Enzymic and in sacco methods to estimate rumen degradation of food protein in cattle

BACKGROUND: Two factorial studies compared enzymic and in sacco methods to estimate degradation of ruminant foods. Enzyme degradation (in vitro = enzyme) was determined from the release of leucine‐equivalent amino acid (LA) crude protein (CP) from sunflower meal (SF), maize gluten meal (MG), distillers' dark grain (DG) and field beans (FB) after their separate incubations with Streptomyces griseus enzyme for 0–24 h. In sacco crude protein (CP) degradation of these foods was estimated during washing (0 h) and rumen incubations in fistulated cows for 2–24 h. The LA data were expressed as g LA per either kg of CP (LACP) or acid‐hydrolysable LA (HLA) of each food and compared with in sacco data. RESULTS: These methods showed comparable degradation with time (P < 0.01). The in sacco and HLA were greater than LACP for all foods except MG where in sacco value was either lower or equal to LACP depending upon the incubation time (P > 0.05 or P < 0.05). Conversely, HLA was significantly (P < 0.01) greater than LACP from 2 h onwards. At 0 h, in sacco values were significantly greater than those of enzyme for SF, DG and FB (P < 0.05) but not for MG. The foods differed significantly for degradation constants (a, b, c) in each method (P < 0.05). CONCLUSIONS: Despite variations between in sacco and enzyme estimates for different foods, the relationships between these estimates suggest that the HLA enzyme method has the potential to estimate food degradation. Copyright © 2007 Society of Chemical Industry     

Enzymatic biodiesel production: Technical and economical considerations

It is well documented in the literature that enzymatic processing of oils and fats for biodiesel is technically feasible. However, with very few exceptions, enzyme technology is not currently used in commercial‐scale biodiesel production. This is mainly due to non‐optimized process design and a lack of available cost‐effective enzymes. The technology to re‐use enzymes has typically proven insufficient for the processes to be competitive. However, literature data documenting the productivity of enzymatic biodiesel together with the development of new immobilization technology indicates that enzyme catalysts can become cost effective compared to chemical processing. This work reviews the enzymatic processing of oils and fats into biodiesel with focus on process design and economy.     

Methacrylic acid–acrylamide-g-poly(ethyleneterephthalate) fibres for urea hydrolysis

This paper presents the results of urease immobilization onto methacrylic acid–acrylamide grafted poly(ethyleneterephthalate) fibres. The graft yield strongly affected the maximum activity of the immobilized enzyme up to a value of 70·2%. Higher grafting caused a decline in urease activity and led to the degradation of the fibres. The minor changes observed in K_m and V_max demonstrated that the conformational changes existed during immobilization were not extensive. However, 70·2% methacrylic acid–acrylamide-g-fibres containing urease were more stable towards acidic and alkaline pH, high temperature and storage conditions compared with free enzyme. Apart from the increase in stability to heat inactivation, the initial enzymatic activity of the urease–fibre system remained almost unchanged even after 40 repeated assays corresponding to 10 h of operation in 4 months, indicating the excellent durability of the system.     

APPLICATION OF A COMMERCIAL ENZYME PREPARATION IN THE BARLEY MALTING PROCESS

This paper presents data on barley micromalting with addition of the CELLUCLAST enzyme complex. This is a commercial, multicarbohydrase with distinct β-glucanase and proteinase activities. The enzyme was added to steeping and germination phases, in different quantities (0.05%; 0.75% and 0.1% of initial barley). The enzyme was added to different malting phases: to the 2nd and 3rd steep water, at the beginning of germination on the 1st day by spraying, on the 2nd day of germination and in combination of addition to 3rd steeping water and in germination start (50% of total quantity of each). CELLUCLAST enzyme had a significant effect on reduction of wort viscosity, extract difference, wort filterability and protein breakdown, depending on the quantity of added enzyme and the malting phase to which it was added. There was no negative effect on other malt quality parameters. The best values of cytolytic breakdown parameters (viscosity, extract difference, filtration rate) were obtained with addition of 0.075% of CELLUCLAST, on the first day of germination.     

