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141.
142.
Initial reaction rates for the hydrolysis of nucleic acids with micrococcal endonuclease (EC 3.1.31.1) insolubilized on Sepharose are strongly influenced by diffusional limitations. Although the absolute values are low, they can be increased substantially by changing particle and pore size of the support, or enzyme concentration in the insoluble derivative. As a result of steric and diffusional limitations, the course of the reaction and selectivity to hydrolysis products for the insoluble derivatives are different to those of the native enzyme; the former produces mainly large and small fragments but few of intermediate size. Because of these differences in course and selectivity of the reaction, diffusional limitations become less important when high initial reaction rates are not required. 相似文献
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A two-step purification procedure for 5-aminolevulinic acid dehydratase (EC 4.2.1.24) from human red blood cells has been developed. It involves one ion exchange and one gel filtration step. The purification is about 1000-fold, and the yield is more than 85%. With the purified enzyme a direct spectrophotometric assay of product formation without subsequent reaction with Ehrlich's reagent is described. 相似文献
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An aldehyde reductase (EC 1.1.1.2) from human liver has been purified to homogeneity. The enzyme is NADPH-dependent, prefers aromatic to aliphatic aldehydes as substrates, and is inhibited by barbiturates and hydantoins. The following physicochemical parameters were determined: molecular weight, 36,200; sedimentation coefficient, 2.9 S; Stokes radius, 2.65 nm; isoelectric point, pH 5.3; extinction coefficient at 280 nm, 54,300 M-1 cm-1. Results from polyacrylamide gel electrophoresis with and without sodium dodecyl sulfate, gel filtration, and ultracentrifugation suggest a monomeric structure. On molecule of NADPH binds to the enzyme causing a red shift of the coenzyme absorption maximum from 340 to 352 nm. The amino acid composition has been determined and a partial specific volume of 0.74 was computed from these data. An alpha-helicity of 7 and 18% was estimated from the ellipticities at 208 and 222 nm, respectively. Combination of the most reactive thiol group with p-mercuribenzoate does not cause loss of catalytic activity. Inactivation occurs when more than one thiol group is modified. The presence of NADPH or NADP+ prevents loss of activity by thiol modification. The comparison of structural features of aldehyde reductase with other monomeric and oligomeric dehydrogenases suggest similarities of aldehyde reductase with octopine dehydrogenase. 相似文献
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Frequency of infection by Cryptosporidium sp. in 1,039 faecal smears stained by Ziehl-Neelsen, obtained from undernourished patients of a Nutritional Recovery Center and an ambulatory undernourished center from the Fifth Region, Chile, were studied. All underwent a coproparasitological examination by the modified Telemann method. Thirty eight (3.7%) patients infected by the parasite were detected, with an overall frequency of 8.5% among patients of the Nutritional Recovery Center and 1.9% among the ambulatory patients; this difference was statistically significant. The highest percentage of positive results were detected among the younger milk feeding infants. Also the percentage of difference among these and the older milk feeding infants (3.7%) was statistically significant. Association of Cryptosporidium sp. and Giardia lamblia was observed in 6 ambulatory patients (2.3%). 相似文献