Global Enzyme Balance (Aspergillus Niger)

Calcium Chloride 0.5M

(100ML)

6 mL 6 mL

A = Enzyme suspended in
Mixing Immobilization
A 360 pearls
buffer phosphate pH 7 (3
mL). Sodium Alginate 3%
w/v (3mL)

Calcium Chloride

Global Enzyme Balance (Commercial Enzyme, Aspergillus Oryzae)

Calcium Chloride 0.5M (100ML)

12 mL C

Mixing Immobilization
C = Enzyme suspended in 536 pearls
buffer phosphate pH 7 (6
mL). Sodium Alginate 3%
w/v (6mL)
Calcium Chloride
 Calcium Chloride 0.5M solution (100mL)

Weight 5.549 g Set on a beaker

Drill to 100 mL
of CaCL2
Sodium Alginate 3% w/v solution (3mL + 3mL of free enzyme) In case of immobilized a bigger
volume, you must do calculous
according to your new parameters.
Example to immobilized 6mL of
enzyme at 3% w/v you must take a
total volume of 12 mL, so the total
sodium alginate weight you must
add will be (0.03)(12mL)=0.36g
Weight 0.18g of Heat 3mL of Mix the alginate with the Mix the alginate solution
sodium alginate distilled water 3mL of distilled water with 3mL of free enzyme
 Immobilization Process

Add the suspension Leave pearls rest

of sodium alginate + for 1 hour in the Wash pearls with distilled
Add drop by drop the
enzyme into a syringe CaCl2 solution water. Then leave it rest in
suspension into the CaCl2
100ml of distilled water and
solution. Keep it mixing.
in refrigeration till it use.
Protein Quantification

Take 3 pearls Add 1.5 mL of Mix in vortex till Take 250 µL set it into a Read absorbance at
and put into a NaOH at 0.4N & pearls get dissolve. microtube with 750 µL λ=595nm. Do a graphic
falcon tube 1.5mL of citrate (20-30 min approx.) of Bradford dye absorbance vs concentration
buffer 0.5M

(Peña Díaz, Arroyo Begovich, & Gómez Puyou, 2004)

Referencias
Peña Díaz, A., Arroyo Begovich, Á., & Gómez Puyou, A. (2004). Bioqiímica. México: Limusa.