Abstract
Amyloglucosidase was immobilized onto granular chicken bone (BIOBONE™) by noncovalent interactions. The amount of activity bound relative to an equal amount of free enzyme was 13.6 ™0.4%. The estimated specific activity for amyloglucosidase decreased from 75.3™0.8 to 43.5 ™9.6 U/mg protein upon immobilization. TheKm value of the bone-immobilized enzyme using glycogen as substrate increased from 3.04™0.38 mg/mL (free) to 9.04™ 1.51 mg/mL (immobilized), butKm showed no change upon immobilization when starches were used as substrates. A decrease in Vmax values occurred upon enzyme immobilization for all substrates, but this largely reflected the percentage of enzyme initially bound to the bone. Immobilization also improved enzyme stability in the presence of various additives (e.g., detergent, KC1, and ethanol) or under low or high pH reaction conditions. Bound amyloglucosidase maintained high activity (>90%) following five cycles of continuous use at moderate (23 ™C) and high (55™C) temperatures. Data derived from Lineweaver-Burk and Arrhenius plots indicated that substrate and product diffusion limitation were minimal.
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Schafhauser, D.Y., Storey, K.B. Immobilization of amyloglucosidase onto granular chicken bone. Appl Biochem Biotechnol 32, 89–109 (1992). https://doi.org/10.1007/BF02922151
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DOI: https://doi.org/10.1007/BF02922151