Enzyme treatment of foods for filtration

14. Preparation of reagents

Tris buffer, 1M

Tris (hydroxymethyl) amino methane 121 g

Dissolve in about 500 ml of distilled water, adjust to desired pH with concentrated HCl and dilute to 1000 ml with distilled water. Store at 4^oC.

Acetate buffer, 1M

Mix 60 ml of glacial acetic acid in about 500 ml of distilled water, adjust to desired pH with 5N NaOH and dilute to 1000 ml with distilled water. Store at 4^oC.

Amylase stock solution

α-Amylase (Sigma No. A1278) >10 g

Tris buffer, pH 7.0 100 ml

Dissolve, warming to 35^oC if required. Filter through Whatman No. 1 paper, then filter-sterilize. Store 1 week at 4^oC or 3 months at -20^oC.

Cellulase stock solution

Cellulase (Sigma No. C7502) 10 g

Acetate buffer, pH 5.0 100 ml

Dissolve, warming to 35^oC if required. Filter through Whatman No. 1 paper, then filter-sterilize. Store 1 week at 4^oC, or 3 months at -20^oC.

Diastase stock solution

Diastase (Sigma No. A6880) 10 g

Tris buffer, pH 7.0 100 ml

Dissolve, warming to 35^oC if required. Filter through Whatman No. 1 paper, then filter-sterilize. Store 1 week at 4^oC or 3 months at -20^oC.

Hemicellulase stock solution

Hemicellulase (Sigma No. H2125) 10 g

Acetate buffer, pH 5.5 100 ml

Dissolve, warming to 35^oC if required. Filter through Whatman No. 1 paper, then filter-sterilize. Store 1 week at 4^oC or 3 months at -20^oC.

Trypsin stock solution

Trypsin (Difco No. 0152) 10 g

Tris buffer, pH 7.6 100 ml

Dissolve, warming to 35^oC if required. Filter through Whatman No. 1 paper, then filter-sterilize. Store 1 week at 4^oC or 3 months at -20^oC.