Enzyme purification and

S. P. Colowick u. N. O. Kaplan, Methods in Enzymology, Vol. XXII, Enzyme Purification and Related Techni-... 

Enzyme purification and characterisation Acetyl esterases were isolated from a Rapidase C-80 preparation according to the scheme shown in Figure 1. The purified acetyl esterases were devoid of relevant side activities, and showed great differences in their specificity towards the different acetylated substrates. 

Volume 104. Enzyme Purification and Related Techniques (Part C)... 

Mao, Q. Schunk, T. Gerber, B. Erni, B. A string of enzymes, purification and characterization of a fusion protein comprising the four subunits of the glucose phosphotransferase system of Escherichia coli. J. Biol. Chem., 270, 18295-18300 (1995)... 

Methods of Enzymatic Analysis, H. Bergmeyer, Editor. Contains methods for enzyme purification and assay, in several volumes. 

W. B. Jakoby, Ed., Enzyme Purification and Related Techniques, Vol. 22 of Methods in Enzymology, Academic Press, New York, 1971. 

Affinity chromatography is one of the most effective procedures for enzyme purification and yet, with the notable exceptions of Picard (PIO) and of La Du and his co-workers (L2), it has received scant attention for the purification of cholinesterase. Its value in purifying acetylcholinesterase from both fish electric organ and erythrocyte membrane is discussed in an excellent review by Rosenberry (R8). In affinity... 

In this context, the use of enzymes has emerged as a practical alternative to chemical synthesis [6-13]. Several examples have been reported that are based on the use of Leloir type-glycosyltransferases, which are mostly membrane-associated and act on nucleotide-activated sugars as donor substrates. However, the use of these enzymes in vitro still remains limited by the difficulty of enzyme purification and by their need for expensive sugar-nucleotides [6]. Re-engineering of microbial cells producing these proteins appears to be promising for the synthesis of specific carbohydrate stmctures [14]. 

Sandmann G (1994) Carotenoid biosynthesis in microorganisms and plants. EurJ Biochem 223 7-24 Sandmann G (1997) High level expression of carotenogenic genes for enzyme purification and biochemical characterization. 

S. viridosporus LiP has been concentrated by ultrafiltration (UF) for further studies on the enzyme purification and the determination of its chemical and biochemical properties [10, 11, 13, 14]. However, there is no report on the UF operational conditions and on the enzyme activity recovery. 

In the course of these reactions, P450 3A4 catalyzes examples of some atypical reactions " including desaturation , oxidative carboxylic acid ester cleavage , and oxidation of a nitrile to an amide . An unexpected reaction encountered in this laboratory was the oxidation of alkylphenyl ether non-ionic detergents, which have been commonly used in enzyme purifications and also have some medical and industrial applications . Methylene hydroxylations yield hemiacetals, which break down to shorten the chains . 

Enzyme purification and assay. NADP-malic enzyme was purified to apparent electrophoretic homogeneity from Zea mays leaves as previously described (3). Enzyme activity was spectrophotometrically assayed at 30°C and pH 7 by following NADP reduction at 340 nm as in (3). One unit (U) of enzyme is defined as the amount that catalyzes the formation of 1 ymol of NADPH per minute. 

Bioengineering enzyme production (including gene expression, enzyme purification, and formulation), fermentation, membrane technology, process systems engineering. 

My mission was to resolve heart muscle aconitase into the two enzymes presumed to be responsible for the two component reactions. This was my first stab at enzyme purification and after a few months work I failed to make any significant progress. I also saw no signs of separation of enzymes responsible for the two reactions aconitase, as others showed later, is a single enzyme. 

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