Modulation of the enzymatic activity of papain by interdomain residues remote from the active site

The two main catalytic residues Cys25 and Hisl59 of the monomericcysteine protease papain are located on different walls of acleft formed by two domains. This topology suggests a possiblerelationship between relative domain organization and catalyticmechanism. The effect on enzymatic parameters of structuralmodifications at various locations of the twodomain interfaceof papain was examined by individual or double replacementsby Ala of pairs of interacting residues. Most modificationshad no effect on enzyme activity. However, the enzyme's substrateturnover (k_cat) decreased following simultaneous alterationof the two most conserved residues, forming an apolar contactlocated 15 Å away from the active site. The pH activityprofile of the double mutant was unchanged, indicating a conservedionization state of the active site thiolate-imidazolium ionpair. This state is strongly dependent on the distance separatingthe two residues, thus suggesting that the active site geometryhas not been significantly altered. Efficient enzymatic activityin papain requires more than a correct active site geometryand is influenced by domain packing properties in a region remotefrom the active site.     

中空纤维酶膜反应器及其动力学初步探讨──Ⅰ．单酶系统中空纤维酶膜反应器

单酶系统中空纤维酶膜反应器是用天津纺织工学院研制的ＴＦＣ－００３型聚砜中空纤维超滤器固定β－淀粉酶，在一定条件下催化可溶性淀粉制取麦芽糖的反应器。试验中，对温度、ｐＨ、酶及底物浓度变化动力学参数进行了初步探讨，试验重复性好。     

Modified Atmosphere Packaging Affects Ascorbic Acid, Enzyme Activity and Market Quality of Broccoli

Reduced ascorbic acid (RAA) content, total chlorophyll and green color retention, enzyme activities and texture changes were followed in broccoli spears packaged in polymeric film and nonpackaged during 96 hr storage (10°C). Concentrations within packages monitored by gas chromatography were (CO₂,) 8% and (0²) 10%. RAA retention, moisture content, total chlorophyll and color retention were greater in packaged broccoli. No differences were found between packaged and nonpackaged broccoli for ascorbate oxidase and peroxidase activities, or texture.     

Fabry Disease and the Heart: A Comprehensive Review

Fabry disease (FD) is an X-linked lysosomal storage disorder caused by mutations of the GLA gene that result in a deficiency of the enzymatic activity of α-galactosidase A and consequent accumulation of glycosphingolipids in body fluids and lysosomes of the cells throughout the body. GB3 accumulation occurs in virtually all cardiac cells (cardiomyocytes, conduction system cells, fibroblasts, and endothelial and smooth muscle vascular cells), ultimately leading to ventricular hypertrophy and fibrosis, heart failure, valve disease, angina, dysrhythmias, cardiac conduction abnormalities, and sudden death. Despite available therapies and supportive treatment, cardiac involvement carries a major prognostic impact, representing the main cause of death in FD. In the last years, knowledge has substantially evolved on the pathophysiological mechanisms leading to cardiac damage, the natural history of cardiac manifestations, the late-onset phenotypes with predominant cardiac involvement, the early markers of cardiac damage, the role of multimodality cardiac imaging on the diagnosis, management and follow-up of Fabry patients, and the cardiac efficacy of available therapies. Herein, we provide a comprehensive and integrated review on the cardiac involvement of FD, at the pathophysiological, anatomopathological, laboratory, imaging, and clinical levels, as well as on the diagnosis and management of cardiac manifestations, their supportive treatment, and the cardiac efficacy of specific therapies, such as enzyme replacement therapy and migalastat.     

Efficient Linear dsDNA Tagging Using Deoxyuridine Excision**

Site-specific strategies for exchanging segments of dsDNA are important for DNA library construction and molecular tagging. Deoxyuridine (dU) excision is an approach for generating 3’ ssDNA overhangs in gene assembly and molecular cloning procedures. Unlike approaches that use a multi-base pair motif to specify a DNA cut site, dU excision requires only a dT→dU substitution. Consequently, excision sites can be embedded in biologically active DNA sequences by placing dU substitutions at non-perturbative positions. In this work, I describe a molecular tagging method that uses dU excision to exchange a segment of a dsDNA strand with a long synthetic oligonucleotide. The core workflow of this method, called deoxyUridine eXcision-tagging (dUX-tagging), is an efficient one-pot reaction: strategically positioned dU nucleotides are excised from dsDNA to generate a 3’ overhang so that additional sequence can be appended by annealing and ligating a tagging oligonucleotide. The tagged DNA is then processed by one of two procedures to fill the 5’ overhang and remove excess tagging oligo. To facilitate its widespread use, all dUX-tagging procedures exclusively use commercially available reagents. As a result, dUX-tagging is a concise and easily implemented approach for high-efficiency linear dsDNA tagging.