Characterization of a novel cell wall hydrolase CwlE involved in Bacillus thuringiensis subsp. israelensis mother cell lysis
Cell wall hydrolases are ubiquitous among spore-form bacteria and essential for mother cell lysis. In this study, a novel cell wall hydrolase gene cwlE involved in mother cell lysis was characterized from Bacillus thuringiensis subsp. israelensis (Bti) strain Bt-59. cwlE was specifically expressed i...
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| Format: | Article |
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Frontiers Media S.A.
2023-09-01
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| Series: | Frontiers in Microbiology |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fmicb.2023.1250542/full |
| _version_ | 1797671798288941056 |
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| author | Lixin Huang Guangjie Han Neil Crickmore Chuanming Li Yang Xia Fuping Song Jian Xu |
| author_facet | Lixin Huang Guangjie Han Neil Crickmore Chuanming Li Yang Xia Fuping Song Jian Xu |
| author_sort | Lixin Huang |
| collection | DOAJ |
| description | Cell wall hydrolases are ubiquitous among spore-form bacteria and essential for mother cell lysis. In this study, a novel cell wall hydrolase gene cwlE involved in mother cell lysis was characterized from Bacillus thuringiensis subsp. israelensis (Bti) strain Bt-59. cwlE was specifically expressed in Bti and located in the large plasmid carrying the insecticidal genes. The encoded CwlE protein consists of a MurNAc-LAA domain and two highly conserved catalytic residues (E26 and E151). The recombinant CwlE-His protein was able to digest the cell wall of Bti, indicating that CwlE is an N-acetylmuramoyl-L-alanine amidase. Transcriptional analysis indicated that cwlE began to express at the early stage of stationary phase and was controlled by SigE. Single mutation of cwlE gene delayed Bti mother cell lysis, while double mutation of cwlE and sigK completely blocked Bti mother cell lysis. After exposure to UV light to deactivate the crystal proteins, the level of decrease of insecticidal activity against mosquito larvae of Bt-59 (ΔcwlE-sigK) was less than that observed for Bt-59. This study elucidates the mechanism of Bti mother cell lysis and provides an effective strategy for mosquito control using Bt products with increased persistence. |
| first_indexed | 2024-03-11T21:20:43Z |
| format | Article |
| id | doaj.art-3774c2633fa548a6b044752ffa065e8b |
| institution | Directory Open Access Journal |
| issn | 1664-302X |
| language | English |
| last_indexed | 2024-03-11T21:20:43Z |
| publishDate | 2023-09-01 |
| publisher | Frontiers Media S.A. |
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| series | Frontiers in Microbiology |
| spelling | doaj.art-3774c2633fa548a6b044752ffa065e8b2023-09-28T05:52:01ZengFrontiers Media S.A.Frontiers in Microbiology1664-302X2023-09-011410.3389/fmicb.2023.12505421250542Characterization of a novel cell wall hydrolase CwlE involved in Bacillus thuringiensis subsp. israelensis mother cell lysisLixin Huang0Guangjie Han1Neil Crickmore2Chuanming Li3Yang Xia4Fuping Song5Jian Xu6Department of Applied Microbiology, Lixiahe District Institute of Agricultural Sciences in Jiangsu/National Agricultural Experimental Station for Agricultural Microbiology in Yangzhou, Yangzhou, ChinaDepartment of Applied Microbiology, Lixiahe District Institute of Agricultural Sciences in Jiangsu/National Agricultural Experimental Station for Agricultural Microbiology in Yangzhou, Yangzhou, ChinaDepartment of Biochemistry, School of Biological Sciences, University of Sussex, Brighton, United KingdomDepartment of Applied Microbiology, Lixiahe District Institute of Agricultural Sciences in Jiangsu/National Agricultural Experimental Station for Agricultural Microbiology in Yangzhou, Yangzhou, ChinaDepartment of Applied Microbiology, Lixiahe District Institute of Agricultural Sciences in Jiangsu/National Agricultural Experimental Station for Agricultural Microbiology in Yangzhou, Yangzhou, ChinaState Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing, ChinaDepartment of Applied Microbiology, Lixiahe District Institute of Agricultural Sciences in Jiangsu/National Agricultural Experimental Station for Agricultural Microbiology in Yangzhou, Yangzhou, ChinaCell wall hydrolases are ubiquitous among spore-form bacteria and essential for mother cell lysis. In this study, a novel cell wall hydrolase gene cwlE involved in mother cell lysis was characterized from Bacillus thuringiensis subsp. israelensis (Bti) strain Bt-59. cwlE was specifically expressed in Bti and located in the large plasmid carrying the insecticidal genes. The encoded CwlE protein consists of a MurNAc-LAA domain and two highly conserved catalytic residues (E26 and E151). The recombinant CwlE-His protein was able to digest the cell wall of Bti, indicating that CwlE is an N-acetylmuramoyl-L-alanine amidase. Transcriptional analysis indicated that cwlE began to express at the early stage of stationary phase and was controlled by SigE. Single mutation of cwlE gene delayed Bti mother cell lysis, while double mutation of cwlE and sigK completely blocked Bti mother cell lysis. After exposure to UV light to deactivate the crystal proteins, the level of decrease of insecticidal activity against mosquito larvae of Bt-59 (ΔcwlE-sigK) was less than that observed for Bt-59. This study elucidates the mechanism of Bti mother cell lysis and provides an effective strategy for mosquito control using Bt products with increased persistence.https://www.frontiersin.org/articles/10.3389/fmicb.2023.1250542/fullBacillus thuringiensis subsp. israelensismother cell lysiscell wall hydrolasetranscriptional regulationencapsulation |
| spellingShingle | Lixin Huang Guangjie Han Neil Crickmore Chuanming Li Yang Xia Fuping Song Jian Xu Characterization of a novel cell wall hydrolase CwlE involved in Bacillus thuringiensis subsp. israelensis mother cell lysis Frontiers in Microbiology Bacillus thuringiensis subsp. israelensis mother cell lysis cell wall hydrolase transcriptional regulation encapsulation |
| title | Characterization of a novel cell wall hydrolase CwlE involved in Bacillus thuringiensis subsp. israelensis mother cell lysis |
| title_full | Characterization of a novel cell wall hydrolase CwlE involved in Bacillus thuringiensis subsp. israelensis mother cell lysis |
| title_fullStr | Characterization of a novel cell wall hydrolase CwlE involved in Bacillus thuringiensis subsp. israelensis mother cell lysis |
| title_full_unstemmed | Characterization of a novel cell wall hydrolase CwlE involved in Bacillus thuringiensis subsp. israelensis mother cell lysis |
| title_short | Characterization of a novel cell wall hydrolase CwlE involved in Bacillus thuringiensis subsp. israelensis mother cell lysis |
| title_sort | characterization of a novel cell wall hydrolase cwle involved in bacillus thuringiensis subsp israelensis mother cell lysis |
| topic | Bacillus thuringiensis subsp. israelensis mother cell lysis cell wall hydrolase transcriptional regulation encapsulation |
| url | https://www.frontiersin.org/articles/10.3389/fmicb.2023.1250542/full |
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