Site-directed chemically-modified magnetic enzymes: fabrication,improvements, biotechnological applications and future prospects |
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Authors: | Ahsan Mushir Shemsi Firdous Ahmad Khanday Ahsanulhaq Qurashi Amjad Khalil Gea Guerriero Khawar Sohail Siddiqui |
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Affiliation: | 1. Center for Environment and Water, Research Institute, King Fahd University of Petroleum and Minerals (KFUPM), Dhahran, Saudi Arabia;2. Life Sciences Department, King Fahd University of Petroleum and Minerals (KFUPM), Dhahran, Saudi Arabia;3. Department of Biotechnology, University of Kashmir, Srinagar, Jammu and Kashmir, India;4. Center of Excellence in Nanotechnology, King Fahd University of Petroleum and Minerals (KFUPM), Dhahran, Saudi Arabia;5. Environmental Research and Innovation (ERIN), Luxembourg Institute of Science and Technology (LIST), Esch/Alzette, Luxembourg |
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Abstract: | Numerous enzymes of biotechnological importance have been immobilized on magnetic nanoparticles (MNP) via random multipoint attachment, resulting in a heterogeneous protein population with potential reduction in activity due to restriction of substrate access to the active site. Several chemistries are now available, where the modifier can be linked to a single specific amino acid in a protein molecule away from the active-site, thus enabling free access of the substrate. However, rarely these site-selective approaches have been applied to immobilize enzymes on nanoparticles. In this review, for the first time, we illustrate how to adapt site-directed chemical modification (SDCM) methods for immobilizing enzymes on iron-based MNP. These strategies are mainly chemical but may additionally require genetic and enzymatic methods. We critically examine each method and evaluate their scope for simple, quick, efficient, mild and economical immobilization of enzymes on MNP. The improvements in the catalytic properties of few available examples of immobilized enzymes are also discussed. We conclude the review with the applications and future prospects of site-selectively modified magnetic enzymes and potential benefits of this technology in improving enzymes, including cold-adapted homologues, modular enzymes, and CO2-sequestering, as well as non-iron based nanomaterials. |
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Keywords: | NP nanoparticles MNP magnetic nanoparticles SDCM site-directed chemical modification VP versatile peroxidase MnP manganese peroxidase LiP lignin peroxidase EPL/IPL expressed (intein-mediated) protein ligation NAA natural amino acid UAA unnatural amino acid NCL native chemical ligation PMT protein methyltransferase PRINT PRotectINcise Tag TTL tubulin tyrosime ligase MOF metal organic framework LDM lynchpin-directed modification Nanobiotechnology Magnetic nanoparticles Site-selective immobilization Genetic modification Native chemical ligation Protein engineering Click chemistry Metal organic framework Catalysis Cold-adapted |
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