Metabolic engineering of ß-oxidation to leverage thioesterases for production of 2-heptanone, 2-nonanone and 2-undecanone.

Medium-chain length methyl ketones are potential blending fuels due to their cetane numbers and low melting temperatures. Biomanufacturing offers the potential to produce these molecules from renewable resources such as lignocellulosic biomass. In this work, we designed and tested metabolic pathways in Escherichia coli to specifically produce 2-heptanone, 2-nonanone and 2-undecanone. We achieved substantial production of each ketone by introducing chain-length specific acyl-ACP thioesterases, blocking the ß-oxidation cycle at an advantageous reaction, and introducing active ß-ketoacyl-CoA thioesterases. Using a bioprospecting approach, we identified fifteen homologs of E. coli ß-ketoacyl-CoA thioesterase (FadM) and evaluated the in vivo activity of each against various chain length substrates. The FadM variant from Providencia sneebia produced the most 2-heptanone, 2-nonanone, and 2-undecanone, suggesting it has the highest activity on the corresponding ß-ketoacyl-CoA substrates. We tested enzyme variants, including acyl-CoA oxidases, thiolases, and bi-functional 3-hydroxyacyl-CoA dehydratases to maximize conversion of fatty acids to ß-keto acyl-CoAs for 2-heptanone, 2-nonanone, and 2-undecanone production. In order to address the issue of product loss during fermentation, we applied a 20% (v/v) dodecane layer in the bioreactor and built an external water cooling condenser connecting to the bioreactor heat-transferring condenser coupling to the condenser. Using these modifications, we were able to generate up to 4.4 g/L total medium-chain length methyl ketones.

Advances in the immunoescape mechanisms exploited by alphaherpesviruses.

Alphaherpesviruses, categorized as viruses with linear DNA composed of two complementary strands, can potentially to induce diseases in both humans and animals as pathogens. Mature viral particles comprise of a core, capsid, tegument, and envelope. While herpesvirus infection can elicit robust immune and inflammatory reactions in the host, its persistence stems from its prolonged interaction with the host, fostering a diverse array of immunoescape mechanisms. In recent years, significant advancements have been achieved in comprehending the immunoescape tactics employed by alphaherpesviruses, including pseudorabies virus (PRV), herpes simplex virus (HSV), varicella-zoster virus (VZV), feline herpesvirus (FeHV), equine herpesvirus (EHV), and caprine herpesvirus type I (CpHV-1). Researchers have unveiled the intricate adaptive mechanisms existing between viruses and their natural hosts. This review endeavors to illuminate the research advancements concerning the immunoescape mechanisms of alphaherpesviruses by delineating the pertinent proteins and genes involved in virus immunity. It aims to furnish valuable insights for further research on related mechanisms and vaccine development, ultimately contributing to virus control and containment efforts.

The Need for Biosecurity Education in Biotechnology Curricula.

The growth of biotechnology in recent decades and the dual-use nature of most bioscience research are making their misuse, or accidental misuse or release, more likely and present as threats to biosecurity. A proactive approach is through educating the next generation of scientists to be more security conscious. However, current educational and professional programs in biosecurity are lacking. In this perspective, we recommend that biosecurity educational opportunities should be implemented and expanded for undergraduate and graduate students who will likely use one or more methods in the field of biotechnology. We then propose that biosecurity education is a key factor in a path toward sustainable and safe research. Finally, a set of 17 biosecurity competencies organized into 6 distinct themes is outlined.

Structure-activity relationship of Cu-based catalysts for the highly efficient CO2 electrochemical reduction reaction.

Electrocatalytic carbon dioxide reduction (CO2RR) is a relatively feasible method to reduce the atmospheric concentration of CO2. Although a series of metal-based catalysts have gained interest for CO2RR, understanding the structure-activity relationship for Cu-based catalysts remains a great challenge. Herein, three Cu-based catalysts with different sizes and compositions (Cu@CNTs, Cu4@CNTs, and CuNi3@CNTs) were designed to explore this relationship by density functional theory (DFT). The calculation results show a higher degree of CO2 molecule activation on CuNi3@CNTs compared to that on Cu@CNTs and Cu4@CNTs. The methane (CH4) molecule is produced on both Cu@CNTs and CuNi3@CNTs, while carbon monoxide (CO) is synthesized on Cu4@CNTs. The Cu@CNTs showed higher activity for CH4 production with a low overpotential value of 0.36 V compared to CuNi3@CNTs (0.60 V), with *CHO formation considered the potential-determining step (PDS). The overpotential value was only 0.02 V for *CO formation on the Cu4@CNTs, and *COOH formation was the PDS. The limiting potential difference analysis with the hydrogen evolution reaction (HER) indicated that the Cu@CNTs exhibited the highest selectivity of CH4 among the three catalysts. Therefore, the sizes and compositions of Cu-based catalysts greatly influence CO2RR activity and selectivity. This study provides an innovative insight into the theoretical explanation of the origin of the size and composition effects to inform the design of highly efficient electrocatalysts.

