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Search indexed NASA NTRS and DOE OSTI research on propulsion, heat transfer, battery materials and energy systems. Follow report and document links to the original sources.

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At least 127 records · Page 7

Aroclor 1254, a developmental neurotoxicant, alters energy metabolism- and intracellular signaling-associated protein networks in rat cerebellum and hippocampus

The vast literature on the mode of action of polychlorinated biphenyls (PCBs) indicates that PCBs are a unique model for understanding the mechanisms of toxicity of environmental mixtures of persistent chemicals. PCBs have been shown to adversely affect psychomotor function and learning and memory in humans. Although the molecular mechanisms for PCB effects are unclear, several studies indicate that the disruption of Ca{sup 2+}-mediated signal transduction plays significant roles in PCB-induced developmental neurotoxicity. Culminating events in signal transduction pathways include the regulation of gene and protein expression, which affects the growth and function of the nervous system. Our previous studies showed changes in gene expression related to signal transduction and neuronal growth. In this study, protein expression following developmental exposure to PCB is examined. Pregnant rats (Long Evans) were dosed with 0.0 or 6.0 mg/kg/day of Aroclor-1254 from gestation day 6 through postnatal day (PND) 21, and the cerebellum and hippocampus from PND14 animals were analyzed to determine Aroclor 1254-induced differential protein expression. Two proteins were found to be differentially expressed in the cerebellum following PCB exposure while 18 proteins were differentially expressed in the hippocampus. These proteins are related to energy metabolism in mitochondria (ATP synthase, sub unit {beta} (ATP5B), creatine kinase, and malate dehydrogenase), calcium signaling (voltage-dependent anion-selective channel protein 1 (VDAC1) and ryanodine receptor type II (RyR2)), and growth of the nervous system (dihydropyrimidinase-related protein 4 (DPYSL4), valosin-containing protein (VCP)). Results suggest that Aroclor 1254-like persistent chemicals may alter energy metabolism and intracellular signaling, which might result in developmental neurotoxicity. -- Highlights: Black-Right-Pointing-Pointer We performed brain proteomic analysis of rats exposed to the neurotoxicant, Aroclor 1254. Black-Right-Pointing-Pointer Cerebellum and hippocampus were analyzed by 2D DIGE and Mass spectrometry. Black-Right-Pointing-Pointer Proteins affected participate in Energy metabolism, calcium signaling and nervous system growth.

60 APPLIED LIFE SCIENCES↗

Polychlorinated biphenyl quinone induces oxidative DNA damage and repair responses: The activations of NHEJ, BER and NER via ATM-p53 signaling axis

Our previous studies demonstrated that polychlorinated biphenyl (PCB) quinone induced oxidative DNA damage in HepG2 cells. To promote genomic integrity, DNA damage response (DDR) coordinates cell-cycle transitions, DNA repair and apoptosis. PCB quinone-induced cell cycle arrest and apoptosis have been documented, however, whether PCB quinone insult induce DNA repair signaling is still unknown. In this study, we identified the activation of DDR and corresponding signaling events in HepG2 cells upon the exposure to a synthetic PCB quinone, PCB29-pQ. Our data illustrated that PCB29-pQ induces the phosphorylation of p53, which was mediated by ataxia telangiectasia mutated (ATM) protein kinase. The observed phosphorylated histone H2AX (γ-H2AX) foci and the elevation of 8-hydroxy-2′-deoxyguanosine (8-OHdG) indicated that DDR was stimulated by PCB29-pQ treatment. Additionally, we found PCB29-pQ activates non-homologous end joining (NHEJ), base excision repair (BER) and nucleotide excision repair (NER) signalings. However, these repair pathways are not error-free processes and aberrant repair of DNA damage may cause the potential risk of carcinogenesis and mutagenesis. - Highlights: • Polychlorinated biphenyl quinone induces oxidative DNA damage in HepG2 cells. • The elevation of γ-H2AX and 8-OHdG indicates the activation of DNA damage response. • ATM-p53 signaling acts as the DNA damage sensor and effector. • Polychlorinated biphenyl quinone activates NHEJ, BER and NER signalings.

