caa a22 4500 605502587 CHVBK 20210128100606.0 cr unu---uuuuu 210128e20151201xx s 000 0 eng 10.1007/s00253-015-6967-5 doi (NATIONALLICENCE)springer-10.1007/s00253-015-6967-5 Domain function dissection and catalytic properties of Listeria monocytogenes p60 protein with bacteriolytic activity [Elektronische Daten] [Minfeng Yu, Jinrong Zuo, Hao Gu, Minliang Guo, Yuelan Yin] The major extracellular protein p60 of Listeria monocytogenes (Lm-p60) is an autolysin that can hydrolyze the peptidoglycan of bacterial cell wall and has been shown to be required for L. monocytogenes virulence. The predicted three-dimensional structure of Lm-p60 showed that Lm-p60 could be split into two independent structural domains at the amino acid residue 270. Conserved motif analysis showed that V30, D207, S395, and H444 are the key amino acid residues of the corresponding motifs. However, not only the actual functions of these two domains but also the catalytic properties of Lm-p60 are unclear. We try to express recombinant Lm-p60 and identify the functions of two domains by residue substitution (V30A, D207A, S395A, and H444A) and peptide truncation. The C-terminal domain was identified as catalytic element and N-terminal domain as substrate recognition and binding element. Either N-terminal domain truncation or C-terminal domain truncation presents corresponding biological activity. The catalytic activity of Lm-p60 with a malfunctioned substrate-binding domain was decreased, while the substrate binding was not affected by a mulfunctioned catalytic domain. With turbidimetric method, we determined the optimal conditions for the bacteriolytic activity of Lm-p60 against Micrococcus lysodeikficus. The assay for the effect of Lm-p60 on the bacteriolytic activity of lysozyme revealed that the combined use of Lm-p60 protein with lysozyme showed a strong synergistic effect on the bacteriolytic activity. Springer-Verlag Berlin Heidelberg, 2015 Domain function nationallicence Listeria monocytogenes p60 protein nationallicence Peptidoglycan hydrolase nationallicence LysM domain nationallicence NlpC/P60 domain nationallicence Yu Minfeng College of Bioscience and Biotechnology, Yangzhou University; Jiangsu Key Laboratory of Zoonosis; Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, 225009, Yangzhou, Jiangsu, People's Republic of China aut Zuo Jinrong College of Bioscience and Biotechnology, Yangzhou University; Jiangsu Key Laboratory of Zoonosis; Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, 225009, Yangzhou, Jiangsu, People's Republic of China aut Gu Hao College of Bioscience and Biotechnology, Yangzhou University; Jiangsu Key Laboratory of Zoonosis; Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, 225009, Yangzhou, Jiangsu, People's Republic of China aut Guo Minliang College of Bioscience and Biotechnology, Yangzhou University; Jiangsu Key Laboratory of Zoonosis; Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, 225009, Yangzhou, Jiangsu, People's Republic of China aut Yin Yuelan College of Bioscience and Biotechnology, Yangzhou University; Jiangsu Key Laboratory of Zoonosis; Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, 225009, Yangzhou, Jiangsu, People's Republic of China aut Applied Microbiology and Biotechnology Springer Berlin Heidelberg 99/24(2015-12-01), 10527-10537 0175-7598 99:24<10527 2015 99 253 https://doi.org/10.1007/s00253-015-6967-5 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/s00253-015-6967-5 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Yu Minfeng College of Bioscience and Biotechnology, Yangzhou University; Jiangsu Key Laboratory of Zoonosis; Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, 225009, Yangzhou, Jiangsu, People's Republic of China aut NATIONALLICENCE 700 1- Zuo Jinrong College of Bioscience and Biotechnology, Yangzhou University; Jiangsu Key Laboratory of Zoonosis; Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, 225009, Yangzhou, Jiangsu, People's Republic of China aut NATIONALLICENCE 700 1- Gu Hao College of Bioscience and Biotechnology, Yangzhou University; Jiangsu Key Laboratory of Zoonosis; Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, 225009, Yangzhou, Jiangsu, People's Republic of China aut NATIONALLICENCE 700 1- Guo Minliang College of Bioscience and Biotechnology, Yangzhou University; Jiangsu Key Laboratory of Zoonosis; Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, 225009, Yangzhou, Jiangsu, People's Republic of China aut NATIONALLICENCE 700 1- Yin Yuelan College of Bioscience and Biotechnology, Yangzhou University; Jiangsu Key Laboratory of Zoonosis; Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, 225009, Yangzhou, Jiangsu, People's Republic of China aut NATIONALLICENCE 773 0- Applied Microbiology and Biotechnology Springer Berlin Heidelberg 99/24(2015-12-01), 10527-10537 0175-7598 99:24<10527 2015 99 253