caa a22 4500 605501513 CHVBK 20210128100601.0 cr unu---uuuuu 210128e20150901xx s 000 0 eng 10.1007/s00253-015-6469-5 doi (NATIONALLICENCE)springer-10.1007/s00253-015-6469-5 Mutational analysis to identify the residues essential for the inhibition of N- acetyl glutamate kinase of Corynebacterium glutamicum [Elektronische Daten] [Yuanyuan Huang, Hao Zhang, Hongming Tian, Cheng Li, Shuangyan Han, Ying Lin, Suiping Zheng] N-acetyl glutamate kinase (NAGK) is a key enzyme in the synthesis of l-arginine that is inhibited by its end product l-arginine in Corynebacterium glutamicum (C. glutamicum). In this study, the potential binding sites of arginine and the residues essential for its inhibition were identified by homology modeling, inhibitor docking, and site-directed mutagenesis. The allosteric inhibition of NAGK was successfully alleviated by a mutation, as determined through analysis of mutant enzymes, which were overexpressed in vivo in C. glutamicum ATCC14067. Analysis of the mutant enzymes and docking analysis demonstrated that residue W23 positions an arginine molecule, and the interaction between arginine and residues L282, L283, and T284 may play an important role in the remote inhibitory process. Based on the results of the docking analysis of the effective mutants, we propose a linkage mechanism for the remote allosteric regulation of NAGK activity, in which residue R209 may play an essential role. In this study, the structure of the arginine-binding site of C. glutamicum NAGK (CgNAGK) was successfully predicted and the roles of the relevant residues were identified, providing new insight into the allosteric regulation of CgNAGK activity and a solid platform for the future construction of an optimized l-arginine producing strain. Springer-Verlag Berlin Heidelberg, 2015 N- acetyl glutamate kinase nationallicence Homology modeling nationallicence Corynebacterium glutamicum nationallicence l -arginine nationallicence Allosteric regulation nationallicence Huang Yuanyuan Guangdong Key Laboratory of Fermentation and Enzyme Engineering, School of Bioscience and Bioengineering, South China University of Technology, 510006, Guangzhou, People's Republic of China aut Zhang Hao Guangdong Key Laboratory of Fermentation and Enzyme Engineering, School of Bioscience and Bioengineering, South China University of Technology, 510006, Guangzhou, People's Republic of China aut Tian Hongming Guangdong Key Laboratory of Fermentation and Enzyme Engineering, School of Bioscience and Bioengineering, South China University of Technology, 510006, Guangzhou, People's Republic of China aut Li Cheng Guangdong Key Laboratory of Fermentation and Enzyme Engineering, School of Bioscience and Bioengineering, South China University of Technology, 510006, Guangzhou, People's Republic of China aut Han Shuangyan Guangdong Key Laboratory of Fermentation and Enzyme Engineering, School of Bioscience and Bioengineering, South China University of Technology, 510006, Guangzhou, People's Republic of China aut Lin Ying Guangdong Key Laboratory of Fermentation and Enzyme Engineering, School of Bioscience and Bioengineering, South China University of Technology, 510006, Guangzhou, People's Republic of China aut Zheng Suiping Guangdong Key Laboratory of Fermentation and Enzyme Engineering, School of Bioscience and Bioengineering, South China University of Technology, 510006, Guangzhou, People's Republic of China aut Applied Microbiology and Biotechnology Springer Berlin Heidelberg 99/18(2015-09-01), 7527-7537 0175-7598 99:18<7527 2015 99 253 https://doi.org/10.1007/s00253-015-6469-5 text/html Onlinezugriff via DOI BK010053 XK010053 XK010000 Metadata rights reserved Springer special CC-BY-NC licence nationallicence 1 research-article jats NATIONALLICENCE NATIONALLICENCE NL-springer NATIONALLICENCE 856 40 https://doi.org/10.1007/s00253-015-6469-5 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Huang Yuanyuan Guangdong Key Laboratory of Fermentation and Enzyme Engineering, School of Bioscience and Bioengineering, South China University of Technology, 510006, Guangzhou, People's Republic of China aut NATIONALLICENCE 700 1- Zhang Hao Guangdong Key Laboratory of Fermentation and Enzyme Engineering, School of Bioscience and Bioengineering, South China University of Technology, 510006, Guangzhou, People's Republic of China aut NATIONALLICENCE 700 1- Tian Hongming Guangdong Key Laboratory of Fermentation and Enzyme Engineering, School of Bioscience and Bioengineering, South China University of Technology, 510006, Guangzhou, People's Republic of China aut NATIONALLICENCE 700 1- Li Cheng Guangdong Key Laboratory of Fermentation and Enzyme Engineering, School of Bioscience and Bioengineering, South China University of Technology, 510006, Guangzhou, People's Republic of China aut NATIONALLICENCE 700 1- Han Shuangyan Guangdong Key Laboratory of Fermentation and Enzyme Engineering, School of Bioscience and Bioengineering, South China University of Technology, 510006, Guangzhou, People's Republic of China aut NATIONALLICENCE 700 1- Lin Ying Guangdong Key Laboratory of Fermentation and Enzyme Engineering, School of Bioscience and Bioengineering, South China University of Technology, 510006, Guangzhou, People's Republic of China aut NATIONALLICENCE 700 1- Zheng Suiping Guangdong Key Laboratory of Fermentation and Enzyme Engineering, School of Bioscience and Bioengineering, South China University of Technology, 510006, Guangzhou, People's Republic of China aut NATIONALLICENCE 773 0- Applied Microbiology and Biotechnology Springer Berlin Heidelberg 99/18(2015-09-01), 7527-7537 0175-7598 99:18<7527 2015 99 253