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Repurposing Photosensitizer Proteins Through Genetic Code Expansion to Facilitate Photo-Biocatalysis

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Genetically Incorporated Non-Canonical Amino Acids

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2676))

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Abstract

Artificial photoenzymes with noncanonical photo-redox cofactors have paved the way for enzyme rational design and the creation of new-to-nature biocatalysts. Genetically encoded photo-redox cofactors endow photoenzymes with enhanced or novel activities that catalyze numerous transformations with high efficiency. Herein, we describe a protocol of repurposing photosensitizer proteins (PSP) through genetic code expansion to facilitate multiple photocatalytic conversions including photo-activated dehalogenation of aryl halides, CO2 to CO and CO2 to formic acid reduction. The methods for expression, purification, and characterization of the PSP are detailed. The installation of the catalytic modules and the utilization of PSP-based artificial photoenzymes for photoenzymatic CO2 reduction and dehalogenation are also described.

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Acknowledgments

The authors are grateful to the National Key R&D Program of China (2021YFA0910802, 2019YFA0904002, 2019YFA0904103, 2020YFA0908503, 2020YFA0907701) and Sanming Project of Medicine in Shenzhen (no. SZSM201811092).

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Author notes

  1. Yan **a and Xuzhen Guo contributed equally to this work

Authors and Affiliations

  1. Key Laboratory of RNA Biology, Institute of Biophysics, Chinese Academy of Sciences, Bei**g, China

    Jiangyun Wang & Yan **a

  2. CAS Key Laboratory of Quantitative Engineering Biology, Institute of Synthetic Biology, Shenzhen Institute of Advanced Technology, Chinese Academy of Sciences, Shenzhen, China

    Jiangyun Wang & Xuzhen Guo

  3. School of Future Technology, University of Chinese Academy of Sciences, Bei**g, China

    Jiangyun Wang & Yan **a

Authors
  1. Jiangyun Wang
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  2. Yan **a
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  3. Xuzhen Guo
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Correspondence to Jiangyun Wang .

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Editors and Affiliations

  1. Institute of Molecular Physiology, Shenzhen Bay Laboratory, Shenzhen, China

    Yu-Hsuan Tsai

  2. Laboratory of Synthetic and Systems Biology, Science for Life Laboratory, Department of Medical Biochemistry and Biophysics, Division of Genome Biology, Karolinska Institutet, Solna, Stockholm, Sweden

    Simon J. Elsässer

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Wang, J., **a, Y., Guo, X. (2023). Repurposing Photosensitizer Proteins Through Genetic Code Expansion to Facilitate Photo-Biocatalysis. In: Tsai, YH., Elsässer, S.J. (eds) Genetically Incorporated Non-Canonical Amino Acids. Methods in Molecular Biology, vol 2676. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-3251-2_3

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  • DOI: https://doi.org/10.1007/978-1-0716-3251-2_3

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-3250-5

  • Online ISBN: 978-1-0716-3251-2

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