Effects of Pulsed Electric Field (PEF) Treatment on Enhancing Activity and Conformation of α-Amylase.

To explore an efficient, safe, and speedy application of pulsed electric field (PEF) technology for enzymatic modification, effects of PEF treatment on the enzymatic activity, property and kinetic parameters of α-amylase were investigated. Conformational transitions were also studied with the aid of circular dichroism (CD) and fluorescence spectra. The maximum enzymatic activity of α-amylase was obtained under 15 kV/cm electric field intensity and 100 mL/min flow velocity PEF treatment, in which the enzymatic activity increased by 22.13 ± 1.14% compared with control. The activation effect could last for 18 h at 4 °C. PEF treatment could widen the range of optimum temperature for α-amylase, however, it barely exerted any effect on the optimum pH. On the other hand, α-amylase treated by PEF showed an increase of Vmax, t1/2 and ΔG, whereas a decrease of Km and k were observed. Furthermore, it can be observed from fluorescence and CD spectra that PEF treatment had increased the number of amino acid residues, especially that of tryptophan, on α-amylase surface with enhanced α-helices by 34.76% and decreased random coil by 12.04% on α-amylase when compared with that of untreated. These changes in structure had positive effect on enhancing α-amylase activity and property.

Microwave and micronization treatments affect dehulling characteristics and bioactive contents of dry beans (Phaseolus vulgaris L.).

BACKGROUND: Heat pretreatment is considered the first step in grain milling. This study therefore evaluated microwave and micronization heat treatments in improving the dehulling characteristics, phenolic composition and antioxidant and α-amylase activities of bean cultivars from three market classes. RESULTS: Heat treatments improved dehulling characteristics (hull yield, rate coefficient and reduced abrasive hardness index) depending on bean cultivar, whereas treatment effects increased with dehulling time. Micronization increased minor phenolic components (tartaric esters, flavonols and anthocyanins) of all beans but had variable effects on total phenolic content depending on market class. Microwave treatment increased α-amylase inhibitor concentration, activity and potency, which were strongly correlated (r² = 0.71, P < 0.0001) with the flavonol content of beans. Heat treatment had variable effects on the phenolic composition of bean hulls obtained by abrasive dehulling without significantly altering the antioxidant activity of black and pinto bean hulls. Principal component analysis on 22 constituents analyzed in this study demonstrated the differences in dehulling characteristics and phenolic components of beans and hulls as major factors in segregating the beneficial heat treatment effects. CONCLUSION: Heat treatment may be useful in developing novel dietary fibers from beans with variable composition and bioactivity with a considerable range of applications as functional food ingredients.

Ultrasonic effect on the desizing efficiency of α-amylase on starch-sized cotton fabrics.

Enzymatic desizing by α-amylase and ultrasound irradiation are the two important clean technologies in the textile industry. In the present work, with the aim of giving a further insight to the influence of ultrasound on α-amylase activity and its desizing efficiency, the ultrasound-based experiments were afforded in two ways: (i) step-wise treatment of α-amylase by ultrasound and then enzymatic desizing, as well as; (ii) simultaneous utilization of ultrasound and α-amylase for the desizing. By the step-wise strategy, it is found that the ultrasound has negative impact on the α-amylase activity using soluble starch as substrate. However, the sonicated α-amylase possesses higher desizing efficiency because there are higher hydrophobic interactions between sonicated α-amylase protein and starch-sized cotton and thus intensifies its catalytic activity. By the simultaneous procedure, the enhancement to desizing efficiency is more pronounced than that by the step-wise procedure. This can be attributed to comprehensive actions of several reasons such as more effective stirring/mixing mechanism, damages or changes to substrate, more effective catalysis to hydrolytic reactions and faster removal of loosened products from the fabric bulk.

Cell phone radiations affect early growth of Vigna radiata (mung bean) through biochemical alterations.

