Purification and Characterization of Esterase 2B of the House Mouse, Mus musculus

Abstract

Esterase 2B was isolated from mouse kidney and purified about 1300‐fold by ion‐exchange chromatography and repeated molecular sieve chromatography. The resulting product is apparently homogeneous by the criteria of gel electrophoresis and immunodiffusion. The molecular weight was found to be 57000. The equivalent weight (54000) was estimated by titration of the catalytic site by bis(p > ‐nitrophenyl) phosphate. The Km constants of seven 4‐nitrophenyl esters of n‐fatty acids (C2–C8) were determined. The logarithm of Km decreases linearly with increasing number of C atoms of the fatty acid moiety. Concurrently, the free enthalpy ΔG′o of the enzyme‐substrate complex increases by about 2.46 kJ/mol CH2 group. Similar results are obtained when a series of esters of 8‐hydroxyquinoline were used as substrates.

Purification and Characterization of Esterase 1F, the Albumin Esterase of the House Mouse (Mus musculus)

“…The data presented in this paper show that esterase 1 can clearly be distinguished biochemically from esterase 2 [8], esterase 7 [9] and esterase 9 [7]. A property especially characteristic for esterase 1 is the wide range of substrate specificity (Table 2).…”

“…The properties of esterase I F were compared with those of purified esterase 2B, prepared according to Lexow et al [8] and. purified esterase 9A, prepared according to Goppinger et al [7].…”

“…Antisera against esterase 1 F, esterase 2B [8] and esterase 9 A [7] were raised in rabbits. Double diffusion experiments were performed according to Ouchterlony [I 51.…”

“…musculus) are genetically the best defined, this species seems to be most suited for the combined genetic and biochemical study of esterases. Three members of the esterase system of the house mouse, esterase 9A [7], esterase 2B [8] and esterase 7B [9] have previously been under investigation. In the present communication the isolation and biochemical characterization of esterase 1 F, the so-called albumin esterase of the house mouse, are described as a further contribution to the subclassification of carboxylesterase isozymes.…”

Purification and Characterization of Esterase 6A, a Trimeric Esterase of the House Mouse (Mus musculus)

“…The procedure was repeated after 6 weeks and the animals were bled after 3 months. Antibodies, were concentrated by ammonium sulphate precipitation and stored in phosphate buffer (0.1 mol/l, pH 7.3, 0.9%, w/v, sodium chloride) at -70 "C. Antisera against esterase IF, 2B and 9A were prepared as described previously [2,4,5]. Double-diffusion experiments were performed according to Ouchterlony [16].…”