Polyhydroxyalkanoates (PHA) production from synthetic waste using Pseudomonas pseudoflava : PHA synthase enzyme activity analysis from P. pseudoflava and P. palleronii

Synthetic wastewater (SW) at various carbon concentrations (5–60 g/l) were evaluated for polyhydroxyalkanoates (PHA) production using the bacteria Pseudomonas pseudoflava. Bacteria showed highest PHA production with 20 g/l (57 ± 5%), and highest carbon removal at 5 g/l (74 ± 6%) concentrations respectively. Structure, molecular weight, and thermal properties of the produced PHA were evaluated using various analytical techniques. Bacteria produced homo-polymer [poly-3-hydroxybutyrate (P3HB)] when only acetate was used as carbon source; and it produced co-polymer [poly-(3-hydroxybutyrate-co-3-hydroxyvalerate) P(3HB-co-3HV)] by addition of co-substrate propionate. PHA synthase, the enzyme which produce PHA was extracted from two bacterial strains i.e., P. pseudoflava and P. palleronii and its molecular weight was analysed using SDS-PAGE. Protein concentration, and PHA synthase enzyme activity of P. pseudoflava and P. palleronii was carried out using spectrophotometer. Results denoted that P. pseudoflava can be used for degradation of organic carbon persistent in wastewaters and their subsequent conversion into PHA

Two-Stage Polyhydroxyalkanoates (PHA) Production from Cheese Whey Using Acetobacter pasteurianus C1 and Bacillus sp. CYR1

Cheese whey (CW) can be an excellent carbon source for polyhydroxyalkanoates (PHA)-producing bacteria. Most studies have used CW, which contains high amounts of lactose, however, there are no reports using raw CW, which has a relatively low amount of lactose. Therefore, in the present study, PHA production was evaluated in a two-stage process using the CW that contains low amounts of lactose. In first stage, the carbon source existing in CW was converted into acetic acid using the bacteria, Acetobacter pasteurianus C1, which was isolated from food waste. In the second stage, acetic acid produced in the first stage was converted into PHA using the bacteria, Bacillus sp. CYR-1. Under the condition of without the pretreatment of CW, acetic acid produced from CW was diluted at different folds and used for the production of PHA. Strain CYR-1 incubated with 10-fold diluted CW containing 5.7 g/L of acetic acid showed the higher PHA production (240.6 mg/L), whereas strain CYR-1 incubated with four-fold diluted CW containing 12.3 g/L of acetic acid showed 126 mg/L of PHA. After removing the excess protein present in CW, PHA production was further enhanced by 3.26 times (411 mg/L) at a four-fold dilution containing 11.3 g/L of acetic acid. Based on Fourier transform infrared spectroscopy (FT-IR), and H-1 and C-13 nuclear magnetic resonance (NMR) analyses, it was confirmed that the PHA produced from the two-stage process is poly-beta-hydroxybutyrate (PHB). All bands appearing in the FT-IR spectrum and the chemical shifts of NMR nearly matched with those of standard PHB. Based on these studies, we concluded that a two-stage process using Acetobacter pasteurianus C1 and Bacillus sp. CYR-1 would be applicable for the production of PHB using CW containing a low amount of lactose

CARMA Survey Toward Infrared-bright Nearby Galaxies (STING) II: Molecular Gas Star Formation Law and Depletion Time Across the Blue Sequence

We present an analysis of the relationship between molecular gas and current star formation rate surface density at sub-kpc and kpc scales in a sample of 14 nearby star-forming galaxies. Measuring the relationship in the bright, high molecular gas surface density (\Shtwo\gtrsim20 \msunpc) regions of the disks to minimize the contribution from diffuse extended emission, we find an approximately linear relation between molecular gas and star formation rate surface density, \nmol\sim0.96\pm0.16, with a molecular gas depletion time \tdep\sim2.30\pm1.32 Gyr. We show that, in the molecular regions of our galaxies there are no clear correlations between \tdep\ and the free-fall and effective Jeans dynamical times throughout the sample. We do not find strong trends in the power-law index of the spatially resolved molecular gas star formation law or the molecular gas depletion time across the range of galactic stellar masses sampled (\mstar $\sim$$10^{9.7}-10^{11.5}$ \msun). There is a trend, however, in global measurements that is particularly marked for low mass galaxies. We suggest this trend is probably due to the low surface brightness CO, and it is likely associated with changes in CO-to-H2 conversion factor.Comment: To appear in ApJ, December 2011; 17 pages; 8 figure

Unveiling the physical properties and kinematics of molecular gas in the Antennae Galaxies (NGC 4038/9) through high resolution CO (J = 3-2) observations

