Physico-chemical properties of a protein-inhibitor of serine proteinases from the potato]

Serine proteases inhibitor with pl-7.3, isolated from potatoe tubers by isoelectric focusing procedure as described previously (V.V. Mosolov et al., Bioorganic Chem., 1, 1449, 1975), was homogeneous under ultracentrifugation, having sedimentation coefficient S20,w 2.8S. Its molecular weight, investigated by sedimentation equilibrium and gel filtration through Sephadex G-100, was found to be 32500 and 31500 respectively. The Stokes radius R and frictional ratio f/fo were found to be 24 A and 1.14. The molecular weight of the inhibitor as determined by sodium dodecylsulfate polyacrylamide electrophoresis was twice as low as determined in ultracentrifuge and by gel filtration procedure. It is suggested that the inhibitor is dimer and consists of two protomers of equal molecular weight.     

Some biochemical aspects of the enzymic transformation of cortisol with Bacillus cereus

The role of a variety of compounds including organic acids, vitamins, growth promoting substances, purines and pyrimidines in the bioconversion of cortisol with Bacillus cereus was investigated. The transformation of cortisol to prednisolone and pregn-4-en-11beta, 17alpha,20beta, 21-tetrol-3-one was affected by these compounds in different manners. The enzymatic delta1-dehydrogenation reaction was greatly induced with fumarate, menadione, and xanthine treatments. However, the enzymic reduction of the 20-carbonyl to the 20beta-ol was specifically stimulated with fumarate, nicotinic acid amide, and uracil treatments.     

Gas chromatographic analysis of acetophenone oxime and its metabolites

A gas-liquid chromatographic (GLC) method has been developed for monitoring the metabolic reduction of acetophenone oxime or oxidative metabolism of the corresponding amine, alpha-methylbenzylamine in liver homogenates. The oxime, amine, n-hydroxy-alpha-methylbenzylamine and acetophenone are quantitatively determined after GLC separation of components with temperature programming on an SP-2401-DB-coated column. The first three compounds were silylated with N,O-bis(trimethylsilyl)-acetamide prior to chromatographic analysis to enhance the stability and improve the chromatographic properties of these components. The effluent gas was monitored with flame ionization detection, and permitted quantitation of components at sub-microgram/ml levels with reproducibility between injections of +/-2%. The optimal composition of enantiomeric mixtures of (R,S)-alpha-methylbenzylamines formed during metabolic reduction of acetophenone oximes were determined by conversion to diastereomeric amides and subsequent GLC analysis.