60 APPLIED LIFE SCIENCES↗

A portable and reusable sensor system based on graphene for real-time and sensitive detection of lead ions in water

Long-term exposure to Pb 2+ can cause irreversible damage to the nervous, cardiovascular, and reproductive systems. Therefore, developing a fast and sensitive detection system capable of monitoring minuscule concentrations of Pb 2+ is essential. In this study, we demonstrated a fully portable sensor system enabling rapid, sensitive, and real-time monitoring of Pb 2+ . The sensor system adopted the remote-gate field-effect transistor (RGFET) detection scheme and was easy to operate, even for non-experts. The sensor system comprised two printed circuit boards (PCBs): a sensor PCB with a remote-gate electrode and an analyzer PCB with a metal-oxide-semiconductor field-effect transistor (MOSFET) transducer and peripheral electronics to manage sensor signals. To achieve a high sensitivity for Pb 2+ , we utilized graphene ink drop-casted on the sensor PCB as a sensing membrane. The graphene film was easy to deposit and remove, enabling the sensor PCB to be reused multiple times. The sensor system was further linked to a smartphone application that instantly monitors the sensor response, allowing for rapid point-of-use detection. The sensor exhibited a high sensitivity of 21.7% when the limit of detection (LOD) value of 1 nM (∼0.2 ppb) was detected, and the typical detection time for each sample was approximately 60 seconds. This portable sensor system advances sensing technologies and could potentially supplement expensive, laborious conventional sensing equipment.

54 ENVIRONMENTAL SCIENCES↗

A novel planar ion funnel design for miniature ion optics

The novel planar ion funnel (PIF) design presented in this article emphasizes simple fabrication, assembly, and operation, making it amenable to extreme miniaturization. Simulations performed in SIMION 8.0 indicate that ion focusing can be achieved by using a gradient of electrostatic potentials on concentric metal rings in a plane. A prototype was fabricated on a 35 × 35 mm custom-designed printed circuit board (PCB) with a center hole for ions to pass through and a series of concentric circular metal rings of increasing diameter on the front side of the PCB. Metal vias on the PCB electrically connected each metal ring to a resistive potential divider that was soldered on the back of the PCB. The PIF was tested at 5.5 × 10⁻⁶ Torr in a vacuum test setup that was equipped with a broad-beam ion source on the front and a micro channel plate (MCP) ion detector on the back of the PIF. The ion current recorded on the MCP anode during testing indicated a 23× increase in the ion transmission through the PIF when electric potentials were applied to the rings. These preliminary results demonstrate the functionality of a 2D ion funnel design with a much smaller footprint and simpler driving electronics than conventional 3D ion funnels. Future directions to improve the design and a possible micromachining approach to fabrication are discussed in the conclusions.

46 INSTRUMENTATION RELATED TO NUCLEAR SCIENCE AND ↗

A compact linear accelerator based on a scalable microelectromechanical-system RF-structure

Here, a new approach for a compact radio-frequency (RF) accelerator structure is presented. The new accelerator architecture is based on the Multiple Electrostatic Quadrupole Array Linear Accelerator (MEQALAC) structure that was first developed in the 1980s. The MEQALAC utilized RF resonators producing the accelerating fields and providing for higher beam currents through parallel beamlets focused using arrays of electrostatic quadrupoles (ESQs). While the early work obtained ESQs with lateral dimensions on the order of a few centimeters, using a printed circuit board (PCB), we reduce the characteristic dimension to the millimeter regime, while massively scaling up the potential number of parallel beamlets. Using Microelectromechanical systems scalable fabrication approaches, we are working on further red ucing the characteristic dimension to the sub-millimeter regime. The technology is based on RF-acceleration components and ESQs implemented in the PCB or silicon wafers where each beamlet passes through beam apertures in the wafer. The complete accelerator is then assembled by stacking these wafers. This approach has the potential for fast and inexpensive batch fabrication of the components and flexibility in system design for application specific beam energies and currents. For prototyping the accelerator architecture, the components have been fabricated using the PCB. In this paper, we present proof of concept results of the principal components using the PCB: RF acceleration and ESQ focusing. Finally, ongoing developments on implementing components in silicon and scaling of the accelerator technology to high currents and beam energies are discussed.

