Hydrogenophaga crocea sp. nov. associated with cyanobacterial mat isolated from farmland mud.

A catalase and oxidase-positive strain BA0156T was isolated from a cyanobacterial mat collected from the farmland mud cultivated with sugarcane from Ahmednagar, India. The 16S rRNA gene of strain BA0156T showed the highest percent sequence similarity with Hydrogenophaga borbori LMG 30805T (98.5%), followed by H. flava DSM 619T (98.3%) and H. intermedia DSM 5680T (98.2%). The strain BA0156T contained the major fatty acids, C16:0 (25.1%) and C17:0 cyclo (3.9%), whereas phosphatidylethanolamine and diphosphatidylglycerol were the major polar lipids. The OrthoANI and dDDH values between strain BA0156T and its closest relative H. borbori LMG 30805T were 84.6% and 28.3%, respectively. The DNA G+C content of strain BA0156T was 69.4 mol %. Furthermore, the biochemical and physiological features of strain BA0156T showed a distinct pattern from their closest phylogenetic neighbours. The phenotypic, genotypic and chemotaxonomic characteristics indicated that the strain BA0156T represents a new species for which the name Hydrogenophaga crocea (type strain BA0156T = MCC 3062T = KCTC 72452T = JCM 34507T) is proposed.

Production of a novel dimeric 4-deoxy-L-erythro-5-hexoseulose uronic acid by a PL-17 exolytic alginate lyase from Hydrogenophaga sp. UMI-18.

Hydrogenopahaga sp. strain UMI-18 is an alginolytic bacterium that can produce poly(3-hydroxybutylate) (PHB) using alginate as its sole carbon source. Genome analysis indicated that this strain harbors both PHB-synthesizing and alginate-assimilating gene clusters. In the present study, we cloned HyAly-I gene that encodes a PL-17 exolytic alginate lyase and investigated its enzymatic properties using recombinant HyAly-I (recHyAly-I) that was produced by Escherichia coli. The recHyAly-I preferably depolymerized poly(ß-D-mannuronate) block of alginate in an exolytic manner at an optimal temperature and a pH at 40 °C and pH 6.0, respectively. It released 4-deoxy-L-erythro-5-hexoseulose uronic acid (DEH) from the non-reducing terminus of polymer and oligomer substrates. Interestingly, recHyAly-I was found to produce a novel unsaturated disaccharide, i.e., dimeric DEH (diDEH), along with monomeric DEH. Production of diDEH was prominent in the degradation of trisaccharides.

Hydrogenophaga borbori sp. nov., isolated from activated sludge.

A novel Gram-stain-negative, yellowish-pigmented bacterial strain, designated LA-38T, was isolated from activated sludge of wastewater treatment plants in Hanam city, South Korea. Cell of LA-38T were rod-shaped, aerobic, motile and non-spore-forming. In phylogenetic analyses based on 16S rRNA genes, LA-38T clustered with species of the genus Hydrogenophaga and appeared closely related to Hydrogenophaga intermedia DSM 5680T (99.2 % similarity), Hydrogenophaga palleronii DSM 63T (98.2 %), Hydrogrenophaga laconesensis KCTC 42478T (98.1 %), Hydrogenophaga. atypica DSM 15342T (98.1 %), Hydrogenophaga defluvii DSM 15341T (98.0 %) and Hydrogenophaga taeniospiralis DSM 2082T (97.2 %). The average nucleotide identities between LA-38T and the closely related strains were 79.3-88.5 %, indicating that LA-38T represents a novel species of the genus Hydrogenophaga. The DNA G+C content of the genomic DNA was 69.9 mol% and ubiquinone Q-8 was the predominant respiratory quinone. The major cellular fatty acids (>5 %) were C16 : 0, cyclo-C19 : 0, C16 : 1ω7c and/or C16 : 1ω6c (summed feature 3), and C18 : 1ω7c and/or C18 : 1ω6c (summed feature 8). The major polar lipids consisted of phosphatidylglycerol, diphosphatidylglycerol and phosphatidylethanolamine, the major polyamines were 2-hydroxyputrescine and putrescine. ANI calculation, physiological and biochemical characteristics indicated that LA-38T represents a novel species of the genus Hydrogenophaga, for which the name Hydrogenophaga borbori sp. nov. is proposed. The type strain is LA-38T (=KACC 19730T=LMG 30805T).

