Open AccessMeasurement of the glycogen synthetic pathway in permeabilized cells of cyanobacteria
Author(s) -
Gómez Casati Diego F.,
Aon Miguel A.,
Cortassa Sonia,
Iglesias Alberto A.
Publication year - 2001
Publication title -
fems microbiology letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.899
H-Index - 151
eISSN - 1574-6968
pISSN - 0378-1097
DOI - 10.1111/j.1574-6968.2001.tb09438.x
Subject(s) - biochemistry , cyanobacteria , glycogen , biology , anabaena , fluorescein , glucan , polysaccharide , enzyme , bacteria , fluorescence , genetics , physics , quantum mechanics
A simple, rapid and reliable procedure for permeabilizing cyanobacterial cells and measuring the glycogen synthetic pathway in situ, is presented. Cells from Anabaena sp. strain PCC 7120 were permeabilized with a mixture of toluene:ethanol (1:4 v/v). Fluorescence microscopy of cells incubated with fluorescein diacetate showed Anabaena non‐permeabilized cells as green fluorescents, whereas permeabilized (viable) cells exhibited the intrinsic red fluorescence. Labelled α‐1,4‐glucan was recovered when permeabilized cells were incubated with the substrates of ADP‐glucose pyrophosphorylase or glycogen synthase. The kinetic and regulatory properties of both enzymes could be reproduced in situ. The simplicity of the procedure and the ability to measure in situ glucan fluxes show the methodology as useful for studying the intracellular regulation of storage polysaccharides in a photosynthetic prokaryote.