43 PARTICLE ACCELERATORS↗

Elucidating the origins of phycocyanobilin biosynthesis and phycobiliproteins

Terrestrial ecosystems and human societies depend on oxygenic photosynthesis, which began to reshape our atmosphere approximately 2.5 billion years ago. The earliest known organisms carrying out oxygenic photosynthesis are the cyanobacteria, which use large complexes of phycobiliproteins as light-harvesting antennae. Phycobiliproteins rely on phycocyanobilin (PCB), a linear tetrapyrrole (bilin) chromophore, as the light-harvesting pigment that transfers absorbed light energy from phycobilisomes to the chlorophyll-based photosynthetic apparatus. Cyanobacteria synthesize PCB from heme in two steps: A heme oxygenase converts heme into biliverdin IXα (BV), and the ferredoxin-dependent bilin reductase (FDBR) PcyA then converts BV into PCB. In the current work, we examine the origins of this pathway. We demonstrate that PcyA evolved from pre-PcyA proteins found in nonphotosynthetic bacteria and that pre-PcyA enzymes are active FDBRs that do not yield PCB. Pre-PcyA genes are associated with two gene clusters. Both clusters encode bilin-binding globin proteins, phycobiliprotein paralogs that we designate as BBAGs (bilin biosynthesis-associated globins). Some cyanobacteria also contain one such gene cluster, including a BBAG, two V4R proteins, and an iron–sulfur protein. Phylogenetic analysis shows that this cluster is descended from those associated with pre-PcyA proteins and that light-harvesting phycobiliproteins are also descended from BBAGs found in other bacteria. We propose that PcyA and phycobiliproteins originated in heterotrophic, nonphotosynthetic bacteria and were subsequently acquired by cyanobacteria.

59 BASIC BIOLOGICAL SCIENCES↗

Operational and Performance Monitoring of East Tennessee Technology Park Ponds - 2024 Results

Performance and operational monitoring are regulatory requirements associated with a non–time-critical removal action for three surface water bodies at the East Tennessee Technology Park (ETTP) in Oak Ridge, Tennessee (Figure 1): the K-1007-P1 Pond, the K-901-A Pond, and the K-720 Slough. This progress report summarizes the monitoring activities and results from FY 2024. The goal of ecological enhancement of the K-1007-P1 and K-901-A Ponds is to reduce risk from polychlorinated biphenyl (PCB) contamination in fish to humans and wildlife. Lowering PCB concentrations in the water column and in the food that fish eat is expected to decrease PCB bioaccumulation in fish. Performance monitoring of the K-1007-P1 Pond focuses on annual PCB trends in water, clams, and fish. PCBs in fish also are sampled annually at the K-720 Slough.

54 ENVIRONMENTAL SCIENCES↗

Is Proxima Centauri b Habitable? A Study of Atmospheric Loss

We address the important question of whether the newly discovered exoplanet, Proxima Centauri b (PCb), is capable of retaining an atmosphere over long periods of time. This is done by adapting a sophisticated multi-species MHD model originally developed for Venus and Mars and computing the ion escape losses from PCb. The results suggest that the ion escape rates are about two orders of magnitude higher than the terrestrial planets of our Solar system if PCb is unmagnetized. In contrast, if the planet does have an intrinsic dipole magnetic field, the rates are lowered for certain values of the stellar wind dynamic pressure, but they are still higher than the observed values for our solar system’s terrestrial planets. These results must be interpreted with due caution since most of the relevant parameters for PCb remain partly or wholly unknown.

