nov., Halomonas sulfidaeris sp. nov., Halomonas axialensis sp. nov. and Halomonas hydrothermalis sp. nov.: halophilic bacteria isolated from deep-sea hydrothermal-vent environments.
International Journal of Systematic and Evolutionary Microbiology, 2004Co-Authors: Jonathan Z Kaye, Antonio Ventosa, M. Carmen Márquez, John A BarossAbstract:To assess the physiological and phylogenetic diversity of culturable halophilic bacteria in deep-sea hydrothermal-vent environments, six isolates obtained from low-temperature hydrothermal fluids, sulfide rock and hydrothermal plumes in North and South Pacific Ocean vent fields located at 1530–2580 m depth were fully characterized. Three strains were isolated on media that contained oligotrophic concentrations of organic carbon (0·002 % yeast extract). Sequencing of the 16S rRNA gene indicated that all strains belonged to the genus Halomonas in the γ-subclass of the Proteobacteria. Consistent with previously described species, the novel strains were slightly to moderately halophilic and grew in media containing up to 22–27 % total salts. The isolates grew at temperatures as low as −1 to 2 °C and had temperature optima of 30 or 20–35 °C. Both the minimum and optimum temperatures for growth were similar to those of Antarctic and sea-ice Halomonas species and lower than typically observed for the genus as a whole. Phenotypic tests revealed that the isolates were physiologically versatile and tended to have more traits in common with each other than with closely related Halomonas species, presumably a reflection of their common deep-sea, hydrothermal-vent habitat of origin. The G+C content of the DNA for all strains was 56·0–57·6 mol%, and DNA–DNA hybridization experiments revealed that four strains (Eplume1T, Esulfide1T, Althf1T and Slthf2T) represented novel species and that two strains (Eplume2 and Slthf1) were related to Halomonas meridiana. The proposed new species names are Halomonas neptunia (type strain Eplume1T=ATCC BAA-805T=CECT 5815T=DSM 15720T), Halomonas sulfidaeris (type strain Esulfide1T=ATCC BAA-803T=CECT 5817T=DSM 15722T), Halomonas axialensis (type strain Althf1T=ATCC BAA-802T=CECT 5812T=DSM 15723T) and Halomonas hydrothermalis (type strain Slthf2T=ATCC BAA-800T=CECT 5814T=DSM 15725T).
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Halomonas neptunia sp. nov., Halomonas sulfidaeris sp. nov., Halomonas axialensis sp. nov. and Halomonas hydrothermalis sp. nov.: halophilic bacteria isolated from deep-sea hydrothermal-vent environments.
International journal of systematic and evolutionary microbiology, 2004Co-Authors: Jonathan Z Kaye, Antonio Ventosa, M. Carmen Márquez, John A BarossAbstract:To assess the physiological and phylogenetic diversity of culturable halophilic bacteria in deep-sea hydrothermal-vent environments, six isolates obtained from low-temperature hydrothermal fluids, sulfide rock and hydrothermal plumes in North and South Pacific Ocean vent fields located at 1530-2580 m depth were fully characterized. Three strains were isolated on media that contained oligotrophic concentrations of organic carbon (0.002 % yeast extract). Sequencing of the 16S rRNA gene indicated that all strains belonged to the genus Halomonas in the gamma-subclass of the Proteobacteria. Consistent with previously described species, the novel strains were slightly to moderately halophilic and grew in media containing up to 22-27 % total salts. The isolates grew at temperatures as low as -1 to 2 degrees C and had temperature optima of 30 or 20-35 degrees C. Both the minimum and optimum temperatures for growth were similar to those of Antarctic and sea-ice Halomonas species and lower than typically observed for the genus as a whole. Phenotypic tests revealed that the isolates were physiologically versatile and tended to have more traits in common with each other than with closely related Halomonas species, presumably a reflection of their common deep-sea, hydrothermal-vent habitat of origin. The G+C content of the DNA for all strains was 56.0-57.6 mol%, and DNA-DNA hybridization experiments revealed that four strains (Eplume1(T), Esulfide1(T), Althf1(T) and Slthf2(T)) represented novel species and that two strains (Eplume2 and Slthf1) were related to Halomonas meridiana. The proposed new species names are Halomonas neptunia (type strain Eplume1(T)=ATCC BAA-805(T)=CECT 5815(T)=DSM 15720(T)), Halomonas sulfidaeris (type strain Esulfide1(T)=ATCC BAA-803(T)=CECT 5817(T)=DSM 15722(T)), Halomonas axialensis (type strain Althf1(T)=ATCC BAA-802(T)=CECT 5812(T)=DSM 15723(T)) and Halomonas hydrothermalis (type strain Slthf2(T)=ATCC BAA-800(T)=CECT 5814(T)=DSM 15725(T)).