The indiscriminate use of wireless technologies, particularly of cell phones, has increased the health risks among living organisms including plants. We investigated the impact of cell phone electromagentic field (EMF) radiations (power density, 8.55 microW cm(-2)) on germination, early growth, proteins and carbohydrate contents, and activities of some enzymes in Vigna radiata. Cell phone EMF radiations significantly reduced the seedling length and dry weight of V radiata after exposure for 0.5, 1, 2, and 4 h. Furthermore, the contents of proteins and carbohydrates were reduced in EMF-exposed plants. However, the activities of proteases, alpha-amylases, beta-amylases, polyphenol oxidases, and peroxidases were enhanced in EMF-exposed radicles indicating their role in providing protection against EMF-induced stress. The study concludes that cell phone EMFs impair early growth of V radiata seedlings by inducing biochemical changes.

Effect of GaAlAs laser irradiation on enzyme activity.

OBJECTIVE: The aim of this study was to determine the influence of laser irradiation on enzyme activity. BACKGROUND DATA: Enzymes are catalysts of extraordinary efficiency, able to accelerate reactions by manifold. Enzyme laser light activation is currently a fast-growing field and a large number of studies have been produced. MATERIALS AND METHODS: Liquid CNPG amylase and control serum (Qualitrol 1H) were used in the experiments. Laboratory analysis of alpha-amylase was performed on two sample groups: (i) E + S and (ii) E + S + L, in six repetitions per irradiation dose. Group 2 was irradiated with gallium-aluminum-arsenide (GaAlAs) 904 nm at doses of 0.01, 0.1, 0.5, and 1 J/cm(2). Enzyme activity was read using a spectrophotometer equipped with a thermostatic chamber capable of precise absorbance measurement at 405 nm. RESULTS: The results were analyzed with the Student's t-test, and the percentage of enzyme activity was determined. Photomodulation of alpha-amylase activity by GaAlAs laser was analyzed following irradiation with different doses. Irradiation doses from 0.01 to 1 J/cm(2) led to differences in enzyme activity: 0.01 J/cm(2) (0.10%), 0.1 J/cm(2) (13.44%), 0.5 J/cm(2) (12.57%), and 1 J/cm(2) (-6.10%). CONCLUSION: Irradiation doses of 0.1 J/cm(2) and 0.5 J/cm(2) led to statistically significant increases in enzyme activity in comparison to the control. The similar curves of the effects of temperature and pH on enzymatic activity observed in this study suggest that laser irradiation also possess an optimum dose to modulate the enzymatic activity. That is, enzymes have an optimum laser dose (or range) at which their activity is maximal, whereas at higher or lower doses activity decreases.

Production and characterization of alpha-amylase from Aspergillus niger JGI 24 isolated in Bangalore.

Five fungal isolates were screened for the production of alpha-amylase using both solid-state and submerged fermentations. The best amylase producer among them, Aspergillus niger JGI 24, was selected for enzyme production by solid-state fermentation (SSF) on wheat bran. Different carbon and nitrogen supplements were used to enhance enzyme production and maximum amount of enzyme was obtained when SSF was carried out with soluble starch and beef extract (1% each) as supplements. Further attempts to enhance enzyme production by UV induced mutagenesis were carried out. Survival rate decreased with increase in duration of UV exposure. Partial purification of the enzyme using ammonium sulphate fractionation resulted in 1.49 fold increase in the enzyme activity. The enzyme showed a molecular weight of 43 kDa by SDS-PAGE. Metal ions Ca2+ and Co2+ increased the enzyme activity. The enzyme was optimally active at 30 degrees C and pH 9.5.

Ultrasonic inactivation of Bacillus alpha-amylase. I. effect of gas content and emitting face of probe.

Sonoinactivation of alpha-amylase from Bacillus amyloliquefacience was studied at a constant frequency of 30 kHz. The effect of sonotrode emitting face and gas content of medium on the efficiency of enzyme inactivation were investigated at different time-temperature combinations and generation of OH free radicals was also monitored. The results showed that ultrasound effectively inactivated alpha-amylase with a minimum overall inactivation rate at 50 degrees C. The tip diameter of the sonotrode and the gas content of the medium both significantly affected the rate of enzyme inactivation. The increase of tip diameter increased the effect of ultrasound on the enzyme, while the removal of dissolved gas adversely influenced the cavitational events and reduced the rate of enzyme inactivation. Calculation of the kinetic and activation parameters revealed that ultrasound decreased the activation energy, E(a), activation enthalpy, DeltaH(#), and the activation Gibbs free energy, DeltaG(#), and strongly reduced the activation entropy, DeltaS(#), down to negative values. This huge reduction in activation entropy was attributed to the different mechanisms of inactivation induced by heat and ultrasound. It is proved in this study that ultrasonically generated OH free radicals and shear forces, which arise from pulsation- or collapse of bubbles, both can destabilize the enzyme, although their contribution to the overall inactivation varies depending on the temperature and the tip diameter of the sonotrode.

