Combination of carbohydrases and essential oils improve dietary performance of feedlot steers on a high-energy diet

Essential oils and enzymes are alternatives to feed additives for ruminants that aim to replace the use of ionophores and improve animal performance, but their mechanisms of action are different. Therefore, the present study aimed to verify if there is a synergistic effect in the combined use of enzymes carbohydrates and essential oils on the performance, ingestive behavior and carcass traits of steers fed a high-energy diet. During the finishing period of 78 days, 40 steers were assigned to four treatments: CON- control; ENZ- enzymatic complex; EO- essential oil blend; ENZ+EO - enzymatic complex combined with essential oil blend. Regardless of the feedlot periods, the ENZ+EO treatment caused a reduction in the dry matter intake (12.48%) compared to the control. The ENZ+EO treatment resulted in the lowest mean fecal output and, consequently, the highest dry matter digestibility (DMD) and starch digestibility (SD), compared to the other treatments. Animals that received EO and ENZ+EO in the diet spent more time in feeding. As for the number of times animals visited the feeding trough, the highest values were presented by the animals in the EO, ENZ and ENZ+EO treatments. For the carcass parameters, only the subcutaneous fat thickness on the rib was significantly different between treatments, with the highest values obtained by adding EO and ENZ+EO (8.80 and 8.10 mm respectively). Thus, the combination of carbohydrate enzymes and essential oils proved to be synergistically beneficial in relation to better use of nutrients and productive performance of feedlot steers.

Nostocyclopeptides as New Inhibitors of 20S Proteasome

Nostocyclopeptides (Ncps) are a small class of bioactive nonribosomal peptides produced solely by cyanobacteria of the genus Nostoc. In the current work, six Ncps were isolated from Nostoc edaphicum strain CCNP1411. The bioactivity of these compounds was tested in vitro against 20S proteasome, a proteolytic complex that plays an important role in maintaining cellular proteostasis. Dysfunction of the complex leads to many pathological disorders. The assays indicated selective activity of specific Ncp variants. For two linear peptide aldehydes, Ncp-A2-L and Ncp-E2-L, the inhibitory effects on chymotrypsin-like activity were revealed, while the cyclic variant, Ncp-A2, inactivated the trypsin-like site of this enzymatic complex. The aldehyde group was confirmed to be an important element of the chymotrypsin-like activity inhibitors. The nostocyclopeptides, as novel inhibitors of 20S proteasome, increased the number of natural products that can be considered potential regulators of cellular processes.

Effect of enzymatic complex in the diet of pirarucu, Arapaima gigas juveniles

ABSTRACT The pirarucu, Arapaima gigas is a native, carnivorous fish species from the Amazon basin. As carnivorous fish have low amylase activity, exogenous enzymes can improve the digestibility of carbohydrates in aquaculture feeds. We evaluated the digestibility of increasing levels of an enzymatic complex in diets of pirarucu juveniles (65.2 ± 0.4 g). The experimental design was randomized with four treatments [diets containing 0.25, 0.50, 0.75, and 1 g kg-1 on-top inclusion of an enzyme complex (Allzyme® SSF®, USA), and a control, with three replications at a density of 5 fish per unit, and a 30-day duration. We quantified apparent digestibility of dry matter, crude protein, and crude energy through nutrient and chromium oxide content in diets and feces. Enzymatic activity, hepatic glycogen and total protein were determined in liver and anterior intestine samples. The diet with 1 g kg-1 of enzyme-complex resulted in an increase in apparent digestibility of crude protein, gross energy, and dry matter, hepatic glycogen, total proteins in liver and in intestine, showing the efficiency of the enzyme complex in pirarucu feeding. A higher accumulation of dry matter, crude energy, and ethereal extract in the carcass indicated weight increase in the fish treated with enzymatic complex. A decrease in the endogenous enzymatic activity (protease, lipase and amylase) suggested an improved efficacy of the digestive process. Our results indicate that the inclusion of 1 g kg-1 enzyme complex in the diet of juvenile pirarucu can be recommended to achieve greater digestibility of nutrients and improvement in productive performance.

Broiler feed formulated with phosphorus, calcium, and energy deficiencies supplemented with exogenous enzymes

