Trehalose will be produced when functional OtsA and OtsB are both present and expressed.
A good trehalose assay kit appears to be the Megazyme Trehalose assay kit:
Other Trehalose Testing Kits:
1. Extract trehalose by boiling cell pellets at 95degrees for 20mins.
2. Convert trehalose in the supernatant to glucose with trehalase obtained from sigma.
3. Measured this by a glucose assay kit from sigma.
4. Determine the preexisting glucose in each sample was determined in a control reaction without trehalase
5. Subtracted this from total glucose
alternatively, use sigma's trehalose test kit
Trehalose is produced after conversion from 2 enzymes, coded for by OtsA and OtsB. These are Trehalose phosphate synthase and Trehalose phosphate phosphatase respectively.
Quantative testing of each enzyme is tested using the following assays:
OtsA (Trehalose–6-Phosphate Synthase)
- quantitative assay - giving units of activity for the enzyme.
- By measuring OD2340
Final output= nmol UDP/min/mg protein
OtsB (Trehalose–6-Phosphate Phosphatase)