Isolation and characterization of 2-butoxyethanol degrading bacterial strains.

A total of 11 bacterial strains capable of completely degrading 2-butoxyethanol (2-BE) were isolated from forest soil, a biotrickling filter, a bioscrubber, and activated sludge, and identified by 16S rRNA gene sequence analysis. Eight of these strains belong to the genus Pseudomonas; the remaining three strains are Hydrogenophaga pseudoflava BOE3, Gordonia terrae BOE5, and Cupriavidus oxalaticus BOE300. In addition to 2-BE, all isolated strains were able to grow on 2-ethoxyethanol and 2-propoxyethanol, ethanol, n-hexanol, ethyl acetate, 2-butoxyacetic acid (2-BAA), glyoxylic acid, and n-butanol. Apart from the only gram-positive strain isolated, BOE5, none of the strains were able to grow on the nonpolar ethers diethyl ether, di-n-butyl ether, n-butyl vinyl ether, and dibenzyl ether, as well as on 1-butoxy-2-propanol. Strains H. pseudoflava BOE3 and two of the isolated pseudomonads, Pseudomonas putida BOE100 and P. vancouverensis BOE200, were studied in more detail. The maximum growth rates of strains BOE3, BOE100, and BOE200 at 30 °C were 0.204 h-1 at 4 mM, 0.645 h-1 at 5 mM, and 0.395 h-1 at 6 mM 2-BE, respectively. 2-BAA, n-butanol, and butanoic acid were detected as potential metabolites during the degradation of 2-BE. These findings indicate that the degradation of 2-BE by the isolated gram-negative strains proceeds via oxidation to 2-BAA with subsequent cleavage of the ether bond yielding glyoxylate and n-butanol. Since Gordonia terrae BOE5 was the only strain able to degrade nonpolar ethers like diethyl ether, the degradation pathway of 2-BE may be different for this strain.

Exploiting Hydrogenophaga pseudoflava for aerobic syngas-based production of chemicals.

Gasification is a suitable technology to generate energy-rich synthesis gas (syngas) from biomass or waste streams, which can be utilized in bacterial fermentation processes for the production of chemicals and fuels. Established microbial processes currently rely on acetogenic bacteria which perform an energetically inefficient anaerobic CO oxidation and acetogenesis potentially hampering the biosynthesis of complex and ATP-intensive products. Since aerobic oxidation of CO is energetically more favorable, we exploit in this study the Gram-negative ß-proteobacterium Hydrogenophaga pseudoflava DSM1084 as novel host for the production of chemicals from syngas. We sequenced and annotated the genome of H. pseudoflava and established a genetic engineering toolbox, which allows markerless chromosomal modification via the pk19mobsacB system and heterologous gene expression on pBBRMCS2-based plasmids. The toolbox was extended by identifying strong endogenous promotors such as PgapA2 which proved to yield high expression under heterotrophic and autotrophic conditions. H. pseudoflava showed relatively fast heterotrophic growth in complex and minimal medium with sugars and organic acids which allows convenient handling in lab routines. In autotrophic bioreactor cultivations with syngas, H. pseudoflava exhibited a growth rate of 0.06 h-1 and biomass specific uptakes rates of 14.2 ±â€¯0.3 mmol H2 gCDW-1 h-1, 73.9 ±â€¯1.8 mmol CO gCDW-1 h-1, and 31.4 ±â€¯0.3 mmol O2 gCDW-1 h-1. As proof of concept, we engineered the carboxydotrophic bacterium for the aerobic production of the C15 sesquiterpene (E)-α-bisabolene from the C1 carbon source syngas by heterologous expression of the (E)-α-bisabolene synthase gene agBIS. The resulting strain H. pseudoflava (pOCEx1:agBIS) produced 59 ±â€¯8 µg (E)-α-bisabolene L-1 with a volumetric productivity Qp of 1.2 ±â€¯0.2 µg L-1 h-1 and a biomass-specific productivity qp of 13.1 ±â€¯0.6 µg gCDW-1 h-1. The intrinsic properties and the genetic repertoire of H. pseudoflava make this carboxydotrophic bacterium a promising candidate for future aerobic production processes to synthesize more complex or ATP-intensive chemicals from syngas.

