Chromobacterium Sp Research Articles

Hydrogen cyanide produced by the soil bacterium Chromobacterium sp. Panama contributes to mortality in Anopheles gambiae mosquito larvae

Mosquito larvae continuously encounter microbes in their aquatic environment, which serve as food and play a critical role in successful development. In previous work, we isolated a Chromobacterium sp. (C.sp_P) with larvicidal activity from the midgut of dengue vector Aedes mosquitoes in Panama. In this study, we found a positive correlation between initial concentrations of C.sp_P and larval mortality rates, and that C.sp_P is more efficient at inducing larval mortality in a high nutrient environment. Multiple Chromobacterium species induce larval mortality with similar efficacy to C.sp_P except for C. subtsugae. We also found that a non-lethal dose of C.sp_P lengthens development time and increases mortality over multiple developmental stages, suggesting persistent effects of exposure. Additionally, we showed that larvicidal activity persists in the larval breeding water after removal of live bacteria, and that the larvicidal factor in C.sp_P-treated water is smaller than 3 kDa, heat resistant to 90 °C, and lost after vacuum centrifugation. We showed that C.sp_P produces hydrogen cyanide in culture and in larval water at concentrations sufficient to kill An. gambiae larvae, and treatment of the larval water with a cyanide antidote eliminated larvicidal activity. We conclude that a potential mechanism by which C.sp_P can induce larval mortality is via production of hydrogen cyanide.

Aminopeptidase secreted by Chromobacterium sp. Panama inhibits dengue virus infection by degrading the E protein.

Dengue virus (DENV) is the most prevalent and burdensome arbovirus transmitted by Aedes mosquitoes, against which there is only a limited licensed vaccine and no approved drug treatment. A Chromobacterium species, C. sp. Panama, isolated from the midgut of A. aegypti is able to inhibit DENV replication within the mosquito and in vitro. Here we show that C. sp. Panama mediates its anti-DENV activity through secreted factors that are proteinous in nature. The inhibitory effect occurs prior to virus attachment to cells, and is attributed to a factor that destabilizes the virion by promoting the degradation of the viral envelope protein. Bioassay-guided fractionation, coupled with mass spectrometry, allowed for the identification of a C. sp. Panama-secreted neutral protease and an aminopeptidase that are co-expressed and appear to act synergistically to degrade the viral envelope (E) protein and thus prevent viral attachment and subsequent infection of cells. This is the first study characterizing the anti-DENV activity of a common soil and mosquito-associated bacterium, thereby contributing towards understanding how such bacteria may limit disease transmission, and providing new tools for dengue prevention and therapeutics.

Chromobacterium spp. mediate their anti-Plasmodium activity through secretion of the histone deacetylase inhibitor romidepsin

The Chromobacterium sp. Panama bacterium has in vivo and in vitro anti-Plasmodium properties. To assess the nature of the Chromobacterium-produced anti-Plasmodium factors, chemical partition was conducted by bioassay-guided fractionation where different fractions were assayed for activity against asexual stages of P. falciparum. The isolated compounds were further partitioned by reversed-phase FPLC followed by size-exclusion chromatography; high resolution UPLC and ESI/MS data were then collected and revealed that the most active fraction contained a cyclic depsipeptide, which was identified as romidepsin. A pure sample of this FDA-approved HDAC inhibitor allowed us to independently verify this finding, and establish that romidepsin also has potent effect against mosquito stages of the parasite’s life cycle. Genomic comparisons between C. sp. Panama and multiple species within the Chromobacterium genus further demonstrated a correlation between presence of the gene cluster responsible for romidepsin production and effective antiplasmodial activity. A romidepsin-null Chromobacterium spp. mutant loses its anti-Plasmodium properties by losing the ability to inhibit P. falciparum HDAC activity, and romidepsin is active against resistant parasites to commonly deployed antimalarials. This independent mode of action substantiates exploring a chromobacteria-based approach for malaria transmission-blocking.

