Enzyme assays
AZCL galactan
This semiquantitative assay was used to identify expressing transformants and galactanase containing fractions. The assay was carried out in a micro titer plate. 20 μL sample (culture broth or elution fractions from purification, or MilliQ water as blank) and 200 μL 4 mg/mL AZCL-galactan (lupin) (from Megazyme International Ireland.) in 0.1 M sodium acetate buffer, pH 4.0, containing 0.01% Triton X-100, was added to each well and incubated for 10 min at room temperature with shaking and then 5 min at room temperature without shaking, to let the insoluble particles settle. The absorbance at 595 nm was then measured on 100 μL of the supernatant without stopping the reaction.