Hydrogenophaga aromaticivorans sp. nov., isolated from a para-xylene-degrading enrichment culture, capable of degrading benzene, meta- and para-xylene.

A benzene, para- and meta-xylene-degrading Gram-stain-negative, aerobic, yellow-pigmented bacterium, designated as D2P1T, was isolated from a para-xylene-degrading enrichment culture. Phylogenetic analyses based on 16S rRNA genes showed that D2P1T shares a distinct phyletic lineage within the genus Hydrogenophaga and shows highest 16S rRNA gene sequence similarity to Hydrogenophaga taeniospiralis NBRC 102512T (99.2 %) and Hydrogenophaga palleronii NBRC 102513T (98.3 %). The draft genome sequence of D2P1T is 5.63 Mb long and the genomic DNA G+C content is 65.5 %. Orthologous average nucleotide identity (OrthoANI) and digital DNA-DNA hybridization (dDDH) analyses confirmed low genomic relatedness to its closest relatives (OrthoANI <86 %; dDDH <30 %). D2P1T contains ubiquinone 8 (Q-8) as the only respiratory quinone and phospholipid, phosphatidylglycerol, phosphatidylethanolamine, diphosphatidylglycerol as major polar lipids. The main whole-cell fatty acids of D2P1T are summed feature 3 (C16 : 1 ω7c/C16 : 1 ω6c), C16 : 0 and summed feature 8 (C18 : 1 ω7c/C18 : 1 ω6c). The polyphasic taxonomic results indicated that strain D2P1T represents a novel species of the genus Hydrogenophaga, for which the name Hydrogenophaga aromaticivorans sp. nov. is proposed. The type strain is D2P1T (=LMG 31780T=NCAIM B 02655T).

Insights into selenate removal mechanism of hydrogen-based membrane biofilm reactor for nitrate-polluted groundwater treatment based on anaerobic biofilm analysis.

The selenate removal mechanism of hydrogen-based membrane biofilm reactor (MBfR) for nitrate-polluted groundwater treatment was studied based on anaerobic biofilm analysis. A laboratory-scale MBfR was operated for over 60 days with electron balance, structural analysis, and bacterial community identification. Results showed that anaerobic biofilm had an excellent removal of both selenate (95%) and nitrate (100%). Reduction of Selenate → Selenite → Se0 with hydrogen was the main pathway of anaerobic biofilm for selenate removal with amorphous Se0 precipitate accumulating in the biofilm. The element selenium was observed to be evenly distributed along the cross-sectional thin biofilm. A part of selenate (3%) was also reduced into methyl-selenide by heterotrophic bacteria. Additionally, Hydrogenophaga bacteria of ß-Proteobacteria, capable of both nitrate and selenate removal, worked as the dominant species (over 85%) in the biofilm and contributed to the stable removal of both nitrate and selenate. With the selenate input, bacteria with a capacity for both selenate and nitrate removal were also developed in the anaerobic biofilm community.

Effects of residual organics in municipal wastewater on hydrogenotrophic denitrifying microbial communities.

Hydrogenotrophic denitrification is promising for tertiary nitrogen removal from municipal wastewater. To reveal the influence of residual organics in municipal wastewater on hydrogenotrophic denitrifiers, we adopted high-throughput 16S rRNA gene amplicon sequencing to examine microbial communities in hydrogenotrophic denitrification enrichments. Using effluent from a municipal wastewater treatment plant as water source, COD, nitrate and pH were controlled the same except for a gradient of biodegradable carbon (i.e., primary effluent (PE), secondary effluent (SE), or combined primary and secondary effluent (CE)). Inorganic synthetic water (IW) was used as a control. Hydrogenophaga, a major facultative autotroph, accounted for 17.1%, 5.3%, 32.7% and 12.9% of the sequences in PE, CE, SE and IW, respectively, implicating that Hydrogenophaga grew well with or without organics. Thauera, which contains likely obligate autotrophic denitrifiers, appeared to be the most dominant genera (23.6%) in IW and accounted for 2.5%, 4.6% and 8.9% in PE, CE and SE, respectively. Thermomonas, which is related to heterotrophic denitrification, accounted for 4.2% and 7.9% in PE and CE fed with a higher content of labile organics, respectively. In contrast, Thermomonas was not detected in IW and accounted for only 0.6% in SE. Our results suggest that Thermomonas are more competitive than Thauera in hydrogenotrophic denitrification with biodegradable organics. Moreover, facultative autotrophic denitrifiers, Hydrogenophaga, are accommodating to residual organic in effluent wastewater, thus we propose that hydrogenotrophic denitrification is amenable for tertiary nitrogen removal.

