Characterization of Maltogenic Amylase Activity Recovery: A Potential Approach for Improving Immobilization

Abstract

Cross-linked enzymes aggregate (CLEA) is a versatile carrier free-immobilization technique that has gained much attention in the development of biocatalyst technology. However, there is no precise and accurate method of using this technique that will lead to an expected outcome that meets the requirements of the industrial standard. Therefore, the objective of this study is to investigate the effect of a few methods of executing cross-linked enzyme aggregates approach using maltogenic amylase by measuring the activity recovery of the developed CLEA. Some factors that are considered in developing the methodologies are the interaction between cross-linkers and enzymes, size of the cross-linked enzyme aggregates and substrate diffusion. The addition of precipitant and the cross-linking agent steps has been manipulated and four different methodologies were developed. Based on the results, Method 2 showed the highest activity recovery (57.9%) whilst Method 4 gave the lowest activity recovery (15.7%). Method 2 is an improvised method that removed supernatant after centrifugation before proceeding to the cross-linking step. The characterization of cross-linked enzyme aggregates such as morphological characterization and Fourier Transform-Infrared Spectroscopy was also determined. In conclusion, the best and most productive preparation method was determined based on the highest activity recovery.