Influence of Fe2O3 and bacterial biofilms on Cu(II) distribution in a simulated aqueous solution: A feasibility study to sediments in the Pearl River Estuary (PR China).

This study investigated physico-chemical interactions among Cu(II), biogenic materials, and Fe2O3 in a continuous-flow biofilm reactor system under a well-controlled environment. The effects of Fe2O3 and bacterial biofilms on the distribution of Cu(II) in a simulated aquatic environment were studied. To control biological and abiotic elements in the marine environment, a biofilm reactor was designed to understand the metal speciation of Cu(II) and its distribution. The reactor consisted of a biofilm chamber equipped with glass slides for biofilms attachment. Due to its ability to grow as biofilm in the medium, Pseudomonas atlantica was cultivated to adsorb trace Cu(II) to attached and suspended cells. It was found that biofilms with 170-285 mequiv chemical oxygen demand (COD) concentration/m2 of total oxidizable materials accelerated the Cu(II) adsorption to the surface of the reactor significantly by a factor of five. A significant inhibition to the bacterial growth took place (p ≤ 0.05; t-test) when Cu(II) concentration was higher than 0.5 mg/L. In the absence of Cu(II), bacterial cells grew normally to 0.075 of optical density (OD). However, at the Cu(II) concentration of 0.2 mg/L, the cells grew to a lower OD of 0.58. The presence of glycine and EDTA substantially reduced the toxicity of Cu(II) on bacterial growth (p ≤ 0.05; paired t-test). Their complexation with Cu(II) rendered the metal ions less available to bacterial cells. This implies that the Fe2O3 and bacterial biofilm affected Cu(II) distribution and speciation in the aquatic environment.

Biosorption of heavy metals from aqueous solutions using activated sludge, Aeromasss hydrophyla, and Branhamella spp based on modeling with GEOCHEM.

This work tests the technical applicability of sewage sludge and isolated dead cells of Aeromasss hydrophyla and Branhamella spp for the elimination of inorganic pollutants such as Zn(II), Pb(II), Cd(II), and/or Cu(II) using synthetic wastewater with their initial concentrations of 100 mg/L, respectively. The sludge samples were collected from local sewage treatment plants. The effects of dose and pH on heavy metals removal were evaluated in batch studies and their removal performances were compared to those of previous studies. Both the Freundlich and the Langmuir models were plotted to study their biosorption using activated sludge and the bacteria. Isotherm data, resulting from the batch studies, were compared to the modeling results of Geochem. It was evident that the activated sludge could achieve 99% of Zn(II), Cd(II), Cu(II) and Pb(II) removal with 100 mg/L of concentration at pH 6.0 and 3 g/L of dose. Under the same conditions, 97% of Cd(II), Cu(II) and/or Pb(II) was removed by Aeromasss hydrophyla and Branhamella spp, as indicated by their adsorption capacities (activated sludge: 99.07 mg Pb2+/g; dewatered sludge: 57.15 mg Pb2+/g; digested sludge: 83.58 mg Pb2+/g; 24.47 mg Cd2+/g; Aeromasss hydrophylla: 71.91 mg Pb2+/g; Branhamella spp: 37.52 mg Cu2+/g). Of the four heavy metals studied, Pb(II) had the highest metal adsorption capacity for all adsorbents studied (Pb2+>Cu2+> Cd2+>Zn2+). The modeling results of the Geochem fitted well with the isotherm data of the batch studies at varying concentrations from 20 to 100 mg/L. The thermodynamic constant at pH 4 were comparable to those obtained from previous works. This indicates a reliable prediction over varying metal concentrations and pHs of the batch studies. In spite of the promising results, the treated effluents still could not meet the required effluent limits set by local legislation. Therefore, it is necessary to subsequently treat the samples using biological processes such as activated sludge.

A bioengineered arginine-depleting enzyme as a long-lasting therapeutic agent against cancer.

