Alteration of immunological properties of bovine serum albumin by covalent attachment of polyethylene glycol.

The location of extracellular enzymes within the soil architecture and their association with the various soil components affects their catalytic potential. A soil fractionation study was carried out to investigate: (a) the distribution of a range of hydrolytic enzymes involved in C, N and P transformations, (b) the effect of the location on their respective kinetics, (c) the effect of long-term N fertilizer management on enzyme distribution and kinetic parameters. Soil (silty clay loam) from grassland which had received 0 or 200 kg N ha(-1) yr(-1) was fractionated, and four particle-size fractions (> 200, 200-63, 63-2 and 0. 1-2 mum) were obtained by a combination of wet-sieving and centrifugation, after low-energy ultrasonication. All fractions were assayed for four carbohydrases (beta-cellobiohydrolase, N-acetyl-beta-glucosammidase, beta-glucosidase and beta-xylosidase), acid phosphatase and leucine-aminopeptidase using a microplate fluorimetric assay based on MUB-substrates. Enzyme kinetics (V-max and K-m) were estimated in three particle-size fractions and the unfractionated soil. The results showed that not all particle-size fractions were equally enzymatically active and that the distribution of enzymes between fractions depended on the enzyme. Carbohydrases predominated in the coarser fractions while phosphatase and leucine-aminopeptidase were predominant in the clay-size fraction. The Michaelis constant (K.) varied among fractions, indicating that the association of the same enzyme with different particle-size fractions affected its substrate affinity. The same values of Km were found in the same fractions from the soil under two contrasting fertilizer management regimes, indicating that the Michaelis constant was unaffected by soil changes caused by N fertilizer management. (C) 2004 Elsevier Ltd. All rights reserved.

Exploring the enzymatic landscape: distribution and kinetics of hydrolytic enzymes in soil particle-size fractions

Proteins and non-protein affinity ligands are covalently bonded to trichloro-s-triazine activated supports. The activated support is prepared by reacting a water-free insoluble solid support with trichloro-s-triazine in a non-aqueous medium and neutralizing HCl generated during the reaction with a tertiary amine which does not form an insoluble complex with the trichloro-s-triazine.

Preparation of trichloro-s-triazine activated supports for coupling ligands

https://onlinelibrary.wiley.com/doi/pdf/10.1002/bit.260150318

The properties of magnetic supports in relation to immobilized enzyme reactors

An overview is given of the preparation and use of model systems for cytochemistry, dealing with quantitative as well as qualitative aspects. Descriptions are given of the various possibilities to prepare cytochemical matrix models, ranging from macroscopic and microscopic films, to models with more cell-like dimensions as agarose beads, artificial cells and erythrocyte ghosts. Such models allow the study of a large variety of cytochemical processes. Their potentialities are demonstrated in a number of specific applications, comprising: the study of the influence of fixation on cellular processes, reaction specificity and reaction kinetics, quality of reagents and biochemical calibration in cytochemical staining; factors influencing localization of the specific endproduct in enzyme cytochemistry; immunocytochemistry and hybridocytochemistry.

G. Kay

Papers

The chemical attachment of chymotrypsin to water-insoluble polymers using 2-amino-4,6-dichloro-s-triazine.