Guo-qiang Chen - One of the best experts on this subject based on the ideXlab platform.
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Reprogramming Halomonas for industrial production of chemicals.
Journal of industrial microbiology & biotechnology, 2018Co-Authors: Xiangbin Chen, Guanqing Qiao, Guo-qiang ChenAbstract:Halomonas spp. are able to grow under a high salt concentration at alkali pH, they are able to resist contamination by other microbes. Development of Halomonas spp. as platform production strains for the next-generation industrial biotechnology (NGIB) is intensively studied. Among Halomonas spp., Halomonas bluephagenesis is the best studied one with available engineering tools and methods to reprogram it for production of various polyhydroxyalkanoates, proteins, and chemicals. Due to its contamination resistance, H. bluephagenesis can be grown under open and continuous processes not just in the labs but also in at least 1000 L fermentor scale. It is expected that NGIB based on Halomonas spp. be able to engineer for production of increasing number of products in a competitive manner.
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Engineering Halomonas species TD01 for enhanced polyhydroxyalkanoates synthesis via CRISPRi
Microbial cell factories, 2017Co-Authors: Wei Tao, Guo-qiang ChenAbstract:Background Clustered regularly interspaced short palindromic repeats interference (CRISPRi) has provided an efficient approach for targeted gene inhibition. A non-model microorganism Halomonas species TD01 has been developed as a promising industrial producer of polyhydroxyalkanoates (PHA), a family of biodegradable polyesters accumulated by bacteria as a carbon and energy reserve compound. A controllable gene repression system, such as CRISPRi, is needed for Halomonas sp. TD01 to regulate its gene expression levels.
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Novel T7-like expression systems used for Halomonas.
Metabolic engineering, 2016Co-Authors: Han Zhao, Haoqian Zhang, Xiangbin Chen, Qi Ouyang, Guo-qiang ChenAbstract:To engineer non-model organisms, suitable genetic parts must be available. However, biological parts are often host strain sensitive. It is therefore necessary to develop genetic parts that are functional regardless of host strains. Here we report several novel phage-derived expression systems used for transcriptional control in non-model bacteria. Novel T7-like RNA polymerase-promoter pairs were obtained by mining phage genomes, followed by in vivo characterization in non-model strains Halomonas spp TD01 and Pseudomonas entomophila. Three expression systems, namely, MmP1, VP4, and K1F, were developed displaying orthogonality (crosstalk 0.94) between Escherichia coli and Halomonas sp. TD01, implying suitability of broad-host range. Three Halomonas TD strains were then constructed based upon these expression systems that enabled interchangeable and controllable gene expression. One of the strains termed Halomonas TD-MmP1 was used to express the cell-elongation cassette (minCD genes) and polyhydroxybutyrate (PHB) biosynthetic pathway, resulting in a 100-fold increase in cell lengths and high levels of PHB production (up to 92% of cell dry weight), respectively. We envision these T7-like expression systems to benefit metabolic engineering in other non-model organisms.
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engineering Halomonas spp as a low cost production host for production of bio surfactant protein phap
Biotechnology Journal, 2016Co-Authors: Luhong Lan, Jin-chun Chen, Han Zhao, Guo-qiang ChenAbstract:Halomonas spp. have been studied as a low cost production host for producing bulk materials such as polyhydroxyalkanoates (PHA) bioplastics, since they are able to grow at high pH and high NaCl concentration under unsterile and continuous conditions without microbial contamination. In this paper, Halomonas strain TD is used as a host to produce a protein named PHA phasin or PhaP which has a potential to be developed into a bio-surfactant. Four Halomonas TD expression strains are constructed based on a strong T7-family expression system. Of these, the strain with phaC deletion and chromosomal expression system resulted in the highest production of PhaP in soluble form, reaching 19% of total cellular soluble proteins and with a yield of 1.86 g/L in an open fed-batch fermentation process. A simple "heat lysis and salt precipitation" method is applied to allow rapid PhaP purification from a mixture of cellular proteins with a PhaP recovery rate of 63%. It clearly demonstrated that Halomonas TD could be used for high yield expression of a bio-surfactant protein PhaP for industrial application in an economical way.
