Whole-Genome Sequencing of a Multidrug-Resistant Strain: Delftia acidovorans B408.

BACKGROUND: Delftia acidovorans is distributed widely in the environment and has the potential to promote the growth of plants and degrade organic pollutants. However, it is also an opportunistic pathogen for human and many reports demonstrated that D. acidovorans has strong resistance to aminoglycosides and polymyxins. OBJECTIVE: The aim of this work was to reveal the antibiotic resistance genes and pathogenic genes in a novel conditional pathogenic strain-D. acidovorans B804, which was isolated from the radiation-polluted soil from Xinjiang Uyghur Autonomous Region, China. METHODS: The antibiotic resistance test was performed according to the Kirby-Bauer disk diffusion method and evaluated by the standards of the Clinical and Laboratory Standards Institute guidelines. The genome of D. acidovorans B804 was sequenced by a PacBio RS II and Illumina HiSeq 4000 platform in Shanghai Majorbio Biopharm Technology Co., Ltd. (Shanghai, China). RESULTS: The multidrug resistance phenotypes of D. acidovorans B804 was experimentally confirmed and its genome was sequenced. The total size of D. acidovorans B804 genome was 6,661,314 bp with a GC content of 66.73%. 403 genes associated with antibiotic resistances were predicted. Meanwhile, 89 pathogenic genes were also predicted and 17 of these genes might be capable of causing diseases to human, such as infections and salmonellosis. CONCLUSIONS: This genomic information can be used as a reference sequence for comparative genomic studies. The results provided more insights regarding the pathogenesis and drug resistance mechanism of D. acidovorans, which will be meaningful for developing more effective therapies toward D. acidovorans-related diseases.

Delftia acidovorans secretes substances that inhibit the growth of Staphylococcus epidermidis through TCA cycle-triggered ROS production.

The proportion of Staphylococcus aureus in the skin microbiome is associated with the severity of inflammation in the skin disease atopic dermatitis. Staphylococcus epidermidis, a commensal skin bacterium, inhibits the growth of S. aureus in the skin. Therefore, the balance between S. epidermidis and S. aureus in the skin microbiome is important for maintaining healthy skin. In the present study, we demonstrated that the heat-treated culture supernatant of Delftia acidovorans, a member of the skin microbiome, inhibits the growth of S. epidermidis, but not that of S. aureus. Comprehensive gene expression analysis by RNA sequencing revealed that culture supernatant of D. acidovorans increased the expression of genes related to glycolysis and the tricarboxylic acid cycle (TCA) cycle in S. epidermidis. Malonate, an inhibitor of succinate dehydrogenase in the TCA cycle, suppressed the inhibitory effect of the heat-treated culture supernatant of D. acidovorans on the growth of S. epidermidis. Reactive oxygen species production in S. epidermidis was induced by the heat-treated culture supernatant of D. acidovorans and suppressed by malonate. Further, the inhibitory effect of the heat-treated culture supernatant of D. acidovorans on the growth of S. epidermidis was suppressed by N-acetyl-L-cysteine, a free radical scavenger. These findings suggest that heat-resistant substances secreted by D. acidovorans inhibit the growth of S. epidermidis by inducing the production of reactive oxygen species via the TCA cycle.

Transcriptomics reveal core activities of the plant growth-promoting bacterium Delftia acidovorans RAY209 during interaction with canola and soybean roots.