79 ASTRONOMY AND ASTROPHYSICS↗

The Ability of AMSTAR Dechlorination Solution to Remove and Degrade PCBs from Contaminated Surfaces

Polychlorinated biphenyls (PCBs) are a group of synthetic aromatic compounds with the general formula C12H(10-x)Cl(-x) that were historically used in industrial paints, caulking material and adhesives, as their properties enhanced structural integrity, reduced flammability and boosted antifungal properties. Although the United States Environmental Protection Agency (USEPA) has banned the manufacture of PCBs since 1979, they have been found in at least 500 of the 1,598 National Priorities List (Superfund) sites identified by the USEPA. Prior to the US EPA's ban on PCB production, PCBs were commonly used as additives in paints and asphalt-based adhesives that were subsequently applied to a variety of structures. Government facilities constructed as early as 1930 utilized PCB-containing binders or PCB-containing paints, which are now leaching into the environment and posing ecological and worker health concerns. In 2006, a commercially available product known as AMSTAR Dechlorination Solution was tested at NASA's Kennedy Space Center for its ability to remove and degrade PCBs from structural materials. This evaluation was requested by the Environmental Security Technology Certification Program (ESTCP) evaluating the ability of NASA's Bimetallic Treatment System (BTS) to remove and degrade PCBs from structural materials. The results of the laboratory testing are to be used to determine if a side-by-side field-scale test comparing BTS to AMSTAR was warranted. A recommended sampling and analysis testing program was submitted to ESTCP that included triplicate screening of AMSTAR's PCB dechlorination capabilities on a variety of surfaces including glass, bare metal, and painted metal coupons. The test procedures, analytical techniques and results obtained are presented in this interim report to ESTCP.

Quinn, Jacqueline E.↗

Space to Space Advanced EMU Radio

The main task for this project was the development of a prototype for the Space to Space Advanced EMU Radio (SSAER). The SSAER is an updated version of the Space to Space EMU Radio (SSER), which is the current radio used by EMUs (Extravehicular Mobility Unit) for communication between suits and with the ISS. The SSER was developed in 1999, and it was desired to update the design used in the system. Importantly, besides replacing out-of-production parts it was necessary to decrease the size of the radio due to increased volume constraints with the updated Portable Life Support System (PLSS) 2.5, which will be attached on future space suits. In particular, it was desired to fabricate a PCB for the front-end of the prototype SSAER system. Once this board was manufactured and all parts assembled, it could then be tested for quality of operation as well as compliancy with the SSER required specifications. Upon arrival, a small outline of the target system was provided, and it was my responsibility to take that outline to a finished, testable board. This board would include several stages, including frequency mixing, amplification, modulation, demodulation, and handled both the transmit and receive lines of the radio. I developed a new design based on the old SSER system and the outline provided to me, and found parts to fit the tasks in my design. It was also important to consider the specifications of the SSER, which included the system noise figure, gain, and power consumption. Further, all parts needed to be impedance matched, and spurious signals needed to be avoided. In order to fulfill these two requirements, it was necessary to perform some calculations using a Smith Chart and excel analysis. Once all parts were selected, I drew the schematics for the system in Altium Designer. This included developing schematic symbols, as well as layout. Once the schematic was finished, it was then necessary to lay the parts out onto a PCB using Altium. Similar to the schematic design, in order to accomplish this it was necessary to develop component land patterns and add component 3D models. All of this was achieved, and the PCB is currently in review. After it is finished being reviewed, this board will be sent out for manufacture. All electronic components used in the PCB have been acquired, and once the board arrives they will be soldered onto the board using a machine in building 44. Finally, the board will be tested for performance on-site. This will likely be accomplished by the end of the internship.