A double-layered liquid metal-based electrochemical sensing system on fabric as a wearable detector for glucose in sweat.

Integrated electrochemical sensing platforms in wearable devices have great prospects in biomedical applications. However, traditional electrochemical platforms are generally fabricated on airtight printed circuit boards, which lack sufficient flexibility, air permeability, and conformability. Liquid metals at room temperature with excellent mobility and electrical conductivity show high promise in flexible electronics. This paper presents a miniaturized liquid metal-based flexible electrochemical detection system on fabric, which is intrinsically flexible, air-permeable, and conformable to the body. Taking advantage of the excellent fluidity and electrical connectivity of liquid metal, a double-layer circuit is fabricated that significantly miniaturizes the size of the whole system. The linear response, time stability, and repeatability of this system are verified by resistance, stability, image characterization, and potassium ferricyanide tests. Finally, glucose in sweat can be detected at the millimolar level using this sensing system, which demonstrates its great potential for wearable and portable detection in biomedical fields, such as health monitoring and point-of-care testing.

Effect of nickel-based electrocatalyst size on electrochemical carbon dioxide reduction: A density functional theory study.

The electrochemical carbon dioxide (CO2) reduction reaction (CO2RR) used for converting higher-value chemicals is a promising solution to mitigate CO2 emissions. Nickel (Ni)-based catalysts have been identified as a potential candidate for CO2 activation and conversion. However, in the CO2RR, the size effect of the Ni-based electrocatalysts has not been well explored. Herein, the single Ni atom and the Ni4 cluster doped nitrogen-doped carbon nanotube (Ni@CNT and Ni4@CNT), and the Ni (110) facet were designed to explore the size effect in the CO2RR by using density functional theory (DFT) calculations. The results show that carbon monoxide (CO) is produced on the Ni@CNT with a free energy barrier of 0.51 eV. The reduction product of CO2 on the Ni4@CNT and Ni(110) facet is methane (CH4) in both cases, via different reaction pathways, and the Ni(110) facet is a more efficient electrocatalyst with a low overpotential of 0.27 V when compared to Ni4@CNT (0.50 V). The rate-determining step (RDS) is the formation of *CHO on the Ni4@CNT (The "*" represents the catalytic surface), while the *COH formation is the RDS on the Ni(110) facet. Meanwhile, the Ni(110) facet also has the highest selectivity of CH4 among the three catalysts. The CO2 reduction product changes from CO to CH4 with the increasing size of the Ni-based catalysts. These results demonstrate that the catalytic activity and selectivity of CO2RR highly depend on the size of the Ni-based catalysts.

Metabolic engineering strategies to produce medium-chain oleochemicals via acyl-ACP:CoA transacylase activity.

Microbial lipid metabolism is an attractive route for producing oleochemicals. The predominant strategy centers on heterologous thioesterases to synthesize desired chain-length fatty acids. To convert acids to oleochemicals (e.g., fatty alcohols, ketones), the narrowed fatty acid pool needs to be reactivated as coenzyme A thioesters at cost of one ATP per reactivation - an expense that could be saved if the acyl-chain was directly transferred from ACP- to CoA-thioester. Here, we demonstrate such an alternative acyl-transferase strategy by heterologous expression of PhaG, an enzyme first identified in Pseudomonads, that transfers 3-hydroxy acyl-chains between acyl-carrier protein and coenzyme A thioester forms for creating polyhydroxyalkanoate monomers. We use it to create a pool of acyl-CoA's that can be redirected to oleochemical products. Through bioprospecting, mutagenesis, and metabolic engineering, we develop three strains of Escherichia coli capable of producing over 1 g/L of medium-chain free fatty acids, fatty alcohols, and methyl ketones.

Morphological background detection and illumination normalization of text image with poor lighting.

In this paper, some morphological transformations are used to detect the unevenly illuminated background of text images characterized by poor lighting, and to acquire illumination normalized result. Based on morphologic Top-Hat transform, the uneven illumination normalization algorithm has been carried out, and typically verified by three procedures. The first procedure employs the information from opening based Top-Hat operator, which is a classical method. In order to optimize and perfect the classical Top-Hat transform, the second procedure, featuring the definition of multi direction illumination notion, utilizes opening by reconstruction and closing by reconstruction based on multi direction structuring elements. Finally, multi direction images are merged to the final even illumination image. The performance of the proposed algorithm is illustrated and verified through the processing of different ideal synthetic and camera collected images, with backgrounds characterized by poor lighting conditions.