The plant growth-promoting rhizobacterium Delftia acidovorans RAY209 is capable of establishing strong root attachment during early plant development at 7 days post-inoculation. The transcriptional response of RAY209 was measured using RNA-seq during early (day 2) and sustained (day 7) root colonization of canola plants, capturing RAY209 differentiation from a medium-suspended cell state to a strongly root-attached cell state. Transcriptomic data was collected in an identical manner during RAY209 interaction with soybean roots to explore the putative root colonization response to this globally relevant crop. Analysis indicated there is an increased number of significantly differentially expressed genes between medium-suspended and root-attached cells during early soybean root colonization relative to sustained colonization, while the opposite temporal pattern was observed for canola root colonization. Regardless of the plant host, root-attached RAY209 cells exhibited the least amount of differential gene expression between early and sustained root colonization. Root-attached cells of either canola or soybean roots expressed high levels of a fasciclin gene homolog encoding an adhesion protein, as well as genes encoding hydrolases, multiple biosynthetic processes, and membrane transport. Notably, while RAY209 ABC transporter genes of similar function were transcribed during attachment to either canola or soybean roots, several transporter genes were uniquely differentially expressed during colonization of the respective plant hosts. In turn, both canola and soybean plants expressed genes encoding pectin lyase and hydrolases - enzymes with purported function in remodelling extracellular matrices in response to RAY209 colonization. RAY209 exhibited both a core regulatory response and a planthost-specific regulatory response to root colonization, indicating that RAY209 specifically adjusts its cellular activities to adapt to the canola and soybean root environments. This transcriptomic data defines the basic RAY209 response as both a canola and soybean commercial crop and seed inoculant.

Urine nitrification with a synthetic microbial community.

During long-term extra-terrestrial missions, food is limited and waste is generated. By recycling valuable nutrients from this waste via regenerative life support systems, food can be produced in space. Astronauts' urine can, for instance, be nitrified by micro-organisms into a liquid nitrate fertilizer for plant growth in space. Due to stringent conditions in space, microbial communities need to be be defined (gnotobiotic); therefore, synthetic rather than mixed microbial communities are preferred. For urine nitrification, synthetic communities face challenges, such as from salinity, ureolysis, and organics. In this study, a synthetic microbial community containing an AOB (Nitrosomonas europaea), NOB (Nitrobacter winogradskyi), and three ureolytic heterotrophs (Pseudomonas fluorescens, Acidovorax delafieldii, and Delftia acidovorans) was compiled and evaluated for these challenges. In reactor 1, salt adaptation of the ammonium-fed AOB and NOB co-culture was possible up to 45mScm-1, which resembled undiluted nitrified urine, while maintaining a 44±10mgNH4+-NL-1d-1 removal rate. In reactor 2, the nitrifiers and ureolytic heterotrophs were fed with urine and achieved a 15±6mg NO3--NL-1d-1 production rate for 1% and 10% synthetic and fresh real urine, respectively. Batch activity tests with this community using fresh real urine even reached 29±3mgNL-1d-1. Organics removal in the reactor (69±15%) should be optimized to generate a nitrate fertilizer for future space applications.

Detecting Gold Biomineralization by Delftia acidovorans Biofilms on a Quartz Crystal Microbalance.

The extensive use of gold in sensing, diagnostics, and electronics has led to major concerns in solid waste management since gold and other heavy metals are nonbiodegradable and can easily accumulate in the environment. Moreover, gold ions are extremely reactive and potentially harmful for humans. Thus, there is an urgent need to develop reliable methodologies to detect and possibly neutralize ionic gold in aqueous solutions and industrial wastes. In this work, by using complementary measurement techniques such as quartz crystal microbalance (QCM), atomic force microscopy, crystal violet staining, and optical microscopy, we investigate a promising biologically induced gold biomineralization process accomplished by biofilms of bacterium Delftia acidovorans. When stressed by Au3+ ions, D. acidovorans is able to neutralize toxic soluble gold by excreting a nonribosomal peptide, which forms extracellular gold nanonuggets via complexation with metal ions. Specifically, QCM, a surface-sensitive transducer, is employed to quantify the production of these gold complexes directly on the D. acidovorans biofilm in real time. Detailed kinetics obtained by QCM captures the condition for maximized biomineralization yield and offers new insights underlying the biomineralization process. To the best of our knowledge, this is the first study providing an extensive characterization of the gold biomineralization process by a model bacterial biofilm. We also demonstrate QCM as a cheap, user-friendly sensing platform and alternative to standard analytical techniques for studies requiring high-resolution quantitative details, which offers promising opportunities in heavy-metal sensing, gold recovery, and industrial waste treatment.