Maicke, Andrew↗

Building Reliable Printed Circuit Boards - the Lessons Learned

"Printed circuit boards (PCBs) are the baseline for electronics manufacturing upon which electronic components are mounted and formed into electronic systems. PCBs are used in a variety of electronic circuits from simple one-transistor amplifiers to large super computers. A PCB serves three main functions: 1) it provides the necessary mechanical support for the components in the circuit 2) it provides the necessary electrical interconnections, and 3) it bears some form of legend which identifies the components it carries. The failure modes on the PCBs can be categorized in a hierarchical structure, in which the mechanisms and causes are site or location dependant. This one day workshop will discuss a variety of failure mechanisms that effect the functionality of PCBs. These mechanisms can be related to how PCB materials are selected, PCBs are designed, manufactured, tested and used in the field conditions. The workshop will begin with an overview of PCB manufacturing, materials and processes. With the help of examples and case studies, a wide range of failure mechanisms will be discussed, case studies are focused on digital circuits, however some failures in analog, double sided boards are also presented. The workshop then provides the guideline for selection of methodologies for identifying potential failure mechanisms based on the failure history and how a systematic root cause failure analysis of the PCB can result in prevention of future issues."

Sood, Bhanu↗

An Overview of Printed Circuit Board Risks and Supplier Capability

This NESC Webinar provides the audience a snapshot of quality variations in printed circuit boards, as assessed, using experiences in processing and risk analysis of PCB structural integrity coupons. The presentation will focus on PCB quality metrics, the relative type and number of non-conformances observed and trend analysis using statistical methods. Trend analysis shows the top five non-conformances observed across suppliers, the associated root cause(s) behind these non-conformance, suggestions of mitigation plans and most importantly – the relation of these non-conformances to end item reliability. The statistical trends will then be matched with the current state of the domestic PCB supplier base, its key challenges and opportunities. The presentation further discusses the methods applied at GSFC for evaluating candidate PCB technologies that promote the adoption of higher throughput and faster processing technology for GSFC missions.

printed cricuit boards, supplier capability, risk ↗

An Overview of Printed Circuit Board Risks and Supplier Capability (Reuse Document ID 20205007963)

This HDPUG Webinar provides the audience a snapshot of quality variations in printed circuit boards, as assessed, using experiences in processing and risk analysis of PCB structural integrity coupons. The presentation will focus on PCB quality metrics, the relative type and number of non-conformances observed and trend analysis using statistical methods. Trend analysis shows the top five non-conformances observed across suppliers, the associated root cause(s) behind these non-conformance, suggestions of mitigation plans and most importantly – the relation of these non-conformances to end item reliability. The statistical trends will then be matched with the current state of the domestic PCB supplier base, its key challenges and opportunities. The presentation further discusses the methods applied at GSFC for evaluating candidate PCB technologies that promote the adoption of higher throughput and faster processing technology for GSFC missions.

printed cricuit boards, supplier capability, risk ↗

The Innovative Universal Separation Process (USEP) for Disposal of Mixed Liquid Waste Via Unconditional Clearance Approach - 17570

Historically, Mixed Liquid Waste (MLW) drums from Canadian nuclear facilities containing Tritium (H-3) and other radionuclides (alpha as well as beta/gamma emitting) were shipped to a licensed US facility for destruction via incineration. This option is relatively complex considering the logistics associated with cross-border shipments and the destruction costs (e.g. H-3 surcharge). Because of this, Kinectrics has developed a novel and flexible process for disposal of oily as well as other (solvents, aqueous etc.) MLW within Canada. This process avoids the need for cross-border shipments and significantly reduces the complexity as well as cost for waste disposal. An overview of the various approaches and activities undertaken by Kinectrics during the period 2009-2013 for successfully processing and disposing of several MLW drums (oils, solvents, aqueous and solids) was reported at the WM-2013 conference. One of the key approaches for management of both oily Polychlorinated Biphenyls (PCB) as well as non-PCB MLW involves treatment of the mixed waste using various well-known unit operations such as filtration, treatment with commercial sorbents, and various proprietary treatment methods developed at Kinectrics. These approaches result in reduction of radionuclide activity levels to below the unconditional clearance criteria and allows disposal of the waste via incineration at a conventional chemical waste facility. A small amount of radioactive secondary waste (the captured radionuclides) is returned to the customer. This approach was successfully employed for processing several drums of PCB and non-PCB MLW. Kinectrics recently completed two projects involving processing for disposal of legacy oily and solvent MLW inventory for a Canadian nuclear customer. Waste treatment processes used previously for unconditional clearance of oily/solvent MLW were ineffective for treatment of these legacy waste drums. Some drums were particularly challenging to process since H-3 was chemically associated with the oil backbone. To address these challenges, Kinectrics developed the innovative Universal Separation (USEP) process for disposal of problematic MLW. This paper presents an overview of the USEP process, the latest results of drum scale (205 L) process demonstration work, and the design concept of a Mobile Processing Unit (MPU) for large scale batch (1500 L) processing of MLW employing the USEP process. (authors)

