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Continuous ATP regeneration process with stable acetate kinase
Journal of Applied Biochemistry
|February 1, 1984
Summary
Immobilized heat-stable acetate kinase (AK) from Bacillus stearothermophilus shows improved stability and can be used for ATP regeneration. A novel immobilization method using an ADP derivative yielded higher enzyme activity.
Area of Science:
- Biochemistry
- Enzyme immobilization
- Biotechnology
Background:
- Acetate kinase (AK) is crucial for ATP regeneration.
- Immobilization enhances enzyme stability and reusability.
- Bacillus stearothermophilus provides a heat-stable AK variant.
Purpose of the Study:
- To immobilize heat-stable acetate kinase (AK) from Bacillus stearothermophilus.
- To evaluate a novel immobilization method using a bifunctional ADP derivative.
- To characterize the properties and kinetics of immobilized AK for ATP regeneration.
Main Methods:
- Covalent immobilization of AK to Sepharose using carbodiimide, hydroxysuccinimide, cyanogen bromide, glutaraldehyde, and a novel ADP derivative spacer.
- Characterization of immobilized AK properties (pH-activity, stability) and kinetics (Michaelis-Menten).
- Assessing immobilized AK performance in continuous operation and as an ATP regeneration system.
Main Results:
- The novel ADP derivative method yielded higher AK activity compared to conventional methods.
- Immobilized AK exhibited similar Michaelis-Menten kinetics and pH-activity profiles to free AK.
- Immobilized AK showed significantly enhanced thermal stability, retaining >80% activity after 1 month at 30°C.
Conclusions:
- Immobilized AK from B. stearothermophilus is a stable and efficient biocatalyst.
- The novel immobilization technique offers advantages for enzyme activity yield.
- Immobilized AK is a viable system for continuous ATP regeneration in bioreactors.