Re-directing bacterial microcompartment systems to enhance recombinant expression of lysis protein E from bacteriophage ϕX174 in Escherichia coli
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Complete conversion of the major sugars of biomass including both the C 5 and C 6 sugars is critical for biofuel production processes. Several inhibitory compounds like acetate, hydroxymethylfurfural (HMF), and furfural are produced from the biomass pretreatment process leading to ‘hydrolysate toxicity,’ a major problem for microorganisms to achieve complete sugar utilization. Therefore, development of more robust microorganisms to utilize the sugars released from biomass under toxic environment is critical. In this study, we use continuous culture methodologies to evolve and adapt the ethanologenic bacterium Zymomonas mobilis to improve its ethanol productivity using corn stover hydrolysate. The results are the following: A turbidostat was used to adapt the Z. mobilis strain 8b in the pretreated corn stover liquor. The adaptation was initiated using pure sugar (glucose and xylose) followed by feeding neutralized liquor at different dilution rates. Once the turbidostat reached 60% liquor content, the cells began washing out and the adaptation was stopped. Several ‘sub-strains’ were isolated, and one of them, SS3 (sub-strain 3), had 59% higher xylose utilization than the parent strain 8b when evaluated on 55% neutralized PCS (pretreated corn stover) liquor. Using saccharified PCS slurry generated by enzymatic hydrolysis from 25% solids loading, SS3 generated an ethanol yield of 75.5% compared to 64% for parent strain 8b. Furthermore, the total xylose utilization was 57.7% for SS3 versus 27.4% for strain 8b. To determine the underlying genotypes in these new sub-strains, we conducted genomic resequencing and identified numerous single-nucleotide mutations (SNPs) that had arisen in SS3. We further performed quantitative reverse transcription PCR (qRT-PCR) on genes potentially affected by these SNPs and identified significant down-regulation of two genes, ZMO0153 and ZMO0776, in SS3 suggesting potential genetic mechanisms behind SS3’s improved performance. In conclusion, we have adapted/evolved Z. mobilis strain 8b for enhanced tolerance to the toxic compounds present in corn stover hydrolysates. The adapted strain SS3 has higher xylose utilization rate and produce more ethanol than the parent strain. We have identified transcriptional changes which may be responsible for these phenotypes, providing foundations for future research directions in improving Z. mobilis as biocatalysts for the production of ethanol or other fuel precursors.
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Photobiological processes are attractive routes to renewable H2 production. With the input of solar energy, photosynthetic microbes such as cyanobacteria and green algae carry out oxygenic photosynthesis, using sunlight energy to extract protons and high energy electrons from water. These protons and high energy electrons can be fed to a hydrogenase system yielding H2. However, most hydrogen-evolving hydrogenases are inhibited by O2, which is an inherent byproduct of oxygenic photosynthesis. The rate of H2 production is thus limited. Certain photosynthetic bacteria are reported to have an O2-tolerant evolving hydrogenase, yet these microbes do not split water, and require other more expensive feedstocks.
Direct-emitting red and amber LEDs based on the AlInGaP material system offer a higher efficacy potential than phosphor-converted (pc) red and amber LEDs due to the absence of Stokes losses. Direct red LEDs (~610-620nm) are increasingly used in color-mixed and color-tunable illumination applications, where they have a significant advantage over pc-red LEDs, with high power conversion efficiency (PCE) and narrower emission enabling higher luminous efficacy of radiation (LER). Still, adoption is limited as the complexities associated with mixing different emitters offset the value of the efficacy gain. Adoption of direct amber LEDs (~580-595nm) in illumination applications is virtually non-existent due to their lower PCE and the availability of very efficient pc-amber emitters, which suffer less from Stokes loss and LER penalties associated with the broader bandwidth than pc-red. Further improvement in PCE of direct red and amber LEDs is needed to achieve wider adoption and ultimately enable more energy savings than possible with pc-LEDs. In this project, Lumileds has developed improved high-power amber and red LEDs incorporating tensile-strain barriers into the active region to improve carrier confinement and thereby increase external quantum efficiency (EQE). This approach is most beneficial for the short (amber) end of the AlInGaP wavelength range where carrier confinement is lowest. An EQE of 22% was achieved for amber LEDs, exceeding the project target of 20%
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Background/Problem Statement • Potential Solution: PAR • Phase I: Preliminary Analyses – Engineering Studies • Thermal Models • Nitric Acid Corrosion Evaluation • Waste Tank Ceiling Temperature Evaluation – Potential Usage in Waste Tanks – Potential Return on Investment • Phase II: Need for In-situ Testing
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Background/Objectives: Control of classical swine fever virus (CSFV) in endemic countries relies on vaccination using live attenuated vaccines (LAVs). Most of these LAVs do not allow for the differentiation of vaccinated animals from infected animals (DIVA) based on their serological response. FlagT4G vaccine is a novel candidate that confers robust protective immunity early after vaccination and shows DIVA capabilities. Methods: This report presents the characterization of FlagT4G virus in terms of the stability of its genomic and attenuated phenotypes assessed by a reversion to virulence protocol, as well as its protective efficacy by determining the minimal protective dose. Results: Results presented here demonstrate that after five consecutive passages in groups of 5-week-old susceptible domestic pigs, FlagT4G virus remains genetically stable, and its attenuated phenotype remains unaltered. In terms of efficacy, FlagT4G virus induced solid protection against the intranasal challenge with 105 tissue culture infectious dose (TCID50) of virulent field isolate Brescia virus, even with a vaccine dose as low as 102 TCID50. Conclusions: Results presented here indicate that the FlagT4G vaccine may be a useful tool for CSFV control.