Cellulose nanofibers for magnetically-separable and highly loaded enzyme immobilization

Hwa Heon Je, Sora Noh, Sung Gil Hong, Youngjun Ju, Jungbae Kim, Dong Soo Hwang

Research output: Contribution to journalArticlepeer-review

35 Citations (Scopus)


Cellulose nanofibers (CNFs) are one of attractive supporting materials for enzyme immobilization due to their unique properties such as high surface area, high porosity and surface carboxyl groups for chemical bonding. In this study, CNFs were prepared via TEMPO-mediated oxidation and physical grinding of cellulose, and further used for the immobilization of α-chymotrypsin (CT) enzyme via four different approaches such as covalent attachment (CA), enzyme coating (EC), enzyme precipitate coating (EPC), and magnetically-separable EPC (Mag-EPC). EPC approach consists of three steps: covalent enzyme attachment, enzyme precipitation and crosslinking, while EC represents a control without the step of enzyme precipitation. Amine-functionalized magnetic nanoparticles were added during the enzyme precipitation and crosslinking steps to produce magnetically-separable EPC. The activities of CA, EC, EPC and Mag-EPC were 0.067, 0.14, 1.3 and 2.6 units per mg CNFs, respectively, representing that the activity of Mag-EPC was 38-, 19- and 2-times higher than those of CA, EC and EPC, respectively. After incubation under shaking (200 rpm) for 30 days, CA, EC, EPC and Mag-EPC maintained 12%, 46%, 77% and 50% of their initial activities, respectively, while free CT showed only 0.2% of its initial activity even after 8 days. Because CT is a tricky enzyme to stabilize due to its inactivation mechanism via autolysis, the present results of stable EPC and Mag-EPC on CNFs have demonstrated the great potential of CNFs as an environmentally-friendly and economical carrier of enzyme immobilization, which allows for magnetic separation as well as high enzyme activity/loading and stability.

Original languageEnglish
Pages (from-to)425-433
Number of pages9
JournalChemical Engineering Journal
Publication statusPublished - 2017

Bibliographical note

Funding Information:
This research was supported by Global Research Laboratory Program (2014K1A1A2043032), the ‘2016, University-Institute Cooperation Program’, and the Basic Core Technology Development Program for the Oceans and the Polar Regions (2016M1A5A1027594) through the National Research Foundation of Korea (NRF) funded by the Ministry of Science, ICT & Future Planning. This research was also supported by the Korea Institute of Energy Technology Evaluation and Planning (KETEP) and the Ministry of Trade, Industry & Energy (MOTIE) of the Republic of Korea (No. 20142020200980). This research was also supported by the Marine Biotechnology program (Marine BioMaterials Research Center) funded by the Ministry of Oceans and Fisheries, Korea (D11013214H480000110).

Publisher Copyright:
© 2017 Elsevier B.V.


  • Alpha-chymotrypsin
  • Cellulose nanofibers
  • Enzyme immobilization
  • Enzyme precipitate coating
  • Enzyme stabilization
  • TEMPO-mediated oxidation

ASJC Scopus subject areas

  • Chemistry(all)
  • Environmental Chemistry
  • Chemical Engineering(all)
  • Industrial and Manufacturing Engineering


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