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Engineering of core promoter regions enables the construction of constitutive and inducible promoters in Halomonas sp
Biotechnology journal, 2015Co-Authors: Qiuyue Wang, Guo-qiang Chen, Haoqian Zhang, Chunbo Lou, Qi OuyangAbstract:Halomonas strain TD01, a newly identified halophilic bacterium, has proven to be a promising low-cost host for the production of chemicals. However, genetic manipulation in Halomonas sp. is still difficult due to the lack of well-characterized and tunable expression systems. In this study, a systematic, efficient method was exploited to construct both a constitutive promoter library and inducible promoters. Porin, a highly expressed protein in Halomonas TD01, was first identified from the Halomonas TD01 proteome. Subsequent study of the intergenic region upstream of porin led to the identification of a core promoter region, including -10 and -35 elements. By randomizing the sequence between the -35 and -10 elements, a constitutive promoter library was obtained with 310-fold variation in transcriptional activity; an inducible promoter with a >200-fold induction was built by integrating a lac operator into the core promoter region. As two complementary expression systems, the constitutive and inducible promoters were then employed to regulate the biosynthetic pathway of poly-3-hydroxybutyrate (PHB) in Halomonas TD01, demonstrating the usefulness of the expression systems, furthermore, they could be applied in future metabolic engineering of Halomonas TD strains, and the systematic method used in this study can be generalized to other less-characterized bacterial strains.
M. Carmen Márquez - One of the best experts on this subject based on the ideXlab platform.
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Description of Kushneria aurantia gen. nov., sp. nov., a novel member of the family Halomonadaceae, and a proposal for reclassification of Halomonas marisflavi as Kushneria marisflavi comb. nov., of Halomonas indalinina as Kushneria indalinina comb.
INTERNATIONAL JOURNAL OF SYSTEMATIC AND EVOLUTIONARY MICROBIOLOGY, 2009Co-Authors: Cristina Sánchez-porro, Rafael R. De La Haba, Nelís Soto-ramírez, M. Carmen Márquez, Rafael Montalvo-rodríguez, Antonio VentosaAbstract:An aerobic, moderately halophilic, Gram-negative, motile, non-sporulating rod-shaped bacterium, designated strain A10T, was isolated from the surface of leaves of the black mangrove Avicennia germinans and was subjected to a polyphasic taxonomic study. Strain A10T was able to grow at NaCl concentrations in the range 5–17.5 % (w/v) with optimum growth at 10 % (w/v) NaCl. Growth occurred at temperatures of 20–40 °C (optimal growth at 37 °C) and pH 5.5–8.5 (optimal growth at pH 7.0–8.0). The major respiratory quinone was ubiquinone 9. The major fatty acids were C16 : 0, C18 : 1 ω7c, C19 : 0 cyclo ω8c and C12 : 0 3-OH. The polar lipids were phosphatidylglycerol, diphosphatidylglycerol, phosphatidylethanolamine, and unidentified phospholipids, glycolipids and an aminoglycolipid. Phylogenetic analysis based on 16S rRNA gene sequence comparisons revealed that strain A10T is closely related to Halomonas avicenniae MW2aT (95.7 % sequence similarity), Halomonas marisflavi SW32T (95.2 %) and Halomonas indalinina GC2.1T (95.0 %). Strain A10T formed a coherent phylogenetic branch with these three species, separated from other species of Halomonas and closely related genera (with 16S rRNA gene sequence similarities below 94.0 %). A complete 23S rRNA gene sequence comparison of strain A10T with closely related species confirmed the phylogenetic position of the novel isolate, forming a branch with the species Halomonas avicenniae, Halomonas indalinina and Halomonas marisflavi, separated from other species of the genera belonging to the family Halomonadaceae (showing sequence similarities below 91.7 %). DNA–DNA hybridization studies between strain A10T and Halomonas avicenniae MW2aT, Halomonas marisflavi DSM 15357T and Halomonas indalinina CG2.1T were 21, 17 and 10 %, respectively. These levels of DNA–DNA relatedness were low enough to classify strain A10T as representing a genotypically distinct species. Overall, the phenotypic, genotypic, chemotaxonomic and phylogenetic results demonstrated that strain A10T represents a new genus and species. The name Kushneria aurantia gen. nov., sp. nov. is proposed, with strain A10T (=CCM 7415T=CECT 7220T) as the type strain. This is the type species of the new proposed genus, which belongs to the family Halomonadaceae. In addition, our data support the placement of the species Halomonas marisflavi, Halomonas indalinina and Halomonas avicenniae within this new genus, as Kushneria marisflavi comb. nov., Kushneria indalinina comb. nov. and Kushneria avicenniae comb. nov., respectively.