L-Arginine (L-Arg) depletion has attracted great attention in cancer therapy. Although two types of arginine-depleting enzymes, arginine deiminase (ADI) and human arginase I, are undergoing clinical trials, random site of PEGylation, low efficacy of heavy metal as co-factor, and immunogenicity limit the performance of these drugs and cause difficulty in a homogeneous production. Here we screened ten catalytic metal ions and have successfully produced a site-specific mono-PEGylated human arginase I mutant by conjugating the Cys45 residue to PEG-maleimide to minimize the decrease in activity and produce a homogeneous product. The catalytic efficiency trend of metal ion-enriched human arginase I mutant (HAI) was Co2+ > Ni2+ ≫ Mn2+. The overall kcat/KM values of Co-HAI and Ni-HAI were higher than Mn-HAI by ~ 8.7- and ~ 5.2-folds, respectively. Moreover, the results of enzyme kinetics and circular dichroism spectrometry demonstrated that the 20 or 40 kDa linear and branched PEG attached on the HAI surface did not affect the enzyme activity and the protein secondary structures. In vitro studies showed that both Co-HAI-PEG20L and Ni-HAI-PEG20L inhibited the growth of eight types of cancer cell lines. The pharmacodynamic study in mice demonstrated that the i.p. administration of Co-HAI-PEG20L at 13 mg/kg and Ni-HAI-PEG20L at 15 mg/kg was able to maintain a L-Arg level below its detection limit for over 120 h after one injection. The body weights of mice could return to normal levels within 5 days after injection, showing that the doses were well-tolerated. Therefore, both the Ni-HAI-PEG20L and Co-HAI-PEG20L are promising candidates for cancer therapy. KEY POINTS: • Mono-PEGylation applied on human arginase I mutant (HAI) successfully. • The catalytic efficiency of Co- and Ni-enriched HAI was higher than the wild type. • At least eight types of cancer cell lines were inhibited by Co- and Ni-HAI-PEG20L. • Co- and Ni-HAI-PEG20L were able to achieve weekly depletion of L-Arg. Graphical abstract.

Recombinant Bacillus caldovelox Arginase Mutant (BCA-M) Induces Apoptosis, Autophagy, Cell Cycle Arrest and Growth Inhibition in Human Cervical Cancer Cells.

With our recent success in developing a recombinant human arginase drug against broad-spectrum cancer cell lines, we have explored the potential of a recombinant Bacillus caldovelox arginase mutant (BCA-M) for human cervical cancer treatment. Our studies demonstrated that BCA-M significantly inhibited the growth of human cervical cancer cells in vitro regardless of argininosuccinate synthetase (ASS) and argininosuccinate lyase (ASL) expression. Drug susceptibilities correlate well with the expressions of major urea cycle genes and completeness of L-arginine regeneration pathways. With the expressions of ASS and ASL genes conferring resistance to L-arginine deiminase (ADI) which is undergoing Phase III clinical trial, BCA-M offers the advantage of a broader spectrum of susceptible cancer cells. Mechanistic studies showed that BCA-M inhibited the growth of human cervical cancer cells by inducing apoptosis and cell cycle arrest at S and/or G2/M phases. Our results also displayed that autophagy served as a protective mechanism, while the growth inhibitory effects of BCA-M could be enhanced synergistically by its combination to the autophagy inhibitor, chloroquine (CQ), on human cervical cancer cells.

Mono-PEGylation of a Thermostable Arginine-Depleting Enzyme for the Treatment of Lung Cancer.

L-arginine (L-Arg) depletion induced by randomly PEGylated arginine deiminase (ADI-PEG20) can treat arginosuccinate synthase (ASS)-negative cancers, and ADI-PEG20 is undergoing phase III clinical trials. Unfortunately, ASS-positive cancers are resistant to ADI-PEG20. Moreover, the yield of ADI production is low because of the formation of inclusion bodies. Here, we report a thermostable arginine-depleting enzyme, Bacillus caldovelox arginase mutant (BCA-M: Ser161->Cys161). An abundant amount of BCA-M was easily obtained via high cell-density fermentation and heat treatment purification. Subsequently, we prepared BCA-M-PEG20, by conjugating a single 20 kDa PEG monomer onto the Cys161 residue via thio-chemistry. Unlike ADI-PEG20, BCA-M-PEG20 significantly inhibited ASS-positive lung cancer cell growth. Pharmacodynamic studies showed that a single intraperitoneal injection (i.p). administration of 250 U/mouse of BCA-M-PEG20 induced low L-Arg level over 168 h. The mono-PEGylation of BCA-M prolonged its elimination half-life from 6.4 to 91.4 h (a 14-fold increase). In an A549 lung cancer xenograft model, a weekly administration of 250 U/mouse of BCA-M-PEG20 suppressed tumor growth significantly. We also observed that BCA-M-PEG20 did not cause any significant safety issue in mouse models. Overall, BCA-M-PEG20 showed excellent results in drug production, potency, and stability. Thereby, it has great potential to become a promising candidate for lung cancer therapy.

