Kinetic Characterization and Effect of Immobilized Thermostable β-Glucosidase in Alginate Gel Beads on Sugarcane Juice

A thermostable β-glucosidase was effectively immobilized on alginate by the method of gel entrapment. After optimization of immobilized conditions, recovered enzyme activity was 60%. Optimum pH, temperature, kinetic parameters, thermal and pH stability, reusability, and storage stability were investigated. The Km and Vmax for immobilized β-glucosidase were estimated to be 5.0 mM and 0.64 U/ml, respectively. When comparing, free and immobilized enzyme, change was observed in optimum pH and temperature from 5.0 to 6.0 and 60°C to 80°C, respectively. Immobilized enzyme showed an increase in pH stability over the studied pH range (3.0–10.0) and stability at temperature up to 80°C. The storage stability and reusability of the immobilized β-glucosidase were improved significantly, with 12.09% activity retention at 30°C after being stored for 25 d and 17.85% residual activity after being repeatedly used for 4 times. The effect of both free and immobilized β-glucosidase enzyme on physicochemical properties of sugarcane juice was also analyzed.

Renal Tubular Damage Evaluation through Enzyme Change Monitoring after Extracorporeal Shock Wave Lithotripsy

To evaluate a possible onset of “biological effects” following ESWL lithotripsy, the Authors measured urinary excretion levels of two enzymes, NAG and GGT. In fact, a rise in the levels of these enzymes is regarded as an early and sensitive sign of renal injury. Fourteen patients with no complicated renal lithiasis were selected; the enzymatic levels before and after a single treatment with Lithostar Plus, a lithotriptor supplied with an electromagnetic impact wave generator, were measured. The results show that a single treatment doesn't produce any significant variation in their excretion. These results were discussed and their significance analysed. Probably, they could be explained by the lower pressure produced in the focal area by the impact wave, due to an electromagnetic generator.

Enzymes of the pathway of purine synthesis in the rat mammary gland. Changes in the lactation cycle and the effects of diabetes

Measurements were made of the activities of the enzymes of the ‘de novo’ and salvage pathways of purine synthesis [phosphoribosyl pyrophosphate amidotransferase (EC 2.4.2.14), adenine phosphoribosyltransferase (EC 2.4.2.7) and hypoxanthine phosphoribosyltranferase (EC 2.4.2.8)] at different stages of the lactation cycle, and the effects of diabetes on the activity of these enzymes in lactation were studied. A distinctive pattern of enzyme change was observed, in which the ‘de novo’ pathway enzyme phosphoribosyl pyrophosphate amidotransferase increased sharply between days 10 and 14 of pregnancy, and then remained sensibly constant until the height of lactation, whereas the enzymes of the salvage pathway increased later in pregnancy and continued to rise during lactation. Diabetes severely depressed the activity of the enzymes of the salvage pathway, but appeared to be without effect on the ‘de novo’ pathway enzyme. These results are discussed in relation to the provision of purine precursors from tissues outside the mammary gland.

THE MOLECULAR BASIS OF DOMINANCE

ABSTRACT The best known genes of microbes, mice and men are those that specify enzymes. Wild type, mutant and heterozygote for variants of such genes differ in the catalytic activity at the step in the enzyme network specified by the gene in question. The effect on the respective phenotypes of such changes in catalytic activity, however, is not defined by the enzyme change as estimated by in vitro determination of the activities obtained from the extracts of the three types. In vivo enzymes do not act in isolation, but are kinetically linked to other enzymes via their substrates and products. These interactions modify the effect of enzyme variation on the phenotype, depending on the nature and quantity of the other enzymes present. An output of such a system, say a flux, is therefore a systemic property, and its response to variation at one locus must be measured in the whole system. This response is best described by the sensitivity coefficient, Z, which is defined by the fractional change in flux over the fractional change in enzyme activity.

Adaptation of the small intestine during pregnancy and lactation in the rat

During pregnancy and lactation in the rat the small intestine in general and the mucosal epithelium in particular gain weight. The specific activities of sucrase, lactate dehydrogenase and succinate-tetrazolium reductase remain constant and those of glucose 6-phosphate dehydrogenase and isocitrate dehydrogenase increase. There is no evidence that the reported decrease in absorption per unit area or weight of mucosal epithelium during pregnancy and lactation is due to decreases in enzyme activities within the epithelium. The pattern of enzyme change shows that the response of the gut to the stimuli of pregnancy and lactation must be a complex one, possibly involving increases in the specific activities of some enzymes.

A mitochondrial lesion in experimental spinal cord trauma

✓ Cytochrome oxidase activities were determined by a microspectrophotometric method on microdissected samples of gray and white matter from frozen-dried sections of dog spinal cord. Experimental animals received 400 gm-cm impact at the T-3 to T-4 level. Sections for enzyme analysis and histological study were taken at the center of the trauma site, at successive 1-mm distances from the center, and at non-traumatized segments. A drop in cytochrome oxidase activities to approximately 50% of normal value was found as early as 15 minutes post-trauma, with greatest decrease at the trauma center and lesser effects at successive distances. This effect was related to known morphological defects in mitochondrial cristae and inner membranes. The enzyme change was significantly correlated with histological estimates of gray matter and neuronal damage, but was also partially independent of these processes. A small but significant increase of enzyme activity in white matter at the trauma site was considered most likely attributable to mitochondrial accumulations due to impaired axoplasmic flow.