Metal tolerance assisted antibiotic susceptibility profiling in Comamonas acidovorans.

Metal ions are known selective agents for antibiotic resistance and frequently accumulate in natural environments due to the anthropogenic activities. However, the action of metals that cause the antibiotic resistance is not known for all bacteria. The present work is aimed to investigate the co-selection of metals and antibiotic resistance in Comamonas acidovorans. Tolerance profile of 16 metals revealed that the strain could tolerate high concentrations of toxic metals i.e., Cr (710 ppm), As (380 ppm), Cd (320 ppm), Pb (305 ppm) and Hg (205 ppm). Additionally, metal tolerant phenotypes were subjected to antibiotic resistance profiling; wherein several metal tolerant phenotypes (Cr 1.35-fold; Co-1.33 fold; Mn-1.29 fold) were resistant, while other metal tolerant phenotypes (Mg 1.32-fold; Hg 1.29-fold; Cu 1.28-fold) were susceptible than control phenotype. Metal accumulation may alter the metabolism of C. acidovorans that activates or inactivates the genes responsible for antibiotic resistance, resulting in the resistance and/or susceptibility pattern observed in metal resistant phenotypes.

Bioconversion of 16-dehydropregnenolone Acetate to Exclusively 4-androstene-3,17-dione by Delftia acidovorans MTCC 3363.

Delftia acidovorans MTCC 3363 was found to convert 16-dehydropregnenolone acetate (16-DPA) exclusively to 4-androstene-3, 17-dione (AD). Addition of 9α-hydroxylase inhibitors was not required for preventing the accumulation of byproducts. The effect of pH, temperature, substrate concentration, surfactants and carrier solvents on this bioconversion has been studied. 16-DPA was maximally converted in buffered medium at pH 7.0, at temperature 30°C and 0.5 mg ml-1 substrate concentration. Detergent addition and temperature above 35°C had deleterious effect on bioconversion. Dioxan was found to be the best carrier solvent for biotransformation of 16-DPA to AD.

A new agent developed by biotransformation of polyphyllin VII inhibits chemoresistance in breast cancer.

Biotransformation by the endophytes of certain plants changes various compounds, and this 'green' chemistry becomes increasingly important for finding new products with pharmacological activity. In this study, polyphyllin VII (PPL7) was biotransformed by endophytes from the medicinal plant Paris polyphylla Smith, var. yunnanensis. This produced a new compound, ZH-2, with pharmacological activity in vitro and in vivo. ZH-2 was more potent than PPL7 in selectively killing more chemoresistant than chemosensitive breast cancer cells. ZH-2 also re-sensitized chemoresistant breast cancer cells, as evidenced by the improved anti-cancer activity of commonly-used chemotherapeutic agent in vitro, in vivo, and in clinical samples. This anti-chemoresistance effect of ZH-2 was associated with inhibiting the epithelial-mesenchymal transition (EMT) pathway. Taken together, our findings are the first one to link biotransformation with a biomedicine. The results provide insights into developing new pharmacologically-active agents via biotransformation by endophytes.

Engineering Delftia acidovorans DSM39 to produce polyhydroxyalkanoates from slaughterhouse waste.

The inexpensive agricultural fatty by-products could be usefully converted to polyhydroxyalkanoates (PHAs) by properly selected and/or developed microbes. Delftia acidovorans DSM39 is a well-known producer of PHAs with high molar fractions of 4-hydroxybutyrate (4HB), but unable to grow on fatty substrates. The aim of this study was to construct a recombinant strain of D. acidovorans DSM39 using fats-containing waste such as udder, lard and tallow, to produce PHAs. The lipC and lipH genes of Pseudomonas stutzeri BT3, proficient lipolytic isolate, were successfully co-expressed into D. acidovorans DSM39 and the resulting recombinant strain displayed high extracellular enzymatic activity on corn oil. The PHAs production from corn oil achieved high levels (26% of cell dry weight, with about 7% of 4HB). Surprisingly, the recombinant strain produced greater values directly from slaughterhouse residues such as udder and lard (43 and 39%, respectively, with almost 7% of 4HB). Moreover, this work proved the ability of the recombinant D. acidovorans strain to produce PHAs with significant percentage of 4HB, without the supplementation of any precursor in the liquid broth. This research paves the way to the efficient one-step conversion of fatty residues into PHAs having valuable properties exploitable in several medical and industrial applications.

