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DC Field | Value | Language |
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dc.citation.endPage | 4454 | - |
dc.citation.number | 9 | - |
dc.citation.startPage | 4447 | - |
dc.citation.title | ANALYTICAL CHEMISTRY | - |
dc.citation.volume | 86 | - |
dc.contributor.author | Oliver-Calixte, Nyoté J. | - |
dc.contributor.author | Uba, Franklin I. | - |
dc.contributor.author | Battle, Katrina N. | - |
dc.contributor.author | Weerakoon-Ratnayake, Kumuditha M. | - |
dc.contributor.author | Soper, Steven A. | - |
dc.date.accessioned | 2023-12-22T02:41:23Z | - |
dc.date.available | 2023-12-22T02:41:23Z | - |
dc.date.created | 2014-05-26 | - |
dc.date.issued | 2014-05 | - |
dc.description.abstract | The process of immobilizing enzymes onto solid supports for bioreactions has some compelling advantages compared to their solution-based counterpart including the facile separation of enzyme from products, elimination of enzyme autodigestion, and increased enzyme stability and activity. We report the immobilization of λ-exonuclease onto poly(methylmethacrylate) (PMMA) micropillars populated within a microfluidic device for the on-chip digestion of double-stranded DNA. Enzyme immobilization was successfully accomplished using 3-(3-dimethylaminopropyl) carbodiimide/N-hydroxysuccinimide (EDC/NHS) coupling to carboxylic acid functionalized PMMA micropillars. Our results suggest that the efficiency for the catalysis of dsDNA digestion using λ-exonuclease, including its processivity and reaction rate, were higher when the enzyme was attached to a solid support compared to the free solution digestion. We obtained a clipping rate of 1.0 × 103 nucleotides s-1 for the digestion of λ-DNA (48.5 kbp) by λ-exonuclease. The kinetic behavior of the solid-phase reactor could be described by a fractal Michaelis-Menten model with a catalytic efficiency nearly 17% better than the homogeneous solution-phase reaction. The results from this work will have important ramifications in new single-molecule DNA sequencing strategies that employ free mononucleotide identification. | - |
dc.identifier.bibliographicCitation | ANALYTICAL CHEMISTRY, v.86, no.9, pp.4447 - 4454 | - |
dc.identifier.doi | 10.1021/ac5002965 | - |
dc.identifier.issn | 0003-2700 | - |
dc.identifier.scopusid | 2-s2.0-84899876589 | - |
dc.identifier.uri | https://scholarworks.unist.ac.kr/handle/201301/4769 | - |
dc.identifier.url | http://www.scopus.com/inward/record.url?partnerID=HzOxMe3b&scp=84899876589 | - |
dc.identifier.wosid | 000335719900054 | - |
dc.language | 영어 | - |
dc.publisher | AMER CHEMICAL SOC | - |
dc.title | Immobilization of lambda exonuclease onto polymer micropillar arrays for the solid-phase digestion of dsDNAs | - |
dc.type | Article | - |
dc.description.isOpenAccess | FALSE | - |
dc.relation.journalWebOfScienceCategory | Chemistry, Analytical | - |
dc.relation.journalResearchArea | Chemistry | - |
dc.description.journalRegisteredClass | scie | - |
dc.description.journalRegisteredClass | scopus | - |
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