A societal transition of MSW management in Xiamen (China) toward a circular economy through integrated waste recycling and technological digitization.

Recently Xiamen (China) has encountered various challenges of municipal solid waste management (MSWM) such as lack of a complete garbage sorting and recycling system, the absence of waste segregation between organic and dry waste at source, and a shortage of complete and clear information about the MSW generated. This article critically analyzes the existing bottlenecks in its waste management system and discusses the way forward for the city to enhance its MSWM by drawing lessons from Hong Kong's effectiveness in dealing with the same problems over the past decades. Solutions to the MSWM problem are not only limited to technological options, but also integrate environmental, legal, and institutional perspectives. The solutions include (1) enhancing source separation and improving recycling system; (2) improving the legislation system of the MSWM; (3) improvement of terminal disposal facilities in the city; (4) incorporating digitization into MSWM; and (5) establishing standards and definitions for recycled products and/or recyclable materials. We also evaluate and compare different aspects of MSWM in Xiamen and Hong Kong SAR (special administrative region) under the framework of 'One Country, Two Systems' concerning environmental policies, generation, composition, characteristics, treatment, and disposal of their MSW. The nexus of society, economics of the MSW, and the environment in the sustainability sphere are established by promoting local recycling industries and the standardization of recycled products and/or recyclable materials. The roles of digitization technologies in the 4th Industrial Revolution for waste reduction in the framework of circular economy (CE) are also elaborated. This technological solution may improve the city's MSWM in terms of public participation in MSW separation through reduction, recycle, reuse, recovery, and repair (5Rs) schemes. To meet top-down policy goals such as a 35% recycling rate for the generated waste by 2030, incorporating digitization into the MSWM provides the city with technology-driven waste solutions.

Biological processes for treatment of landfill leachate.

This review presents an overview with critical analysis of the technical applicability of biological treatments for landfill leachate. A particular focus is given to activated sludge (AS), sequencing batch reactors (SBR), aerated lagoons (AL), and upflow anaerobic sludge blankets (UASB). Their advantages and limitations in application are evaluated. Selected information is presented such as pH, hydraulic retention time (HRT), organic loading rate (OLR), characteristics of leachate and treatment performance. It is evident from the literature survey of 188 papers (1976-2010) that none of the individual biological treatments presented is universally applicable for removing recalcitrant contaminants from leachate. Among the biological treatments reviewed, AS, SBR and UASB are the most frequently applied. These treatments are effective not only to remove over 90% of COD with a concentration ranging from 3500-26 000 mg L(-1), but also to achieve 80% of NH(3)-N removal with a concentration ranging from 100-1000 mg L(-1). A combination of physico-chemical and biological treatment into an integrated process is effective for leachate treatment. Almost complete removal of COD and NH(3)-N was reported for combined reverse osmosis (RO) and UASB with an initial COD concentration of 35 000 mg L(-1) and NH(3)-N concentration of 1600 mg L(-1). Integrated Fenton's oxidation and AS could achieve about 98% and 99% of COD and NH(3)-N removal, respectively, with initial COD and NH(3)-N concentrations of 7000 mg L(-1) and 1800 mg L(-1). Overall, the selection of the most suitable treatment for leachate depends on its characteristics, technical applicability and potential constraints, effluent limit required, cost-effectiveness, regulatory requirements and long-term environmental impacts.

Pegylated derivatives of recombinant human arginase (rhArg1) for sustained in vivo activity in cancer therapy: preparation, characterization and analysis of their pharmacodynamics in vivo and in vitro and action upon hepatocellular carcinoma cell (HCC).