Effects of outer membrane vesicle formation, surface-layer production and nanopod development on the metabolism of phenanthrene by Delftia acidovorans Cs1-4.

Nanopods are extracellular structures arising from the convergence of two widely distributed bacterial characteristics: production of outer membrane vesicles (OMV) and formation of surface layers (S-layers). Nanopod production is driven by OMV formation, and in Delftia acidovorans Cs1-4 growth on phenanthrene induces OMV/nanopod formation. While OMV production has been associated with many functions, particularly with pathogens, linkage to biodegradation has been limited to a membrane stress response to lipophilic compounds. The objectives of this study were to determine: 1.) Whether induction of nanopod formation was linked to phenanthrene metabolism or a non-specific membrane stress response, and 2.) The relative importance of OMV/nanopod formation vs. formation of the S-layer alone to phenanthrene utilization. Membrane stress response was investigated by quantifying nanopod formation following exposure to compounds that exceeded phenanthrene in membrane stress-inducing potential. Naphthalene did not induce nanopod formation, and toluene was a weak inducer compared to phenanthrene (two- vs. six-fold increase, respectively). Induction of nanopod formation by growth on phenanthrene was therefore linked to phenanthrene metabolism and not a membrane stress response. Impacts on phenanthrene biodegradation of OMV/nanopod production vs. S-layer formation were assessed with D. acidovorans Cs1-4 mutants deficient in S-layer formation or OMV/nanopod production. Both mutants had impaired growth on phenanthrene, but the loss of OMV/nanopod production was more significant than loss of the S-layer. The S-layer of D. acidovorans Cs1-4 did not affect phenanthrene uptake, and its primary role in phenanthrene biodegradation process appeared to be enabling nanopod development. Nanopods appeared to benefit phenanthrene biodegradation by enhancing cellular retention of metabolites. Collectively, these studies established that nanopod/OMV formation was an essential characteristic of the D. acidovorans Cs1-4 phenanthrene degradation process. This report thus established a new dimension in the area of biodegradation, namely, the involvement of extracellular structures as elements supporting metabolic processes underlying biodegradation.

Characterization of SDS-degrading Delftia acidovorans and in situ monitoring of its temporal succession in SDS-contaminated surface waters.

Incomplete removal of sodium dodecyl sulfate (SDS) in wastewater treatment plants may result in SDS residues escaping and finding their way into receiving water bodies like rivers, lakes, and sea. Introduction of effective microorganisms into the aerobic treatment facilities can reduce unpleasant by-products and SDS residues. Selecting effective microorganisms for SDS treatment is a big challenge. Current study reports the isolation, identification, and in situ monitoring of an effective SDS-degrading isolate from detergent-polluted river waters. Screening was carried out by the conventional enrichment culture technique and the isolate was tentatively identified by using fatty acid methyl ester and 16S ribosomal RNA (rRNA) sequence analyses. Fatty acids produced by the isolate investigated were assumed as typical for the genus Comamonas. 16S rRNA sequence analysis also confirmed that the isolate had 95% homology with Delftia acidovorans known as Comamonas or Pseudomonas acidovorans previously. D. acidovorans exhibited optimum growth at SDS concentration of 1 g l(-1) but tolerated up to 10 g l(-1) SDS. 87% of 1.0 g l(-1) pure SDS was degraded after 11 days of incubation. The temporal succession of D. acidovorans in detergent-polluted river water was also monitored in situ by using Comamonas-specific fluorescein-labeled Cte probe. Being able to degrade SDS and populate in SDS-polluted surface waters, D. acidovorans isolates seem to be very helpful in elimination of SDS.

