The Metabolic Flux Probe (MFP)—Secreted Protein as a Non-Disruptive Information Carrier for 13C-Based Metabolic Flux Analysis

Novel cultivation technologies demand the adaptation of existing analytical concepts. Metabolic flux analysis (MFA) requires stable-isotope labeling of biomass-bound protein as the primary information source. Obtaining the required protein in cultivation set-ups where biomass is inaccessible due to low cell densities and cell immobilization is difficult to date. We developed a non-disruptive analytical concept for 13C-based metabolic flux analysis based on secreted protein as an information carrier for isotope mapping in the protein-bound amino acids. This “metabolic flux probe” (MFP) concept was investigated in different cultivation set-ups with a recombinant, protein-secreting yeast strain. The obtained results grant insight into intracellular protein turnover dynamics. Experiments under metabolic but isotopically nonstationary conditions in continuous glucose-limited chemostats at high dilution rates demonstrated faster incorporation of isotope information from labeled glucose into the recombinant reporter protein than in biomass-bound protein. Our results suggest that the reporter protein was polymerized from intracellular amino acid pools with higher turnover rates than biomass-bound protein. The latter aspect might be vital for 13C-flux analyses under isotopically nonstationary conditions for analyzing fast metabolic dynamics.

The Metabolic Flux Probe (MFP) - Secreted Protein as a Non-Disruptive Information Carrier for 13C-Based Metabolic Flux Analysis

Novel cultivation technologies demand the adaptation of existing analytical concepts. Metabolic flux analysis (MFA) requires stable-isotope labeling of biomass-bound protein as the primary information source. Obtaining the required protein in cultivation set-ups where biomass is scarce or inaccessible due to low cell densities and cell immobilization is difficult to date. We developed a non-disruptive analytical concept for 13C-based metabolic flux analysis based on secreted protein as an information carrier for isotope mapping in the protein-bound amino acids. This "metabolic flux probe" (MFP) concept was investigated in different cultivation set-ups with a recombinant, protein-secreting yeast strain. The obtained results grant insight into intracellular protein turnover dynamics. Experiments under metabolic but isotopically nonstationary conditions in continuous glucose-limited chemostats at high dilution rates demonstrated faster incorporation of isotope information from labeled glucose into the recombinant reporter protein than in biomass-bound protein. Our results suggest that the reporter protein was polymerized from intracellular amino acid pools with higher turnover rates than biomass-bound protein. The latter aspect might be vital for 13C-flux analyses under isotopically nonstationary conditions for analyzing fast metabolic dynamics.

Stable sub-micrometre high-flux probe for soft X-ray ARPES using a monolithic Wolter mirror

A focusing optics that can provide a sub-micrometre high-flux probe for soft X-ray micrometre-scale angle-resolved photoemission spectroscopy (ARPES) is proposed. A monolithic Wolter-type mirror with a large acceptance, achromatism and small comatic aberration was designed and evaluated. A focused beam size of 0.4 µm (vertical) × 4 µm (horizontal), a high throughput of 59% and a high tolerance of 1.6 mrad to the pitching error were realized at a photon energy of 1000 eV. A Wolter-type mirror can be practically employed as a stable sub-micrometre focusing mirror with high throughput in ARPES applications.

Design and Implementation of an Automated Gamma Probe for Jet Boring Uranium Mining

A proof-of-concept detector prototype capable of collecting and storing radiometric data in the jet boring system (JBS) during pilot hole drilling at the Cigar Lake uranium mine is presented. Cigar Lake is the world’s second highest known grade uranium mine and is located in northern Saskatchewan, Canada. Variant design is used to design, develop, test, and implement the detector’s firmware, software, and hardware. The battery-powered detector is attached inside a JBS drill rod to collect radiometric data through the drilling cycle. A readout box is used to initiate the detector, recharge the battery, and download radiometric data after the pilot hole drilling cycle is complete. Functional testing results are presented, and comparative test results between the JBS gamma probe and the AlphaNUCLEAR Hi-Flux probe are evaluated. Field data collected from a pilot hole are plotted against the pilot hole’s driving layout and jetting recipe to show the accuracy of the readings collected.

In-furnace temperature and heat flux mapping in a kraft recovery boiler

Gas temperatures, incident heat flux, and surface temperatures were measured in a large kraft recovery boiler. The measurements were a part of an extensive campaign planned and carried out to support validation of models based on computational fluid dynamics. The gas temperatures were measured with three different techniques: infrared (IR) pyrometer, suction pyrometer, and unshielded thermocouples. In addition to the temperature measurements, the radiative heat flux was measured in a number of locations in the boiler using a portable heat flux probe, and the surface temperatures inside the boiler were measured using a portable single-band IR camera.