Optimization of the IPP-bypass mevalonate pathway and fed-batch fermentation for the production of isoprenol in Escherichia coli.

Isoprenol (3-methyl-3-buten-1-ol) is a drop-in biofuel and a precursor for commodity chemicals. Biological production of isoprenol via the mevalonate pathway has been developed and optimized extensively in Escherichia coli, but high ATP requirements and isopentenyl diphosphate (IPP) toxicity have made it difficult to achieve high titer, yield, and large-scale production. To overcome these limitations, an IPP-bypass pathway was previously developed using the promiscuous activity of diphosphomevalonate decarboxylase, and enabled the production of isoprenol at a comparable yield and titer to the original pathway. In this study, we optimized this pathway, substantially improving isoprenol production. A titer of 3.7 g/L (0.14 g isoprenol per g glucose) was achieved in batch conditions using minimal medium by pathway optimization, and a further optimization of the fed-batch fermentation process enabled an isoprenol titer of 10.8 g/L (yield of 0.105 g/g and maximum productivity of 0.157 g L-1 h-1), which is the highest reported titer for this compound. The substantial increase in isoprenol titer via the IPP-bypass pathway in this study will facilitate progress toward commercialization

Photoreduction of Shewanella oneidensis Extracellular Cytochromes by Organic Chromophores and Dye-Sensitized TiO2.

The transfer of photoenergized electrons from extracellular photosensitizers across a bacterial cell envelope to drive intracellular chemical transformations represents an attractive way to harness nature's catalytic machinery for solar-assisted chemical synthesis. In $\textit{Shewanella oneidensis}$ MR-1 (MR-1), trans-outer-membrane electron transfer is performed by the extracellular cytochromes MtrC and OmcA acting together with the outer-membrane-spanning porin$\cdot$cytochrome complex (MtrAB). Here we demonstrate photoreduction of solutions of MtrC, OmcA, and the MtrCAB complex by soluble photosensitizers: namely, eosin Y, fluorescein, proflavine, flavin, and adenine dinucleotide, as well as by riboflavin and flavin mononucleotide, two compounds secreted by MR-1. We show photoreduction of MtrC and OmcA adsorbed on Ru$^{\text{II}}$-dye-sensitized TiO$_2$ nanoparticles and that these protein-coated particles perform photocatalytic reduction of solutions of MtrC, OmcA, and MtrCAB. These findings provide a framework for informed development of strategies for using the outer-membrane-associated cytochromes of MR-1 for solar-driven microbial synthesis in natural and engineered bacteria.This work was supported by the UK Biotechnology and Biological Sciences Research Council (grants BB/K009753/1, BB/K010220/1, BB/K009885/1, and BB/K00929X/1), the Engineering and Physical Sciences Research Council (EP/M001989/1, PhD studentship 1307196 to E.V.A.), a Royal Society Leverhulme Trust Senior Research Fellowship to J.N.B., the Christian Doppler Research Association, and OMV group

A Decaheme Cytochrome as a Molecular Electron Conduit in Dye-Sensitized Photoanodes.

In nature, charge recombination in light-harvesting reaction centers is minimized by efficient charge separation. Here, it is aimed to mimic this by coupling dye-sensitized TiO2 nanocrystals to a decaheme protein, MtrC from Shewanella oneidensis MR-1, where the 10 hemes of MtrC form a ≈7-nm-long molecular wire between the TiO2 and the underlying electrode. The system is assembled by forming a densely packed MtrC film on an ultra-flat gold electrode, followed by the adsorption of approximately 7 nm TiO2 nanocrystals that are modified with a phosphonated bipyridine Ru(II) dye (RuP). The step-by-step construction of the MtrC/TiO2 system is monitored with (photo)electrochemistry, quartz-crystal microbalance with dissipation (QCM-D), and atomic force microscopy (AFM). Photocurrents are dependent on the redox state of the MtrC, confirming that electrons are transferred from the TiO2 nanocrystals to the surface via the MtrC conduit. In other words, in these TiO2/MtrC hybrid photodiodes, MtrC traps the conduction-band electrons from TiO2 before transferring them to the electrode, creating a photobioelectrochemical system in which a redox protein is used to mimic the efficient charge separation found in biological photosystems.This work was supported by the BBSRC (grants BB/K009753/1, BB/K010220/1, and BB/K009885/1), the EPSRC (EP/H00338X/2; PhD studentship to Emma Ainsworth), the Christian Doppler Research Association and the OMV Group. The authors appreciate Dr. Liang Shi (PNNL) and Dr. Marcus Edwards (UEA) for providing the S. oneidensis strain and the protocol allowing for purification of MtrC.This is the final published version of the article. It was originally published in Advanced Functional Materials (Hwang ET, Sheikh K, Orchard KL, Hojo D, Radu V, Lee C-Y, Ainsworth E, Lockwood C, Gross MA, Adschiri T, Reisner E, Butt JN, Jeuken LJC, Advanced Functional Materials 2015, 25, 2308–2315, doi: 10.1002/adfm.201404541) http://dx.doi.org/10.1002/adfm.201404541

Towards compartmentalized photocatalysis: multihaem proteins as transmembrane molecular electron conduits.