Adaptation of Delftia acidovorans for degradation of 2,4-dichlorophenoxyacetate in a microfluidic porous medium.

Delftia acidovorans MC1071 can productively degrade R-2-(2,4-dichlorophenoxy)propionate (R-2,4-DP) but not 2,4-dichlorophenoxyacetate (2,4-D) herbicides. This work demonstrates adaptation of MC1071 to degrade 2,4-D in a model two-dimensional porous medium (referred to here as a micromodel). Adaptation for 2,4-D degradation in the 2 cm-long micromodel occurred within 35 days of exposure to 2,4-D, as documented by substrate removal. The amount of 2,4-D degradation in the adapted cultures in two replicate micromodels (~10 and 20 % over 142 days) was higher than a theoretical maximum (4 %) predicted using published numerical simulation methods, assuming instantaneous biodegradation and a transverse dispersion coefficient obtained for the same pore structure without biomass present. This suggests that the presence of biomass enhances substrate mixing. Additional evidence for adaptation was provided by operation without R-2,4-DP, where degradation of 2,4-D slowly decreased over 20 days, but was restored almost immediately when R-2,4-DP was again provided. Compared to suspended growth systems, the micromodel system retained the ability to degrade 2,4-D longer in the absence of R-2,4-DP, suggesting slower responses and greater resilience to fluctuations in substrates might be expected in the soil environment than in a chemostat.

Gold biomineralization by a metallophore from a gold-associated microbe.

Microorganisms produce and secrete secondary metabolites to assist in their survival. We report that the gold resident bacterium Delftia acidovorans produces a secondary metabolite that protects from soluble gold through the generation of solid gold forms. This finding is the first demonstration that a secreted metabolite can protect against toxic gold and cause gold biomineralization.

Enantioselective stable isotope analysis (ESIA) of polar herbicides.

Assessing the environmental fate of chiral micropollutants such as herbicides is challenging. The complexity of aquatic systems often makes it difficult to obtain hydraulic mass balances, which is a prerequisite when assessing degradation based on concentration data. Elegant alternatives are concentration-independent approaches like compound-specific isotope analysis or enantiospecific concentration analysis. Both detect degradation-induced changes from ratios of molecular species, either isotopologues or enantiomers. A combination of both-enantioselective stable isotope analysis (ESIA)-provides information on (13)C/(12)C ratios for each enantiomer separately. Recently, Badea et al. demonstrated for the first time ESIA for the insecticide α-hexachlorocyclohexane. The present study enlarges the applicability of ESIA to polar herbicides such as phenoxy acids: 4-CPP ((RS)-2-(4-chlorophenoxy)-propionic acid), mecoprop (2-(4-chloro-2-methylphenoxy)-propionic acid), and dichlorprop (2-(2,4-dichlorophenoxy)-propionic acid). Enantioselective gas chromatography-isotope ratio mass spectrometry was accomplished with derivatization prior to analysis. Precise carbon isotope analysis (2σ ≤ 0.5‰) was obtained with ≥7 ng C on column. Microbial degradation of dichlorprop, 2-(2,4-dichlorophenoxy)-propionic acid by Delftia acidovorans MC1 showed pronounced enantiomer fractionation, but no isotope fractionation. In contrast, Badea et al. observed isotope fractionation, but no enantiomeric fractionation. Hence, the two lines of evidence appear to complement each other. They may provide enhanced insight when combined as ESIA.

(R,S)-dichlorprop herbicide in agricultural soil induces proliferation and expression of multiple dioxygenase-encoding genes in the indigenous microbial community.

