Biocatalysis pp 83-112 | Cite as

Cross-linked Enzyme Aggregates: Current Developments and Applications

  • Rubia Noori
  • Mohammad Perwez
  • Meryam SardarEmail author


Enhanced performance of enzyme is the major challenge which restricts their applications in industrial processes. Different strategies have been proposed to increase their lifetime/performance, they are (1) isolation of novel enzymes which can function under extreme conditions, (2) modifications of enzyme structure to increase their resistance i.e. protein engineering and (3) modification of the solvent environment. However, immobilization, an old technique is still considered as a very powerful tool to improve enzyme properties. In this review, we are presenting a detailed discourse of cross-linked enzyme aggregate (CLEA), as a new method of immobilization. CLEA technology involves precipitation of enzymes using organic solvents followed by cross-linking the precipitated enzymes using a crosslinker. Several additives like starch, bovine serum albumin and polyamines are used to provide additional cross-linking sites for better stability. An appropriate precipitant is required for CLEA formation for a selected enzyme, therefore screening of precipitant is needed which depends on the nature of the enzymes. Optimization of organic solvents, cross-linking concentration and incubation time is a must for optimum performance of CLEA. It shows high stability, reusability and acts as a better catalyst in terms of performance. Furthermore, this technology can be used for the preparation of combi-CLEA using two or more enzymes. It can be a great tool for stabilizing enzymes to improve their overall performance which can be exploited in industries for chemical transformations.


CLEA combi-CLEA Glutaraldehyde Cross linker Precipitant Magnetic-CLEA Immobilization Biocatalyst 




Degree Celsius




Bovine serum albumin


Candida antarctica B Cross-linked enzyme aggregates


CLEA-based enzyme microreactor


Cross linked enzymes


Cross-linked enzyme aggregates


Cross linked enzyme crystals


Direct Black-38


Davallia tyermannii Hydroxynitrile lyase Cross-linked enzyme aggregates


Endocrine disrupting chemicals


Fatty acid ethyl esters


Fatty acid propyl esters


Glucose oxidase


Horseradish peroxidase




MagneticCross-linked enzyme aggregates






Magnetic nanoparticles


Nicotinamide adenine dinucleotide


Phenylalanine ammonia lyase- cross-linked enzyme aggregates


Pseudomonassp. Lipase Cross-linked enzyme aggregates


Sodium dodecyl sulphate


Scanning Electron Microscopy


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© Springer Nature Switzerland AG 2019

Authors and Affiliations

  1. 1.Department of BiosciencesJamia Millia IslamiaNew DelhiIndia

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