Lag Time Spectrophotometric Assay for Studying Transport Limitation in Immobilized Enzymes

Matteo Grattieri, David P. Hickey, Han Sol Kim, Vanesa Teijeiro Seijas, Jungbae Kim, Shelley D. Minteer

Research output: Contribution to journalArticlepeer-review

1 Citation (Scopus)


Enzymes are promising catalysts for bioprocessing. For instance, the enzymatic capture of CO2 using carbonic anhydrase (CA) is a carbon capture approach that allows obtaining bicarbonate (HCO3 -) with no high-energy input required. However, application in a commercially viable biotechnology requires sufficient enzymatic lifetime. Although enzyme stabilization can be achieved by different immobilization techniques, most of them are not commercially viable because of transport limitations induced by the immobilization method. Therefore, it is necessary to develop assays for evaluating the role of immobilization on transport limitations. Herein, we describe the development of a fast and reproducible assay for screening immobilized CA by means of absorbance measurement using a computer-controlled microplate reader in stop-flow format. The automated assay allowed minimizing the required volume for analysis to 120 μL. We validated the assay by determining lag times and activities for three immobilization techniques (modified Nafion, hydrogels, and enzyme precipitates), of which linear polyethyleneimine hydrogel showed outstanding performance for CA immobilization.

Original languageEnglish
Pages (from-to)11945-11949
Number of pages5
JournalACS Omega
Issue number9
Publication statusPublished - 2018 Sept 30

Bibliographical note

Funding Information:
This work was 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 work was also supported by the Global Research Laboratory Program (2014K1A1A2043032) through the National Research Foundation of Korea (NRF) grants funded by the Korea government Ministry of Science and ICT (MSIT).

Publisher Copyright:
Copyright © 2018 American Chemical Society.

ASJC Scopus subject areas

  • General Chemistry
  • General Chemical Engineering


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