The mosquito adulticidal Chromobacterium sp. Panama causes transgenerational impacts on fitness parameters and elicits xenobiotic gene responses

BackgroundVector control is critical in reducing the disease burden caused by mosquitoes, and insecticides are an effective tool to control vector populations. Resistance to common insecticides is now widespread, and novel classes of insecticides are needed. In previous work, we described the mosquitocidal activity of Chromobacterium sp. Panama (C.sp_P), a bacterium found in association with mosquitoes in natural populations. In the current work, we further explored the effects of exposure to the bacterium on mosquito fitness and mosquito physiology.ResultsWe found that C.sp_P has mosquitocidal activity against a broad range of mosquito taxa. When exposed to C.sp_P as adults, female An. gambiae suffered reduced longevity, but experienced no change in fecundity. The offspring of these females, however, had higher mortality as larvae and were slower to develop compared to offspring of control females. We also found that the mosquitocidal activity of C.sp_P was retained after removal of live cells from biofilm culture media, suggesting the bacteria secrete mosquitocidal compound(s) into the media during growth. Exposure to this cell-free C.sp_P-conditioned media caused female midgut transcriptional changes comprising detoxification, xenobiotic response, and stress response genes, suggesting the physiological response to C.sp_P is similar to that of insecticide exposure. Finally, we found that multiple members of the Chromobacterium genus had mosquitocidal activity, but this activity was highest in mosquitoes treated with C.sp_P.ConclusionsOur findings suggest that C.sp_P produces factor(s) with strong effects on mosquito longevity and fitness, which may be of interest for mosquitocide development. More generally, they indicate that further exploration of mosquito-associated and environmental microbes for novel insecticidal compounds or biocontrol agents is warranted.

KEBERADAAN BAKTERI PELARUT FOSFAT DAN AKTIVITAS ENZIM FOSFATASE TANAH DAERAH PERAKARAN TANAMAN OBAT DARI KEBUN RAYA CIBODAS

A study was undertaken to investigate to occurance of phosphate solubilizing bacteria from rhizosphere soil samples of medicine plants in Cibodas Botanical Garden. 13 soil samples of medicine plants are collected randomly The result shows that 71 isolates of phosphate solubilizing bacteria were isolated, and 10 species of these organism was identified as Azotobacter sp, Bacillus sp, Chromobacterium sp, C.violaceum, Citrobacter sp. , Enterobacter sp., E. liquefaciens. Nitrosomonas sp., Serratia rubidaea, Sphaerotillus natans. Azotobacter sp. And Bacillus sp. Are found in all of soil tested. Conversely, Serratia rubidaea is only in the sample from rhizosphere of Plantago mayor The activity of acid alkaline phosphatase in soil tested ranged from 0.78 – 60,18 ugp nitrophenole/g/h, with the higest values being recorded in soil sample from rhizosphere of “Lavender”.Keywords : phosphate solubilizing bacteria, soil enzyme phosphatase

Structure of polyhydroxyalkanoate (PHA) synthase PhaC from Chromobacterium sp. USM2, producing biodegradable plastics

Polyhydroxyalkanoate (PHA) is a promising candidate for use as an alternative bioplastic to replace petroleum-based plastics. Our understanding of PHA synthase PhaC is poor due to the paucity of available three-dimensional structural information. Here we present a high-resolution crystal structure of the catalytic domain of PhaC from Chromobacterium sp. USM2, PhaCCs-CAT. The structure shows that PhaCCs-CAT forms an α/β hydrolase fold comprising α/β core and CAP subdomains. The active site containing Cys291, Asp447 and His477 is located at the bottom of the cavity, which is filled with water molecules and is covered by the partly disordered CAP subdomain. We designated our structure as the closed form, which is distinct from the recently reported catalytic domain from Cupriavidus necator (PhaCCn-CAT). Structural comparison showed PhaCCn-CAT adopting a partially open form maintaining a narrow substrate access channel to the active site, but no product egress. PhaCCs-CAT forms a face-to-face dimer mediated by the CAP subdomains. This arrangement of the dimer is also distinct from that of the PhaCCn-CAT dimer. These findings suggest that the CAP subdomain should undergo a conformational change during catalytic activity that involves rearrangement of the dimer to facilitate substrate entry and product formation and egress from the active site.

Introducing inducible fluorescent split cholesterol oxidase to mammalian cells

Cholesterol oxidase (COase) is a bacterial enzyme catalyzing the first step in the biodegradation of cholesterol. COase is an important biotechnological tool for clinical diagnostics and production of steroid drugs and insecticides. It is also used for tracking intracellular cholesterol; however, its utility is limited by the lack of an efficient temporal control of its activity. To overcome this we have developed a regulatable fragment complementation system for COase cloned from Chromobacterium sp. The enzyme was split into two moieties that were fused to FKBP (FK506-binding protein) and FRB (rapamycin-binding domain) pair and split GFP fragments. The addition of rapamycin reconstituted a fluorescent enzyme, termed split GFP-COase, the fluorescence level of which correlated with its oxidation activity. A rapid decrease of cellular cholesterol induced by intracellular expression of the split GFP-COase promoted the dissociation of a cholesterol biosensor D4H from the plasma membrane. The process was reversible as upon rapamycin removal, the split GFP-COase fluorescence was lost, and cellular cholesterol levels returned to normal. These data demonstrate that the split GFP-COase provides a novel tool to manipulate cholesterol in mammalian cells.

