Engineering and characterization of hybrid carboxylic acid reductases

Levi Kramer, Xuan Le, Erome D. Hankore, Mark A. Wilson, Jiantao Guo, Wei Niu

Research output: Contribution to journalArticle

Abstract

Carboxylic acid reductases (CARs) are valuable biocatalysts due to their ability to reduce a broad range of carboxylate substrates into the corresponding aldehyde products. CARs are multi-domain enzymes with separate catalytic domains for the adenylation and the subsequent reduction of substrates. Inter-domain dynamics are crucial for the catalytic activities of CARs. In this work, hybrid enzymes that contain domains from four bacterial CARs and one fungal CAR were constructed based on domain boundaries that were defined using a combination of bioinformatics and structural analysis. Hybrid CARs were characterized in both steady-state and transient kinetics studies using aromatic and straight-chain (C3-C5) carboxylate substrates. Kinetic data support that the inter-domain interactions play an important role in the function of both wild-type and hybrid CARs and further lead to the hypothesis that reduction is the rate-determining step in CAR catalysis. Our results provide both fundamental insights into CAR catalysis and rationale for hybrid CAR engineering.

Original languageEnglish (US)
Pages (from-to)52-56
Number of pages5
JournalJournal of Biotechnology
Volume304
DOIs
StatePublished - Oct 10 2019

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Carboxylic acids
Catalysis
Substrates
Enzymes
Oxidoreductases
carboxylic acid reductase
Biocatalysts
Kinetics
Bioinformatics
Computational Biology
Aldehydes
Structural analysis
Catalyst activity
Catalytic Domain

Keywords

  • Carboxylic acid reductase
  • Hybrid enzymes
  • Inter-domain interaction
  • Kinetics

ASJC Scopus subject areas

  • Biotechnology
  • Bioengineering
  • Applied Microbiology and Biotechnology

Cite this

Engineering and characterization of hybrid carboxylic acid reductases. / Kramer, Levi; Le, Xuan; Hankore, Erome D.; Wilson, Mark A.; Guo, Jiantao; Niu, Wei.

In: Journal of Biotechnology, Vol. 304, 10.10.2019, p. 52-56.

Research output: Contribution to journalArticle

Kramer, Levi ; Le, Xuan ; Hankore, Erome D. ; Wilson, Mark A. ; Guo, Jiantao ; Niu, Wei. / Engineering and characterization of hybrid carboxylic acid reductases. In: Journal of Biotechnology. 2019 ; Vol. 304. pp. 52-56.
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