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Application 1028

Enzyme immobilization screening using rotating bed reactors

Finding the optimal chemistry and solid-phase material for immobilization of enzymes relies heavily on trial and error. The right resin will ensure satisfactory immobilization yield, as well as high activity and stability of the enzyme.

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Immobilizing enzymes can be a challenge due to the difficulty in achieving a balance between enzyme activity, stability and yield. Achieving high immobilization yields can be difficult due to the hydrophobicity of the enzyme, as well as the potential for competing nonspecific binding of the enzyme to the support material. In addition, the enzyme activity may be affected by the immobilization process, or may be lost during reuse of the enzyme-resin system due to leaching of the enzyme from the support material.

The rotating bed reactor (RBR) offers several advantages for enzyme immobilization screening, such as automation, time and labor efficiency in sampling and monitoring. Its easy handling also makes it a great choice for enzyme immobilization screening. The RBR also offers the flexibility of using different types of resins, which can be packed in different amounts, making it suitable for a variety of applications.

In this application a screening kit with six different enzyme carrier resins was used in parallel to immobilize lipase CalB.

Residual activity in supernatant after 1h of immobilization.

Graphs showing CalB residual enzyme activity in the supernatant after 1 h, as percentages of initial enzyme activity. The enzyme solution was prepared for each type of resin according to the protocols outlined in the SpinChem guidelines for enzyme immobilization.

ECR8806F was determined to be the best candidate, and it was estimated that 97% of the total enzyme amount was immobilized within 1 h. Next, the biocatalytic activity of the beads is assessed.

Further reading:

Industries Pharmaceutical

Topics Biocatalysis, Reaction Screening

Products

Details

Conditions: The enzyme solution was prepared for each type of resin according to the protocols outlined in the SpinChem guidelines for enzyme immobilization (available at www.spinchem.com/support). Each MagRBR contained 0.5 mL of ECR carrier resin, and was spun in enzyme supernatant at 500 rpm for 24 h. To monitor the immobilization process, enzymatic activity in the supernatant was measured at five points during a 24 h timespan to determine the amount of residual enzyme in the solution. The enzymatic activity was determined using a lipase activity test based on the transformation of p-nitrophenylbutyrate to p-nitrophenol and butyric acid. The reaction was quantified by the use of a spectrometer through an increase in absorbance at 450 nm due to the conversion into the products.