Viscometric Determination of p-Glucanase and Endoxylanase Activity in Feed

Engelen, Adrianus J; Randsdorp, Peter H G

doi:10.1093/jaoac/79.5.1019

Cited by 3 publications

(3 citation statements)

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“…The level of enzyme supplementation was based on recommendations by the manufacturer (BASF AG, Ludwigshafen, Germany): xylanase (endo‐1,4‐ β ‐xylanase; EC 3.2.1.8) and phospholipase (phospholipase A2; EC 3.1.1.4) were added at levels of 5600 EXU/kg and 1000 IU/kg diet respectively. One endoxylanase unit (EXU) is defined as the amount of enzyme required to liberate 4.53 μ mol reducing sugars (measured as xylose equivalents) per min from a 0.5% arabinoxylan solution at pH 3.5 and 40 °C (Engelen et al., 1996). One IU phospholipase activity is defined as the amount of enzyme producing 1 μ mol of free fatty acids per min under standard conditions (egg yolk substrate, 0.4% phospholipids, pH 8, 40 °C, 6 mm Ca 2+ ) as described by Beudeker and Kies (2000).…”

Section: Methodsmentioning

confidence: 99%

Supplementation of xylanase and phospholipase to wheat‐based diets for weaner pigs

Diebold

Mosenthin

Sauer

et al. 2005

Animal Physiology Nutrition

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The effects of supplementing a wheat-based diet for weaner pigs with exogenous xylanase and phospholipase on ileal and faecal nutrient digestibilities and on the level of microbial metabolites in ileal digesta were examined. Fourteen piglets, weaned at 11 days, were fitted with a simple T-cannula at the distal ileum. The pigs were offered a control diet or diets supplemented with xylanase and phospholipase individually or in combination, in a two period crossover design. The combination of xylanase and phospholipase tended to increase the ileal recovery of the amino sugar galactosamine, whereas the concentration expressed in mg/kg dry matter intake of glucosamine was slightly decreased (p < 0.10). There was neither an effect of enzyme supplementation on ileal and faecal digestibility of the other nutrients and energy, nor was there an effect on pH and on the level of microbial metabolites in ileal digesta. However, an increase in ileal and faecal nutrient and energy digestibility with increasing age was observed. The ileal and faecal digestibility coefficients (except for ether extract) were significantly (p < 0.05) higher in experimental period I than in period II. These higher values may be attributed to a lower feed intake during period I. Since a lower level of feed intake is generally associated with a slower rate of passage and a longer retention time of digesta, a positive impact on digestion and absorption of nutrients can be assumed, which, on the other hand, limits the potential of additional enzyme effects.

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Section: Methodsmentioning

confidence: 99%

Supplementation of xylanase and phospholipase to wheat‐based diets for weaner pigs

Diebold

Mosenthin

Sauer

et al. 2005

Animal Physiology Nutrition

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“…The level of enzyme supplementation was based on recommendations by the manufacturer (BASF AG, Ludwigshafen, Germany): xylanase (endo-1,4-β-xylanase; EC 3.2.1.8) and phospholipase (phospholipase A2; EC 3.1.1.4) were added at levels of 5,600 EXU/kg and 1,000 IU/kg diet, respectively. One EXU (endo-xylanase unit) is defined as the amount of enzyme required to liberate 4.53 mol/min of reducing sugars (measured as xylose equivalents) from a 0.5% arabinoxylan solution at pH 3.5 and 40°C (Engelen et al, 1996). One IU of phospholipase activity is defined as the amount of enzyme producing 1 mol/min of FFA under standard conditions (egg yolk substrate, 0.4% phospholipids, pH 8, 40°C, 6 mM Ca 2+ ) as described by Beudeker and Kies (2000).…”

Section: Experimental Design and Dietary Treatmentsmentioning

confidence: 99%

Effect of supplementation of xylanase and phospholipase to a wheat-based diet for weanling pigs on nutrient digestibility and concentrations of microbial metabolites in ileal digesta and feces1

