Tunneling of redox enzymes to design nano-probes for monitoring NAD+ dependent bio-catalytic activity.
IR@CFTRI: CSIR-Central Food Technological Research Institute, Mysore
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| Relation |
http://ir.cftri.com/12735/
http://dx.doi.org/10.1016/j.bios.2016.05.006 |
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| Title |
Tunneling of redox enzymes to design nano-probes for monitoring
NAD+ dependent bio-catalytic activity.
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| Creator |
Akshath, U. S.
Praveena, Bhatt |
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| Subject |
16 Enzyme Chemistry
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| Description |
Monitoring of bio-catalytic events by using nano-probes is of immense interest due to unique optical properties of metal nanoparticles. In the present study, tunneling of enzyme activity was achieved using redox cofactors namely oxidized cytochrome-c (Cyt-c) and Co-enzyme-Q (Co-Q) immobilized on Quantum dots (QDs) which acted as a bio-probe for NAD(+) dependent dehydrogenase catalyzed reaction. We studied how electron transfer from substrate to non-native electron acceptors can differentially modify photoluminescence properties of CdTe QDs. Two probes were designed, QD-Ox-Cyt-c and QD-Ox-Co-Q, which were found to quench the fluorescence of QDs. However, formaldehyde dehydrogenase (FDH) catalyzed reduction of Cyt-c and Co-Q on the surface of QDs lead to fluorescence turn-on of CdTe QDs. This phenomenon was successfully used for the detection of HCHO in the range of 0.01-100,000ng/mL (LOD of 0.01ng/mL) using both QD-Ox-Cyt-c (R(2)=0.93) and QD-Ox-Co-Q (R(2)=0.96). Further probe performance and stability in samples like milk, wine and fruit juice matrix were studied and we could detect HCHO in range of 0.001-100,000ng/mL (LOD of 0.001ng/mL) with good stability and sensitivity of probe in real samples (R(2)=0.97). Appreciable recovery and detection sensitivity in the presence of metal ions suggests that the developed nano-probes can be used successfully for monitoring dehydrogenase based bio-catalytic events even in the absence of NAD(+). Proposed method is advantageous over classical methods as clean up/ derivatization of samples is not required for formaldehyde detection.
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| Date |
2016
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| Type |
Article
PeerReviewed |
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| Format |
pdf
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| Language |
en
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| Identifier |
http://ir.cftri.com/12735/1/Biosensors%20and%20Bioelectronics%2085%282016%29%20240%E2%80%93246.pdf
Akshath, U. S. and Praveena, Bhatt (2016) Tunneling of redox enzymes to design nano-probes for monitoring NAD+ dependent bio-catalytic activity. Biosensors and Bioelectronics, 85. pp. 240-246. ISSN 0956-5663 |
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