Chapter 5
Protein-Engineering Approach
for Improvement of DET-Type
Bioelectrocatalytic Performance
Abstract Recent advances in the area of protein-engineering have facilitated the
production of mutated proteins predesigned to enhance the electronic coupling with
electrodes. In this chapter, we will start from the motivation of protein engineering
and introduce typical protein-engineering strategies for improvement of DET-type
bioelectrocatalysis.
Keywords Protein-engineering · Mutants · Direct electron transfer ·
Bioelectrocatalysis
5.1 Motivation of Protein-Engineering
As discussed in Chap. 1, redox enzymes that are capable of direct electron transfer
with electrodes is very limited in number, because the redox active sites are in many
cases embedded by insulating peptides of the enzymes. Increasing attention in the
field of electrochemistry of redox enzymes is driven by designing efficient interfaces
for electron transfer between electrode surfaces and redox enzymes. Because a DETtype bioelectrocatalysis proceeds between an enzyme and an electrode, strategies
to improve the performance should be just considered in views of two directions:
enzymes and electrodes. The past decades have witnessed the development of various
electrode materials, such as carbon nanotubes, mesoporous carbon materials, and
metal nanoparticles, for improving the performance of DET-type bioelectrocatalysis
[1–6]. Such nanomaterials improved the interfacial electron transfer kinetics of redox
enzymes thanks to their nanostructures that increased the effective enzyme loading
[7, 8]. Besides, rationally designed electrode surfaces with special properties are also
widely studied to achieve productive orientation or attachment of enzymes [9–11].
In this chapter, we will focus on the protein-engineering approach for improvement of DET-type bioelectrocatalysis. Enzymes can be modified by several techniques of the protein-engineering. Remarkable progress has been made in proteinengineering of redox enzymes in the past decades and several reviews related to
the protein-engineering for bioelectrocatalysis and bioelectrochemistry are found in
the literature [12–15]. Protein-engineering through rational design, direct evolutions,
and combined approaches, has been successfully utilized to reform the enzymes with
© The Author(s), under exclusive license to Springer Nature Singapore Pte Ltd. 2021
K. Kano et al., Enzymatic Bioelectrocatalysis,
https://doi.org/10.1007/978-981-15-8960-7_5
93
Protein-Engineering Approach
for Improvement of DET-Type
Bioelectrocatalytic Performance
Abstract Recent advances in the area of protein-engineering have facilitated the
production of mutated proteins predesigned to enhance the electronic coupling with
electrodes. In this chapter, we will start from the motivation of protein engineering
and introduce typical protein-engineering strategies for improvement of DET-type
bioelectrocatalysis.
Keywords Protein-engineering · Mutants · Direct electron transfer ·
Bioelectrocatalysis
5.1 Motivation of Protein-Engineering
As discussed in Chap. 1, redox enzymes that are capable of direct electron transfer
with electrodes is very limited in number, because the redox active sites are in many
cases embedded by insulating peptides of the enzymes. Increasing attention in the
field of electrochemistry of redox enzymes is driven by designing efficient interfaces
for electron transfer between electrode surfaces and redox enzymes. Because a DETtype bioelectrocatalysis proceeds between an enzyme and an electrode, strategies
to improve the performance should be just considered in views of two directions:
enzymes and electrodes. The past decades have witnessed the development of various
electrode materials, such as carbon nanotubes, mesoporous carbon materials, and
metal nanoparticles, for improving the performance of DET-type bioelectrocatalysis
[1–6]. Such nanomaterials improved the interfacial electron transfer kinetics of redox
enzymes thanks to their nanostructures that increased the effective enzyme loading
[7, 8]. Besides, rationally designed electrode surfaces with special properties are also
widely studied to achieve productive orientation or attachment of enzymes [9–11].
In this chapter, we will focus on the protein-engineering approach for improvement of DET-type bioelectrocatalysis. Enzymes can be modified by several techniques of the protein-engineering. Remarkable progress has been made in proteinengineering of redox enzymes in the past decades and several reviews related to
the protein-engineering for bioelectrocatalysis and bioelectrochemistry are found in
the literature [12–15]. Protein-engineering through rational design, direct evolutions,
and combined approaches, has been successfully utilized to reform the enzymes with
© The Author(s), under exclusive license to Springer Nature Singapore Pte Ltd. 2021
K. Kano et al., Enzymatic Bioelectrocatalysis,
https://doi.org/10.1007/978-981-15-8960-7_5
93