BACKGROUND: Protein used in medicine, e.g. interferon, are immunogenic and quickly broken down by the body. Pegylation is a recognized way of preserving their integrity and reducing immune reactions, and works well with enzymes used to degrade amino acids, a recent focus of attention in controlling cancer growth. Of the two arginine-degrading enzymes being explored clinically, arginine deiminase is a decidedly foreign mycoplasm-derived enzyme, whereas human arginase 1 is a native liver enzyme. Both have been pegylated, the former with adjuncts of 20 kD, the latter with 5 kD PEG. Pegylation is done by several different methods, not all of which are satisfactory or desirable. METHODS: The preparation of novel polyethylene glycol (PEG) derivatives for modifying proteins is described, but directed specifically at pegylation of recombinant human arginase 1 (rhArg1). rhArg1 expressed in Escherichia coli was purified and coupled in various ways with 5 different PEG molecules to compare their protective properties and the residual enzyme activity, using hepatocellular cell lines both in vitro and in vivo. RESULTS: Methoxypolyethylene glycol-succinimidyl propionate (mPEG-SPA 5,000) coupled with very high affinity under mild conditions. The resulting pegylated enzyme (rhArg1-peg5,000 mw) had up to 6 PEG chains of 5K length which not only protected it from degradation and any residual immunogenicity, but most importantly let it retain >90% of its native catalytic activity. It remained efficacious in depleting arginine in rats after a single ip injection of 1,500 U of the conjugate as the native enzyme, plasma arginine falling to >0.05 microM from approximately 170 microM within 20 min and lasting 6 days. The conjugate had almost the same efficacy as unpegylated rhArg1 on 2 cultured human liver cancer (HCC) cell lines. It was considerably more effective than 4 other pegylated conjugates prepared. CONCLUSION: Valuable data on the optimization of the pegylation procedure and choice of ligand that best stabilizes the enzyme arginase 1 are presented, a protocol that should equally fit many other enzymes and proteins. It is a long lasting arginine-depleting enzyme in vivo which will greatly improve its use in anti-cancer therapy.

Pegylated recombinant human arginase (rhArg-peg5,000mw) inhibits the in vitro and in vivo proliferation of human hepatocellular carcinoma through arginine depletion.

Hepatocellular carcinoma (HCC) is believed to be auxotrophic for arginine through the lack of expression of argininosuccinate synthetase (ASS). The successful use of the arginine-depleting enzyme arginine deiminase (ADI) to treat ASS-deficient tumors has opened up new possibilities for effective cancer therapy. Nevertheless, many ASS-positive HCC cell lines are found to be resistant to ADI treatment, although most require arginine for proliferation. Thus far, an arginine-depleting enzyme for killing ASS-positive tumors has not been reported. Here, we provide direct evidence that recombinant human arginase (rhArg) inhibits ASS-positive HCCs. All the five human HCC cell lines we used were sensitive to rhArg but ADI had virtually no effect on these cells. They all expressed ASS, but not ornithine transcarbamylase (OTC), the enzyme that converts ornithine, the product of degradation of arginine with rhArg, to citrulline, which is converted back to arginine via ASS. Transfection of HCC cells with OTC resulted in resistance to rhArg. Thus, OTC expression alone may be sufficient to induce rhArg resistance in ASS-positive HCC cells. This surprising correlation between the lack of OTC expression and sensitivity of ASS-positive HCC cells shows that OTC-deficient HCCs are sensitive to rhArg-mediated arginine depletion. Therefore, pretreatment tumor gene expression profiling of ASS and OTC could aid in predicting tumor response to arginine depletion with arginine-depleting enzymes. We have also shown that the rhArg native enzyme and the pegylated rhArg (rhArg-peg(5,000mw)) gave similar anticancer efficacy in vitro. Furthermore, the growth of the OTC-deficient Hep3B tumor cells (ASS-positive and ADI-resistant) in mice was inhibited by treatment with rhArg-peg(5,000mw), which is active alone and is synergistic in combination with 5-fluorouracil. Thus, our data suggest that rhArg-peg(5,000mw) is a novel agent for effective cancer therapy.

Comparisons of low-cost adsorbents for treating wastewaters laden with heavy metals.

