Analysis ofα-glucosidase enzyme activity used in a rapid test for steam sterilization assurance

Abstract

The results of this work provide new insight into the science behind a rapid test for steam sterilization assurance.

Effects of steam autoclave treatment on Geobacillus stearothermophilus spores

“…; Setlow et al . ). The spores were free (>98%) of growing or sporulating cells or germinated spores, and were stored frozen at −80°C in water.…”

“…One millilitre aliquots of the supernatant and pellet fractions were then autoclaved, spore viability determined, and α‐glucosidase activity was assayed at 56°C in 10 mmol l −1 NaPO 4 buffer (pH 7·4) −210 μ g ml −1 methylumbelliferyl‐α‐D‐glucoside (α‐MUG), by measuring release of fluorescent methylumbelliferone (MU) (Setlow et al . ). In a second experiment, after spore viability was determined, washed spores (1·5 ml; initial O.D.…”

A disclosure gel for visual detection of liveBacillus anthracisspores

“…It may be that the aglucosidase is simply a cytoplasmic enzyme in the mother cell that becomes trapped in the outer layers of the spore before its release. However, Albert et al (1998) described spore coat-associated a-glucosidase present in Geobacillus stearothermophilus spores, which was supported by Setlow et al (2016). The authors predicted that this enzyme may have a role in early germination and outgrowth, which may also be the case for B. anthracis.…”

“…The increase in α‐glucosidase activity may be due to new enzyme synthesis as a result of initiating germination, as seen previously (Chandrapati and Woodson ) or release of α‐glucosidases from within the inner layers of the spore (Setlow et al . ).…”

“…() described spore coat‐associated α‐glucosidase present in Geobacillus stearothermophilus spores, which was supported by Setlow et al . (). The authors predicted that this enzyme may have a role in early germination and outgrowth, which may also be the case for B. anthracis .…”