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Biomaterials Science Vol -2

Biomaterials Science Vol -1

Enhancement of deoxyribozyme activity by cationic copolymers

Content Provider	Royal Society of Chemistry (RSC)
Author	Shimada, Naohiko Gao, Jueyuan Maruyama, Atsushi
Copyright Year	2015
Abstract	Deoxyribozymes, or DNAzymes, are DNA molecules with enzymatic activity. DNAzymes with ribonuclease activity have various potential applications in biomedical and bioanalytical fields; however, most constructs have limited turnover despite optimization of reaction conditions and DNAzyme structures. A cationic comb-type copolymer accelerates DNA hybridization and strand exchange rates, and we hypothesized that the copolymer would enhance deoxyribozyme activity by promoting turnover. The copolymer did not change DNAzyme activity under single-turnover conditions, suggesting that the copolymer affects neither the folding structure of DNAzyme nor the association of a divalent cation, a catalytic cofactor, to DNAzyme. The copolymer enhanced activity of the evaluated DNAzyme over a wide temperature range under multiple-turnover conditions. The copolymer increased the DNAzyme kcat/KM by fifty-fold at 50 °C, the optimal temperature for the DNAzyme in the absence of the copolymer. The acceleration effect was most significant when the reaction temperature was slightly higher than the melting temperature of the enzyme/substrate complex; acceleration of two orders of magnitude was observed. We concluded that the copolymer accelerated the turnover step without influencing the chemical cleavage step. In contrast to the copolymer, a cationic surfactant, CTAB, strongly inhibited the DNAzyme activity under either single- or multiple-turnover conditions.
Starting Page	308
Ending Page	316
Page Count	9
File Format	HTM / HTML PDF
ISSN	20474830
Volume Number	3
Issue Number	2
Journal	Biomaterials Science
DOI	10.1039/c4bm00256c
Language	English
Publisher	Royal Society of Chemistry
Access Restriction	Subscribed
Subject Keyword	Enzyme DNA Nucleic acid hybridization Specificity constant Cofactor (biochemistry) Ion Ribonuclease Copolymer Surfactant CTAB Mathematical optimization Deoxyribozyme
Content Type	Text
Resource Type	Article
Subject	Materials Science Biomedical Engineering

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