Genetically Expanded Reactive-Oxygen-Tolerant Alcohol Dehydrogenase II

Ashok Kumar Bhagat; Hadar Buium; Guy Shmul; Lital Alfonta

doi:10.1021/acscatal.9b03739

Genetically Expanded Reactive-Oxygen-Tolerant Alcohol Dehydrogenase II

Ashok Kumar Bhagat, Hadar Buium, Guy Shmul, Lital Alfonta

Department of Life Sciences

Research output: Contribution to journal › Article › peer-review

11 Scopus citations

Abstract

Herein, we report the site-specific incorporation of l-3,4-dihydroxyphenylalanine as a promising method to engineer an oxygen-tolerant alcohol dehydrogenase II. The engineered mutant alcohol dehydrogenase II binds Zn²⁺ with high binding affinity and is functional under aerobic and oxidative conditions for a longer time than the wild-type, Fe²⁺-binding alcohol dehydrogenase II. Overall, the mutant enzyme demonstrated electrochemical activity toward both acetaldehyde reduction and ethanol oxidation reactions. This enzyme could have a potential use in efficient biofuel production under aerobic conditions in photosynthetic organisms despite the inherent oxygen evolution reaction by photosystem II.

Original language	English
Pages (from-to)	3094-3102
Number of pages	9
Journal	ACS Catalysis
Volume	10
Issue number	5
DOIs	https://doi.org/10.1021/acscatal.9b03739
State	Published - 6 Mar 2020

Keywords

alcohol dehydrogenase II
ethanol biosynthesis
genetic code expansion
l-DOPA
oxygen tolerance
zinc-binding metalloproteins

ASJC Scopus subject areas

Catalysis
General Chemistry

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

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10.1021/acscatal.9b03739

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title = "Genetically Expanded Reactive-Oxygen-Tolerant Alcohol Dehydrogenase II",

abstract = "Herein, we report the site-specific incorporation of l-3,4-dihydroxyphenylalanine as a promising method to engineer an oxygen-tolerant alcohol dehydrogenase II. The engineered mutant alcohol dehydrogenase II binds Zn2+ with high binding affinity and is functional under aerobic and oxidative conditions for a longer time than the wild-type, Fe2+-binding alcohol dehydrogenase II. Overall, the mutant enzyme demonstrated electrochemical activity toward both acetaldehyde reduction and ethanol oxidation reactions. This enzyme could have a potential use in efficient biofuel production under aerobic conditions in photosynthetic organisms despite the inherent oxygen evolution reaction by photosystem II.",

keywords = "alcohol dehydrogenase II, ethanol biosynthesis, genetic code expansion, l-DOPA, oxygen tolerance, zinc-binding metalloproteins",

author = "Bhagat, {Ashok Kumar} and Hadar Buium and Guy Shmul and Lital Alfonta",

note = "Funding Information: We would like to gratefully acknowledge the help of Prof. Jeremy Mills, Prof. Raz Zarivach, and Dr. Sofia Kolusheva for helpful discussions and Prof. P. G. Schultz for a generous gift of the pAC-DHPheRS6TRN plasmid. We also thank Dr. Rambabu Dandela, Dr. Orr Schlesinger, Yonatan Chemla, Eden Ozer, Michael Shaferman, Richa Amiya, and Itay Algov for their technical assistance and advice. The support of parts of this study by an ISF-NCSF grant number 2920/19 is gratefully acknowledged. Publisher Copyright: Copyright {\textcopyright} 2020 American Chemical Society.",

year = "2020",

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doi = "10.1021/acscatal.9b03739",

language = "English",

volume = "10",

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AU - Bhagat, Ashok Kumar

AU - Buium, Hadar

AU - Shmul, Guy

AU - Alfonta, Lital

N1 - Funding Information: We would like to gratefully acknowledge the help of Prof. Jeremy Mills, Prof. Raz Zarivach, and Dr. Sofia Kolusheva for helpful discussions and Prof. P. G. Schultz for a generous gift of the pAC-DHPheRS6TRN plasmid. We also thank Dr. Rambabu Dandela, Dr. Orr Schlesinger, Yonatan Chemla, Eden Ozer, Michael Shaferman, Richa Amiya, and Itay Algov for their technical assistance and advice. The support of parts of this study by an ISF-NCSF grant number 2920/19 is gratefully acknowledged. Publisher Copyright: Copyright © 2020 American Chemical Society.

PY - 2020/3/6

Y1 - 2020/3/6

N2 - Herein, we report the site-specific incorporation of l-3,4-dihydroxyphenylalanine as a promising method to engineer an oxygen-tolerant alcohol dehydrogenase II. The engineered mutant alcohol dehydrogenase II binds Zn2+ with high binding affinity and is functional under aerobic and oxidative conditions for a longer time than the wild-type, Fe2+-binding alcohol dehydrogenase II. Overall, the mutant enzyme demonstrated electrochemical activity toward both acetaldehyde reduction and ethanol oxidation reactions. This enzyme could have a potential use in efficient biofuel production under aerobic conditions in photosynthetic organisms despite the inherent oxygen evolution reaction by photosystem II.

AB - Herein, we report the site-specific incorporation of l-3,4-dihydroxyphenylalanine as a promising method to engineer an oxygen-tolerant alcohol dehydrogenase II. The engineered mutant alcohol dehydrogenase II binds Zn2+ with high binding affinity and is functional under aerobic and oxidative conditions for a longer time than the wild-type, Fe2+-binding alcohol dehydrogenase II. Overall, the mutant enzyme demonstrated electrochemical activity toward both acetaldehyde reduction and ethanol oxidation reactions. This enzyme could have a potential use in efficient biofuel production under aerobic conditions in photosynthetic organisms despite the inherent oxygen evolution reaction by photosystem II.

KW - alcohol dehydrogenase II

KW - ethanol biosynthesis

KW - genetic code expansion

KW - l-DOPA

KW - oxygen tolerance

KW - zinc-binding metalloproteins

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DO - 10.1021/acscatal.9b03739

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JO - ACS Catalysis

JF - ACS Catalysis

IS - 5

ER -

Genetically Expanded Reactive-Oxygen-Tolerant Alcohol Dehydrogenase II

Abstract

Keywords

ASJC Scopus subject areas

UN SDGs

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