Diebold

Mosenthin

Piepho

et al. 2004

Journal of Animal Science

110

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The objective of this study was to determine the effect of supplementing a wheat-based diet with xylanase and phospholipase either alone or in combination on the ileal and fecal digestibilities of nutrients and energy in early-weaned pigs. In addition, the concentrations of ammonia, lactate, and VFA were measured in ileal digesta and feces. The experiment was carried out with 16 barrows weaned at the age of 11 d with an average initial BW of 4.1 kg. On d 4 and 5 postweaning, the piglets were fitted with a simple T-cannula at the distal ileum. The experiment was designed as a balanced incomplete block design with three periods. The piglets received the basal diet with or without supplementation of either xylanase or phospholipase or a combination of these. There was a positive (P = 0.005 to 0.018) effect on the digestibility values of GE, OM, CP, crude fiber (CF), and NDF with xylanase supplementation. Apart from lysine, threonine, cysteine, glycine, and proline, the digestibility values of all AA were improved (P = 0.001 to 0.024). Phospholipase supplementation had a positive effect on CP (P = 0.047) and CF (P = 0.002) digestibilities, but no effect on ether extract (EE) digestibility. Supplementation of both enzymes showed the largest response in nutrient digestibilities, except that EE digestibility was not affected. No differences were found in D-/L- lactate, and ammonia concentrations among treatments. Acetate and propionate concentrations tended to increase when xylanase was supplemented and were highest for the combination of both enzymes. Despite the positive effects on ileal nutrient and energy digestibilities, there was no effect of xylanase or the combined enzyme supplementation on the fecal digestibilities of OM, CP, EE, CF, NDF, ADF, or GE, and on fecal concentrations of VFA. Phospholipase alone slightly decreased the total-tract nutrient and energy digestibilities (P < 0.05). In conclusion, the combination of both enzymes generally led to the highest increases in ileal digestibilities, which were of small numerical magnitude (approximately 2%). However, on a relative basis, this increase of 2% represents approximately 13% of the remaining diet that was available for digestion based on the fact that approximately 15% of the diet was not digested in the control pigs. Thus, the potential benefits in the nutrition of weanling pigs from combinations of enzymes should be validated under practical conditions.

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“…However, as there is no unified standard endoglucanase activity assay method, declared activity of standard enzyme depends on the manufacturer (including the method for the measurement of enzyme activity and specificity). Therefore, in our opinion, viscometry, of the two approaches, is the more promising for the standardization of endoglucanase activity research methods and for obtaining comparable results, and is used in a number of laboratories [14][15][16]. The activity unit used in viscosimetric studies is the amount of enzyme which increases the relative substrate flow rate equal to 1 min  1 under standard -glucan hydrolysis conditions.…”

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confidence: 99%

Viscosimetric Assay of Endoglucanase Activity in Enzyme Fodder Additives

Комаров¹,

Telishevskaya²,

Sarkhanova³

et al. 2016

S-h. biol.

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Worldwide a large number of enzymes, including those with endo-β-glucanase activity are produced by different companies as fodder additives. In Russia, there is no uniform method for endo-β-glucanase activity analysis that causes difficulties in assessing quality of the enzyme preparations. The aim of the study was to develop such uniform method. In this, we optimized a viscometric determination based on change of the relative viscosity of a substrate mixed with enzyme preparation using Ostwald capillary viscometer. The following regime has been accepted: the diameter of the capillary is 0.73 mm; the temperature of hydrolysis is 30 C; the solvent is the acetate buffer (pH 4.7); time of incubation is 5, 10, 15 min in accordance with the kinetics of the enzymatic reaction; and βglucane (0.4 %) was used as substrate. The carboxymethyl cellulose is shown not to be relevant so far as it gives results incomparable with those obtained with β-glucane due to estimation of the cellulase but not the β-glucanase activity, that does not correspond to the objectives of forage production. The unit used in the developed method is the amount of enzyme which increases the relative substrate flow rate in 1 min-1 under standard conditions of hydrolysis. The measurement of accuracy of the method was carried out according to GOST R ISO 5725-2002 (the state standards). In this, 5 certified samples with different activity were prepared from a commercial sample of β-glucanase (Sigma-Aldrich, USA) with known activity (GLA/g). The activity of the standard was expressed in GLA/g as glucose equivalents corresponding to the amount of enzyme which acts on -glucane, releasing 1 µmol of sugars in terms of glucose. Certified samples were analyzed under intermediate precision conditions with three factors changed (i.e., operator, matrix, reagents). The matrices were mineral fillers, the calcium carbonate and zeolite. To calculate the accuracy of the method the viscosity of certified samples was expressed in GLA units using the calibration curves. Expanded uncertainty of measurements ranged within 10-22 % for the coverage factor k = 2.

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Viscometric Determination of p-Glucanase and Endoxylanase Activity in Feed

Cited by 3 publications

References 0 publications

Supplementation of xylanase and phospholipase to wheat‐based diets for weaner pigs

Supplementation of xylanase and phospholipase to wheat‐based diets for weaner pigs

Effect of supplementation of xylanase and phospholipase to a wheat-based diet for weanling pigs on nutrient digestibility and concentrations of microbial metabolites in ileal digesta and feces1

Viscosimetric Assay of Endoglucanase Activity in Enzyme Fodder Additives

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