Improvement of the Streptomyces griseus method for degradable protein in ruminant feeds

Licitra, G.; Lauria, F.; Carpino, S.; Schadt, Iris; Sniffen, C.J.; Soest, P.J. Van

doi:10.1016/s0377-8401(97)00178-8

Cited by 35 publications

(27 citation statements)

References 9 publications

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“…Intestinal digestibility of RUP was estimated via a three‐step enzymatic in vitro method (Hippenstiel, Kivitz, Benninghoff, & Südekum, ). Basically, the method corresponds to Calsamiglia and Stern (), except that ruminal incubation was replaced by application of Streptomyces griseus protease (Licitra et al., ). True protein content was determined according to Licitra et al.…”

Section: Methodsmentioning

confidence: 99%

Protection of protein from ruminal degradation by alkali‐induced oxidation of chlorogenic acid in sunflower meal

Bongartz

Böttger

Wilhelmy

et al. 2017

Animal Physiology Nutrition

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SummaryLactating ruminants require an adequate supply of absorbable amino acids for the synthesis of milk protein from two sources, that is crude protein (CP) synthesized microbially in the rumen and ruminally undegraded CP (RUP) from feed which can both be digested in the small intestine. Several chemical and physical methods have been identified as being effective in increasing the proportion of RUP of total CP of a feedstuff, yet there is a continuing need for developing and establishing methods which protect feed protein from ruminal degradation with acceptable expenditure of labour and other costs. The objective of this study was to identify and quantify effects of and interactions between chlorogenic acid and protein in solvent-extracted sunflower meal (SFM) as induced by alkali treatment. Response surface methodology was employed to investigate the influence of pH, reaction time and drying temperature on the resulting SFM and, subsequently, its protein value for ruminants estimated from laboratory values. For this purpose, alkali-treated SFM was subjected to a fractionation of feed CP according to the Cornell net carbohydrate and protein system as a basis for estimating RUP at different assumed ruminal passage rates (K p ). To estimate the intestinal digestibility of the treated SFM and its RUP, a three-step enzymatic in vitro procedure was applied. Alkaline treatment of SFM increased RUP values with factors ranging from approximately 3 (K p =.08/hr) to 12 (K p =.02/hr). Furthermore, the intestinal digestibility of the alkali-treated SFM was enhanced by approximately 10% compared to untreated SFM. Increasing pH and reaction time led to both increasing RUP values and intestinal digestibility. In conclusion, a targeted alkaline treatment of naturally occurring compounds in feedstuffs might be a promising approach to provide high-RUP feeds for ruminants which, at the same time, have improved intestinal digestibility values. K E Y W O R D Sinteractions, polyphenols, protected protein, protein degradation, rumen

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Section: Methodsmentioning

confidence: 99%

Protection of protein from ruminal degradation by alkali‐induced oxidation of chlorogenic acid in sunflower meal

Bongartz

Böttger

Wilhelmy

et al. 2017

Animal Physiology Nutrition

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“…The in vitro protease procedures used in this study were similar to those described by Krishnamoorthy et al (1983), Licitra et al (1998), and Coblentz et al (1999) Streptomyces griseus protease (P‐5147; Sigma Chemical Co., St. Louis, MO) contained 4.5 enzyme activity units per milligram of solid, where one activity unit of enzyme was able to hydrolyze casein to produce color equivalent to 1.0 μmol (181μg) of tyrosine per minute at pH 7.5 and 37°C. Sample size for all incubations was set on the basis of a common N content (15 mg) within each incubation flask; therefore, the actual sample weight was adjusted for CP concentration, and varied somewhat across forages.…”

Section: Methodsmentioning

confidence: 99%

“…Currently, the in situ procedure (Vanzant et al, 1998), corrected for microbial contaminant N, is the most common method used for evaluating relative proportions of RDP and RUP in ruminant feedstuffs; however, in vitro procedures that utilize semipurified proteolytic enzymes have been developed as routine laboratory techniques for the estimation of these protein fractions in forages (Krishnamoorthy et al, 1983; Licitra et al, 1998; Coblentz et al, 1999). Generally, single‐endpoint enzymatic techniques can more easily accommodate the sample numbers generated from plot‐type studies than full time‐course kinetic evaluations by in situ methodologies.…”

mentioning

confidence: 99%

Harvest Timing Effects on Estimates of Rumen Degradable Protein from Alfalfa Forages