12 MANAGEMENT OF RADIOACTIVE AND NON-RADIOACTIVE W↗

Characterization of a contaminated facility and associated soils in support of D and D and subsequent soil remediation - 15292

Before starting any D and D field work at a United States Department of Energy (DOE) facility such as the Savannah River Site (SRS), it is recommended that a complete review and understanding of the full nature and extent of any contamination located in or previously associated with that facility is known. An integral aspect of this review and research entails a complete characterization of that facility, including, but not limited to, Toxic Substances Control Act (TSCA) limits for Polychlorinated Biphenyl (PCB), radiological, Resource Conservation and Recovery Act of 1976 (RCRA) and other hazardous chemicals, etc. A thorough characterization of a facility and its underlying soils supports: - establishment of safe working conditions to protect human health and environment - Deactivation and Decommissioning (D and D) methodology - cost estimates to support future budget planning for D and D and soil cleanup A prime example of this is documented in the characterization of the 690-N, Process Heat Exchanger Repair Facility that had a history of PCB and radioactive contamination. Characterization of 690-N included the sampling and analysis of concrete and underlying soils for radiological, PCB and hazardous contaminants. In addition, a sump, the walls of the facility, expansion joint material and the metal base of a drill press were characterized. Results of this comprehensive characterization of the 690-N Building are provided in a detailed report. Analytical results indicate minimal radiological, PCB, or RCRA hazardous contamination is present in the concrete floor slab (industrial exposure levels) and on the steel hut walls. Soils below the concrete floor slab show only trace amounts of radiological contamination and are well below TSCA and RCRA limits. Subsequent decommissioning/demolition activities can be performed safely without harming people or the environment, and cost estimates to perform building demolition can be developed for future budget planning. (authors)

12 MANAGEMENT OF RADIOACTIVE AND NON-RADIOACTIVE W↗

Remedial effectiveness of a pond biomanipulation: Habitat value and concentrations of polychlorinated biphenyls in fish

The fish and plant communities in a pond contaminated with polychlorinated biphenyls (PCBs) in East Tennessee, USA, were manipulated to reduce ecological and human-health risk associated with exposure to the chemical contaminants. We evaluated the success of the remedial action using a habitat valuation approach, as well as measuring PCB concentrations in fish. Risk reduction objectives included: alter the fish community to favor fish that do not resuspend, bioaccumulate, or biomagnify PCBs; stabilize contaminated sediments to improve water quality; and stabilize shoreline soils and enhance riparian habitat. Fish targeted for removal included gizzard shad, largemouth bass, and nonnative carp. Reduced PCB concentrations in fish have characterized the new bluegill-dominated community, although a weir-overtopping event led to the need for additional removals of gizzard shad and largemouth bass. Sunfish abundance is high, as was intended. Moreover, amphibian and waterbird diversities have increased in the years following biomanipulation, possibly owing to improvements in the riparian zone and increased structural (vegetation) complexity in both the aquatic and terrestrial environment. Thus, the remedial action has improved aspects of habitat value, and PCB concentrations in sunfish have dropped below the remediation level (risk-based target value) for this pond (1 µg/g in fish fillets or 2.3 µg/g in whole body fish).