In this article, the removal performance and cost-effectiveness of various low-cost adsorbents derived from agricultural waste, industrial by-product or natural material are evaluated and compared to those of activated carbon for the removal of heavy metals (Cd(II), Cr(III), Cr(VI), Cu(II), Ni(II) and Zn(II)) from metals-contaminated wastewater. To highlight their technical applicability, selected information on pH, dose required, initial metal concentration, adsorption capacity and the price of the adsorbents is presented. It is evident from the survey of 102 published studies (1984-2005) that low cost adsorbents derived from agricultural waste have demonstrated outstanding capabilities for the removal of heavy metal (Cr(VI): 170 mg/g of hazelnut shell activated carbon, Ni(II): 158 mg/g of orange peel, Cu(II): 154.9 mg/g of soybean hull treated with NaOH and citric acid, Cd(II): 52.08 mg/g of jackfruit), compared to activated carbon (Cd(II): 146 mg/g, Cr(VI): 145 mg/g, Cr(III): 30 mg/g, Zn(II): 20 mg/g). Therefore, low-cost adsorbents can be viable alternatives to activated carbon for the treatment of metals-contaminated wastewater. It is important to note that the adsorption capacities presented in this paper vary, depending on the characteristics of the individual adsorbent, the extent of surface modification and the initial concentration of the adsorbate. In general, technical applicability and cost-effectiveness are the key factors that play major roles in the selection of the most suitable adsorbent to treat inorganic effluent.

Degradation of recalcitrant compounds from stabilized landfill leachate using a combination of ozone-GAC adsorption treatment.

Laboratory experiments were undertaken to investigate the treatment performances of ozonation alone and/or its combination with granular activated carbon (GAC) adsorption for raw leachate from the NENT landfill (in Hong Kong). To improve its removal of recalcitrant contaminants from the leachate, the surface of GAC was oxidized with ozone prior to treatment. With respect to ozone dose and pH, the removal of COD and/or NH(3)-N from ozonation alone and combined ozone-GAC adsorption were evaluated and compared to those of other physico-chemical treatments in some reported studies. The removal mechanism of recalcitrant compounds by ozone-GAC adsorption treatment was presented. Among the various treatments studied, the combination of ozone-GAC adsorption using ozone-modified GAC had the highest removal for COD (86%) and/or NH(3)-N (92%) compared to ozonation alone (COD: 35%; NH(3)-N: 50%) at the same initial COD and/or NH(3)-N concentrations of 8000 and 2620 mg/L, respectively. Although the integrated treatment was more effective than ozonation alone for treating stabilized leachate, the results suggested that it could not generate treated effluent that complied with the COD limit of lower than 200 mg/L and the NH(3)-N discharge standard of less than 5 mg/L. Therefore, further biological treatments to complement the degradation of the leachate are still required to meet the environmental legislation.

Physico-chemical treatments for removal of recalcitrant contaminants from landfill leachate.

In this paper, the technical applicability and treatment performance of physico-chemical techniques (individual and/or combined) for landfill leachate are reviewed. A particular focus is given to coagulation-flocculation, chemical precipitation, ammonium stripping, membrane filtration and adsorption. The advantages and limitations of various techniques are evaluated. Their operating conditions such as pH, dose required, characteristics of leachate in terms of chemical oxygen demand (COD) and NH3-N concentration and treatment efficiency are compared. It is evident from the survey of 118 papers (1983-2005) that none of the individual physico-chemical techniques is universally applicable or highly effective for the removal of recalcitrant compounds from stabilized leachate. Among the treatments reviewed in this article, adsorption, membrane filtration and chemical precipitation are the most frequently applied and studied worldwide. Both activated carbon adsorption and nanofiltration are effective for over 95% COD removal with COD concentrations ranging from 5690 to 17,000 mg/L. About 98% removal of NH3-N with an initial concentration ranging from 3260 to 5618 mg/L has been achieved using struvite precipitation. A combination of physico-chemical and biological treatments has demonstrated its effectiveness for the treatment of stabilized leachate. Almost complete removal of COD and NH3-N has been accomplished by a combination of reverse osmosis (RO) and an upflow anaerobic sludge blanket (UASB) with an initial COD concentration of 35,000 mg/L and NH3-N concentration of 1600 mg/L and/or RO and activated sludge with an initial COD concentration of 6440 mg/L and NH3-N concentration of 1153 mg/L. It is important to note that the selection of the most suitable treatment method for landfill leachate depends on the characteristics of landfill leachate, technical applicability and constraints, effluent discharge alternatives, cost-effectiveness, regulatory requirements and environmental impact.

Construction of recombinant Bacillus subtilis for production of polyhydroxyalkanoates.

