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Ultrafast sonochemical synthesis of protein-inorganic nanoflowers.

Batule BS, Park KS, Kim MI, Park HG - Int J Nanomedicine (2015)

Bottom Line: With this technique, we synthesized nanoflowers containing laccase as a model protein and copper phosphate within 5 minutes at room temperature.The resulting laccase nanoflowers yielded greatly enhanced activity, stability, and reusability, and their usefulness was successfully demonstrated by applying them in the colorimetric detection of epinephrine.The strategy developed could be used to rapidly synthesize nanoflowers for various applications in biosensor and enzyme catalysis and would expand the utilization of nanoflowers in diverse fields of biotechnology.

View Article: PubMed Central - PubMed

Affiliation: Department of Chemical and Biomolecular Engineering (BK21+ Program), Korea Advanced Institute of Science and Technology (KAIST), Daejeon, Republic of Korea.

ABSTRACT
We developed a simple but efficient method to synthesize protein-inorganic hybrid nanostructures with a flower-like shape (nanoflowers), which relies on sonication to facilitate the synthesis of the nanoflowers. With this technique, we synthesized nanoflowers containing laccase as a model protein and copper phosphate within 5 minutes at room temperature. The resulting laccase nanoflowers yielded greatly enhanced activity, stability, and reusability, and their usefulness was successfully demonstrated by applying them in the colorimetric detection of epinephrine. The strategy developed could be used to rapidly synthesize nanoflowers for various applications in biosensor and enzyme catalysis and would expand the utilization of nanoflowers in diverse fields of biotechnology.

No MeSH data available.


Enhanced performance of laccase nanoflowers synthesized by sonication-based method. (A) Measurement of laccase activity by employing 40 μM syringaldazine as a substrate in phosphate-buffered saline (PBS) buffer (pH 7.4) at 25°C. (B) Visual detection of epinephrine by using sonicated laccase nanoflowers and free laccase. (C) Stability of sonicated laccase nanoflowers and free laccase for detecting epinephrine in PBS (pH 7.4) at 25°C. (D) Reusability of the sonicated laccase nanoflowers for detecting epinephrine.Note: In (B), 25.0, 12.5, 5.0, 2.5, and 1.0 μg·mL−1 of epinephrine in PBS (pH 7.4) are denoted by 1, 2, 3, 4, and 5, respectively.Abbreviations: F, free laccase (30 μg·mL−1); N, sonicated laccase nanoflowers.
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f5-ijn-10-137: Enhanced performance of laccase nanoflowers synthesized by sonication-based method. (A) Measurement of laccase activity by employing 40 μM syringaldazine as a substrate in phosphate-buffered saline (PBS) buffer (pH 7.4) at 25°C. (B) Visual detection of epinephrine by using sonicated laccase nanoflowers and free laccase. (C) Stability of sonicated laccase nanoflowers and free laccase for detecting epinephrine in PBS (pH 7.4) at 25°C. (D) Reusability of the sonicated laccase nanoflowers for detecting epinephrine.Note: In (B), 25.0, 12.5, 5.0, 2.5, and 1.0 μg·mL−1 of epinephrine in PBS (pH 7.4) are denoted by 1, 2, 3, 4, and 5, respectively.Abbreviations: F, free laccase (30 μg·mL−1); N, sonicated laccase nanoflowers.

Mentions: The catalytic activity of the sonicated laccase nanoflowers synthesized under the optimal condition was then evaluated. In a typical activity assay using syringaldazine as a substrate, the sonicated laccase nanoflowers showed ~150% activity compared to the free laccase (Figure 5A), confirming that the synthesizing of laccase nanoflowers by simple sonication also effectively enhanced their catalytic activity like the conventional method.11 The enhanced catalytic activity of sonicated laccase nanoflowers could be a very distinctive advantage considering that the immobilization of enzymes generally leads to lowered activity.23 We also confirmed that the sonication treatment of free laccase for 5 minutes did not deteriorate the enzyme activity (Figure 5A).


