Effects of rumen fluid pre-incubation on in vitro proteolytic activity of enzymatic extracts from rumen microorganisms

datacite.alternateIdentifier.citationAnimal Feed Science and Technology, Vol. 162, Nº3 abr., 75-82, 2010es
datacite.alternateIdentifier.doi10.1016/j.anifeedsci.2010.09.003es
datacite.creatorVelásquez, A.
datacite.creatorPichard, Gastón
datacite.date2010
datacite.date.issued2012-03-03
datacite.subjectBovinoes
datacite.subjectExtractos enzimáticoses
datacite.subjectZimogramaes
datacite.titleEffects of rumen fluid pre-incubation on in vitro proteolytic activity of enzymatic extracts from rumen microorganismses
dc.date.accessioned2012-03-03T22:02:17Z
dc.date.available2012-03-03T22:02:17Z
dc.description.abstractEnzymatic extracts were prepared from rumen fluid (RF) either directly after collection or after incubation in enriched media and their proteolytic activities were measured and compared. The objective of RF in vitro cultivation was to augment enzymatic activity by using diverse incubation media containing different feed substrates to selectively induce microorganisms, with essentially unrestricted sources of energy and N. The effects on protein degradation in bovine serum albumin (BSA) and soybean meal, rapeseed meal, dehydrated alfalfa meal and perennial ryegrass were determined. Enzymatic extracts were characterized by means of protease zymograms. Two adult Holstein Friesian rumen fistulated cows were used as RF donors. Standard anaerobic incubation techniques in buffer-mineral media were used, and protein degradation was measured as residual true protein after enzymatic incubations. Results reveal an effect of RF pre-incubation on protein degradation (P<0.0001) measured in BSA and in feed substrates, and also on the hydrolysis rate (P<0.001), measured with native substrates only. The enzymatic extracts showed a high potential for proteolytic degradation, reaching 785 mg/g crude protein (CP) degraded in BSA after 12 h of incubation, and 830 mg/g CP degraded in soybean meal with 48 h of incubation, compared to only 654 mg/g and 414 mg/g in controls. The zymograms of these extracts revealed six zones of proteolytic activity, with the highest concentration of peptidases in the zone of high molecular weight (117 and 130 kDa). In vitro pre-incubation of rumen fluid in culture media enriched with energy and N sources allowed collection of enzymatic extracts with higher proteolytic activity than extracts obtained directly from fresh rumen fluid. © 2010 Elsevier B.V.es
dc.formatPDFes
dc.identifier.urihttps://repositoriodigital.uct.cl/handle/10925/837
dc.language.isoenes
dc.sourceAnimal Feed Science and Technologyes
oaire.resourceTypeArtículo de Revistaes
uct.carreraAgronomíaes
uct.carreraMedicina Veterinariaes
uct.catalogadorpopes
uct.comunidadRecursos Naturaleses
uct.facultadFacultad de Recursos Naturaleses
uct.indizacionISIes
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