Hydrogenophaga aquatica sp. nov., isolated from a hot spring.

A polyphasic approach was used to characterize an aerobic, Gram-negative, rod-shaped bacterium (designated strain CC-KL-3T) isolated from a hot spring. Phylogenetic analyses based on 16S rRNA genes indicated that strain CC-KL-3T showed highest sequence similarity to Hydrogenophaga bisanensis (97.7 %) and Hydrogenophaga atypica (97.6 %) and lower sequence similarity to other species (less than 97.6 %). The levels of DNA-DNA relatedness between strain CC-KL-3T, H. bisanensis and H. atypica were estimated to be 13.0 and 8.7 % (the reciprocal value was 14.7 and 6.3 %). Strain CC-KL-3T was non-motile, without apparent flagella and able to grow between 15-42 °C (optimal 30 °Ð¡), pH 6.0-8.0 (optimal 7.0) and 0-2 % (w/v) NaCl (optimal 0 %). The DNA G+C content was 61.4 mol% and the major quinone system was ubiquinone (Q-8). The polyamine profile revealed the predominance of 2-hydroxyputrescine and putrescine and the dominant cellular fatty acids were C16 : 0 (28.9 %), C16 : 1ω7c/C16 : 1ω6c (41.4 %) and C18 : 1ω7c/C18 : 1ω6c (11.9 %). These data corroborated the affiliation of strain CC-KL-3T to the genus Hydrogenophaga. Based on the distinct phylogenetic, phenotypic and chemotaxonomic traits, and the results of comparative 16S rRNA gene sequence analysis, strain CC-KL-3T is considered to represent a novel species of the genus Hydrogenophaga, affiliated to the family Comamonadaceae, for which the name Hydrogenophaga aquatica sp. nov. is proposed. The type strain is CC-KL-3T (=BCRC 80937T=JCM 31216T).

Hydrogenophaga soli sp. nov., isolated from rice field soil.

A novel Gram-stain-negative bacterial strain, designated strain S10T, was isolated from soil collected in a rice field in Goyang, South Korea. Cells of strain S10T were strictly aerobic, motile and rod-shaped. Colonies were round, convex, smooth and white. The strain grew optimally at 37 °C, pH 7.0 and 0 % (w/v) NaCl. Phylogenetic analysis of the 16S rRNA gene sequence of strain S10T revealed that the bacterium belongs to the family Comamonadaceae and is related to members of the genus Hydrogenophaga, with Hydrogenophaga caeni EMB71T being its closest relative (97.9 % sequence similarity). The DNA G+C content of strain S10T was 68.2 mol%. Strain S10T contained phosphatidylethanolamine and diphosphatidylglycerol as the major polar lipids. The major fatty acids were C16 : 0 and summed feature 3 (C16 : 1ω7c and/or iso-C15 : 0 2-OH). The predominant respiratory quinone was ubiquinone Q-8. DNA-DNA hybridization values of strain S10T with Hydrogenophaga caeni KCTC 12613T, Hydrogenophaga atypica DSM 15342T and Hydrogenophaga defluvii DSM 15341T were 16.1±4.8, 49.0±3.2 and 21.9±8.8 %, respectively. Based on phylogenetic distinctiveness, DNA-DNA hybridization and specific physiological and biochemical characteristics, strain S10T (=KCTC 52520T=JCM 31711T) is classified as a novel species of the genus Hydrogenophaga, for which the name Hydrogenophagasoli sp. nov. is proposed.

