scholarly journals A proposed methodology to standardize the determination of enzymic activities present in enzyme additives used in ruminant diets

2003 ◽  
Vol 83 (3) ◽  
pp. 559-568 ◽  
Author(s):  
D. Colombatto and K. A. Beauchemin

There is increasing interest in using enzymes that degrade plant cell walls in ruminant diets to enhance production efficiency. Despite strong evidence from several studies suggesting a beneficial effect of enzyme supplementation on nutrient utilization and animal performance, overall the results have been somewhat inconsistent. One of the main problems faced by researchers is the lack of adequate biochemical characterization of the products used, which leads to a poor understanding of their mode of action. Of these biochemical characteristics, enzyme activities are the most important, but they are not always evaluated prior to use. Furthermore, as many arbitrary units of expression for these activities coexist, direct comparisons among studies are essentially impossible. In this paper, we propose a methodology that we feel accounts for the requirements of accuracy, simplicity and safety of use. In addition, a rationale for the standardization of the assays as a function of the conditions under which the enzymes are expected to act is presented. The standardization of these assays will benefit researchers, the feed industry, regulatory organizations, and ultimately the consumer, as it will result in the development of better, safer and more consistent enzyme additives for use in ruminant diets. Key words: Cellulase, enzyme additives, methodology, ruminants, xylanase

Author(s):  
WILLIAM S. YORK ◽  
ALAN G. DARVILL ◽  
MICHAEL MCNEIL ◽  
THOMAS T. STEVENSON ◽  
PETER ALBERSHEIM

Author(s):  
William S. York ◽  
Alan G. Darvill ◽  
Michael McNeil ◽  
Thomas T. Stevenson ◽  
Peter Albersheim

1994 ◽  
Vol 64 (1) ◽  
pp. 63-65 ◽  
Author(s):  
Gordon J Provan ◽  
Lorraine Scobbie ◽  
Andrew Chesson

1997 ◽  
Vol 75 (11) ◽  
pp. 1957-1964 ◽  
Author(s):  
T. H. Koh ◽  
L. D. Melton ◽  
R. H. Newman

Ripe and unripe cell walls isolated from the cortical tissues of strawberry (Fragaria × ananassa Duchesne cv. Yolo) using HEPES-buffered phenol of pH 6.5 were analysed using solid-state 13C nuclear magnetic resonance. Changes in cell wall components during ripening were investigated by separating the spectra, using proton spin relaxation editing, into three subspectra based on the mobility of the molecules. The subspectra can be assigned to rigid material (cellulose), semirigid components (primarily polygalacturonic acid) and semimobile (other detectable noncellulosic substances). The results show that, with ripening, separation between the semirigid and semimobile domains became more distinct. Associated with this, the ratio of noncellulosic material (i.e., pectins and hemicelluloses) to rigid cellulose decreased from 2.3 for unripe to 1.9 for ripe. The crystallinity of the cellulose molecules remained unaltered throughout ripening. Furthermore, our work indicates that the basic cellulose crystallite of strawberry cell walls appeared exceptionally small compared with other systems studied thus far. Key words: solid-state CP-MAS 13C NMR, cellulose, plant cell walls, strawberry, fruit ripening.


2001 ◽  
Vol 67 (2) ◽  
pp. 733-741 ◽  
Author(s):  
Cécile J. B van der Vlugt-Bergmans ◽  
Mariët J. van der Werf

ABSTRACT A monoterpene ɛ-lactone hydrolase (MLH) from Rhodococcus erythropolis DCL14, catalyzing the ring opening of lactones which are formed during degradation of several monocyclic monoterpenes, including carvone and menthol, was purified to apparent homogeneity. It is a monomeric enzyme of 31 kDa that is active with (4R)-4-isopropenyl-7-methyl-2-oxo-oxepanone and (6R)-6-isopropenyl-3-methyl-2-oxo-oxepanone, lactones derived from (4R)-dihydrocarvone, and 7-isopropyl-4-methyl-2-oxo-oxepanone, the lactone derived from menthone. Both enantiomers of 4-, 5-, 6-, and 7-methyl-2-oxo-oxepanone were converted at equal rates, suggesting that the enzyme is not stereoselective. Maximal enzyme activity was measured at pH 9.5 and 30°C. Determination of the N-terminal amino acid sequence of purified MLH enabled cloning of the corresponding gene by a combination of PCR and colony screening. The gene, designated mlhB(monoterpene lactone hydrolysis), showed up to 43% similarity to members of the GDXG family of lipolytic enzymes. Sequencing of the adjacent regions revealed two other open reading frames, one encoding a protein with similarity to the short-chain dehydrogenase reductase family and the second encoding a protein with similarity to acyl coenzyme A dehydrogenases. Both enzymes are possibly also involved in the monoterpene degradation pathways of this microorganism.


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