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Use of central composite design to optimize working conditions of Streptomyces griseus enzymatic method in estimating in vitro rumen undegraded crude protein of feedstuffs

Published online by Cambridge University Press:  24 January 2018

A. Gallo ,
P. Fortunati ,
S. Bruschi ,
G. Giuberti  and
F. Masoero
A. Gallo*
Affiliation:
Departement of Animal Science, Food and Nutrition, Faculty of Agricultural, Food and Environmental Sciences, Università Cattolica del Sacro Cuore, 29122 Piacenza, Italy
P. Fortunati
Affiliation:
Departement of Animal Science, Food and Nutrition, Faculty of Agricultural, Food and Environmental Sciences, Università Cattolica del Sacro Cuore, 29122 Piacenza, Italy
S. Bruschi
Affiliation:
Departement of Animal Science, Food and Nutrition, Faculty of Agricultural, Food and Environmental Sciences, Università Cattolica del Sacro Cuore, 29122 Piacenza, Italy
G. Giuberti
Affiliation:
Departement of Animal Science, Food and Nutrition, Faculty of Agricultural, Food and Environmental Sciences, Università Cattolica del Sacro Cuore, 29122 Piacenza, Italy
F. Masoero
Affiliation:
Departement of Animal Science, Food and Nutrition, Faculty of Agricultural, Food and Environmental Sciences, Università Cattolica del Sacro Cuore, 29122 Piacenza, Italy
*
Author for correspondence: A. Gallo, E-mail: antonio.gallo@unicatt.it
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Abstract

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.

Keywords

In vitro assayproteaseproteinrumen
Type
Animal Research Paper
Information
The Journal of Agricultural Science , Volume 156 , Issue 1 , January 2018 , pp. 100 - 109
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Copyright
Copyright © Cambridge University Press 2018 

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