Why was the exo-1,3-β-Glucanase plus β-Glucosidase removed from the K-YBGL procedure for total glucan hydrolysis?

The method has been refined and improved. The following changes were implemented:

1. The temperature of the preliminary acid incubation was increased to 30°C (previously required an ice bath).
2. The solubilisation step duration was reduced to 60 minutes (previously 2 hours).
3. The calculation was updated to include a correction factor to account for glucose degradation under the new conditions.

Under these improved acid-hydrolysis conditions the results were consistent across all samples tested, with or without the inclusion of exo-1,3-β-Glucanase plus β-Glucosidase. Consequently, the enzyme step was eliminated from the procedure.

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