Identification and characterization of a phosphinothricin N-acetyltransferase from Enterobacter LSJC7

Feng Zhu; Yu Yan; Xi-Mei Xue; Rui-Lian Yu; Jun Ye

doi:10.1016/j.pestbp.2023.105464

Identification and characterization of a phosphinothricin N-acetyltransferase from Enterobacter LSJC7

Pestic Biochem Physiol. 2023 Jun:193:105464. doi: 10.1016/j.pestbp.2023.105464. Epub 2023 May 10.

Authors

Feng Zhu¹, Yu Yan², Xi-Mei Xue³, Rui-Lian Yu⁴, Jun Ye⁵

Affiliations

¹ Department of Environmental Science and Engineering, Huaqiao University, Xiamen 361021, China; Key Lab of Urban Environment and Health, Institute of Urban Environment, Chinese Academy of Sciences, Xiamen 361021, China.
² Department of Environmental Science and Engineering, Huaqiao University, Xiamen 361021, China.
³ Key Lab of Urban Environment and Health, Institute of Urban Environment, Chinese Academy of Sciences, Xiamen 361021, China.
⁴ Department of Environmental Science and Engineering, Huaqiao University, Xiamen 361021, China. Electronic address: ruiliany@hqu.edu.cn.
⁵ School of Life Sciences, Institute of Life Sciences and Green Development, Hebei University, Baoding 071002, China. Electronic address: jye@hbu.edu.cn.

PMID: 37247996
DOI: 10.1016/j.pestbp.2023.105464

Abstract

Phosphinothricin (PPT) is a widely used and non-selective herbicide. PPT-resistance genes, especially PPT N-acetyltransferase genes, have been used in the development of transgenic PPT-resistant crops. However, there are only a limited number of available PPT-resistance genes for use in plant biotechnology. In this study, we found that Enterobacter LSJC7 is highly resistant to PPT and can acetylate PPT to N-acetyl phosphinothricin (Ac-PPT). Furthermore, a novel PPT N-acetyltransferase gene, named LsarsN, was identified from LSJC7. When LsarsN was expressed in E. coli AW3110, it confered resistance to PPT. Ac-PPT was detected in both the culture medium and cells of AW3110 expressing the LsarsN-pET22b plasmid. The purified LsArsN protein also showed strong N-acetylation ability in vitro, and its enzymatic kinetic curve was fitted with the Michaelis-Mentan equation. Compared with wild-type LsArsN, both R72A and R74A mutants showed significantly lower PPT N-acetylation ability. In summary, our results systematically characterized LsArsN with strong ability for PPT N-acetylation, which lays the groundwork for future research into the use of this novel gene, LsarsN, to create PPT-resistant crops.

Keywords: Herbicide; LSJC7; N-acetyltasnferase; Phosphinothricin.

MeSH terms

Acetyltransferases / genetics
Acetyltransferases / metabolism
Aminobutyrates* / metabolism
Escherichia coli* / genetics
Plants, Genetically Modified / metabolism

Substances

phosphinothricin
phosphinothricin N-acetyltransferase
Aminobutyrates
Acetyltransferases