Improved Membrane Permeability via Hypervesiculation for In Situ Recovery of Lycopene in Escherichia coli.

Lycopene biosynthesis is frequently hampered by downstream processing hugely due to its inability to be secreted out from the producing chassis. Engineering cell factories can resolve this issue by secreting this hydrophobic compound. A highly permeable E. coli strain was developed for a better release rate of lycopene. Specifically, the heterologous mevalonate pathway and crtEBI genes from Corynebacterium glutamicum were overexpressed in Escherichia coli BL21 (DE3) for lycopene synthesis. To ensure in situ lycopene production, murein lipoprotein, lipoprotein NlpI, inner membrane permease protein, and membrane-anchored protein in TolA-TolQ-TolR were deleted for improved membrane permeability. The final strain, LYC-8, produced 438.44 ± 8.11 and 136.94 ± 1.94 mg/L of extracellular and intracellular lycopene in fed-batch fermentation. Both proteomics and lipidomics analyses of secreted outer membrane vesicles were perfect indicators of hypervesiculation. Changes in the ratio of saturated fatty acids, unsaturated fatty acids, and cyclopropane fatty acids coupled with the branching and acyl chain lengths altered the membrane fatty acid composition. This ensured membrane fluidity and permeability for in situ lycopene release. The combinatorial deletion of these genes altered the cellular morphology. The structural and morphological changes in cell shape, size, and length were associated with changes in the mechanical strength of the cell envelope. The enhanced lycopene production and secretion mediated by improved membrane permeability established a cell lysis-free system for an efficient releasing rate and downstream processing, demonstrating the importance of vesicle-associated membrane permeability in efficient lycopene production.

Engineering Escherichia coli BL21 (DE3) for high-yield production of germacrene A, a precursor of ß-elemene via combinatorial metabolic engineering strategies.

ß-elemene is one of the most commonly used antineoplastic drugs in cancer treatment. As a plant-derived natural chemical, biologically engineering microorganisms to produce germacrene A to be converted to ß-elemene harbors great expectations since chemical synthesis and plant isolation methods come with their production deficiencies. In this study, we report the design of an Escherichia coli cell factory for the de novo production of germacrene A to be converted to ß-elemene from a simple carbon source. A series of systematic approaches of engineering the isoprenoid and central carbon pathways, translational and protein engineering of the sesquiterpene synthase, and exporter engineering yielded high-efficient ß-elemene production. Specifically, deleting competing pathways in the central carbon pathway ensured the availability of acetyl-coA, pyruvate, and glyceraldehyde-3-phosphate for the isoprenoid pathways. Adopting lycopene color as a high throughput screening method, an optimized NSY305N was obtained via error-prone polymerase chain reaction mutagenesis. Further overexpression of key pathway enzymes, exporter genes, and translational engineering produced 1161.09 mg/L of ß-elemene in a shake flask. Finally, we detected the highest reported titer of 3.52 g/L of ß-elemene and 2.13 g/L germacrene A produced by an E. coli cell factory in a 4-L fed-batch fermentation. The systematic engineering reported here generally applies to microbial production of a broader range of chemicals. This illustrates that rewiring E. coli central metabolism is viable for producing acetyl-coA-derived and pyruvate-derived molecules cost-effectively.

Wavelength conversion using fiber cross-phase modulation driven by two pump waves.

Wavelength conversion using all-optical phase modulation in a fiber driven by two pump waves is investigated. The operation features are analyzed using an all-optical phase modulation model with two parallel-/cross-polarized pump waves to generate a phase-preserving copy of the optical signal at an exact frequency up-/down-shifted by the two-pump detuning. The conversion efficiency is experimentally verified using a 300-m highly-nonlinear fiber. The results agree well with a theoretical prediction. The conversion bandwidth over 4 THz is achieved and error-free wavelength conversion for a 32-GBd polarization-division multiplexed 16QAM signal is demonstrated. The technique's applicability to a large-capacity wavelength-division multiplexed signals is also discussed.

1.024 Tb/s wavelength conversion in a silicon waveguide with reverse-biased p-i-n junction.

We experimentally show an all-optical wavelength conversion of 8 × 32-GBd single-polarization 16QAM signals using a silicon nano-rib waveguide. The application of reverse biasing of the p-i-n junction of the waveguide allows a conversion efficiency of -8.5 dB with a measured 3-dB optical bandwidth of about 40 nm. Using digital coherent reception, it is shown that the receiver optical signal-to-noise ratio penalty, at a bit-error ratio of 1 × 10-3, of the wavelength-converted signals over all eight channels was less than 0.6 dB with reference to their respective back-to-back signal channels.

Kerr nonlinearity mitigation in 5 × 28-GBd PDM 16-QAM signal transmission over a dispersion-uncompensated link with backward-pumped distributed Raman amplification.

We present experimental and numerical investigations of Kerr nonlinearity compensation in a 400-km standard single-mode fiber link with distributed Raman amplification with backward pumping. A dual-pump polarization-independent fiber-based optical parametric amplifier is used for mid-link spectral inversion of 5 × 28-GBd polarization-multiplexed 16-QAM signals. Signal quality factor (Q-factor) improvements of 1.1 dB and 0.8 dB were obtained in the cases of a single-channel and a five-channel wavelength-division multiplexing (WDM) system, respectively. The experimental results are compared to numerical simulations with good agreement. It is also shown with simulations that a maximum transmission reach of 2400 km enabled by the optical phase conjugator is possible for the WDM signal.