Carbohydrase Enzymes Research Articles

Effect of dietary ideal amino acid ratios, and supplemental carbohydrase in hulless-bar ley-based diets on pig performance and nitrogen excretion in manure

Two experiments were conducted, using a total of 224 (112 barrows + 112 gilts) crossbred pigs, to determine the effect of dietary ideal amino acid ratios, and supplemental carbohydrase in hulless barley (HB) diets, on pig performance and N excretion in manure. The four experimental grower and finisher diets were 1) a covered-barley (CB) control diet, 2) a HB (cv. Condor) diet, 3) same as diet 2 except that the soybean meal was replaced with supplemental lysine, threonine, methionine and tryptophan, to provide ideal amino acid ratios of 0.65 for threonine to lysine and 0.30 for methionine to lysine, and 4) same as diet 3 with supplemental carbohydrase (RonozymeTM W) enzyme at 350 g t–1. All diets were fed ad libitum as pellets with free access to drinking water. The average daily gain (ADG) was not different (P > 0.10) between diets 1 and 2, or among diets 2, 3 and 4 during the grower or finisher periods. The feed conversion efficiency was higher (P < 0.01) for diets 3 and 4 than for diet 2 during the grower period, but it was higher (P < 0.05) only for diet 4 than diet 2 during the finisher and combined grower-finisher periods. Backfat thickness, carcass index and dressing % were not different (P > 0.10) among the diets. The absorption of dry matter (88.9 vs. 83.7%) and energy (87.8 vs. 84.1%) was greater (P < 0.05) for diet 2 than diet 1. The excretion of faecal dry matter was 31.6% lower (P < 0.01) for diet 2 than diet 1 during the finisher period. The N excretion was 22.0% lower in pigs fed diet 3 than diet 2. Ammonia, carbon dioxide and hydrogen sulphide concentration in stored manure were not different (P > 0.10) among the diets, but ammonia production was positively correlated with urinary N excretion. These results indicated that feeding HB diets with supplemental amino acids minimises the excretion of both faecal dry matter and N with no adverse effect on the performance of lean-genotype pigs. Key words: Pigs, hulless-barley, amino acids, carbohydrase, nitrogen, manure

Effect of xylanase addition in feed containing either pre-characterised wheat or wheat byproducts on the performance of growing pigs

Wheat and wheat by-products vary in their available energy and protein content (Batterham et al., 1980) leading to unpredictable growth rate and feed use efficiency. Carbohydrase enzymes, targeting the non-starch polysaccharides in these raw materials, may improve their nutrient availability. However, questions remain over the relationship between the quality of the dietary raw materials and the level of exogenous enzyme activity required in the feed to elicit a response from pigs. Hence, the response of pigs to the use of a fungal xylanase, produced by Trichoderma longibrachiatum, was studied in diets containing wheat and wheat by-products.

Digestibility of canola protein products derived from the physical, enzymatic and chemical processing of commercial canola meal in rainbow trout Oncorhynchus mykiss (Walbaum) held in fresh water

This study was undertaken to assess the relative importance of nutrient composition and levels of antinutritional factors present in various canola protein products derived from the processing of commercial canola meal (CCM) on digestibility in rainbow trout held in 9.9–11°C fresh water. Laboratory sieving of CCM (SCM) reduced levels of crude fibre, cellulose and hemicellulose by 31.6%, 16.6% and 56.4%, respectively. Aqueous methanol–ammonia washing of sieved canola meal (ACM) increased neutral detergent fibre (NDF, insoluble dietary fibre) and phytate, and dramatically decreased levels of total glucosinolates (> 88%) and phenolic compounds (≈ 59%) relative to CCM. Further treatment of ACM with phytase in acidified (pH 5.0) aqueous media (PCM), followed by filtration, increased levels of crude protein, crude fibre and NDF, and decreased levels of phytate (33%), phenolic compounds (81%) and glucosinolates (below detection limit) relative to CCM. Additional treatment of PCM with carbohydrase enzymes (SP-249 and/or Alpha Gal, an oligosaccharide-degrading enzyme) further elevated levels of NDF, cellulose, protein (SP-249 only) and indispensable amino acids (SP-249/Alpha Gal), and concurrently decreased levels of phytate relative to PCM. An industrial source of canola protein isolate (CPI), produced by first extracting CCM with aqueous salt solution and then recovering the protein fraction, had the best nutrient composition and lowest levels of the antinutritional factors mentioned above. The processing protocols had slight or marked negative effects (ACM and SP-249/Alpha Gal) on apparent dry matter digestibility. Protein digestibility coefficients were unaltered in the laboratory-processed products, except for the products treated with SP-249 and SP-249/Alpha-Gal. In these cases, the protein digestibility coefficients (77.4% and 79.5%) were significantly below that found for CCM (88.1%). With two of the treatments, ACM and SP-249/Alpha-Gal, processing depressed DE. In contrast, the DE level in CPI (20.6 MJ kg−1) and the digestibility coefficients for dry matter (77.1%) and protein (97.6%) were high. We conclude that high levels of fibre, either alone or together with phytate, have the greatest adverse effects on the digestibility of canola protein products for rainbow trout. The high digestibility of CPI and its low content of antinutritional factors may also enable this protein product to be an excellent substitute for premium quality fish meal in diets for salmonids.

