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Synthetic biology is an interdisciplinary science that involves using principles from disciplines such as engineering, molecular biology, cell biology, and systems biology. It involves remodeling existing organisms from nature or constructing completely new synthetic organisms for applications such as protein or enzyme production, bioremediation, value-added macromolecule production, and the addition of desirable traits to crops, to name a few.
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Solid Phase 11C-Methylation, Purification and Formulation for the Production of PET Tracers
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Totally synthetic microperoxidase-11.

Junichi Tanabe1, Koji Nakano1, Ryutaro Hirata1

  • 1Department of Applied Chemistry, Faculty of Engineering, Kyushu University, 744 Motooka, Nishi-ku, Fukuoka 819-0395, Japan.

Royal Society Open Science
|June 13, 2018
PubMed
Summary

Researchers synthesized a fully artificial microperoxidase-11 (MP-11) using peptide synthesis and click chemistry. This synthetic MP-11 demonstrates comparable catalytic activity and electrochemical properties to its natural counterpart, offering a nature-independent preparation method.

Keywords:
electrocatalytic reactionenzymatic assayhigh-speed atomic force microscopymicroperoxidasespectroelectrochemistrytotal synthesis

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Area of Science:

  • Biochemistry
  • Synthetic Chemistry
  • Biomaterials

Background:

  • Microperoxidase-11 (MP-11) is a heme-containing peptide with peroxidase activity.
  • Natural MP-11 is typically obtained from cytochrome c, limiting its availability and modification.
  • Developing synthetic routes for MP-11 is crucial for broader applications and mutant studies.

Purpose of the Study:

  • To report the successful synthesis of a fully synthetic microperoxidase-11 (MP-11).
  • To characterize the catalytic activity and electrochemical properties of the synthetic MP-11.
  • To establish a convenient, nature-independent method for preparing MP-11 and its analogs.

Main Methods:

  • Solid-phase peptide synthesis of the undecapeptide VQKCAQCHTVE.
  • Thiol-ene click reaction for haemin reconstitution.
  • High-speed atomic force microscopy (HS-AFM) for structural confirmation.
  • Enzymatic assays (TMB oxidation) and electrochemical measurements (cyclic voltammetry).
  • Computer simulation for electrocatalytic reaction parameter analysis.

Main Results:

  • Successful synthesis and reconstitution of MP-11 using chemical methods.
  • Synthetic MP-11 exhibited significant catalytic activity (27% of natural MP-11) in TMB oxidation.
  • A point mutation (H8M) drastically reduced catalytic activity (2.7%).
  • Electrochemical studies showed direct electron transfer and catalytic current-voltage behavior comparable to natural MP-11.
  • Experimental data were well-reproduced by computer simulations, yielding consistent electrocatalytic parameters.

Conclusions:

  • A totally synthetic MP-11 has been successfully prepared via solid-phase peptide synthesis and haemin reconstitution.
  • The synthetic MP-11 displays comparable catalytic and electrochemical performance to the natural enzyme.
  • This synthetic approach provides a versatile and reliable method for producing MP-11 and its mutants without relying on natural sources.