Marine-derived chromopeptide A, a novel class I HDAC inhibitor, suppresses human prostate cancer cell proliferation and migration.

Histone deacetylases (HDACs), especially HDAC1, 2, 3 and 4, are abundantly expressed and over-activated in prostate cancer that is correlated with the poor prognosis. Thus, inhibition of HDAC activity has emerged as a potential alternative option for prostate cancer therapy. Chromopeptide A is a depsipeptide isolated from the marine sediment-derived bacterium Chromobacterium sp. HS-13-94; it has a chemical structure highly similar to FK228, a class I HDAC inhibitor that is approved by FDA for treating T-cell lymphoma. In this study, we determined whether chromopeptide A, like FK228, acted as a class I HDAC inhibitor, and whether chromopeptide A could inhibit the growth and migration of human prostate cancer in vitro and in vivo. HDAC enzyme selectivity and kinetic analysis revealed that chromopeptide A selectively inhibited the enzymatic activities of HDAC1, 2, 3 and 8 in a substrate non-competitive manner with comparable IC50 values for each HDAC member as FK228 in vitro. Importantly, chromopeptide A dose-dependently suppressed the proliferation of human prostate cancer cell lines PC3, DU145 and LNCaP with IC50 values of 2.43±0.02, 2.08±0.16, and 1.75±0.06 nmol/L, respectively, accompanied by dose-dependent inhibition of HDAC enzymatic activity in PC3 and DU145 cells. Chromopeptide A (0.2-50 nmol/L) caused G2/M phase arrest and induced apoptosis in the prostate cancer cell lines. Moreover, chromopeptide A dose-dependently inhibited the migration of PC3 cells. In mice bearing PC3 prostate cancer xenografts, intravenous injection of chromopeptide A (1.6, 3.2 mg/kg, once a week for 18 d) significantly suppressed the tumor growth, which was associated with increased expression levels of Ac-H3 and p21 in tumor tissues. Our results identify chromopeptide A as a novel class I HDAC inhibitor and provide therapeutic strategies that may be implemented in prostate cancer.

Rhizosphere mediated biodegradation of 1,4-dichlorobenzene by plant growth promoting rhizobacteria of Jatropha curcas

The aim of this study was to investigate the ability of rhizospheric isolates of Jatropha curcas to degrade Dichlorobenzene (DCB) in addition to their plant growth promotion activity, for effective rhizoremediation. DCB degrading rhizobacteria were isolated from Jatropha curcas rhizosphere and eight isolates were selected on the basis of superior 1,4 dichlorobenzene (0.125mM) degradation potential. These isolates showed more than 97% of DCB degradation in vitro condition, while highest degradation was recorded with Chromobacterium sp. B5 (99.46%), as estimated by HPLC analysis. The isolates also showed plant growth promoting (PGP) attributes including phosphate solubilization, siderophores production, IAA release and increased seed germination. Effect of bacterial inoculations on plant growth and seed germination was checked at different DCB concentrations. The uninoculated control seeds had less germination index and biomass production than the seeds inoculated with the rhizospheric isolates. Plant growth promoting rhizobacteria (PGPR) interaction with Jatropha plants resulted in more efficient degradation of 1,4-dichlorobenzene along with an increase in germination and growth of Jatropha curcas plants. In DCB contaminated soil, Flavobacterium (B7), Serratia (B8), Pasteurella (B1) and Azotobacter (B6) resulted in 118%, 117%, 84.11% and 90% increase, in seed vigor index, respectively as compared to control.