The effects of protease and carbohydrase supplementation on the nutritive value of canola meal for poultry: In vitro and in vivo studies

An in vitro incubation system was used to assess a variety of enzyme preparations regarding activities toward protein and carbohydrate components in canola meal with the objective of identifying those preparations that possessed potential for improvement of the nutritive value of canola meal. Promising preparations were evaluated further in 2-week growth trials utilizing 4-day-old broiler chickens. Enhanced protein hydrolysis was demonstrated for several of the protease enzyme preparations studied in the in vitro incubation system. Protein hydrolysis was most effective when either pancreatin or pronase were included in the incubation medium along with the protease enzyme and for the most effective protease preparations values for percent of total protein hydrolyzed exceeded those for pancreatin or pronase acting alone. The most effective protease preparation also resulted in improved broiler chick growth performance when added to semipurified canola meal diets. While some carbohydrase enzyme preparations were shown to be effective in the in vitro test of cell wall polysaccharide solubilization, only a trend toward improved growth performance was noted when broiler chickens were fed semipurified canola meal diets containing these enzyme preparations. A synergistic response in growth of young (4–11 days of age) broiler chickens was noted when phytase, carbohydrase and protease enzymes were added to a wheat/canola meal based diet deficient in available phosphorus.

Enzyme‐aidedvs. two‐stage processing of canola: Technology, product quality and cost evaluation

AbstractThe investigation focussed on the use of carbohydrase enzymes to enhance oil extraction during pressing in a laboratory expeller. Enzyme‐treated seeds at 6% moisture were pressed in the expeller set at full‐press conditions. Control seeds were pressed at wider choke openings but at the same barrel pressures as enzyme‐treated samples. Time of pressing and temperature and pressure inside the expeller barrel were used to calculate throughput and energy requirements per unit weight of processed material. Treatment with enzymes improved throughput of the expeller, increased oil flow rate and oil recovery. Material throughput was increased by 30–50%, depending on canola variety. The recovery of the oil was increased from 72% of the seed oil for control samples, to 90–93% for enzyme‐treated samples. The average residual oil content in presscakes from enzyme‐treated seeds was 7.4%. The oil quality was inferior to cold‐pressed control but was much better than has been reported for solvent‐extracted oil.

Mimosine, a nonprotein amino acid, inhibits growth and enzyme systems in Tribolium castaneum

The potency of mimosine is much greater in inhibiting growth and development of first-instar larvae of Tribolium castaneum than of fourth-instar larvae as expressed in suppression of larval weight gain and retardation in pupation. Approximately 70% suppression in weight gain of first-instar larvae was obtained at 0.06% mimosine and close to 100% at 0.08%. A dose-dependent decrease in the activity of trehalase, invertase, and amylase of fourth-instar larvae was obtained in vivo with the increase in mimosine concentration. At 0.3% dietary concentration, reductions of 58, 63, and 25% in trehalase, invertase, and amylase activity, respectively, were obtained after 2 days' feeding on treated diet, and of 65, 83, and 35%, respectively, after 5 days' feeding. On the other hand, increases of 27, 77, and 93% in protease activity were obtained in larvae fed for 5 days on diet containing 0.1%, 0.2%, and 0.3% minosine, respectively. The reduced activity of the trehalase enzyme might hamper the energy supply needed for growth and development, and that of the invertase and amylase enzymes for the feeding process. At concentrations of 10 −4 and 10 −3 M, mimosine had no effect on any of the carbohydrase enzymes in vitro. Hence, the observed effect on these enzymes in vivo could result from disturbances in protein synthesis leading to alteration in enzyme activity. Our study indicates that mimosine is a potent suppressor of growth and development of T. castaneum and may promote the development of a new group of insecticides.

