Abstract
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 language | English |
|---|---|
| Pages (from-to) | 11945-11949 |
| Number of pages | 5 |
| Journal | ACS Omega |
| Volume | 3 |
| Issue number | 9 |
| DOIs | |
| Publication status | Published - 2018 Sep 30 |
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ASJC Scopus subject areas
- Chemistry(all)
- Chemical Engineering(all)
Cite this
Lag Time Spectrophotometric Assay for Studying Transport Limitation in Immobilized Enzymes. / Grattieri, Matteo; Hickey, David P.; Kim, Han Sol; Seijas, Vanesa Teijeiro; Kim, Jungbae; Minteer, Shelley D.
In: ACS Omega, Vol. 3, No. 9, 30.09.2018, p. 11945-11949.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Lag Time Spectrophotometric Assay for Studying Transport Limitation in Immobilized Enzymes
AU - Grattieri, Matteo
AU - Hickey, David P.
AU - Kim, Han Sol
AU - Seijas, Vanesa Teijeiro
AU - Kim, Jungbae
AU - Minteer, Shelley D.
PY - 2018/9/30
Y1 - 2018/9/30
N2 - 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.
AB - 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.
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U2 - 10.1021/acsomega.8b01527
DO - 10.1021/acsomega.8b01527
M3 - Article
AN - SCOPUS:85054200474
VL - 3
SP - 11945
EP - 11949
JO - ACS Omega
JF - ACS Omega
SN - 2470-1343
IS - 9
ER -