We present a ~ 1" (100 pc) resolution 12CO (3-2) map of the nearby intermediate stage interacting galaxy pair NGC 4038/9 (the Antennae galaxies) obtained with the Submillimeter Array. We find that half the CO (3-2) emission originates in the overlap region where most of the tidally induced star formation had been previously found in shorter wavelength images, with the rest being centered on each of the nuclei. The gross distribution is consistent with lower resolution single dish images, but we show for the first time the detailed distribution of the warm and dense molecular gas across this galaxy pair at resolutions comparable to the size of a typical giant molecular complex. While we find that 58% (33/57) of the spatially resolved Giant Molecular Associations (GMAs; a few x 100 pc) are located in the overlap region, only \leqq 30% spatially coincides with the optically detected star clusters, suggesting that the bulk of the CO (3-2) emission traces the regions with very recent or near future star formation activity. The spatial distribution of the CO (3-2)/CO (1-0) integrated brightness temperature ratios mainly range between 0.3 and 0.8, which suggests that on average the CO (3-2) line in the Antennae is not completely thermalized and similar to the average values of nearby spirals. A higher ratio is seen in both nuclei and the southern complexes in the overlap region. Higher radiation field associated with intense star formation can account for the nucleus of NGC 4038 and the overlap region, but the nuclear region of NGC 4039 show relatively little star formation or AGN activities and cannot be easily explained. We show kinematical evidence that the high line ratio in NGC 4039 is possibly caused by gas inflow into the counter-rotating central disk.Comment: 20 pages, 12 figures, 4 tables, accepted for publication in Astrophysical Journa

Polyglutamine tracts regulate beclin 1-dependent autophagy

Nine neurodegenerative diseases are caused by expanded polyglutamine (polyQ) tracts in different proteins, such as huntingtin in Huntington's disease and ataxin 3 in spinocerebellar ataxia type 3 (SCA3). Age at onset of disease decreases with increasing polyglutamine length in these proteins and the normal length also varies. PolyQ expansions drive pathogenesis in these diseases, as isolated polyQ tracts are toxic, and an N-terminal huntingtin fragment comprising exon 1, which occurs $\textit{in vivo}$ as a result of alternative splicing, causes toxicity. Although such mutant proteins are prone to aggregation, toxicity is also associated with soluble forms of the proteins. The function of the polyQ tracts in many normal cytoplasmic proteins is unclear. One such protein is the deubiquitinating enzyme ataxin 3 (refs 7, 8), which is widely expressed in the brain. Here we show that the polyQ domain enables wild-type ataxin 3 to interact with beclin 1, a key initiator of autophagy. This interaction allows the deubiquitinase activity of ataxin 3 to protect beclin 1 from proteasome-mediated degradation and thereby enables autophagy. Starvation-induced autophagy, which is regulated by beclin 1, was particularly inhibited in ataxin-3-depleted human cell lines and mouse primary neurons, and $\textit{in vivo}$ in mice. This activity of ataxin 3 and its polyQ-mediated interaction with beclin 1 was competed for by other soluble proteins with polyQ tracts in a length-dependent fashion. This competition resulted in impairment of starvation-induced autophagy in cells expressing mutant huntingtin exon 1, and this impairment was recapitulated in the brains of a mouse model of Huntington's disease and in cells from patients. A similar phenomenon was also seen with other polyQ disease proteins, including mutant ataxin 3 itself. Our data thus describe a specific function for a wild-type polyQ tract that is abrogated by a competing longer polyQ mutation in a disease protein, and identify a deleterious function of such mutations distinct from their propensity to aggregate.We thank the Wellcome Trust (Principal Research Fellowship to D.C.R. (095317/Z/11/Z), Wellcome Trust Strategic Grant to Cambridge Institute for Medical Research (100140/Z/12/Z)), National Institute for Health Research Biomedical Research Centre at Addenbrooke’s Hospital, and Addenbrooke’s Charitable Trust and Federation of European Biochemical Societies (FEBS Long-Term Fellowship to A.A.) for funding; R. Antrobus for mass spectrometry analysis; S. Luo for truncated HTT constructs; M. Jimenez-Sanchez and C. Karabiyik for assistance with the primary mouse cell cultures; and J. Lim and Z. Ignatova for reagents

Fabrication of Surface Plasmon Resonance Sensor Surface with Control of the Non-Specific Adsorption and Affinity for the Detection of 2,4,6-Trinitrotoluene Using an Antifouling Copolymer