Effect of Glucocorticoids and Limiting Nursing on the Carbohydrate Digestive Capacity and Growth Rate of Piglets

The influence of glucocorticoid administration and limited nursing on piglet carbohydrase enzyme development and subsequent growth was examined in three experiments using 371 piglets. Treatments in the first two experiments were formed by the factorial arrangement of hydrocortisone (-HYD or +HYD) and limited nursing (-LN or +LN) imposed form d 14 to weaning (d 28). Hydrocortisone was replaced by adrenocorticotropic hormone (ACTH) in the third experiment. Growth rates were severely depressed by HYD (P less than .01), LN (P less than .001) and to a lesser extent (P less than .06) by ACTH during the last 2 wk of lactation. During the first 14 d postweaning, piglets continued to grow more slowly following HYD treatment (P less than .01), whereas LN piglets grew more rapidly than those allowed to suckle normally. Although piglets were smaller at weaning after HYD injection (P less than .01), relative weights of liver, pancreas and small intestine were increased (P less than .05). Only adrenal weights were increased by ACTH (P less than .09). Pancreatic and intestinal amylase activities were increased two- to three-fold by HYD injection (P less than .05) but were unaffected by ACTH or LN (P greater than .10). Sucrase and maltase activity increased linearly with age (P less than .001). This rate of increase was numerically enhanced by glucocorticoid treatment and LN. The normal decrease in lactase activity was accelerated by LN and HYD injection, with the greatest depression caused by the combination of LN and either HYD or ACTH administration (P less than .05). Glucocorticoid administration to nursing piglets can evoke premature elevation of the carbohydrase enzymes necessary for initiating the hydrolysis of starch.

Saccharification of untreated agrowastes during mycelial growth of mushroom Termitomyces clypeatu on solid beds

Low-cost production of fermentable sugar from renewable cellulosic biomass is a highly desired technology for future planning. Although a number of active cellulase systems have been studied from various microorganism~,'-~ in vitro saccharification of agrowastes by these enzymes has not yet been found economical. Mushroom fruit bodies which grow excellently on cellulosic wastes have received little attention in this context. Chang and Steinkraus have only recently reported saccharification of agrowaste bed during mycelial growth of mushroom Volvariella v~lvacea.~ A few mushroom strains5-' growing under submerged conditions in our laboratory have been reported as potent producers of different carbohydrase enzyme^.^,^ The present study describes the in vivo saccharification of cellulosic biomass during the mycelial growth of mushroom Termitomyces clypeatus on various agrowastes.

The presence and partial characterization of carbohydrase enzymes in the gut ofCallosobruchus maculatus

Carbohydrase activity has been demonstrated in homogenates of the alimentary tract ofC. maculatus: β-D galactosidase > α-D glucosidase > α-D galactosidase > β-D glucosidase activity. The effects of pH, temperature and substrate concentration on β-D galactosidase, α-D glucosidase and α-D galactosidase activities are described.

The secretions and fine structure of bivalve crystalline style sacs

Abstract The style sac epithelia of the eulamellibranch bivalves Macomona Liliana and Amphidesma australe were examined to determine the sites of Ctystalline style secretion. In both species, Band D cell types were the main secretory cells. In Amphidesma other cell types could play minor roles in secretion. The basal few millimeters of the style sac in both species was found to differ from the upper region. Relatively more secretion was produced by this basal section. Aspirochaete, Cristispira sp. was found in the styles of Amphidesma, and observations suggested that this organism could produce extracellular carbohydrase enzymes. The study did not produce any evidence to support the view that the style sac had absorptive functions.

The role of aminopeptidases in inflammatory and neoplastic tissues

The role of leucine andalanine aminopeptidases is stidued in three different biologic systems: experimental wound healing in the rat, experimental carrageenan induced intraderman granulomas in the rat, and human laryngeal carcinomas. The wound healing experiments indicate that the proliferating granulation tissue has high quantities of aminopeptidases activity which is residing primarily intracellularly in granulocytes, macrophages, mast cells fibroblasts, and new budding vessels. The quantititave levels of tissue aminopeptidases correlate positively with the degree of cellularity of the wound and fibroblastic activity. Some aminopeptidases (isoenzymes) are secreted or released by the fibroblasts in be blood serum. Starch gel electrophoretic analysis demonstrates thses with different concentrations and migration rates. Two are probably released or secreted into the serum, and th third is membranous bound in the cytoplasm (lysosomal). The carrageenan intradermal granuloma demonstrates a different inflammatory reaction which is rich in macrophages and produces a different pattern of aminopeptidases activity. Macrophages produce a high tissue level of aminopeptidase activity which is intracellular bound and not readily leached out into the serum. Gel electrophoresis studies of aminopeptidases produced by the granuloma demonstrate that the tissue bound enzyme is the main component. In addition, there appears to be a mechanism, which is not understood, for specific induction or activation of lysosomal proteolytic and carbohydrase enzymes. This mechanism is dependent on the nature of composition of the injuring agent. Laryngeal carcinomas demonstrate high tissue homogenate levels and normal serum levels of aminopeptidase activities. These enzymes are located in the tumor stroma. They are not directly related to tumor invasiveness but to the degree of stromal proliferation. The tumor stroma behaves as though it were a non-healing wound constantly secreted proteolytic enzymes (aminopeptidases). A major problem that needs to be resolved is to find the operating mechanism by which malignant cells interact with their constantly proliferating fibroblastic stroma.