63 RADIATION, THERMAL, AND OTHER ENVIRON. POLLUTAN↗

Two-Year Mortality After Angioplasty of the Femoro-Popliteal Artery with Uncoated Balloons and Paclitaxel-Coated Balloons—A Pooled Analysis of Four Randomized Controlled Multicenter Trials

Purpose: In view of a recent meta-analysis reporting increased mortality following angioplasty with paclitaxel-coated devices in peripheral arteries, we performed a patient-level 2-year mortality analysis based on pooled original data of four randomized controlled trials (THUNDER, FEMPAC, PACIFIER and CONSEQUENT). Methods and Results: Clinical data of four randomized controlled trial were pooled to assess 2-year mortality following paclitaxel-coated balloon (PCB) angioplasty compared to angioplasty without paclitaxel (control group). A logistic regression model was applied to identify potential predictors of mortality. At two years, 13 of 185 (7.0%) patients had died in the control group and 16/184 (8.7%) in the PBC group, p = 0.55. Kaplan–Meier analysis revealed no significant difference from all-cause death at 2 years (log rank p = 0.54). Causes of death were well balanced between the groups with no pattern or trend in favour of any specific causes in the PBC group. Logistic regression revealed that treatment groups (controls or PBC) were not a predictor of 2-year mortality. The only predictor for mortality was patient age ≥ 75 years. The delivered paclitaxel doses per patient were not significantly different in patients that died and those who did not die during the 24-month follow-up (5.300 ± 4.224 μg vs. 6.248 ± 4.629 μg, p = 0.433). Conclusions: Based on original patient-level data of four pooled randomized controlled trials, we found no increase in 2-year mortality in patients treated with PCB compared to control patients treated with uncoated balloons. Causes of death were well balanced between PCB and control patients.

62 RADIOLOGY AND NUCLEAR MEDICINE↗

Metabolism of chlorobiphenyls by a variant biphenyl dioxygenase exhibiting enhanced activity toward dibenzofuran

Highlights: Black-Right-Pointing-Pointer Burkholderia xenovorans LB400 biphenyl dioxygenase (BphAE{sub LB400}) metabolizes PCBs. Black-Right-Pointing-Pointer Asn338Gln/Leu409Phe double mutation speeds up electron transfer of enzyme reaction. Black-Right-Pointing-Pointer We tested how the mutations affect the PCB-degrading abilities of BphAE{sub LB400} variants. Black-Right-Pointing-Pointer The same mutations also broaden the PCB substrate range of BphAE{sub LB400} variants. -- Abstract: The biphenyl dioxygenase of Burkholderia xenovorans LB400 (BphAE{sub LB400}) catalyzes the dihydroxylation of biphenyl and of several polychlorinated biphenyls (PCBs) but it poorly oxidizes dibenzofuran. In this work we showed that BphAE{sub RR41}, a variant which was previously found to metabolize dibenzofuran more efficiently than its parent BphAE{sub LB400}, metabolized a broader range of PCBs than BphAE{sub LB400}. Hence, BphAE{sub RR41} was able to metabolize 2,6,2 Prime ,6 Prime -, 3,4,3 Prime ,5 Prime - and 2,4,3 Prime ,4 Prime -tetrachlorobiphenyl that BphAE{sub LB400} is unable to metabolize. BphAE{sub RR41} was obtained by changing Thr335Phe336Asn338Ile341Leu409 of BphAE{sub LB400} to Ala335Met336Gln338Val341Phe409. Site-directed mutagenesis was used to create combinations of each substitution, in order to assess their individual contributions. Data show that the same Asn338Glu/Leu409Phe substitution that enhanced the ability to metabolize dibenzofuran resulted in a broadening of the PCB substrates range of the enzyme. The role of these substitutions on regiospecificities toward selected PCBs is also discussed.

60 APPLIED LIFE SCIENCES↗