Polyhydroxyalkanoates (PHAs) are polyesters of hydroxyalkanoates synthesized by numerous bacteria as intracellular carbon and energy storage compounds and accumulated as granules in the cytoplasm of cells. In this work, we constructed two recombinant plasmids, pBE2C1 and pBE2C1AB, containing one or two PHA synthse genes, respectively. The two plasmids were inserted into Bacillus subtilis DB104 to generate modified strains, B. subtilis/pBE2C1 and B. subtilis/pBE2C1AB. The two recombinants strains were subjected to fermentation and showed PHA accumulation, the first reported example of mcl-PHA production in B. subtilis. Gas Chromatography analysis identified the compound produced by B. subtilis/pBE2C1 to be a hydroxydecanoate-co-hydroxydodecanoate (HD-co-HDD) polymer whereas that produced by B. subtilis/pBE2C1AB was a hydroxybutyrate-co-hydroxydecanoate-co-hydroxydodecanoate (HB-HD-HDD) polymer.

Air-stable Ir-(P-Phos) complex for highly enantioselective hydrogenation of quinolines and their immobilization in poly(ethylene glycol) dimethyl ether (DMPEG).

An air-stable catalyst system Ir-(P-Phos) catalyst was found to be highly effective in the asymmetric hydrogenation of quinoline derivatives. The catalyst immobilized in DMPEG was efficiently recovered and reused eight times, retaining reactivity and enantioselectivity.

Sphaerotilus natans isolated from activated sludge and its production of poly(3-hydroxybutyrate-co-3-hydroxyvalerate).

Sphaerotilus natans is a sheathed bacterium existing in the activated sludge of wastewater treatment plants. It is one of the filamentous bacteria causing the bulking and foaming difficulties of activated sludge. Isolating the strain and culturing it in an axenic environment could not only provide the metabolic knowledge of the strains that would be useful in the development of wastewater treatment methods, but also could enable us to gain an understanding of the mechanism by which poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (poly[3-HB-co-3-HV]) is produced by this strain. This article reports the screening and isolation of the strain from the activated sludge using the Nile blue staining method together with Fourier transform infrared analysis. We investigated the ability of the selected strain to produce poly(3-HB-co-3-HV) copolymer using glucose and peptone, or by adding valeric acid or sodium propionate as precursor. Proper precursor feeding could dramatically enhance its 3HV content in the copolymer P(3HB-co-3HV). By controlling the different feeding times in fed-batch fermentation, different desired copolymers were obtained with 15, 40, and 70% 3HV mole fraction of the copolymer. Polymer properties were analyzed by gas chromatography, differential scanning calorimetry, thermo-gravimetry, and nuclear magnetic resonance analysis.

Production of specific copolymers of polyhydroxyalkanoates from industrial waste.

Polyhydroxyalkanoates, biodegradable plastics with the desired physical and chemical properties of conventional synthetic plastics, are extensively investigated. In this study, specific bacterial strains produced specific copolymers from food waste. Copolymers of HB and HV (poly[3-hydroxybutyrate-co-3-hydroxyvalerate]) were obtained using various ratios of butyric acid (C4) and valeric acid (C5) as carbon sources. The C4 to C5 ratio affected the melting points of the copolymers. Melting and glass transition temperatures and many other thermal properties are important parameters relative to in-service polymer applications. Higher ratios of butyrate to valerate gave higher melting points. When a mixed culture of activated sludge was employed to produce copolymers using food wastes as nutrients, the obtained copolymers showed various monomer compositions. Copolymers with a higher portion of HV were obtained using soy waste; copolymers with less HV were obtained using malt wastes. Pure strains, (i.e., Alcaligenes latus DSM 1122, and DSM 1124, Staphylococcus spp., Klebsiella spp.) produced specific copolymers from food waste. Only Klebsiella spp. produced different copolymers; the ratios of HB:HV were 93:7 and 79:21 from malt waste and soy waste, respectively. The other strains produced polymers of 100% HB. Selecting industrial food wastes as carbon sources can further reduce the cost of producing copolymers.

Construction of recombinant Escherichia coli strains for production of poly-(3-hydroxybutyrate-co-3-hydroxyvalerate).