We investigated the effect of (R,S)-dichlorprop herbicide addition to soil microcosms on the degrading indigenous microbial community by targeting multiple α-ketoglutarate-dependent (α-KG) dioxygenase-encoding genes (rdpA, sdpA and tfdA group I) at both gene and transcript level. The soil microbial community responded with high growth of potential degraders as measured by the abundance of dioxygenase-encoding genes using quantitative real-time PCR (qPCR). rdpA DNA was not detectable in unamended soil but reached over 106 copies g⁻¹ soil after amendment. sdpA and tfdA were both present prior to amendment at levels of ~5 × 104 and ~ 10² copies g⁻¹ soil, respectively, and both reached over 105copies g⁻¹ soil. While expression of all three target genes was detected during two cycles of herbicide degradation, a time-shift occurred between maximum expression of each gene. Gene diversity by denaturing gradient gel electrophoresis (DGGE) uncovered a diversity of sdpA and tfdA genes at the DNA level while rdpA remained highly conserved. However, mRNA profiles indicated that all transcribed tfdA sequences were class III genes while rdpA transcripts shared 100% identity to rdpA of Delftia acidovorans MC1 and sdpA transcripts shared 100% identity to sdpA from Sphingomonas herbicidovorans MH. This is the first report to describe expression dynamics of multiple α-KG dioxygenase-encoding genes in the indigenous microbial community as related to degradation of a phenoxypropionate herbicide in soil.

Declining capacity of starving Delftia acidovorans MC1 to degrade phenoxypropionate herbicides correlates with oxidative modification of the initial enzyme.

Bioremediation relies on the stability of enzymatic activities, particularly when bioavailable contaminant concentrations do not permit much renewal of microbial biomass. Starving Delftia acidovorans MC1 were found to lose specific degradation activity, while accumulating variants of the alpha-ketoglutarate-dependent dioxygenase RdpA, the enzyme initiating the degradation of (RS)-2-(2,4-dichlorophenoxy)propionate. These variants differed in their pI and originated from post-translational modification, since there is only one rdpA gene in the genome. It was tested if RdpA modification resulted from carbonylation by reactive oxygen species, known side products of dioxygenase reactions. Carbonylated amino acids in proteins of starved cells were specifically derivatized with 2,4-dinitrophenylhydrazine. Subsequent immunolabeling of the resulting hydrazones and mass spectrometry of tryptic digests confirmed different levels of carbonylation of RdpA.

Abundance and expression of enantioselective rdpA and sdpA dioxygenase genes during degradation of the racemic herbicide (R,S)-2-(2,4-dichlorophenoxy)propionate in soil.

The rdpA and sdpA genes encode two enantioselective alpha-ketoglutarate-dependent dioxygenases catalyzing the initial step of microbial degradation of the chiral herbicide (R,S)-2-(2,4-dichlorophenoxy)propionate (R,S-dichlorprop). Primers were designed to assess abundance and transcription dynamics of rdpA and sdpA genes in a natural agricultural soil. No indigenous rdpA genes were detected, but sdpA genes were present at levels of approximately 10(3) copies g of soil(-1). Cloning and sequencing of partial sdpA genes revealed a high diversity within the natural sdpA gene pool that could be divided into four clusters by phylogenetic analysis. BLASTp analysis of deduced amino acids revealed that members of cluster I shared 68 to 69% identity, cluster II shared 78 to 85% identity, cluster III shared 58 to 64% identity, and cluster IV shared 55% identity to their closest SdpA relative in GenBank. Expression of rdpA and sdpA in Delftia acidovorans MC1 inoculated in soil was monitored by reverse transcription quantitative real-time PCR (qPCR) during in situ degradation of 2 and 50 mg kg(-1) of (R,S)-dichlorprop. (R,S)-Dichlorprop amendment created a clear upregulation of both rdpA and sdpA gene expression during the active phase of (14)C-labeled (R,S)-dichlorprop mineralization, particularly following the second dose of 50 mg kg(-1) herbicide. Expression of both genes was maintained at a low constitutive level in nonamended soil microcosms. This study is the first to report the presence of indigenous sdpA genes recovered directly from natural soil and also comprises the first investigation into the transcription dynamics of two enantioselective dioxygenase genes during the in situ degradation of the herbicide (R,S)-dichlorprop in soil.