The high quantum efficiency of natural photosynthesis has inspired chemists for solar fuel synthesis. In photosynthesis, charge recombination in photosystems is minimized by efficient charge separation across the thylakoid membrane. Building on our previous bioelectrochemical studies of electron transfer between a light-harvesting nanoparticle (LHNP) and the decahaem subunit MtrC, we demonstrate photo-induced electron transfer through the full transmembrane MtrCAB complex in liposome membranes. Successful photoelectron transfer is demonstrated by the decomposition of a redox dye, Reactive Red 120 (RR120), encapsulated in MtrCAB proteoliposomes. The photoreduction rates are found to be dependent on the identity of the external LHNPs, specifically, dye-sensitized TiO2, amorphous carbon dots (a-CD) and graphitic carbon dots with core nitrogen doping (g-N-CDs). Agglomeration or aggregation of TiO2 NPs likely reduces the kinetics of RR120 reductive decomposition. In contrast, with the dispersed a-CD and g-N-CDs, the kinetics of the RR120 reductive decomposition are observed to be faster with the MtrCAB proteoliposomes and we propose that this is due to enhancement in the charge-separated state. Thus, we show a proof-of-concept for using MtrCAB as a lipid membrane-spanning building block for compartmentalised photocatalysis that mimics photosynthesis. Future work is focused on incorporation of fuel generating redox catalysts in the MtrCAB proteoliposome lumen.This work was supported by the BBSRC (DTP studentship 1827308; grants BB/ K009753/1, BB/K010220/1 and BB/K009885/1) and the Engineering and Physical Sciences Research Council (PhD studentship 1307196)

Management of the poultry red mite Dermanyssus gallinae with physical control methods by inorganic material and future perspectives

ABSTRACT: Poultry red mite (PRM), the ectoparasitic mite Dermanyssus gallinae found in laying hen farms, is a significant threat to poultry production and human health worldwide. It is a suspected disease vector and attacks hosts’ other than chickens, including humans, and its economic importance has increased greatly. Different strategies to control PRM have been widely tested and investigated. In principle, several synthetic pesticides have been applied to control PRM. However, recent alternative control methods to avoid the side effects of pesticides have been introduced, although many remain in the early stage of commercialization. In particular, advances in material science have made various materials more affordable as alternatives for controlling PRM through physical interactions between PRM. This review provides a summary of PRM infestation, and then includes a discussion and comparison of different conventional approaches: 1) organic substances, 2) biological approaches, and 3) physical inorganic material treatment. The advantages of inorganic materials are discussed in detail, including the classification of materials, as well as the physical mechanism-induced effect on PRM. In this review, we also consider the perspective of using several synthetic inorganic materials to suggest novel strategies for improved monitoring and better information regarding treatment interventions

A Visual Interface for Scripting Virtual Behaviors

. The storyboarding stage in filmmaking is necessary for communicating overall scenarios rapidly through approximate visual drawings. In contrast, most 3D animation systems produce final products through repeated precise modeling. The interaction suggested in this paper promote the maximum transfer of the users' knowledge of physical human actions into virtual environments, so the users can rapidly generate virtual human behaviors such as running, walking, grasping, and other motions. To achieve this, users visually describe the behaviors of virtual human, then the interface parses the visual scripting and finally achieves the semantics. In this approach, users draw only curves, which describe synchronization among motions as well as geometric motion paths of the human, in the same three-dimensional space using 3D devices such as a spaceball. Keywords: 3-D interfaces, visual programming, animation, and interaction models 1. Introduction Despite the growing popularity and the..

Visual Scripting for Virtual Behaviors

. We suggest an interactive method that visually describes the behaviors of virtual objects, parses the visual scripting and finally achieves the semantics. In this approach, users draw only curves, which describe synchronization among motions as well as geometric motion paths of virtual characters, in the same three-dimensional space. This approach promotes the maximum transfer of the users' knowledge of behaviors of physical objects and actions into virtual environments, so the users can rapidly generate virtual characters' behaviors such as running, walking, grasping, and other motions. 1. INTRODUCTION Despite the growing popularity and the existence of a number of successful 3D graphics applications, learning how to implement or use 3D graphics software is still extremely laborious and it's still an open problem which is the most effective ways for humans to interact with synthetic 3D environments. While systems employing WIMP-style (Windows, Icons, Menus and Pointers) inte..

Incorporating co-presence in distributed virtual music environment

In this paper, we present “PODIUM (POstech Distributed virtUal Music environment)”, a distributed virtual environment that allows users to participate in a shared space and play music with other participants in a collaborative manner. In addition to playing virtual instruments, users can communicate and interact in various ways to enhance the collaboration and, thus, the quality of the music played together. Musical messages are generated note by note through interaction with the keyboard, mouse, and other devices, and transmitted through an IP-multicasting network among participants. In addition to such note-level information, additional messages for visualization, and interaction are supported. Real world based visualization has been chosen, against, for instance, abstract music world based visualization, to promote “co-presence ” (e.g. recognize and interact with other players), which is deemed important for collaborative music production. In addition to the entertainment purpose, we hope that DVME will find great use in casual practice sessions for even professional performers/orchestras/bands. Since even a slight interruption in the flow of the music or out-of-synch graphics and sound would dramatically decrease utility of the system, we employ various techniques to minimize the network delay. An adapted server-client architecture and UDP’s are used to ensure fast packet deliveries and reduce the data bottleneck problem. Time-critical messages such as MIDI messages are multicasted among clients, and the less time-critical and infrequently updated messages are sent through the server. Predefined animations of avatars are invoked by interpreting the musical messages. Using the latest graphics and sound processing hardware, and by maintaining a