Abstract
In nature, enzymes often form multienzyme complexes to enhance their catalytic efficiencies and, moreover, evolve into genetically fused multidomain enzymes. Inspired by a natural fusion cytochrome P450 (P450) containing a monooxygenase domain and a reductase domain, we have developed a heterotrimeric protein-utilized method to form a multienzyme complex composed of a bacterial P450 and its catalytically essential two redox proteins. Three distinct proliferating cell nuclear antigens (PCNAs) from Sulfolobus solfataricus, each of which can be separately expressed, spontaneously form a heterotrimer. Fusion to the PCNAs enables complex formation of a bacterial P450 and two redox proteins through the self-assembling of the PCNAs and enhances the activity due to efficient electron transfer in the complex. This PCNA-mediated multienzyme complex formation will be available for other multienzyme reactions.
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Acknowledgements
We gratefully acknowledge the support from a Grant-in-Aid for Young Scientists (B) from the Ministry of Education, Culture, Sports, Science and Technology (MEXT) and the support from the Center for NanoBio Integration (Research and Development in a New Converting Field Based on Nanotechnology and Materials Science, MEXT).
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Hirakawa, H., Nagamune, T. (2013). Use of Sulfolobus solfataricus PCNA Subunit Proteins to Direct the Assembly of Multimeric Enzyme Complexes. In: Samuelson, J. (eds) Enzyme Engineering. Methods in Molecular Biology, vol 978. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-293-3_11
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DOI: https://doi.org/10.1007/978-1-62703-293-3_11
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Publisher Name: Humana Press, Totowa, NJ
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Online ISBN: 978-1-62703-293-3
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