A novel microbial BOD biosensor developed by the immobilization of P. Syringae in micro‐cellular polymers

BACKGROUND: A biochemical oxygen demand (BOD) sensor, based on an immobilized Pseudomonas syringae in highly porous micro‐cellular polymer (MCP) in combination with a dissolved oxygen electrode, has been developed for the analysis of biodegradable organic compounds in aqueous samples. Microorganisms were immobilized in a molded MCP disk and a wastewater sample was injected into the biocomposite disk by a flow injection system. Dissolved oxygen (DO) changes as a measure of soluble BOD was read with a DO probe placed into a flow cell carrying biocatalytically activated disk. RESULTS: Optimal response of the MCP BOD sensor was obtained at pH 6.8 and 25 °C with a typical response time of 3–5 min for a 2 mm thick molded polymeric disk. The sensor showed detection linearity over the range 5–100 mg L^?1 BOD₅ (r² > 0.99) at a flow rate of 0.6 mL min^?1. The repeatability and reproducibility of the sensor response were found to be 3.08% and 7.77%, respectively. BOD values produced with this biosensor for various municipal and industrial wastewaters correlated well with those determined by the conventional 5‐day BOD test. CONCLUSION: This new biosensor was different from present amperometric BOD biosensor configurations in which the biocatalyst (microbial/enzymatic) is placed between cellulose and Teflon membranes installed on a DO probe. The use of a molded MCP disk coniainng microbial activity offers better stability and lifetime for commercial use in environmental monitoring. Copyright © 2008 Society of Chemical Industry     

Effect of temperature on the performance of a biofilter inoculated withPseudomonas putida to treat waste-air containing ethanol

The microbes ofPseudomonas putida (KCTC1768) were fixed on the biofilter-packing media comprising an equivolume mixture of granular activated carbon (GAC) and compost, by recycling the liquid medium containing incubatedPseudomonas putida (KCTC1768). A biofilter experiment was performed to observe its transient behavior under the operating condition of 2,180 ppmv of ethanol-inlet concentration and 158 g/m³/h of ethanol-inlet load for the five consecutive temperature-stages of operation ranging from 25 °C to 40 °C. For the five temperaturestages of operation their removal efficiencies were measured and were compared with each other. The optimum operating temperature of the biofilter turned out to beca. 30 °C, which was consistent with the previous experimental result of Lim and Park. However, the optimum incubation-temperatures ofPseudomonas putida (KCTC1768) and the equivalent (i.e., NCIMB8858) were announced to be of 26 °C and 25 °C by Korea Collection for Type Cultures (KCTC) and National Collections of Industrial, Food and Marine Bacteria (NCIMB), respectively. It was also confirmed by the experiment in which the microbes were incubated in the same liquid medium as in the previous work of Lim and Park at temperature ranging from 20 °C to 40 °C and their growth rates were subsequently measured. Thus, the optimum operating temperature of a biofilter inoculated withPseudomonas putida (KCTC 1768) was proved to be 30 °C, which was higher than its optimum incubation-temperature byca. 5 °C     

Bioproduction of cis‐(1S,2R)‐indandiol,a chiral pharmaceutical intermediate,using a solid–liquid two‐phase partitioning bioreactor for enhanced removal of inhibitors

BACKGROUND: A solid‐liquid two‐phase partitioning bioreactor (TPPB) was used in the biotransformation of indene to cis‐(1S,2R)‐indandiol by Pseudomonas putida 421‐5 (ATCC 55687). Metered substrate feeding in single‐phase operation, or delivery from an immiscible liquid, have previously been employed to regulate the exposure of the biocatalyst to inhibitory concentrations of the substrate. In contrast, the solid‐liquid platform provided in situ substrate addition (ISSA) as well as simultaneous it in situ product removal (ISPR) as a means of overcoming substrate and product toxicity. Three different modes of operation were compared for their effects on the performance of this biotransformation: single‐phase, fed‐batch operation was carried out as a benchmark in 2.75 L aqueous medium, and subsequently with the inclusion of either 700 g liquid silicone oil or 700 g solid polymer beads. RESULTS: Biphasic modes achieved a 3‐fold productivity improvement with respect to single‐phase (30 to 90 mg L^?1 h^?1), and solid‐liquid productivity was similar to liquid‐liquid operation while achieving more extensive removal of inhibitory compounds resulting in a slightly higher product titer (1.29 vs 1.16 g L^?1). The operability of the reactor was improved by the phase stability of the solid polymer beads relative to immiscible organic solvents, preventing emulsion formation and facilitating analytics. CONCLUSION: Solid polymer beads replaced the immiscible liquid auxiliary phase for substrate delivery while performing simultaneous inhibitory molecule sequestration. Copyright © 2011 Society of Chemical Industry     

Inactivation of E. coli and Pseudomonas aeruginosa by electrochloration under bipolar pulsed polarization

This work is devoted to the biocides effects by electrochlorination under a pulsated bipolar polarization (PBP) of low potentials (− 1 V/cm to + 1.6 V/cm). Two working electrodes were used, one in platinum and the other in AISI 430 stainless steel, in two different electrolytes: sterilized distilled water and 0,9N sterilized NaCl solution, contaminated by E. Coli or by Pseudomonas Aeruginosa. The inactivation of these bacteria was established under the action of three biocide treatments: E.F (Electric Field); the combination of E.F and Active Chlorine (A.C) and the combination of E.F, A.C and Ions in Solutions (I.S). The synergistic effect of these treatments leads to a high efficiency in disinfection. The results showed that the total inactivation of the E. Coli stain is reached in half a minute, while that of P. aeruginosa is reached in 20 min. The use of an economic stainless steel electrode allowed us to observe that, for pulsate frequencies higher than 10³ s^− 1, the formation of active chlorine increased whereas the corrosion speed decreased.     