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Description of Kushneria aurantia gen. nov., sp. nov., a novel member of the family Halomonadaceae, and a proposal for reclassification of Halomonas marisflavi as Kushneria marisflavi comb. nov., of Halomonas indalinina as Kushneria indalinina comb.
International journal of systematic and evolutionary microbiology, 2009Co-Authors: Cristina Sánchez-porro, Rafael R. De La Haba, Nelís Soto-ramírez, M. Carmen Márquez, Rafael Montalvo-rodríguez, Antonio VentosaAbstract:An aerobic, moderately halophilic, Gram-negative, motile, non-sporulating rod-shaped bacterium, designated strain A10(T), was isolated from the surface of leaves of the black mangrove Avicennia germinans and was subjected to a polyphasic taxonomic study. Strain A10(T) was able to grow at NaCl concentrations in the range 5-17.5 % (w/v) with optimum growth at 10 % (w/v) NaCl. Growth occurred at temperatures of 20-40 degrees C (optimal growth at 37 degrees C) and pH 5.5-8.5 (optimal growth at pH 7.0-8.0). The major respiratory quinone was ubiquinone 9. The major fatty acids were C(16 : 0), C(18 : 1)omega7c, C(19 : 0) cyclo omega8c and C(12 : 0) 3-OH. The polar lipids were phosphatidylglycerol, diphosphatidylglycerol, phosphatidylethanolamine, and unidentified phospholipids, glycolipids and an aminoglycolipid. Phylogenetic analysis based on 16S rRNA gene sequence comparisons revealed that strain A10(T) is closely related to Halomonas avicenniae MW2a(T) (95.7 % sequence similarity), Halomonas marisflavi SW32(T) (95.2 %) and Halomonas indalinina GC2.1(T) (95.0 %). Strain A10(T) formed a coherent phylogenetic branch with these three species, separated from other species of Halomonas and closely related genera (with 16S rRNA gene sequence similarities below 94.0 %). A complete 23S rRNA gene sequence comparison of strain A10(T) with closely related species confirmed the phylogenetic position of the novel isolate, forming a branch with the species Halomonas avicenniae, Halomonas indalinina and Halomonas marisflavi, separated from other species of the genera belonging to the family Halomonadaceae (showing sequence similarities below 91.7 %). DNA-DNA hybridization studies between strain A10(T) and Halomonas avicenniae MW2a(T), Halomonas marisflavi DSM 15357(T) and Halomonas indalinina CG2.1(T) were 21, 17 and 10 %, respectively. These levels of DNA-DNA relatedness were low enough to classify strain A10(T) as representing a genotypically distinct species. Overall, the phenotypic, genotypic, chemotaxonomic and phylogenetic results demonstrated that strain A10(T) represents a new genus and species. The name Kushneria aurantia gen. nov., sp. nov. is proposed, with strain A10(T) (=CCM 7415(T)=CECT 7220(T)) as the type strain. This is the type species of the new proposed genus, which belongs to the family Halomonadaceae. In addition, our data support the placement of the species Halomonas marisflavi, Halomonas indalinina and Halomonas avicenniae within this new genus, as Kushneria marisflavi comb. nov., Kushneria indalinina comb. nov. and Kushneria avicenniae comb. nov., respectively.
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Halomonas neptunia sp. nov., Halomonas sulfidaeris sp. nov., Halomonas axialensis sp. nov. and Halomonas hydrothermalis sp. nov.: halophilic bacteria isolated from deep-sea hydrothermal-vent environments.