Description of Hydrogenophaga laconesensis sp. nov. isolated from tube well water.

A light cream color colony was isolated, using oligotrophic LB agar medium, from a water sample collected from a tube well and designated as HWB-10(T). Cells of strain HWB-10(T) were Gram-negative, motile rods, non-spore forming, positive for catalase, oxidase, nitrate reduction and esculin. The predominant fatty acids were C16:0, summed feature 4 (C16:1 ω7c/iso-C15:0 2-OH) and summed feature 7 (C18:1 ω7c, C18:1 ω9t and/or C18:1 ω12t), and the major ubiquinone was Q-8. NCBI-BLAST- and EzTaxon-based 16S rRNA gene sequence similarity search identified strain HWB-10(T) as a member of the genus Hydrogenophaga and H. atypica DSM 15342(T), H. defulvi DSM 15341(T), H. palleronii LMG2366(T) and H. taeniospiralis LMG7170(T) being the nearest phylogenetic species with a similarity (%) of 99.3, 99.1, 98.4 and 98.2, respectively, while the similarity was <98.0 % with other species of the genus. However, DNA-DNA similarities between HWB-10(T) and H. atypica DSM 15342(T) and H. defulvi DSM 15341(T) were 37.0 and 43.0 %, respectively, indicating that strain HWB-10(T) is a novel species. Further, the DNA fingerprinting, based on BOX-, ERIC-, (GTG)5- and REP-PCR amplifications, distinguished strain HWB-10(T) from its closest species, H. atypica DSM 15342(T) and H. defulvi DSM 15341(T) with similarity coefficients of 0.45 and 0.37, respectively, a value sufficient to establish the species status within the genus Hydrogenophaga. In addition, strain HWB-10(T) exhibited several phenotypic differences with its closely related species. Based on the above cumulative differentiating characteristics, strain HWB-10(T) was identified as a new species of the genus Hydrogenophaga and proposed as Hydrogenophaga laconesensis sp. nov. with strain HWB-10(T) (KTCC 42478(T) = LMG 28681(T)) as its type strain.

Aerobic biodegradation of cycloalkanes in non-aqueous extracted oil sands tailings.

Management of growing volumes of fluid fine tailings (FFT) is a significant challenge for oil sands industry. A potential alternative non-aqueous solvent extraction (NAE) process uses cycloalkane solvent such as cyclohexane or cyclopentane with very little water and generates smaller volumes of 'dry' solids (NAES) with residual solvent. Here we investigate remediation of NAES in a simulated bench-scale upland reclamation scenario. In the first study, microcosms with nutrient medium plus FFT as inoculum were amended with cyclohexane and incubated for ∼1 year, monitoring for cyclohexane biodegradation under aerobic conditions. Biodegradation of cyclohexane occurred under aerobic conditions with no metabolic intermediates detected. A second study using NAES mixed with FFT spiked with cyclohexane and cyclopentane, with or without additional nutrients (nitrogen and phosphorus), showed complete and rapid aerobic biodegradation of both cycloalkanes in NAES inoculated with FFT and supplemented with nutrients. 16S rRNA gene sequencing revealed dominance of Rhodoferax and members of Burkholderiaceae during aerobic cyclohexane biodegradation in FFT, and Hydrogenophaga, Acidovorax, Defluviimonas and members of Porticoccaceae during aerobic biodegradation of cyclohexane and cyclopentane in NAES inoculated with FFT and supplemented with nutrients. The findings indicate that biodegradation of cycloalkanes from NAES is possible under aerobic condition, which will contribute to the successful reclamation of oil sands tailings for land closure.

High nitrite accumulation in hydrogenotrophic denitrification at low temperature: Transcriptional regulation and microbial community succession.

