Immobilization of cross‐linked lipase aggregates onto magnetic beads for enzymatic degradation of polycaprolactone

Abstract

Candida rugosa lipase was immobilized on amino-functionalized magnetic supports via cross-linked enzyme aggregates (CLEA) and used to enhance the enzymatic degradation of polycaprolactone (PCL). The maximum amounts of lipase immobilized on the magnetic beads using glutaraldehyde as a coupling agent were determined to be 33.7 mg/g of beads with an 81% recovery of activity after immobilization. Compared to the free enzyme, the immobilized lipase showed the optimum pH at 1 unit higher (pH 8.0) and also retained its enzymatic activity at higher temperatures. There was 62.9% retention of lipase activity after 30 consecutive reuses, indicating its stability and reusability in aqueous media. Moreover, the immobilized lipase maintained more than 80% of its initial activity during 30 days storage period, while the free lipase lost all under same condition. In addition, the immobilized lipase showed a more than 6-fold increase in biodegradability over the free lipase when the immobilized lipase was used to degrade PCL in a batch system. Higher thermal and storage stability, as well as good durability after repeated use of the immobilized lipase CLEA, highlights its potential applicability as large scale continuous systems for the enzymatic degradation of PCL.

Crosslinked aggregates of Rhizopus oryzae lipase as industrial biocatalysts: Preparation, optimization, characterization, and application for enantioselective resolution reactions

“…A pH gradient between the immobilized lipase particles' domains and the external solution is formed during the enzymatic hydrolysis reaction shifting the optimum pH toward the alkaline region. Similar pH shifts to the alkaline region through positively charged groups for immobilized enzymes have been reported previously by many researchers 44, 46–49…”

Analysis of α-amylase inhibitor from corni fructus by coupling magnetic cross-linked enzyme aggregates of α-amylase with HPLC–MS

“…According to the affinity interaction between enzyme and inhibitor, MCLEAs have promise to be useful materials for specific analysis of active compounds from complex natural products. Besides some reports on the synthesis of MCLEAs and their catalytic activity, there was no report on the screening and analysis of inhibitors using MCLEAs so far [12,[15][16][17].…”

Synthesis of magnetic nanoparticles with an IDA or TED modified surface for purification and immobilization of poly-histidine tagged proteins

“…[1][2][3][4] Current approaches for immobilizing enzymes include embedding, adsorption, crosslinking, covalent bonding. [5][6][7][8] For example, glucose sensitivity was greatly improved by using glucose oxidase (GOx) and horseradish peroxidase (HRP) co-embedded nanoowers. 5 Cui et al prepared an electrochemical sensor for the detection of organophosphorus pesticides (OPs) by adsorbing acetylcholinesterase (AChE) to (CSTiO 2 )-Ti-@-CS/rGO.…”

“…6 Kim et al used glutaraldehyde as a cross-linking agent to immobilize Candida rugosa lipase to amino-modied magnetic beads to degrade polycaprolactone (PCL). 7 Hosseini et al covalently immobilized cellulase with epoxy polymer modied Fe 3 O 4 , which improved enzyme activity, thermal stability and reusability. 8 Reasonable quantity, quality, purity, activity, as well as stability of enzymes are always important issues in immobilizing enzymes.…”