Three experiments were carried out to verify the effects of the enzyme phytase, alone or combined with an enzyme complex, in diets deficient in available phosphorus (AP), calcium (Ca), and metabolizable energy (ME) on broiler performance, ME, and dietary amino acid digestibility. A total of 1,538 male Cobb 500 broilers were allocated to the three experiments, each of which consisted of five treatments: positive control (PC; basal ration); negative control 1 (NC1; PC minus 0.15% of AP, 0.16% of Ca, and 68 kcal kg-1 ME); negative control 2 (NC2; PC minus 0.15% of AP, 0.16% of Ca, and 101 kcal kg-1 of ME); NC1 plus phytase; and NC2 plus phytase plus enzymatic complex. Body weight gain (WG) and feed intake were measured from 1-21 days and from 1-42 days, and the corrected feed conversion rate (FCR) for mortality was calculated. In the second and third experiments, the apparent ME corrected for nitrogen balance (AMEn) and standardized digestibility of amino acids, respectively, were determined, for the diets supplemented with phytase and the enzymatic complex. In the first experiment, enzyme supplementation increased (p < 0.05) WG at 21 days and 42 days relative to the negative controls. Phytase inclusion improved (p < 0.05) FCR at the initial phase compared to the NC1 diet. In the second experiment, enzyme supplementation did not affect (p > 0.05) AMEn. In the third experiment, both enzyme treatments improved (p < 0.05) the digestibility of amino acids in the supplemented diets compared to the deficient diets. Supplementation with phytase and carbohydrases preserves the performance of broilers fed diets deficient in AP, Ca, and ME and improves amino acid digestibility.

MACRO-ASPARTATE AMINOTRANSFERASE

Measurement of serum enzyme activity is one of the most common laboratory tests. Increased activity may be caused by abnormal enzymes with a high molecular mass, the so-called macroenzymes. Macroenzymes may be seen in healthy subjects, but can also be related to disease. Macro-aspartate aminotransferase (macro-AST) is a macroenzyme that results from an enzymatic complex consisting of AST linked to serum immunoglobulin (IgA, IgG or both). MacroAST persistence is a rare benign condition. Macro-AST is generally characterized by increased serum AST activity. The article contains analysis of literature data on patients with macro-AST.

Production of enzymatic complex from agro-industrial biomass and its application in combustible ethanol

Waste biomass and agro-industrial by-products, for production ethanol, will meet much of the great demand for this product. To reduce costs and optimize production, this study investigated solid-state fermentation (SSF) to obtain crude enzyme complex (CEC) from different agro-industrial biomasses (sugarcane bagasse, corn peel bran, rice straw bran and roasting and ground coffee residue) using cellulolytic fungi. The most promising CEC were evaluated in simultaneous hydrolysis and fermentation (SHF) for ethanol production by Saccharomyces cerevisiae in a culture broth containing sugarcane bagasse treated by steam explosion, and roast and ground coffee residue. In SSF with bioreactor volume of 0.25 L, containing 40 g of the biomass mixture and 40 g of sterile water with resuspended cells (1.0 x108 spores/g of solid medium) and temperature of 30±2 ºC, the strains Trichoderma reesei and Penicilium oxalicum provided the best enzyme activity. The CEC of T. reesei provided a concentration of 7.5 g L-1 of ethanol in a substrate containing treated sugarcane bagasse (60%) and roast and ground coffee residue (40%), under SHF conditions (pH 4.5, 35±2 °C, 48 h). The results obtained in this study show a promising alternative for correct disposal and use of residues and agro-industrial by-products by use in the production of enzymes and lignocellulosic ethanol.

A novel and specific regulator of neuronal V-ATPase in Drosophila

The V-ATPase is a highly conserved enzymatic complex that ensures appropriate levels of organelle acidification in virtually all eukaryotic cells. While the general mechanisms of this proton pump have been well studied, little is known about the specific regulations of neuronal V-ATPase. Here, we studied CG31030, a previously uncharacterized Drosophila protein predicted from its sequence homology to be part of the V-ATPase family. We found that this protein is essential and apparently specifically expressed in neurons, where it is addressed to synaptic terminals. We observed that CG31030 co-immunoprecipitated with V-ATPase subunits, in particular with ATP6AP2, and that synaptic vesicles of larval motoneurons were not properly acidified in CG31030 knockdown context. This defect was associated with a decrease in quantal size at the neuromuscular junction, severe locomotor impairments and shortened lifespan. Overall, our data provide evidence that CG31030 is a specific regulator of neuronal V-ATPase that is required for synaptic vesicle acidification and neurotransmitter release.

The nuclear DICER–circular RNA complex drives the deregulation of the glioblastoma cell microRNAome

The assortment of cellular microRNAs (“microRNAome”) is a vital readout of cellular homeostasis, but the mechanisms that regulate the microRNAome are poorly understood. The microRNAome of glioblastoma is substantially down-regulated in comparison to the normal brain. Here, we find malfunction of the posttranscriptional maturation of the glioblastoma microRNAome and link it to aberrant nuclear localization of DICER, the major enzymatic complex responsible for microRNA maturation. Analysis of DICER’s nuclear interactome reveals the presence of an RNA binding protein, RBM3, and of a circular RNA, circ2082, within the complex. Targeting of this complex by knockdown of circ2082 results in the restoration of cytosolic localization of DICER and widespread derepression of the microRNAome, leading to transcriptome-wide rearrangements that mitigate the tumorigenicity of glioblastoma cells in vitro and in vivo with correlation to favorable outcomes in patients with glioblastoma. These findings uncover the mechanistic foundation of microRNAome deregulation in malignant cells.