Using multiconformation continuum electrostatics to compare chloride binding motifs in alpha-amylase, human serum albumin, and Omp32.

Ions are a ubiquitous component of the cellular environment, transferring into cells through membrane-embedded proteins. Ions bind to proteins to regulate their charge and function. Here, using multiconformation continuum electrostatics (MCCE), we show that the changes of chloride binding to alpha-amylase, human serum albumin (HSA) and Omp32 with pH, and of alpha-amylase with mutation agree well with experimental data. The three proteins represent three different types of binding. In alpha-amylase, chloride is bound in a specific buried site. Chloride binding is strongly coupled to the protonation state of a nearby lysine. MCCE calculates an 11-fold change in chloride affinity between the wild-type alpha-amylase and the K300R mutant, in good agreement with the measured 10-fold change.Without considering the coupled protonation reaction, the calculated affinity change would be more than 10(6)-fold. In HSA, chlorides are distributed on the protein surface. Although HSA has a negative net charge, it binds more anions than cations. There are no highly occupied binding sites in HSA. Rather, there are many partially occupied sites near clusters of basic residues. The relative affinity of bound ions of different charges is shown to depend on the distribution of charged residues on the surface rather than the overall net charge of the protein. The calculated strong pH dependence of the number of chlorides bound and the anion selectivity agree with those of previous experiments. In Omp32, chlorides are stabilized in an anion-selective transmembrane channel in a pH-independent manner. The positive electrostatic potential in Omp32 results in about two chlorides and no cations bound in the transmembrane region of this anion-selective channel. The studies here show that with the ability to sample multiple binding sites and coupled protein protonation states, MCCE provides a powerful tool to analyze and predict ion binding. The calculations overestimate the affinity of surface chloride in HSA and Omp32 relative to the buried ion in amylase. Differences between ion-solvent interactions for buried and surface ions will be discussed.

Mutagenic and clastogenic characterization of poststerilized poly(3-hydroxybutyrate-co-4-hydroxybutyrate) copolymer biosynthesized by Delftia acidovorans.

Polyhydroxyalkanoates (PHA) are naturally occurring biopolyesters that have great potential in the medical field. However, the leachables resulting from sterilization process of the biomaterials may exert toxic effect including genetic damage. Here, we demonstrate that although gamma-irradiation of poly(3-hydroxybutyrate-co-50 mol % 4-hydroxybutyrate) [P(3HB-co-4HB)] did not cause any change in the morphology by scanning electron microscopy, there was a significant degradation of this copolymer where the molecular weight was reduced by 37% after sterilization indicating the generation of leachables. Therefore, further investigation on the ability of the extract of this poststerilized copolymer to induce mutagenic effect was performed using Ames test (S. typhimurium strains TA1535 and TA1537) and umu test (S. typhimurium strain TA1535/pSK1002). Additionally, the capability of the extract to induce clastogenic effect was determined using Chinese hamster lung V79 fibroblast cells. Our results showed that with and without the presence of S9 metabolic activation, no mutagenic effects were observed in both Ames and umu tests when treated with P(3HB-co-4HB) extract. Similarly, treatment of P(3HB-co-4HB) extract in V79 fibroblast cells showed no significant production of micronuclei when compared with the positive control (Mitomycin C). Together, these results indicate that leachables of poststerilized P(3HB-co-4HB) cause no mutagenic and clastogenic effects.

Biodegradation of monoethylene glycol on continuous bench scale reactor by bacterial consortia.

Wastewater containing monoethylene glycol (MEG) was investigated for biodegradation on a continuous bench scale bioreactor applying an activated sludge process. The bacterial consortia were prepared using three isolated bacterial strains and process conditions, viz. HRT, biomass concentration and air requirements were optimized to achieve efficient biodegradation. The results show 89% COD, 95% BOD reduction at a HRT of 10 h and organic loading of 0.0136 kg/day. The reduction in COD/BOD was related to removal of MEG concentration. The metabolic pathway was investigated in a batch mode and confirmed the formation of acetaldehyde and acetic acid as intermediates.