REVIEW Enzyme immobilization: an update Ahmad Abolpour Homaei & Reyhaneh Sariri & Fabio Vianello & Roberto Stevanato Received: 5 June 2013 /Accepted: 31 July 2013 /Published online: 29 August 2013 # Springer-Verlag Berlin Heidelberg 2013 Abstract Compared to free enzymes in solution, immobilized enzymes are more robust and more resistant to environmental changes. More importantly, the heterogeneity of the immo- bilized enzyme systems allows an easy recovery of both en- zymes and products, multiple re-use of enzymes, continuous operation of enzymatic processes, rapid termination of reac- tions, and greater variety of bioreactor designs. This paper is a review of the recent literatures on enzyme immobilization by various techniques, the need for immobilization and different applications in industry, covering the last two decades. The most recent papers, patents, and reviews on immobilization strategies and application are reviewed. Keywords Enzyme immobilization . Biocatalyst . Enzyme reuse Abbreviations ALG Alginate AuNPs Gold nanoparticles CLEAs Cross-linked enzyme aggregates CLECs Cross-linked enzyme crystals CLIO Cross-linked iron oxide CS Chitosan DEAE Dimethylaminoethyl EDC 1-Ethyl-3-(3-dimethylaminopropyl) carbodiimide FDA Food and Drug Association HPLC High-performance liquid chromatography IMAC Immobilized metal affinity chromatography K d Dissociation constant K M Michaelis constant LC–MS Liquid chromatography–mass spectrometry LCST Low critical solution temperature MIONs Monocrystalline iron oxide nanoparticles NHS N -Hydroxysuccinimide Ni-NTA Nickel nitrilotriacetic acid PLA Poly(lactic acid) PLGA Poly(lactic-co-glycolic acid) polyNIPAM Poly-N -isopropylacrylamide PVA Polyvinyl alcohol T m Transition temperature USPIO Ultrasmall superparamagnetic iron oxide A. A. Homaei (*) Department of Biology, Faculty of Science, University of Hormozgan, Bandarabbas, Iran e-mail: A.Homaei@Hormozgan.ac.ir R. Sariri (*) Reyhaneh Sariri, Department of Microbiology, Lahijan Branch, Islamic Azad University, Lahijan, Iran e-mail: sariri@guilan.ac.ir F. Vianello Department of Comparative Biomedicine and Food Science, University of Padua, Padua, Italy R. Stevanato (*) Department of Molecular Sciences and Nanosystems, University of Venice, Venice, Italy e-mail: rstev@unive.it J Chem Biol (2013) 6:185–205 DOI 10.1007/s12154-013-0102-9