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DC Field | Value | Language |
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dc.citation.endPage | 533 | - |
dc.citation.number | 4 | - |
dc.citation.startPage | 528 | - |
dc.citation.title | JOURNAL OF BIOSCIENCE AND BIOENGINEERING | - |
dc.citation.volume | 123 | - |
dc.contributor.author | Park, Eul-Soo | - |
dc.contributor.author | Park, Sunghoon | - |
dc.contributor.author | Shin, Jong-Shik | - |
dc.date.accessioned | 2023-12-21T22:18:48Z | - |
dc.date.available | 2023-12-21T22:18:48Z | - |
dc.date.created | 2017-06-14 | - |
dc.date.issued | 2017-04 | - |
dc.description.abstract | Glycerol dehydratase (GDHt) is a pivotal enzyme for fermentative utilization of glycerol by catalyzing radical-mediated conversion of glycerol into 3-hydroxypropionaldehyde (3-HPA). Precise and sensitive monitoring of cellular GDHt activity during the fermentation process is a prerequisite for reliable metabolic analysis to afford efficient cellular engineering and process optimization. Here we report a new spectrophotometric assay for the sensitive measurement of the GDHt activity with a sub-nanomolar limit of detection (LOD). The assay method employs aldehyde dehydrogenase (ALDH) as a reporter enzyme, so the readout of the GDHt activity is recorded at 340 nm as an increase in UV absorbance which results from NADH generation accompanied by oxidation of 3-HPA to 3-hydroxypropionic acid (3-HP). The GDHt assay was performed under the reaction conditions where the ALDH activity overwhelms the GDHt activity (i.e., 50-fold higher activity of ALDH relative to GDHt activity), affording sensitive detection of GDHt with 360 pM LOD. The ALDH-coupled assay was used to determine kinetic parameters of GDHt for glycerol, leading to K-M = 0.73 +/- 0.09 mM and k(cat) = 400 +/- 20 s(-1) which are in reasonable agreements with the previous reports. Our assay method allowed measurement of even a 10(4)-fold decrease in the cellular GDHt activity during fermentative production of 3-HP, which demonstrates the detection sensitivity much higher than the previous methods. | - |
dc.identifier.bibliographicCitation | JOURNAL OF BIOSCIENCE AND BIOENGINEERING, v.123, no.4, pp.528 - 533 | - |
dc.identifier.doi | 10.1016/j.jbiosc.2016.12.002 | - |
dc.identifier.issn | 1389-1723 | - |
dc.identifier.scopusid | 2-s2.0-85008402643 | - |
dc.identifier.uri | https://scholarworks.unist.ac.kr/handle/201301/22227 | - |
dc.identifier.url | http://www.sciencedirect.com/science/article/pii/S1389172316302699 | - |
dc.identifier.wosid | 000401048800018 | - |
dc.language | 영어 | - |
dc.publisher | SOC BIOSCIENCE BIOENGINEERING JAPAN | - |
dc.title | Spectrophotometric assay for sensitive detection of glycerol dehydratase activity using aldehyde dehydrogenase | - |
dc.type | Article | - |
dc.description.journalRegisteredClass | scie | - |
dc.description.journalRegisteredClass | scopus | - |
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