PMMA‐multigraft copolymers derived from linseed oil,soybean oil,and linoleic acid: Protein adsorption and bacterial adherence

Synthesis of Poly(methyl methacrylate), PMMA‐multigraft copolymers derived from linseed oil, soybean oil, and linoleic acid PMMA‐g‐polymeric oil/oily acid‐g‐poly(3‐hydroxy alkanoate) (PHA), and their protein adsorption and bacterial adherence have been described. Polymeric oil/oily acid peroxides [polymeric soybean oil peroxide (PSB), polymeric linseed oil peroxide (PLO), and polymeric linoleic acid peroxide (PLina)] initiated the copolymerization of MMA and unsaturated PHA‐soya to yield PMMA–PLO–PHA, PMMA–PSB–PHA, and PMMA–PLina–PHA multigraft copolymers. PMMA–PLina–PHA multigraft copolymers were completely soluble while PMMA–PSB–PHA and PMMA–PLO–PHA multigraft copolymers were partially crosslinked. Crosslinked parts of the PLO‐ and PSB‐multigraft copolymers were isolated by the sol gel analysis and characterized by swelling measurements in CHCl₃. Soluble part of the PLO‐ and PSB‐multigraft copolymers and completely soluble PLina‐multigraft copolymers were obtained and characterized by spectroscopic, thermal, gel permeation chromatography (GPC), and scanning electron microscopy (SEM) techniques. In the mechanical properties of the PHA–PLina–PMMA, the elongation at break is reduced up to ～ 9%, more or less preserving the high stress values at its break point (48%) when compared to PLina‐g‐PMMA. The solvent casting film surfaces were studied by means of adsorption of blood proteins and bacterial adhesion. Insertion of the PHA into the multigraft copolymers caused the dramatic increase in bacterial adhesion on the polymer surfaces. PHA insertion into the graft copolymers also increased the protein adsorption. © 2007 Wiley Periodicals, Inc. J Appl Polym Sci 2007     

伤寒Vi多糖与不同载体蛋白制备的结合物的特性分析

目的比较以白喉类毒素(diphtheria toxoid,DT)和重组铜绿假单胞菌外毒素A(recombinant Pseudomonas aeruginosa exotoxin A,r EPA)为载体蛋白,分别与伤寒Vi多糖(typhoid Vi polysaccharide,STVi)偶联的结合物的理化特性、抗原性及免疫效应。方法分别以DT、r EPA为载体蛋白,以己二酰肼(adipyl dihydrazide,ADH)为连接剂,使Vi多糖与载体蛋白共价偶联制备多糖蛋白结合物STVi-DT和STVi-r EPA,检测其理化特性指标、抗原性,再将其于0、14、28 d免疫NIH小鼠,经眼眶采血,分离血清,采用间接ELISA法检测小鼠血清抗体水平,分析其免疫原性、免疫记忆效应及免疫持久性。结果 STVi-DT、STVi-r EPA理化特性指标相似。STVi-DT针对STVi、DT抗血清均具有明显抗原性,而针对r EPA抗血清无抗原性,STVi-r EPA针对STVi、r EPA抗血清均具有明显抗原性,而针对DT抗血清无抗原性。Vi多糖和Vi多糖蛋白结合物均具有动物免疫原性,STVi蛋白结合物较STVi具有更强的免疫原性,STVi-DT的动物免疫原性试验结果略优于STVi-r EPA;免疫2剂STVi-DT和STVi-r EPA后,小鼠血清GMT明显升高,且STViDT组明显高于STVi-r EPA组(P0.05),具有较强的免疫记忆效应;免疫STVi-DT和STVi-r EPA后,小鼠血清GMT随免疫剂次增加和时间延长持续升高,直至稳定在一定水平,免疫持久性较好。结论 STVi-DT、STVi-r EPA理化特性相似,均具有显著的免疫原性、免疫记忆效应及免疫持久性,而STVi-DT免疫原性、免疫记忆效应更强。     

苯酚降解菌CN-6的分离及其生长特性

从台湾某纺织厂污泥中分离得到一株能够将苯酚作为唯一碳源和能源生长的细菌。根据其形态和生理生化反应结合16S rDNA测定分析,该菌被鉴定为假单胞杆菌(Pseudomonas sp.)。降解特性研究表明,该菌降解苯酚的最适pH和温度分别为8.5和30℃,在此条件下,浓度高达500mg/L的苯酚能被该菌在17 h内将完全降解。在动力学研究中,由于高浓度苯酚对CN-6菌株的降解过程存在底物抑制现象,故采用Haldane非竞争性底物抑制模型,计算确定了模型参数μmax、KS和KI分别为0.452 h-12、8.617 mg/L7、82.4 mg/L,该动力学方程对实验数据能很好拟合。