Are sonochemically prepared alpha-amylase protein microspheres biologically active?

Using high-intensity ultrasound, we have synthesized alpha-amylase microspheres. The paper presented characterization as well as catalytic experiments of the sonochemically-produced microspheres. It also provided an estimate of the efficiency of the sonochemical process in converting the native protein to microspheres. These microspheres showed a very good enzymatic activity compared with the native alpha-amylase. The enzymatic activity of the amylase microspheres was 27% of that of the native protein after a short reaction time (3 min), while over a longer reaction time (1 h) it reached 56% of the activity of the native protein.

The alterations in the activity of amylase and its salivary isoenzyme in the serum of patients with ovarian carcinoma, submitted to radiotherapy.

Total activity of alpha-amylase and its salivary isoenzyme in the serum of patients with ovarian carcinoma of various types were evaluated before radiotherapy, in the middle of radiotherapy period, in the last day and 2 months after radiotherapy. Before radiotherapy the activity of these enzymes were significantly higher in patients with ovarian serous cystadenocarcinoma. It was found that irradiation resulted in a decrease of both total amylase activity and its salivary isoenzyme in the serum. The proportional participation of salivary isoenzymes in total amylase activity was normalized. It is suggested that the assay of salivary alpha-amylase activity may be useful in the evaluation of radiotherapy effectiveness.

[Enzyme activity in rat intestines after exposure to x-irradiation]. / Aktyvnist' travnykh fermentiv kyshky shchuriv pislia diï renthenivs'koho vyprominiuvannia.

It is established that 24 hours after total X-ray irradiation by doses of 0.5, 1 and 2 Gy activity of intestine enzymes (saccharose and alpha-amylase) increases. A dose of 3 and above induces an increase of saccharose activity and inhibition of alpha-amylase. The functional state of basic phosphatase under action of the doses investigated remains practically unchanged.

Alpha-amylase circadian rhythm of young rat parotid gland: an endogenous rhythm with maternal coordination.

The circadian rhythm of alpha-amylase, E.C. 3.2.1.1. alpha-1,4-glucan-4-glucanohydrolase) in the parotid glands of 25-day-old rats were studied under different experimental designs (fasting, reversed photoperiod, constant lighting conditions and treatment with reserpine and alpha-methyl-p-tyrosine). The rhythm of fasted rats did not change. There were modifications in the rhythm of rats submitted to a reversed photoperiod or treated with reserpine or alpha-methyl-p-tyrosine. The rhythm was present, with changes in the acrophase, in parotids of rats kept during their gestation and postnatal life in constant light or dark. Results suggest that the circadian rhythm of alpha-amylase in parotid gland of young rats is endogenous, synchronized by the photoperiod, and with maternal coordination.

Enzyme immobilization by radiation-induced polymerization of 2-hydroxyethyl methacrylate at low temperatures.

Enzyme immobilization by radiation-induced polymerization of hydrophilic glass-forming monomers, such as 2-hydroxyethyl methacrylate, was studied. Enzyme radiation damage could be sufficiently retarded at low temperatures. The immobilized enzyme activity yield was markedly higher at low temperature than at higher temperature polymerization. At low temperatures the polymerized composite had a porous structure owing to ice crystallization which depends on the monomer concentration. It was deduced that the enzyme was partially trapped on the polymer surface, partially isolated in the pore, and partially occluded inside the polymer matrix. A decrease in activity caused by enzyme leakage was observed with repeated use in enzyme reactions where the composites had a large porosity. The activity yield showed a maximum at certain optimum porosities, i.e., at optimum monomer concentrations. Continuous enzyme reaction was preferably carried out using immobilized enzyme columns.