High Pressure Hydrogenotrophic Denitrification (HPHD) provided a promising alternative for efficient and clean nitrate removal. In particular, the denitrification rates at low temperature could be compensated by elevated H2 partial pressure. However, nitrite reduction was strongly inhibited while nitrate reduction was barely affected at low temperature. In this study, the nitrate reduction gradually recovered under long-term low temperature stress, while nitrite accumulation increased from 0.1 to 41.0 mg N/L. The activities of the electron transport system (ETS), nitrate reductase (NAR), and nitrite reductase (NIR) decreased by 45.8 %, 27.3 %, and 39.3 %, respectively, as the temperature dropped from 30 °C to 15 °C. Real time quantitative PCR analysis revealed that the denitrifying gene expression rather than gene abundance regulated nitrogen biotransformation. The substantial nitrite accumulation was attributed to the significant up-regulation by 54.7 % of narG gene expression and down-regulation by 73.7 % of nirS gene expression in hydrogenotrophic denitrifiers. In addition, the nirS-gene-bearing denitrifiers were more sensitive to low temperature compared to those bearing nirK gene. The dominant populations shifted from the genera Paracoccus to Hydrogenophaga under long-term low temperature stress. Overall, this study revealed the microbial mechanism of high nitrite accumulation in hydrogenotrophic denitrification at low temperature.

Case report: isolation of Hydrogenophaga from septic blood culture following near-death drowning in lakewater.

A patient suffered a non-fatal wet drowning in a freshwater lake and developed bacteraemia several days later. Blood culture grew a Gram-negative rod that could not be identified by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS). 16S ribosomal RNA sequencing of the isolate identified the microbe as Hydrogenophaga laconesensis - an environmental microbe commonly found in freshwater. The recovery of multiple pathogenic micro-organisms (although not H. laconesensis ) from culture of respiratory specimens prompted the initiation of antibiotic therapy with cefepime and, later, vancomycin. The patient's clinical course gradually improved over the course of 2 weeks and she was ultimately discharged home with minimal sequelae. To our knowledge, this is the first evidence of human infection with bacteria in the genus Hydrogenophaga . Hydrogenophaga may be considered in cases of freshwater near-drowning, and MALDI-TOF MS databases should be updated to include H. laconesensis .

Biosynthesis and properties of polyhydroxyalkanoates synthesized from mixed C5 and C6 sugars obtained from hardwood hydrolysis.

Glucose and xylose are fermentable sugars readily available from lignocellulosic biomass, and are a sustainable carbon substrate supporting industrial biotechnology. Three strains were assessed in this work - Paraburkholderia sacchari, Hydrogenophaga pseudoflava, and Bacillus megaterium - for their ability to uptake both C5 and C6 sugars contained in a hardwood hydrolysate produced via a thermomechanical pulping-based process with concomitant production of poly(3-hydroxyalkanoate) (PHA) biopolymers. In batch conditions, B. megaterium showed poor growth after 12 h, minimal uptake of xylose throughout the cultivation, and accumulated a maximum of only 25 % of the dry biomass as PHA. The other strains simultaneously utilized both sugars, although glucose uptake was faster than xylose. From hardwood hydrolysate, P. sacchari accumulated 57 % of its biomass as PHA within 24 h, whereas H. pseudoflava achieved an intracellular PHA content of 84 % by 72 h. The molecular weight of the PHA synthesized by H. pseudoflava (520.2 kDa) was higher than that of P. sacchari (265.5 kDa). When the medium was supplemented with propionic acid, the latter was rapidly consumed by both strains and incorporated as 3-hydroxyvalerate subunits into the polymer, demonstrating the potential for production of polymers with improved properties and value. H. pseudoflava incorporated 3-hydroxyvalerate subunits with at least a 3-fold higher yield, and produced polymers with higher 3-hydroxyvalerate content than P. sacchari. Overall, this work has shown that H. pseudoflava can be an excellent candidate for bioconversion of lignocellulosic sugars to PHA polymers or copolymers as part of an integrated biorefinery.

Thiamine-Mediated Microbial Interaction between Auxotrophic Rhodococcus ruber ZM07 and Prototrophic Cooperators in the Tetrahydrofuran-Degrading Microbial Community H-1.

