Year: 1962
Type: article
Abstract: invertase refer to enzyme preparations obtained from plants, bacteria, fungi, protozoa or the honey bee (Edelman, 1956; Allen & Bacon, 1956; Pavlinova & Kursanov, 1956; Christie & Porteous, 1957; Carnie & Porteous, 1959a; Howard, 1959).Borgstrom & Dahlqvist (1958), Dahlqvist & Borgstrom (1959) and Dahlqvist (1958, 1959a-c, 1960a-f, 1961) have provided the only recent systematic accounts of the properties of an invertase obtained from an animal source, namely hog intestine.The characterization of the invertase activity of the mucosal cells of rabbit small intestine is now described.A brief account of some of the work has been published (Carnie & Porteous, 1959b).EXPERIMENTAL Animals.Dutch rabbits of either sex and weighing 1.5-3-0 kg. were kept on a diet of cabbage, rabbit cake and water for several days before an experiment.For the 24 hr.before an experiment all solid food was withheld, the cages were cleaned of faeces twice during the starvation period, but water was allowed ad libitum.The primary purpose of starvation was to free the small intestine of partially digested food.Isolation of mucosal cells of small intestine.A single rabbit was used for each experiment and killed under light chloroform anaesthesia by a sharp blow across the neck.The complete small intestine was rapidly excised and transferred to chipped ice.The chilled intestine was cut into approximately 15 cm.lengths, fat and omentum were trimmed away and the lumen was washed out with 0-9% NaCl or with 0 25M-sucrose at 20.Each length of trimmed and washed intestine was cut open longitudinally, laid mucosal side uppermost on a cold glass plate and the mucosal layer removed with the aid of a glass slide into a tared beaker immersed in chipped ice.All subsequent manipulations were carried out in pre-cooled vessels immersed in chipped ice or contained in centrifuges operated at 2-5 .Cell homogenization.The weighed mucosal cell preparation (usually 30 g.) was suspended in 4 vol. of ice-cold freshly prepared 0 25M-sucrose.This suspension was treated in a Waring Blendor at approximately 7500 rev./ min.for 1 min., then for four 1 min.periods at approximately 15 000 rev./min.with 15 sec.intervals between each maceration.
Source: Biochemical Journal
DOI: 10.1042/bj0850450
Authors: JA CARNIE, JW PORTEOUS
Institutions: University of Aberdeen, University of Manchester
Language: English
Subfield: Genetics
Domain: Life Sciences
Open Access status: bronze
Funder: Medical Research Council
OpenAlex ID: W168809651