Rumen protein degradation rates estimated by non-linear regression analysis of Michaelis–Menten in vitro data

Abstract

An in vitro method applying Michaelis–Menten saturation kinetics was developed as an alternative approach for estimating protein degradation rates in the rumen. Non-linear regression (NLR) analysis of the integrated Michaelis–Menten equation yielded fractional degradation rates,kd, from direct estimates of the maximum velocity: Michaelis constant ratio (kd=Vmax:Km). Degradation rates obtained using data from a series of 2 h inhibitor in vitro incubations were respectively 0.989, 0.134, and 0.037 /h for casein, solvent soya-bean meal (SSBM) and expeller soya-bean meal (ESBM). Degradation rates obtained from 2 h incubations had lower standard errors than those obtained using 1 h incubations; 2 h rates were not significantly different from 1 h rates, suggesting end-product inhibition was not significant at 2 h. The NLR Michaelis–Menten method was used to determine degradation rates for twelve protein sources: casein, bovine serum albumin, two samples of lucerne (Medicago sativa) hay, and four samples each of SSBM and ESBM. Statistical analysis of NLR results revealed significant differences among the twelve protein sources. Casein was degraded most rapidly (0.827 /h), and the four ESBM samples most slowly (0.050–0.098 /h). Degradation rate for serum albumin was 0.135 /h; rates for SSBM and lucerne hays ranged from 0.160 to 0.208 /h. Degradation rates estimated using the NLR method were more rapid than those obtained with a limited substrate approach; NLR rates were more consistent with in vivo estimates of rumen protein escape. Greater concentrations of slowly degraded proteins were needed with the NLR method to define curvilinearity of the degradation curve more accurately.Protein degradation rate: Rumen protein escape: Michaelis–Menten kinetics: Non-linear regression