Bacteriological and physicochemical analysis of waste water from fish ponds

Rearing of fish especially catfish is very common in many communities in Nigeria and the wastewater from these fish ponds are often discharged into the surrounding drains. Assessment of the bacteriological and physicochemical characteristics of ten freshwater fish ponds stocked with African catfish (Clarias gariepinus) was conducted within Ilorin metropolis. The results of physicochemical properties of the water samples showed that the pH, total hardness, salinity, and suspended solid ranged from 6.77 – 8.43, 65 – 124 mg/l, 7.02 – 10.062 g/l, and 0.012 – 0.128g/100ml respectively. The bacterial count, total coliform, and faecal coliform count of the wastewater ranged from 4.0 x 103 – 6.7 x 105cfu/ml, nil to 2 x 104cfu/ml, and nil to 1 x 103cfu/ml respectively. The bacterial species isolated from the ponds were Escherichia coli, Staphylococcus sp., Staphylococcus aureus, Aeromonas hydrophila, Staphylococcus saprophyticus, Chromobacterium sp, Micrococcus sp., Micrococcus kristinae, Providencia sp., and Listeria sp. The antibiotics susceptibility testing of the bacterial isolates were determined using the disc method. All the isolates exhibited multiple antibiotic resistance (MAR). It can be concluded from this study that there is need to monitor the quality of wastewater from the fish ponds before being discharged into the environment since potential pathogens were isolated and most of these isolates were multiple antibiotic resistant. This study is of socioeconomic significance in the sense that while the farmers are able to raise their fish in the ponds and derive their livelihood, they are being cautioned on the need to treat their waste water before being discharged into the receiving water body.Keywords: Wastewater, Fish pond, Catfish, Assessment, Antibiotic resistance

EKSPLORASI DAN UJI SENYAWA BIOAKTIF BAKTERI AGENSIA HAYATI UNTUK PENGENDALIAN PENYAKIT KRESEK PADA PADI

Exploration of bacterial biocontrol agent and its potential bioactive compound to control rice bacterial leaf blight. The research aims were to obtain bacterial isolates which were potential as biological control agent of X. oryzae pv. oryzae, the causal agent of rice bacterial blight and to assess the effectiveness of their bioactive compounds, and to identify of the potential isolates. The research steps included bacterial isolation, screening based on antibiosis activity and pathogenicity test, characterization based on chitinolytic enzyme production, siderophores, and phosphate dissolution test, effectiveness test of bioactive compounds and molecular identification of potential isolates. Out of 156 bacterial isolates from rice crop tested, 11 isolates showed to be non plant pathogenic and to have activity as biological agents against X. oryzae pv. oryzae pathotype III, IV and VIII. Further characterization of 11 isolates resulted in 2 isolates that showrd ability to produce chitinase (isolates T5-1118 and R7-1018), phosphatase (isolates T5-1105 and T6-1109), and siderophores (isolates T5-1118 and T6- 1109). The test of bioactive compound effectiveness of 4 isolates to the growth of X. oryzae pv. oryzae showed thatT5-1118, T5-1105, T6-1109 and R7-1018 have ability to inhibit X. oryzae pv. oryzae at 48 hours after inoculation of 66,61%, 62,4%, 23,97% and 12,40%, respectively. Identification of 4 bacterial biocontrol isolates with partial sequencing of 16S rRNA gene showed that those bacteria are close to Bacillus nealsonii strain F22 (R7-1018), Chromobacterium sp. MWU328 (T5-1118), Streptomyces sp. Antag 1 (T5-1105) and Kitasatospora nipponensis strains H2-4 (T6-1109).

Study of Class I and Class III Polyhydroxyalkanoate (PHA) Synthases with Substrates Containing a Modified Side Chain.

Polyhydroxyalkanoates (PHAs) are carbon and energy storage polymers produced by a variety of microbial organisms under nutrient-limited conditions. They have been considered as an environmentally friendly alternative to oil-based plastics due to their renewability, versatility, and biodegradability. PHA synthase (PhaC) plays a central role in PHA biosynthesis, in which its activity and substrate specificity are major factors in determining the productivity and properties of the produced polymers. However, the effects of modifying the substrate side chain are not well understood because of the difficulty to accessing the desired analogues. In this report, a series of 3-(R)-hydroxyacyl coenzyme A (HACoA) analogues were synthesized and tested with class I synthases from Chromobacterium sp. USM2 (PhaCCs and A479S-PhaCCs) and Caulobacter crescentus (PhaCCc) as well as class III synthase from Allochromatium vinosum (PhaECAv). It was found that, while different PHA synthases displayed distinct preference with regard to the length of the alkyl side chains, they could withstand moderate side chain modifications such as terminal unsaturated bonds and the azide group. Specifically, the specific activity of PhaCCs toward propynyl analogue (HHxyCoA) was only 5-fold less than that toward the classical substrate HBCoA. The catalytic efficiency (kcat/Km) of PhaECAv toward azide analogue (HABCoA) was determined to be 2.86 × 10(5) M(-1) s(-1), which was 6.2% of the value of HBCoA (4.62 × 10(6) M(-1) s(-1)) measured in the presence of bovine serum albumin (BSA). These side chain modifications may be employed to introduce new material functions to PHAs as well as to study PHA biogenesis via click-chemistry, in which the latter remains unknown and is important for metabolic engineering to produce PHAs economically.