We fabricated a surface plasmon resonance (SPR) sensor using a hydrophilic polymer for the highly sensitive detection of 2,4,6-trinitrotoluene (TNT). The hydrophilic polymer was made from mono-2-(methacryloyloxy)ethylsuccinate (MES) and 2-hydroxyethylmethacrylate (HEMA) by surface-initiated atom transfer radical polymerization. The detection of TNT was carried out by displacement assay with the SPR measurement. In displacement assay, the affinity between anti-TNT antibody and the sensor surface, affects to the sensitivity. In the SPR measurement, non-specific adsorption should be controlled because SPR sensor cannot discriminate between specific and non-specific adsorption. Therefore, the affinity and non-specific adsorption were controlled by changing the ratio of HEMA to MES. A detection limit of 0.4 ng/ml (ppb) for TNT was achieved using a sensor surface with the lowest affinity without non-specific adsorption

Sensitive Detection of Capsaicinoids Using a Surface Plasmon Resonance Sensor with Anti-Homovanillic Acid Polyclonal Antibodies

Recently, highly functional biosensors have been developed in preparation for possible large-scale terrorist attacks using chemical warfare agents. Practically applicable sensors are required to have various abilities, such as high portability and operability, the capability of performing rapid and continuous measurement, as well as high sensitivity and selectivity. We developed the detection method of capsaicinoids, the main component of some lachrymators, using a surface plasmon resonance (SPR) immunosensor as an on-site detection sensor. Homovanillic acid, which has a vanillyl group similar to capsaicinoids such as capsaicin and dihydrocapsaicin, was bound to Concholepas concholepas hemocyanin (CCH) for use as an immunogen to generate polyclonal antibodies. An indirect competitive assay was carried out to detect capsaicinoids using SPR sensor chips on which different capsaicin analogues were immobilized. For the sensor chip on which 4-hydroxy-3-methoxybenzylamine hydrochloride was immobilized, a detection limit of 150 ppb was achieved. We found that the incubation time was not required and the detection can be completed in five minutes

Two-Stage Polyhydroxyalkanoates (PHA) Production from Cheese Whey Using Acetobacter pasteurianus C1 and Bacillus sp. CYR1

Cheese whey (CW) can be an excellent carbon source for polyhydroxyalkanoates (PHA)-producing bacteria. Most studies have used CW, which contains high amounts of lactose, however, there are no reports using raw CW, which has a relatively low amount of lactose. Therefore, in the present study, PHA production was evaluated in a two-stage process using the CW that contains low amounts of lactose. In first stage, the carbon source existing in CW was converted into acetic acid using the bacteria, Acetobacter pasteurianus C1, which was isolated from food waste. In the second stage, acetic acid produced in the first stage was converted into PHA using the bacteria, Bacillus sp. CYR-1. Under the condition of without the pretreatment of CW, acetic acid produced from CW was diluted at different folds and used for the production of PHA. Strain CYR-1 incubated with 10-fold diluted CW containing 5.7 g/L of acetic acid showed the higher PHA production (240.6 mg/L), whereas strain CYR-1 incubated with four-fold diluted CW containing 12.3 g/L of acetic acid showed 126 mg/L of PHA. After removing the excess protein present in CW, PHA production was further enhanced by 3.26 times (411 mg/L) at a four-fold dilution containing 11.3 g/L of acetic acid. Based on Fourier transform infrared spectroscopy (FT-IR), and 1H and 13C nuclear magnetic resonance (NMR) analyses, it was confirmed that the PHA produced from the two-stage process is poly-β-hydroxybutyrate (PHB). All bands appearing in the FT-IR spectrum and the chemical shifts of NMR nearly matched with those of standard PHB. Based on these studies, we concluded that a two-stage process using Acetobacter pasteurianus C1 and Bacillus sp. CYR-1 would be applicable for the production of PHB using CW containing a low amount of lactose

Rational Design of Peptide-Functionalized Surface Plasmon Resonance Sensor for Specific Detection of TNT Explosive

In this study, a rationally-designed 2,4,6-trinitrotoluene (TNT) binding peptide derived from an amino acid sequence of the complementarity-determining region (CDR) of an anti-TNT monoclonal antibody was used for TNT detection based on a maleimide-functionalized surface plasmon resonance (SPR) sensor. By antigen-docking simulation and screening, the TNT binding candidate peptides were obtained as TNTHCDR1 derived from the heavy chain of CDR1, TNTHCDR2 derived from CDR2, and TNTHCDR3 from CDR3 of an anti-TNT antibody. The binding events between candidate peptides and TNT were evaluated using the SPR sensor by direct determination based on the 3-aminopropyltriethoxysilane (APTES) surface. The TNT binding peptide was directly immobilized on the maleimide-functionalized sensor chip surface from N-γ-maleimidobutyryl-oxysuccinimide ester (GMBS). The results demonstrated that peptide TNTHCDR3 was identified and selected as a TNT binding peptide among the other two candidate peptides. Five kinds of TNT analogues were also investigated to testify the selectivity of TNT binding peptide TNTHCDR3. Furthermore, the results indicated that the APTES-GMBS-based SPR sensor chip procedure featured a great potential application for the direct detection of TNT