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Issue 2, 2015
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Enhancement of deoxyribozyme activity by cationic copolymers

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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.

Graphical abstract: Enhancement of deoxyribozyme activity by cationic copolymers

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Publication details

The article was received on 23 Jul 2014, accepted on 11 Sep 2014 and first published on 03 Oct 2014


Article type: Paper
DOI: 10.1039/C4BM00256C
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Citation: Biomater. Sci., 2015,3, 308-316
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    Enhancement of deoxyribozyme activity by cationic copolymers

    J. Gao, N. Shimada and A. Maruyama, Biomater. Sci., 2015, 3, 308
    DOI: 10.1039/C4BM00256C

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