Chromobacterium spp. harbour Ambler class A β-lactamases showing high identity with KPC.

The origin of KPC is unknown. The aim of this study was to detect progenitors of KPC in silico and to functionally verify their β-lactam hydrolysis activity. The sequence of KPC-2 was used to mine the NCBI protein sequence database. The best non-KPC hits were analysed by amino acid (aa) alignment and phylogenetic tree construction. Genes encoding KPC-2 homologues were expressed in Escherichia coli. The carbapenemase activities of the recombinant strains were characterized by the CarbaNP test and UV spectrophotometry and MICs of selected β-lactams were determined. Genes encoding the closest KPC-2 homologues were identified on the chromosome of Chromobacterium piscinae strain ND17 (CRP-1, 76% aa identity), Chromobacterium sp. C-61 (CRS-1, 70% aa identity) and Chromobacterium haemolyticum DSM19808 (CRH-1, 69% aa identity). All three Chromobacterium β-lactamases were phylogenetically more related to KPC than to other Ambler class A β-lactamases. The 27 bp region preceding the start codon of blaCRP-1 displayed high nucleotide identity to the corresponding region upstream from blaKPC (74%). Heterologous expression of blaCRP-1 and to a lesser extent of blaCRH-1 in E. coli significantly increased the MICs of meropenem and most cephalosporins. The CarbaNP test was positive for both recombinant strains, but spectrophotometric analysis confirmed higher carbapenemase activity for CRP-1-producing clones. The recovery of three class A β-lactamases with up to 76% aa identity to KPC from distinct Chromobacterium species is highly indicative of the role played by this genus in the evolution of KPC.

Antifungal activity of violacein purified from a novel strain of Chromobacterium sp. NIIST (MTCC 5522).

A novel strain of Chromobacterium sp. NIIST (MTCC 5522) producing high level of purple blue bioactive compound violacein was isolated from clay mine acidic sediment. During 24 h aerobic incubation in modified Luria Bertani medium, around 0.6 g crude violacein was produced per gram of dry weight biomass. An inexpensive method for preparing crystalline, pure violacein from crude pigment was developed (12.8 mg violacein/L) and the pure compound was characterized by different spectrometric methods. The violacein prepared was found effective against a number of plant and human pathogenic fungi and yeast species such as Cryptococcus gastricus, Trichophyton rubrum, Fusarium oxysporum, Rhizoctonia solani, Aspergillus flavus, Penicillium expansum, and Candida albicans. The best activity was recorded against Trichophyton rubrum (2 -g/ml), a human pathogen responsible for causing athlete-s foot infection. This is the first report of antifungal activity of purified violacein against pathogenic fungi and yeast.

Efficient poly(3-hydroxypropionate) production from glycerol using Lactobacillus reuteri and recombinant Escherichia coli harboring L. reuteri propionaldehyde dehydrogenase and Chromobacterium sp. PHA synthase genes

Poly(3-hydroxypropionate), P(3HP), is a polymer combining good biodegradability with favorable material properties. In the present study, a production system for P(3HP) was designed, comprising conversion of glycerol to 3-hydroxypropionaldehyde (3HPA) as equilibrium mixture with 3HPA-hydrate and -dimer in aqueous system (reuterin) using resting cells of native Lactobacillus reuteri in a first stage followed by transformation of the 3HPA to P(3HP) using recombinant Escherichia coli strain co-expressing highly active coenzyme A-acylating propionaldehyde dehydrogenase (PduP) from L. reuteri and polyhydroxyalkanoate synthase (PhaCcs) from Chromobacterium sp. P(3HP) content of up to 40% (w/w) cell dry weight was reached, and the yield with respect to the reuterin consumed by the cells was 78%. Short biotransformation period (4.5h), lack of additives or expensive cofactors, and use of a cheap medium for cultivation of the recombinant strain, provides a new efficient and potentially economical system for P(3HP) production.

