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- Title
枯草杆菌芽孢皮层裂解酶CwlJ的 重组表达及分离纯化.
- Authors
马慧娇; 郭洪伟; 曾朝玮; 郭家俊; 章中
- Abstract
[Objective] Cortical lysozyme is a specific enzyme for the hydrolysis of cortical peptidoglycan in spores. The hydrolysis of pepti-doglvcan is an important cause of spore death. Isolation and extraction of CwlJ, a spore cortical lysozyme, provides raw materials for promoting CwlJ to hydrolyze peptidoglycan and kill spores and for food sterilization. [Method]The CwlJ gene of cortical lyase was introduced into plasmid pCZN-1 , and the genetic engineering bacteria were successfully constructed. IPTG was used to induce recombinant bacteria to express CwlJ, SDS-PAGE and Western-Blot to detect tbe expression of recombinant cortical lyase, and the recombinant protein CwlJ was purified by affinity chromatography. [Result] IPTG of 0. 5 mM could express CwlJ gene in large quantities. The molecular weight of purified recombinant cortical lyase CwlJ was 17. 9 KD, the optimum temperature was 40 °C , the optimum pH was 5, and the specific enzyme activity was 146.67 U/mg. [Conclusion] Compared with the cortical lyase extracted from natural spores, the activity of recombinant cortical lyase CwlJ increased two times, which ensured the further hydrolysis of peptidoglycan by cortical lyase CwlJ.
- Subjects
BACTERIAL genetic engineering; RECOMBINANT proteins; AFFINITY chromatography; MOLECULAR weights; LYSOZYMES; SPORES
- Publication
Southwest China Journal of Agricultural Sciences, 2019, Vol 32, Issue 11, p2564
- ISSN
1001-4829
- Publication type
Article
- DOI
10.16213/j.cnki.scjas.2019.11.013