A novel thermostable chimeric beta-galactosidase was constructed by fusing a poly-His tag to the N-terminal region of the beta-galactosidase from Thermus sp. strain T2 to facilitate its overexpression in Escherichia coli and its purification by immobilized metal-ion affinity chromatography (IMAC). The poly-His tag fusion did not affect the activation, kinetic parameters, and stability of the be...

introduction: oxalate oxidase (ec.1, 2, 3, 4), an acute phase protein of plants, is used widely in agriculture, food industries, medical diagnosis of kidney stones and oxaluria. in this study oxalate oxidase from barley was purified and partially characterized. materials & methods: roots and sprouts of barley cultured in hydroponic states were separated and extracted. ion exchange resins of the...

The enzyme kynureninase converts kynurenine and hydroxykynurenine to alanine and anthranilic acid and to alanine and hydroxyanthranilic acid, respectively14>. Previously, Knox 51, Jakoby et al. 4>, a~d Wiss et al. 6> have described the purification and some properties of kynureninase. According to their reports, this enzyme obtained from bacteria was activated with magnesium ions, while it from...

Recently we have identified a mitogen-activated S6 kinase from Swiss 3T3 cells (Jenö, P., Ballou, L. M., Novak-Hofer, I., and Thomas, G. (1988) Proc. Natl. Acad. Sci. U.S.A. 85, 406-410). Here we describe the detailed purification of this enzyme from high-speed supernatants (400,000 x g) of vanadate-treated cell extracts. The enzyme is purified through six sequential steps including cation- and...

A convenient and rapid method for the purification of Streptococcus mutans dextransucrase is described. Affinity chromatography, on a column containing insoluble dextran purified from a culture of S. mutans 6715-49, gave an almost 300-fold purification, with 76% recovery of enzyme. Subsequent hydrophobic chromatography on butyl-agarose increased the overall enzyme purification to more than 1,00...

The practical work described here was designed in the aim of combining several periods that were previously carried-out independently during the academic year and to more appropriately mimic a "research" environment. It illustrates several fundamental biochemical principles as well as experimental aspects and important techniques including spectrophotometry, chromatography, centrifugation, and ...

Cloning and expression of the L-phenylalanine dehydrogenase gene, from B. sphaericus in E. coli were done. The gene was cloned in the vector pET16b and transformed into E. coli BL21 (DE3). The functional form of the L-phenylalanine dehydrogenase enzyme was purified by affinity purification techniques, taking advantage of the ability of this enzyme to bind to the nucleotide site affinity dye, Re...

During the purification of acetyl coenzyme A synthetase from bovine heart mitochondria it was noted that cruder preparations catalyzed appreciable butyrate-dependent disappearance of coenzyme A; activity toward butyrate disappeared from the acetate enzyme during advanced stages of the fractionation procedure (1). The present report describes the purification of a butyryl-CoA synthetase from bov...

An enzyme was extensively purified from jack bean leaves (Canavalia ensiformis L.) which produced o-ureidohomoserine from l-canaline and carbamyl phosphate. The most highly purified preparations catalyzed both this reaction and citrulline synthesis from ornithine and carbamyl phosphate, and the ratio of the two activities remained nearly constant during purification. When hydrated jack bean see...

<p><strong>Training on Making Eco-Enzyme Multipurpose Liquids from OrganicWastes and How to Use in Gondangmanis Village, Bae, Kudus. </strong>Eco-Enzyme (EE) is a multifunctional liquid produced anaerobic fermentation of organic waste, especially fresh fruit vegetable wastes. Until now, most Indonesian citizens have not known about this enzyme, including the partner, residents...