As a crucial growth factor, thiamine can regulate functional microbial communities; however, our current understanding of its effect on bioremediation is lacking. Using metatranscriptome and 16S rRNA gene sequencing, we explored the mechanism of response of an efficient tetrahydrofuran (THF)-degrading microbial culture, designated H-1, to exogenous thiamine. Rhodococcus ruber ZM07, a strain performing the THF degradation function in H-1, is a thiamine-auxotrophic bacterium. Furthermore, thiamine affected the microbial community structure of H-1 by altering resource and niche distributions. A microbial co-occurrence network was constructed to help us identify and isolate the cooperators of strain ZM07 in the microbial community. Based on the prediction of the network, two non-THF-degrading bacteria, Hydrogenophaga intermedia ZM11 and Pigmentiphaga daeguensis ZM12, were isolated. Our results suggest that strain ZM11 is a good cooperator of ZM07, and it might be more competitive than other cooperators (e.g., ZM12) in cocultured systems. Additionally, two dominant strains in our microbial culture displayed a "seesaw" pattern, and they showed completely different responses to exogenous thiamine. The growth of the THF degrader ZM07 was spurred by additional thiamine (with an increased relative abundance and significant upregulation of most metabolic pathways), while the growth of the cooperator ZM11 was obviously suppressed under the same circumstances. This relationship was the opposite without thiamine addition. Our study reveals that exogenous thiamine can affect the interaction patterns between THF- and non-THF-degrading microorganisms and provides new insight into the effects of micronutrients on the environmental microbial community. IMPORTANCE Auxotrophic microorganisms play important roles in the biodegradation of pollutants in nature. Exploring the interspecies relationship between auxotrophic THF-degrading bacteria and other microbes is helpful for the efficient utilization of auxotrophic functional microorganisms. Herein, the thiamine-auxotrophic THF-degrading bacterium ZM07 was isolated from the microbial culture H-1, and the effect of thiamine on the structure of H-1 during THF bioremediation was studied. Thiamine may help ZM07 occupy more niches and utilize more resources, thus improving THF degradation efficiency. This research provides a new strategy to improve the THF or other xenobiotic compound biodegradation performance of auxotrophic functional microorganisms/microbial communities by artificially adding special micronutrients. Additionally, the "seesaw" relationship between the thiamine-auxotrophic strain ZM07 and its prototrophic cooperator ZM11 during THF bioremediation could be changed by exogenous thiamine. This study reveals the effect of micronutrients on microbial interactions and provides an effective way to regulate the pollutant biodegradation efficiency of microbial communities.

Simultaneous removal of nitrogen and arsenite by heterotrophic nitrification and aerobic denitrification bacterium Hydrogenophaga sp. H7.

Introduction: Nitrogen and arsenic contaminants often coexist in groundwater, and microbes show the potential for simultaneous removal of nitrogen and arsenic. Here, we reported that Hydrogenophaga sp. H7 was heterotrophic nitrification and aerobic denitrification (HNAD) and arsenite [As(III)] oxidation bacterium. Methods: The appearance of nitrogen removal and As(III) oxidation of Hydrogenophaga sp. H7 in liquid culture medium was studied. The effect of carbon source, C/N ratio, temperature, pH values, and shaking speeds were analyzed. The impact of strains H7 treatment with FeCl3 on nitrogen and As(III) in wastewater was assessed. The key pathways that participate in simultaneous nitrogen removal and As(III) oxidation was analyzed by genome and proteomic analysis. Results and discussion: Strain H7 presented efficient capacities for simultaneous NH4 +-N, NO3 --N, or NO2 --N removal with As(III) oxidation during aerobic cultivation. Strikingly, the bacterial ability to remove nitrogen and oxidize As(III) has remained high across a wide range of pH values, and shaking speeds, exceeding that of the most commonly reported HNAD bacteria. Additionally, the previous HNAD strains exhibited a high denitrification efficiency, but a suboptimal concentration of nitrogen remained in the wastewater. Here, strain H7 combined with FeCl3 efficiently removed 96.14% of NH4 +-N, 99.08% of NO3 --N, and 94.68% of total nitrogen (TN), and it oxidized 100% of As(III), even at a low nitrogen concentration (35 mg/L). The residues in the wastewater still met the V of Surface Water Environmental Quality Standard of China after five continuous wastewater treatment cycles. Furthermore, genome and proteomic analyses led us to propose that the shortcut nitrification-denitrification pathway and As(III) oxidase AioBA are the key pathways that participate in simultaneous nitrogen removal and As(III) oxidation.

