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Selective Release (selective + release)

Distribution by Scientific Domains
Life Sciences50%

Selected Abstracts

Use of Glycol Ethers for Selective Release of Periplasmic Proteins from Gram-Negative Bacteria

BIOTECHNOLOGY PROGRESS, Issue 5 2007
Jeffrey R. Allen
Genetic modification of Gram-negative bacteria to express a desired protein within the cellapos;s periplasmic space, located between the inner cytoplasmic membrane and the outer cell wall, can offer an attractive strategy for commercial production of therapeutic proteins and industrial enzymes. In certain applications, the product expression rate is high, and the ability to isolate the product from the cell mass is greatly enhanced because of the productapos;s compartmentalized location within the cell. Protein release methods that increase the permeability of the outer cell wall for primary recovery, but avoid rupturing the inner cell membrane, reduce contamination of the recovered product with other host cell components and simplify final purification. This article reports representative data for a new release method employing glycol ether solvents. The example involves the use of 2-butoxyethanol (commonly called ethylene glycol n -butyl ether or EB) for selective release of a proprietary biopharmaceutical protein produced in the periplasmic space of Pseudomonas fluorescens. In this example, glycol ether treatment yielded ,65% primary recovery with ,80% purity on a protein-only basis. Compared with other methods including heat treatment, osmotic shock, and the use of surfactants, the glycol ether treatment yielded significantly reduced concentrations of other host cell proteins, lipopolysaccharide endotoxin, and DNA in the recovered protein solution. The use of glycol ethers also allowed exploitation of temperature-change-induced phase splitting behavior to concentrate the desired product. Heating the aqueous EB extract solution to 55 °C formed two liquid phases: a glycol ether-rich phase and an aqueous product phase containing the great majority of the product protein. This phase-splitting step yielded an approximate 10-fold reduction in the volume of the initial product solution and a corresponding increase in the productapos;s concentration. [source]

Selective release of D and 13C from insoluble organic matter of the Murchison meteorite by impact shock

METEORITICS & PLANETARY SCIENCE, Issue 3 2007
Koichi Mimura
We also performed shock experiments on type III kerogen and compared the results of these experiments with the experimental results regarding IOM. The shock selectively released D and 13C from the IOM, while it preferably released H and 12C from the kerogen. The release of these elements from IOM cannot be explained in terms of the isotope effect, whereas their release from kerogen can be explained by this effect. The selective release of heavier isotopes from IOM would be due to its structure, in which D and 13C-enriched parts are present as an inhomogeneity and are weakly attached to the main network. Shock gave rise to a high release of D even at a lower degree of dehydrogenation compared with the stepwise heating of IOM. This effective release of D is probably an inherent result of shock, in which a dynamic high-pressure and high-temperature condition prevails. Thus, shock would effectively control the hydrogen isotope behavior of extraterrestrial organic matter during the evolution of the solar nebula. [source]

Cover Picture: Electrophoresis 14/2008

ELECTROPHORESIS, Issue 14 2008
Article first published online: 23 JUL 200
Issue 14 is a regular issue including an Emphasis Section offering a series of 9 papers on ,Microfluidics and Miniaturization". These 9 research papers report on various topics including studying single DNA molecules, selective release of intracellular molecules on the single cell level, isoelectric focusing of proteins in an ordered micropillar array, sample stream focusing in a microchip, integrated microfluidic system for sensing infectious viral disease, EOF in annulus and rectangular channels, confinement effects on monolith morphology, accumulation and filtering of nanoparticles in microchannels, and carbon nanotubes disposable detectors. [source]

MscL of Bacillus subtilis prevents selective release of cytoplasmic proteins in a hypotonic environment

PROTEINS: STRUCTURE, FUNCTION AND BIOINFORMATICS, Issue 4 2009
Thijs R. H. M. Kouwen
Abstract Bacillus subtilis serves as an excellent model to study protein secretion at a proteomic scale. Most of the extracellular proteins are exported from the cytoplasm via the secretory (Sec) pathway. Despite extensive studies, the secretion mechanisms of about 25% of the extracellular proteins are unknown. This suggests that B. subtilis makes use of alternative mechanisms to release proteins into its environment. In search for novel pathways, which contribute to biogenesis of the B. subtilis exoproteome, we investigated a possible role of the large conductance mechanosensitive channel protein MscL. We compared protein secretion by MscL deficient and proficient B. subtilis cells. MscL did not contribute to secretion under standard growth conditions. Unexpectedly, we discovered that under hypo-osmotic shock conditions specific, normally cytoplasmic proteins were released by mscL mutant cells. This protein release was selective since not all cytoplasmic proteins were equally well released. We established that this protein release by mscL mutant cells cannot be attributed to cell death or lysis. The presence of MscL, therefore, seems to prevent the specific release of cytoplasmic proteins by B. subtilis during hypo-osmotic shock. Our unprecedented findings imply that an unidentified system for selective release of cytoplasmic proteins is active in B. subtilis. [source]

Use of Glycol Ethers for Selective Release of Periplasmic Proteins from Gram-Negative Bacteria

BIOTECHNOLOGY PROGRESS, Issue 5 2007
Jeffrey R. Allen
Genetic modification of Gram-negative bacteria to express a desired protein within the cellapos;s periplasmic space, located between the inner cytoplasmic membrane and the outer cell wall, can offer an attractive strategy for commercial production of therapeutic proteins and industrial enzymes. In certain applications, the product expression rate is high, and the ability to isolate the product from the cell mass is greatly enhanced because of the productapos;s compartmentalized location within the cell. Protein release methods that increase the permeability of the outer cell wall for primary recovery, but avoid rupturing the inner cell membrane, reduce contamination of the recovered product with other host cell components and simplify final purification. This article reports representative data for a new release method employing glycol ether solvents. The example involves the use of 2-butoxyethanol (commonly called ethylene glycol n -butyl ether or EB) for selective release of a proprietary biopharmaceutical protein produced in the periplasmic space of Pseudomonas fluorescens. In this example, glycol ether treatment yielded ,65% primary recovery with ,80% purity on a protein-only basis. Compared with other methods including heat treatment, osmotic shock, and the use of surfactants, the glycol ether treatment yielded significantly reduced concentrations of other host cell proteins, lipopolysaccharide endotoxin, and DNA in the recovered protein solution. The use of glycol ethers also allowed exploitation of temperature-change-induced phase splitting behavior to concentrate the desired product. Heating the aqueous EB extract solution to 55 °C formed two liquid phases: a glycol ether-rich phase and an aqueous product phase containing the great majority of the product protein. This phase-splitting step yielded an approximate 10-fold reduction in the volume of the initial product solution and a corresponding increase in the productapos;s concentration. [source]