Ruminal digestion of chloroplasts and the protection of protein by glutaraldehyde treatment

Abstract

SummaryChloroplasts, 50–80% intact, were prepared from kale (Brassica oleracea L. var. Marrowstem) by rapid homogenization (3·5 sec) of deveined leaves into partially frozen buffer pH 7·4 made 0·4 M with sucrose, and separated by differential centrifugation. Chloroplasts from 10 kg leaves were administered to the rumen of a sheep and the degradation followed by analysis of the rumen fluid which was collected continuously. Particulate matter of the rumen fluid was separated on discontinuous sucrose density gradients, 8 ml each of 35, 46, 50, 65 and 75% (w/v), centrifuging at 300 g for 1 h at + 2 °C. Chloroplasts were rapidly ingested by entodiniomorphid protozoa and digested with no detectable increase in soluble nitrogen in the rumen fluid. No net production of ammonia occurred but δ-amino-valeric acid increased indicating that amino acid transformations had occurred. Chlorophyll in the ingested chloroplasts was rapidly degraded with a half-life of 50–60 min. In contrast a similar amount of soluble casein added to the rumen degraded with a half-life of 13–14 min with large increase in soluble nitrogen consisting of peptides, amino acids, ammonia and δ-amino-valeric acid. Glutaraldehyde treated chloroplasts were also ingested by protozoa, chlorophyll was rapidly degraded but there was no increase in the soluble nitrogen of the rumen fluid, including ammonia and δ-amino-valeric acid. Water disrupted chloroplasts released soluble protein and in the rumen behaved like casein, producing high proportions of ammonia and δ-amino-valeric acid. The significance of these experiments in the protection of soluble leaf protein from degradation in the rumen of pasture-fed animals is discussed.