Chromopeptide A, a highly cytotoxic depsipeptide from the marine sediment-derived bacterium Chromobacterium sp. HS-13-94

A bicyclic depsipeptide, chromopeptide A (1), was isolated from a deep-sea-derived bacterium Chromobacterium sp. HS-13-94. Its structure was determined by extensive spectroscopic analysis and by comparison with a related known compound. The absolute configuration of chromopeptide A was established by X-ray diffraction analysis employing graphite monochromated Mo Kα radiation (λ=0.71073Å) with small Flack parameter 0.03. Chromopeptide A suppressed the proliferation of HL-60, K-562, and Ramos cells with average IC50 values of 7.7, 7.0, and 16.5nmol/L, respectively.

Biosynthesis of P(3HB-co-3HV-co-3HHp) terpolymer by Cupriavidus necator PHB-4 transformant harboring the highly active PHA synthase gene of Chromobacterium sp. USM2

Aims: This study evaluates potentials of Cupriavidus necator PHB  4 transformant harboring the highly active polyhydroxyalkanoate synthase gene (phaC) of a locally isolated Chromobacterium sp. USM2 for its ability to incorporate 3-hydroxyheptanoate (3HHp) monomer. Methodology and results: A mixture of fructose and sodium heptanoate fed to the culture gave rise to poly(3hydroxybutyrate-co-3-hydroxyvalerate-co-3-hydroxyheptanoate), [P(3HB-co-3HV-co-3HHp)] terpolymer synthesis, with traces of 3HHp monomers confirmed through gas chromatography (GC), proton ( 1 H) and carbon ( 13 C) NMR spectra. Conclusion, significance and impact of study: This study has revealed that the PHA synthase of Chromobacterium sp. USM2 has a broad range of substrate specificity. The synthase is able to polymerize 3-hydroxyalkanoate monomers having 4–7 carbon atoms.

In vitro and in vivo antitumor activity of crude extracts obtained from Brazilian Chromobacterium sp isolates

Natural products produced by microorganisms have been an important source of new substances and lead compounds for the pharmaceutical industry. Chromobacterium violaceum is a Gram-negative β-proteobacterium, abundant in water and soil in tropical and subtropical regions and it produces violacein, a pigment that has shown great pharmaceutical potential. Crude extracts of five Brazilian isolates of Chromobacterium sp (0.25, 2.5, 25, and 250 µg/mL) were evaluated in an in vitro antitumor activity assay with nine human tumor cells. Secondary metabolic profiles were analyzed by liquid chromatography and electrospray ionization mass spectrometry resulting in the identification of violacein in all extracts, whereas FK228 was detected only in EtCE 308 and EtCE 592 extracts. AcCE and EtCE 310 extracts showed selectivity for NCI/ADR-RES cells in the in vitro assay and were evaluated in vivo in the solid Ehrlich tumor model, resulting in 50.3 and 54.6% growth inhibition, respectively. The crude extracts of Chromobacterium sp isolates showed potential and selective antitumor activities for certain human tumor cells, making them a potential source of lead compounds. Furthermore, the results suggest that other compounds, in addition to violacein, deoxyviolacein and FK228, may be involved in the antitumor effect observed.

Effect of different recovery strategies of P(3HB-co-3HHx) copolymer from Cupriavidus necator recombinant harboring the PHA synthase of Chromobacterium sp. USM2

Five strategies (Experiment A, B, C, D and E) for recovery of PHA were carried out in 10L bioreactor using cells which contained 68wt% PHA and 3mol% 3HHx. NaCl (8g/L), NaOH (0.1M) and ethanol (20% v/v) were used in the treatments. Separated treatments of NaCl and NaOH with ethanol (Experiment D) was found as the best strategy with high PHA recovery (95wt%) and purity (90wt%). Although, combined treatments (Experiment B) obtained the highest PHA purity (97wt%) severe degradation of polymer was detected. The molecular weight (Mw) was reduced to almost half (2.8×105Da) compared to polymer extracted using chloroform (4.6×105Da). In addition, pre-treatment using NaCl was found to increase the PHA recovery and purity to 97.7wt% and 97.5wt%, respectively. Also, no significant changes in the PHA recovery and purity were observed when lyophilized cell or wet cell was used. Further characterizations of recovered polymers were carried out using GPC, DSC, TGA and tensile machine.

Draft Genome Sequence of the Biocontrol Bacterium Chromobacterium sp. Strain C-61

Chromobacterium sp. strain C-61 is a plant-associated bacterium with proven capacities to suppress plant diseases. Here, we report the draft genome sequence and automatic annotation of strain C-61. A comparison of this sequence to the sequenced genome of Chromobacterium violaceum ATCC 12472 indicates the novelty of C-61 and a subset of gene functions that may be related to its biocontrol activities.