Plastic wastes constitute a worldwide environmental problem, and the demand for biodegradable plastics has become high. One of the most important characteristics of microbial polyesters is that they are thermoplastic with environmentally degradable properties. In this study, pUC19/PHA was cloned and transformed into three different Escherichia coli strains. Among the three strains that were successfully expressed in the production of polyhydroxyalkanoates (PHA), E. coli HMS174 had the highest yield in the production of poly-(3-hydroxybutyrate-co-3-hydroxyvalerate) (P[HB-HV]). The cell dry weight and PHA content of recombinant HMS174 reached as high as 10.27 g/L and 43% (w/w), respectively, in fed-batch fermentor culture. The copolymer of PHA, P(HB-HV), was found in the cells, and the biopolymers accumulated were identified and analyzed by gas chromatography, proton nuclear magnetic resonance spectroscopy, and differential scanning calorimetry. We demonstrated clearly that the E. coli host for PHA production has to be carefully selected to obtain a high yield. The results obtained indicated that a superior E. coli with high PHA production can be constructed with a desirable ratio of P(HB-HV), which has potential applications in industry and medicine.

Biosorption and desorption of copper (II) ions by Bacillus sp.

Batch biosorption experiments were conducted to investigate the removal of Cu2+ ions from aqueous solutions by a series of bacterial strains isolated from a local activated sludge process. The characteristics of 12 isolates were identified and examined for their ability to bind Cu2+ ions from aqueous solution. Among the isolates, two species exhibited biosorption capacity >40 mg of Cu/g of dry cell. Isotherms for the biosorption of copper on bacterial cells were developed and compared, and the equilibrium data fitted well to the Langmuir and Freundlich isotherm models. The biosorption of copper increased significantly with increasing pH from 2.0 to 6.0 regardless of the species. More than 90% of copper sorbed on the cells of Bacillus sp. could be recovered by washing with 0.1 M HNO3 for 5 min. The performance of two different desorption processes was also tested and compared. The results show that five biosorption and desorption cycles are a better operation process than five successive biosorptions followed by one desorption to remove and recover copper from aqueous solution. The biosorbent could be used for at least five biosorptions and desorption cycles without loss of copper removal capacity. It can be concluded that the activated sludge or sludge-isolated bacteria could be a potential biosorbent for copper removal.

An Amyloid-Fibril-Based Colorimetric Nanosensor for Rapid and Sensitive Chromium(VI) Detection.

Appearing with a trace! A new colorimetric nanosensor for CrVI from the amyloid fibrils of hen lysozyme has been developed. This nanosensor, which makes use of hen lysozyme fibrils as CrVI binders and acidified diphenylcarbazide (DPC) as the colorimetric probe, can specifically detect CrVI at the ppb level without sample pretreatment and the use of advanced instruments.

Deprivation of arginine by recombinant human arginase in prostate cancer cells.

BACKGROUND: Recombinant human arginase (rhArg) has been developed for arginine deprivation therapy in cancer, and is currently under clinical investigation. During pre-clinical evaluation, rhArg has exhibited significant anti-proliferative activity in cancer cells deficient in the expression of ornithine carbamoyl transferase (OCT). Interestingly, a variety of cancer cells such as melanoma and prostate cancer deficient in argininosuccinate synthetase (ASS) are sensitive to arginine deprivation by arginine deiminase. In this study, we investigated levels of gene expression of OCT and ASS, and the effects of rhArg in human prostate cancer cells: LNCaP (androgen-dependent), PC-3 and DU-145 (both androgen-independent). RESULTS: Quantitative real-time PCR showed minimal to absent gene expression of OCT, but ample expression of ASS expression in all 3 cell lines. Cell viability assay after 72-h exposure of rhArg showed all 3 lines had half maximal inhibitory concentration less than or equal to 0.02 U/ml. Addition of ornithine to cell culture media failed to rescue these cells from rhArg-mediated cytotoxicity.Decreased phosphorylation of 4E-BP1, a downstream effector of mammalian target of rapamycin (mTOR), was noted in DU-145 and PC-3 after exposure to rhArg. Moreover, there was no significant apoptosis induction after arginine deprivation by rhArg in all 3 prostate cancer cell lines. CONCLUSION: rhArg causes significant cytotoxicity in LNCaP, DU-145 and PC-3 prostate cancer cells which all demonstrate decreased OCT expression. Inhibition of mTOR manifested by hypophosphorylation of 4E-BP1 suggests autophagy is involved as alternative cell death mechanism. rhArg demonstrates a promising novel agent for prostate cancer treatment.