Assay and Biochemical Properties of the Proteinase Inhibitor-inducing Factor, a Wound Hormone

An assay has been developed for the proteinase inhibitor-inducing factor (PIIF), a wound hormone. PIIF is present in tomato (Lycopersicum esculentum var. Bonnie Best) leaf extracts and induces accumulation of proteinase Inhibitor I when the extracts are supplied briefly to excised leaves that are subsequently incubated in water under constant light. An active water-soluble crude PIIF solution was conveniently prepared from autoclaved and lyophilized tomato leaves. Accumulation of Inhibitor I, induced by crude PIIF, is linear, commencing at about 8 to 10 hours after feeding and continues for several hours. Evidence is presented that the PIIF-induced accumulation of Inhibitor I, determined immunologically, is accompanied by the accumulation of other trypsin and chymotrypsin inhibitors, determined enzymatically. The accumulation of Inhibitor I is inhibited by actinomycin D and cycloheximide but not by chloramphenicol or rifampin. PIIF cannot be replaced by traumatin, indoleacetic acid, gibberellic acid, kinetin, ethylene, or abscisic acid. PIIF activity was not destroyed by incubation with a number of proteolytic, carbohydrase, phosphatase, or pyrophosphatase enzymes. The active substance is insoluble in lipid solvents.

A SEASONAL STUDY OF OXYGEN UPTAKE OF SOME PASTURE SOILS AND ACTIVITIES OF ENZYMES HYDROLYSING SUCROSE AND STARCH

SummaryRates of oxygen uptake and activities of enzymes hydrolysing sucrose and starch were determined in soils under pastures of grasses and clovers at bimonthly intervals at eight sites in New Zealand with different climates. Differences between sampling periods were significant in most soils. Values of oxygen uptake and enzyme activities were not generally significantly correlated between themselves nor with the moisture contents of the soils nor their prior rainfall history. It has not been possible to suggest an optimal sampling period for different soils. The oxygen uptakes of all soils increased two‐ to threefold in the presence of glucose. Mean annual values of oxygen uptake of the eight soils were significantly correlated with their contents of organic carbon, reducing sugars produced on incubation in buffer, and rates of hydrolysis of starch, but not of sucrose. Climatic factors and the composition of the pastures probably influence the activities of carbohydrase enzymes in soil. The ratios of the activities of enzymes hydrolysing sucrose to those hydrolysing starch were greatest in the driest soils under predominantly grass pastures at sites with the lowest mean annual temperatures.

The development of the digestive system of the young animal III. Carbohydrase enzyme development in the young lamb

1. Twelve lambs varying in age from 5 to 35 days in age were slaughtered and their carbohydrate-digesting enzymes studied for any change in activity with increase in age.2. Results are given for the changes in weight of different parts of the digestive organs and their contents, and for the content of NaCl and of total acidity in the abomasum contents.3. Pancreatic and small intestine amylase showed only a slight increase in activity up to 5 weeks of age.4. Lactase and maltase activity in the small intestine were almost constant from 1 week to 5 weeks of age.5. No sucrase activity was measurable in any of the lambs slaughtered in either the small intestine wall or its contents.6. A comparison was made between the carbohydrase activity of the young pig and the lamb.7. Calculations based on the enzyme activities of the tissues showed the theoretical amounts of different carbohydrates which can be digested by lambs and young pigs of varying age groups.8. The results suggest that the young lamb is dependent on the early development of its rumen fauna and flora for the utilization of all other carbohydrates except lactose and glucose.

The development of the digestive system of the young animal II. Carbohydrase enzyme development in the young pig

1. Forty Large White pigs varying in age from birth to 5 weeks were slaughtered and their carbohydrate digesting enzymes studied for any change in activity with increase in age.2. Pancreatic amylase, sucrase and maltase increased from low levels at birth to high levels at 5 weeks of age.3. The total amount of lactase present at birth and of small intestine amylase at 1-week-old was maintained at a constant level up to 5 weeks of age. The increased weight of the intestine with the growth of the pig made the quantity of these enzymes appear to decrease with age when the values were expressed per kg. body weight or per g. dry tissue.4. Anaemia had no marked effect on the activity of the enzymes though the rate of enzyme development appeared to be retarded in the case of maltase.5. The quantity of pancreatic amylase present in the pancreas was sufficient at all ages to digest the amounts of starch likely to be included in a synthetic diet. Maltase, on the other hand, appeared to be limiting in early life.