et al. 2008

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Alfalfa (Medicago sativa L.) proteins ingested by dairy cows typically degrade at rapid rates and exhibit extensive ruminal degradability. Although the effects of conservation method (hay or silage) on these characteristics have been evaluated extensively, agronomic factors, such as harvest timing, have not. Our objective was to quantify rumen degradable protein (RDP) for ‘Affinity’ alfalfa harvested over a range of ages (0, 5, 10, 15, and 20 d following Stage 2) within each of four harvest periods (spring, early and late summer, and fall). For 2004, there were no interactions (P ≥ 0.372) between harvest period and days within harvest period for any protein component. Crude protein (CP), neutral‐detergent soluble CP (NDSCP; g kg−1 dry matter [DM]), and RDP (g kg−1 DM) declined in a quadratic (P ≤ 0.026) relationship with days following Stage 2. A quadratic (P = 0.002) pattern also was observed for rumen undegradable protein (RUP), but the overall range was small (60.4–66.5 g kg−1 DM). On a CP basis, RDP declined linearly (P < 0.001) from 720 to 659 g kg−1 CP during 2004. For 2005, there were interactions (P ≤ 0.020) of harvest period and days within period for all protein‐related response variables, but trends over time within each harvest period generally were similar to those observed in 2004. Overall, RDP declined as alfalfa plants aged within harvest period, but these responses were due pri marily to reduced concentrations of CP within the cell‐soluble fraction.

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“…However, quadratic terms of the main effects tested also influenced the enzymatic RUP determination within different feed categories. Concerning CONC, it was decided to test increasing concentrations of S. griseus protease and to express them as U for ml in 10 ml working solution, in line with previous approaches (Krishnamoorthy et al 1983;Licitra et al 1998Licitra et al , 1999Coblentz et al 1999). Thus, final enzyme concentrations in 50 ml buffer solution ranged from 0.016 to 0.160 U/ml.…”

Section: Discussionmentioning

confidence: 99%

Use of central composite design to optimize working conditions ofStreptomyces griseusenzymatic method in estimatingin vitrorumen undegraded crude protein of feedstuffs

et al. 2018

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The aim was to identify optimized combinations of Streptomyces griseus protease concentration (CONC), incubation length (TIME), or amount of crude protein (CP) incubated in buffered enzymatic solution (CPW) to predict the in vitro rumen-undegraded feed CP (RUP) of 26 different feeds (soybean, rapeseed or sunflower meals, wheat bran, distillers dried grains with solubles, maize co-products and alfalfa hay). Different levels of CONC (0.08, 0.19, 0.44, 0.69 and 0.80 enzymatic units [U] of S. griseus protease/ml), TIME (6, 10, 18, 26 and 30 h) and CPW (69, 118, 235, 353 and 401 mg CP) were tested in agreement with a central composite design (CCD) with four replications of the central point to calculate second-order polynomial equations of main tested effects. The RUP was estimated by incubating samples in a buffered rumen fluid for 16 h or by adopting different enzymatic approaches as planned a priori in CCD. Differences between rumen and enzymatic RUP (ΔRUP) were estimated and regression terms of second-order polynomial equations for estimating ΔRUP were calculated between and within feeds. These equations were optimized using the non-linear generalized reduced gradient method with the objective set at ΔRUP equal to 0. The adoption of CCD permitted identification of optimized enzymatic combinations of CONC (0.12 U of S. griseus protease/ml), TIME (18 h) and CPW (from 233 to 458 mg CP for distillers dried grains with solubles and soft white wheat bran, respectively) to predict RUP accurately in all feed categories except for soybean meal, where optimized combinations were 0.47 U of S. griseus protease/ml, 18 h and 435 mg CP.

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Improvement of the Streptomyces griseus method for degradable protein in ruminant feeds

Cited by 35 publications

References 9 publications

Protection of protein from ruminal degradation by alkali‐induced oxidation of chlorogenic acid in sunflower meal

Protection of protein from ruminal degradation by alkali‐induced oxidation of chlorogenic acid in sunflower meal

Harvest Timing Effects on Estimates of Rumen Degradable Protein from Alfalfa Forages

Use of central composite design to optimize working conditions ofStreptomyces griseusenzymatic method in estimatingin vitrorumen undegraded crude protein of feedstuffs

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