Ultrafast sonochemical synthesis of protein-inorganic nanoflowers.

Batule BS, Park KS, Kim MI, Park HG - Int J Nanomedicine (2015)

Enhanced performance of laccase nanoflowers synthesized by sonication-based method. (A) Measurement of laccase activity by employing 40 μM syringaldazine as a substrate in phosphate-buffered saline (PBS) buffer (pH 7.4) at 25°C. (B) Visual detection of epinephrine by using sonicated laccase nanoflowers and free laccase. (C) Stability of sonicated laccase nanoflowers and free laccase for detecting epinephrine in PBS (pH 7.4) at 25°C. (D) Reusability of the sonicated laccase nanoflowers for detecting epinephrine.Note: In (B), 25.0, 12.5, 5.0, 2.5, and 1.0 μg·mL−1 of epinephrine in PBS (pH 7.4) are denoted by 1, 2, 3, 4, and 5, respectively.Abbreviations: F, free laccase (30 μg·mL−1); N, sonicated laccase nanoflowers.
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f5-ijn-10-137: Enhanced performance of laccase nanoflowers synthesized by sonication-based method. (A) Measurement of laccase activity by employing 40 μM syringaldazine as a substrate in phosphate-buffered saline (PBS) buffer (pH 7.4) at 25°C. (B) Visual detection of epinephrine by using sonicated laccase nanoflowers and free laccase. (C) Stability of sonicated laccase nanoflowers and free laccase for detecting epinephrine in PBS (pH 7.4) at 25°C. (D) Reusability of the sonicated laccase nanoflowers for detecting epinephrine.Note: In (B), 25.0, 12.5, 5.0, 2.5, and 1.0 μg·mL−1 of epinephrine in PBS (pH 7.4) are denoted by 1, 2, 3, 4, and 5, respectively.Abbreviations: F, free laccase (30 μg·mL−1); N, sonicated laccase nanoflowers.
Mentions: The catalytic activity of the sonicated laccase nanoflowers synthesized under the optimal condition was then evaluated. In a typical activity assay using syringaldazine as a substrate, the sonicated laccase nanoflowers showed ~150% activity compared to the free laccase (Figure 5A), confirming that the synthesizing of laccase nanoflowers by simple sonication also effectively enhanced their catalytic activity like the conventional method.11 The enhanced catalytic activity of sonicated laccase nanoflowers could be a very distinctive advantage considering that the immobilization of enzymes generally leads to lowered activity.23 We also confirmed that the sonication treatment of free laccase for 5 minutes did not deteriorate the enzyme activity (Figure 5A).

Bottom Line: With this technique, we synthesized nanoflowers containing laccase as a model protein and copper phosphate within 5 minutes at room temperature.The resulting laccase nanoflowers yielded greatly enhanced activity, stability, and reusability, and their usefulness was successfully demonstrated by applying them in the colorimetric detection of epinephrine.The strategy developed could be used to rapidly synthesize nanoflowers for various applications in biosensor and enzyme catalysis and would expand the utilization of nanoflowers in diverse fields of biotechnology.

View Article: PubMed Central - PubMed

Affiliation: Department of Chemical and Biomolecular Engineering (BK21+ Program), Korea Advanced Institute of Science and Technology (KAIST), Daejeon, Republic of Korea.

ABSTRACT
We developed a simple but efficient method to synthesize protein-inorganic hybrid nanostructures with a flower-like shape (nanoflowers), which relies on sonication to facilitate the synthesis of the nanoflowers. With this technique, we synthesized nanoflowers containing laccase as a model protein and copper phosphate within 5 minutes at room temperature. The resulting laccase nanoflowers yielded greatly enhanced activity, stability, and reusability, and their usefulness was successfully demonstrated by applying them in the colorimetric detection of epinephrine. The strategy developed could be used to rapidly synthesize nanoflowers for various applications in biosensor and enzyme catalysis and would expand the utilization of nanoflowers in diverse fields of biotechnology.

No MeSH data available.