Characterization and identification of three novel aldo-keto reductases from Lodderomyces elongisporus for reducing ethyl 4-chloroacetoacetate

Chenxi Ning; Erzheng Su; Dongzhi Wei

doi:10.1016/j.abb.2014.10.007

Characterization and identification of three novel aldo-keto reductases from Lodderomyces elongisporus for reducing ethyl 4-chloroacetoacetate

Arch Biochem Biophys. 2014 Dec 15:564:219-28. doi: 10.1016/j.abb.2014.10.007. Epub 2014 Oct 22.

Authors

Chenxi Ning¹, Erzheng Su², Dongzhi Wei³

Affiliations

¹ State Key Laboratory of Bioreactor Engineering, New World Institute of Biotechnology, East China University of Science and Technology, Shanghai 200237, PR China.
² Enzyme and Fermentation Technology Laboratory, College of Light Industry Science and Engineering, Nanjing Forestry University, Nanjing 210037, PR China. Electronic address: ezhsu@njfu.edu.cn.
³ State Key Laboratory of Bioreactor Engineering, New World Institute of Biotechnology, East China University of Science and Technology, Shanghai 200237, PR China. Electronic address: dzhwei@ecust.edu.cn.

PMID: 25447817
DOI: 10.1016/j.abb.2014.10.007

Abstract

Lodderomyces elongisporus LH703 isolated from soil samples contained three novel aldo-keto reductases (AKRs) (LEAKR 48, LEAKR 49, and LEAKR 50). The three enzymes were cloned, expressed, and purified to homogeneity for characterization. These three AKRs shared <40% amino acid identity with each other. LEAKR 50 was identified as a member of AKR3 family, whereas the other two LEAKRs were identified as members of two novel AKR families, respectively. All the three AKRs required nicotinamide adenine dinucleotide phosphate as a cofactor. However, they showed diverse characteristics, including optimum catalyzing conditions, resistance to adverse reaction conditions, and substrate specificity. LEAKR 50 was estimated to be a promising biocatalyst that could reduce ethyl 4-chloroacetoacetate with high enantiomeric excess (98% e. e.) and high activity residue under adverse conditions.

Keywords: Aldo–keto reductase; Characterization; Ethyl 4-chloroacetoacetate; Identification; Lodderomyces elongisporus.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Acetoacetates / chemistry*
Aldehyde Reductase / chemistry*
Aldehyde Reductase / genetics
Aldo-Keto Reductases
Amino Acid Sequence
Cloning, Molecular
Fungal Proteins / chemistry*
Fungal Proteins / genetics
Molecular Sequence Data
NAD / chemistry*
Saccharomycopsis / enzymology*
Saccharomycopsis / genetics

Substances

Acetoacetates
Fungal Proteins
NAD
ethyl 4-chloro-3-oxobutanoate
Aldo-Keto Reductases
Aldehyde Reductase