| Title |
A DNA-scaffold platform enhances a multi-enzymatic cycling reaction
|
|---|---|
| Published in |
Biotechnology Techniques, January 2018
|
| DOI | 10.1007/s10529-018-2517-4 |
| Pubmed ID | |
| Authors |
Yasumasa Mashimo, Masayasu Mie, Eiry Kobatake |
| Abstract |
We explored the co-localization of multiple enzymes on a DNA backbone via a DNA-binding protein, Gene-A* (A*-tag) to increase the efficiency of cascade enzymatic reactions. Firefly luciferase (FLuc) and pyruvate orthophosphate dikinase (PPDK) were genetically fused with A*-tag and modified with single-stranded (ss) DNA via A*-tag. The components were assembled on ssDNA by hybridization, thereby enhancing the efficiency of the cascading bioluminescent reaction producing light emission from pyrophosphate. The activity of A*-tag in each enzyme was investigated with dye-labeled DNA. Co-localization of the enzymes via hybridization was examined using a gel shift assay. The multi-enzyme complex showed significant improvement in the overall efficiency of the cascading reaction in comparison to a mixture of free enzymes. A*-tag is highly convenient for ssDNA modification of versatile enzymes, and it can be used for construction of functional DNA-enzyme complexes. |
X Demographics
The data shown below were collected from the profiles of 2 X users who shared this research output. Click here to find out more about how the information was compiled.
Geographical breakdown
| Country | Count | As % |
|---|---|---|
| Unknown | 2 | 100% |
Demographic breakdown
| Type | Count | As % |
|---|---|---|
| Members of the public | 2 | 100% |
Mendeley readers
The data shown below were compiled from readership statistics for 12 Mendeley readers of this research output. Click here to see the associated Mendeley record.
Geographical breakdown
| Country | Count | As % |
|---|---|---|
| Unknown | 12 | 100% |
Demographic breakdown
| Readers by professional status | Count | As % |
|---|---|---|
| Student > Ph. D. Student | 3 | 25% |
| Student > Master | 2 | 17% |
| Professor | 1 | 8% |
| Student > Doctoral Student | 1 | 8% |
| Student > Bachelor | 1 | 8% |
| Other | 1 | 8% |
| Unknown | 3 | 25% |
| Readers by discipline | Count | As % |
|---|---|---|
| Chemistry | 3 | 25% |
| Biochemistry, Genetics and Molecular Biology | 2 | 17% |
| Unknown | 7 | 58% |
Attention Score in Context
This research output has an Altmetric Attention Score of 1. This is our high-level measure of the quality and quantity of online attention that it has received. This Attention Score, as well as the ranking and number of research outputs shown below, was calculated when the research output was last mentioned on 01 September 2018.
All research outputs
#20,663,600
of 25,382,440 outputs
Outputs from Biotechnology Techniques
#2,354
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Outputs of similar age
#344,091
of 450,227 outputs
Outputs of similar age from Biotechnology Techniques
#9
of 18 outputs
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