International Journal of Systematic and Evolutionary Microbiology, 2004Co-Authors: Jonathan Z Kaye, Antonio Ventosa, M. Carmen Márquez, John A BarossAbstract:To assess the physiological and phylogenetic diversity of culturable halophilic bacteria in deep-sea hydrothermal-vent environments, six isolates obtained from low-temperature hydrothermal fluids, sulfide rock and hydrothermal plumes in North and South Pacific Ocean vent fields located at 1530–2580 m depth were fully characterized. Three strains were isolated on media that contained oligotrophic concentrations of organic carbon (0·002 % yeast extract). Sequencing of the 16S rRNA gene indicated that all strains belonged to the genus Halomonas in the γ-subclass of the Proteobacteria. Consistent with previously described species, the novel strains were slightly to moderately halophilic and grew in media containing up to 22–27 % total salts. The isolates grew at temperatures as low as −1 to 2 °C and had temperature optima of 30 or 20–35 °C. Both the minimum and optimum temperatures for growth were similar to those of Antarctic and sea-ice Halomonas species and lower than typically observed for the genus as a whole. Phenotypic tests revealed that the isolates were physiologically versatile and tended to have more traits in common with each other than with closely related Halomonas species, presumably a reflection of their common deep-sea, hydrothermal-vent habitat of origin. The G+C content of the DNA for all strains was 56·0–57·6 mol%, and DNA–DNA hybridization experiments revealed that four strains (Eplume1T, Esulfide1T, Althf1T and Slthf2T) represented novel species and that two strains (Eplume2 and Slthf1) were related to Halomonas meridiana. The proposed new species names are Halomonas neptunia (type strain Eplume1T=ATCC BAA-805T=CECT 5815T=DSM 15720T), Halomonas sulfidaeris (type strain Esulfide1T=ATCC BAA-803T=CECT 5817T=DSM 15722T), Halomonas axialensis (type strain Althf1T=ATCC BAA-802T=CECT 5812T=DSM 15723T) and Halomonas hydrothermalis (type strain Slthf2T=ATCC BAA-800T=CECT 5814T=DSM 15725T).
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Halomonas neptunia sp. nov., Halomonas sulfidaeris sp. nov., Halomonas axialensis sp. nov. and Halomonas hydrothermalis sp. nov.: halophilic bacteria isolated from deep-sea hydrothermal-vent environments.
International journal of systematic and evolutionary microbiology, 2004Co-Authors: Jonathan Z Kaye, Antonio Ventosa, M. Carmen Márquez, John A BarossAbstract:To assess the physiological and phylogenetic diversity of culturable halophilic bacteria in deep-sea hydrothermal-vent environments, six isolates obtained from low-temperature hydrothermal fluids, sulfide rock and hydrothermal plumes in North and South Pacific Ocean vent fields located at 1530-2580 m depth were fully characterized. Three strains were isolated on media that contained oligotrophic concentrations of organic carbon (0.002 % yeast extract). Sequencing of the 16S rRNA gene indicated that all strains belonged to the genus Halomonas in the gamma-subclass of the Proteobacteria. Consistent with previously described species, the novel strains were slightly to moderately halophilic and grew in media containing up to 22-27 % total salts. The isolates grew at temperatures as low as -1 to 2 degrees C and had temperature optima of 30 or 20-35 degrees C. Both the minimum and optimum temperatures for growth were similar to those of Antarctic and sea-ice Halomonas species and lower than typically observed for the genus as a whole. Phenotypic tests revealed that the isolates were physiologically versatile and tended to have more traits in common with each other than with closely related Halomonas species, presumably a reflection of their common deep-sea, hydrothermal-vent habitat of origin. The G+C content of the DNA for all strains was 56.0-57.6 mol%, and DNA-DNA hybridization experiments revealed that four strains (Eplume1(T), Esulfide1(T), Althf1(T) and Slthf2(T)) represented novel species and that two strains (Eplume2 and Slthf1) were related to Halomonas meridiana. The proposed new species names are Halomonas neptunia (type strain Eplume1(T)=ATCC BAA-805(T)=CECT 5815(T)=DSM 15720(T)), Halomonas sulfidaeris (type strain Esulfide1(T)=ATCC BAA-803(T)=CECT 5817(T)=DSM 15722(T)), Halomonas axialensis (type strain Althf1(T)=ATCC BAA-802(T)=CECT 5812(T)=DSM 15723(T)) and Halomonas hydrothermalis (type strain Slthf2(T)=ATCC BAA-800(T)=CECT 5814(T)=DSM 15725(T)).