Novel pathway and acetate-facilitated complete atenolol degradation by Hydrogenophaga sp. YM1 isolated from activated sludge.

Atenolol is a widely prescribed beta-blocker that has been detected in wastewater at concentrations up to 300 µg/L. The parent compound and its transformation products pose risks to aquatic organisms. Efficient atenolol degrading microorganism has yet to be identified, and its biodegradation pathway is unknown. In this study, Hydrogenophaga sp. YM1 isolated from activated sludge can degrade atenolol efficiently (286.1 ± 4.0 µg/g dry wt/h in actual wastewater), where atenolol acid, and four newly detected products (4-hydroxyphenylacetic acid, 3-(isopropylamino)-1,2-propanediol, 3-amino-1,2-propanediol and 4-(1-amino-2-hydroxy-3-propoxy) benzeneacetic acid) were the main intermediates. Key genes involved in atenolol degradation were proposed based on RNA-seq and validated by RT-qPCR. The ether bond cleavage of atenolol acid was the rate-limiting step likely catalyzed by the α-ketoglutarate dependent 2,4-dichlorophenoxyacetate dioxygenase. The further degradation of 4-hydroxyphenylacetic acid followed the homoprotocatechuate degradation pathway, enabling complete conversion to CO2. Acetate addition (39-156 mg COD/L) under aerobic condition enhanced atenolol degradation by 29-37% and decreased the accumulation of atenolol acid, likely because acetate oxidation provided α-ketoglutarate and additional reducing power. Activated sludge core microorganisms have limited atenolol mineralization potentials. Enriching Hydrogenophaga-like populations and/or providing such as acetate can drive more complete conversion of atenolol in natural and engineered biosystems.

Polyhydroxyalkanoate Production from Fruit and Vegetable Waste Processing.

Traditional plastics represent a tremendous threat to the environment because of increases in polluting manufacturing as well as their very extended degradation time. Polyhydroxyalkanoates (PHAs) are polymers with similar performance to plastic but are compostable and synthesizable from renewable sources and therefore could be a replacement for fossil-based plastics. However, their production costs are still too high, thus demanding the investigation of new and cheap substrates. In this sense, agricultural wastes are attractive because they are inexpensive and largely available. Specifically, fruit and vegetables are rich in sugars that could be fermented into PHAs. In this work two strains, Cupriavidus necator DSM 545 and Hydrogenophaga pseudoflava DSM 1034, well-known PHA-producing microbes, were screened for their ability to grow and accumulate PHAs. Ten different fruit and vegetable processing waste streams, never before reported in combination with these strains, were tested. Residues from red apple and melon were found to be the most suitable feedstocks for PHA production. Under specific selected conditions, C. necator DSM 545 accumulated up to 7.4 and 4.3 g/L of 3-hydroxybutyrate (3HB) from red apple and melon, respectively. Copolymer production was also obtained from melon. These results confirm the attractiveness of food processing waste as a promising candidate for PHA production. Ultimately, these novel substrates draw attention for future studies on process optimization and upscaling with C. necator.

Exploiting Aerobic Carboxydotrophic Bacteria for Industrial Biotechnology.

Aerobic carboxydotrophic bacteria are a group of microorganisms which possess the unique trait to oxidize carbon monoxide (CO) as sole energy source with molecular oxygen (O2) to produce carbon dioxide (CO2) which subsequently is used for biomass formation via the Calvin-Benson-Bassham cycle. Moreover, most carboxydotrophs are also able to oxidize hydrogen (H2) with hydrogenases to drive the reduction of carbon dioxide in the absence of CO. As several abundant industrial off-gases contain significant amounts of CO, CO2, H2 as well as O2, these bacteria come into focus for industrial application to produce chemicals and fuels from such gases in gas fermentation approaches. Since the group of carboxydotrophic bacteria is rather unknown and not very well investigated, we will provide an overview about their lifestyle and the underlying metabolic characteristics, introduce promising members for industrial application, and give an overview of available genetic engineering tools. We will point to limitations and discuss challenges, which have to be overcome to apply metabolic engineering approaches and to utilize aerobic carboxydotrophs in the industrial environment.