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Updated: Mar 10, 2026
Defining Substrate Specificities for Lipase and Phospholipase Candidates
Published on: November 23, 2016
Identification of lysophospholipase protein from Spiroplasma eriocheiris and verification of its function
Huanxi Zhu1,2, Peng Liu2, Jie Du2
1Laboratory of Animal Improvement and Reproduction, Institute of Animal Science, Jiangsu Academy of Agricultural Sciences, Nanjing 210014, PR China.
Abstract:
Spiroplasma eriocheiris is known to cause tremor disease in the Chinese mitten crab Eriocheir sinensis; however, the molecular characterization of this pathogen is still unclear. S. eriocheiris has the ability to invade and survive within mouse 3T6 cells. The invasion process may require causing damage to the host cell membrane by chemical, physical or enzymatic means. In this study, we systematically characterized a novel lysophospholipase (lysoPL) of S. eriocheiris TDA-040725-5T. The gene that encodes lysoPL in S. eriocheiris (SE-LysoPL) was cloned, sequenced and expressed in Escherichia coli BL21 (DE3). Enzymatic assays revealed that the purified recombinant SE-LysoPL hydrolysed long-chain acyl esterases at pH 7 and 30 °C. SE-LysoPL was detected in the membrane and cytoplasmic protein fractions using the SE-LysoPL antibody in Western blot. The virulence ability of S. eriocheiris was effectively reduced at the early stage of infection (m.o.i.=100) by the SE-LysoPL antibody neutralization test. To the best of our knowledge, this is the first study to identify and characterize a gene from S. eriocheiris encoding a protein exhibiting lysoPL and esterase activities. Our findings indicate that SE-LysoPL plays important roles in the pathogenicity of S. eriocheiris.
Insights
Spiroplasma eriocheiris causes tremor disease in crabs. Researchers identified and characterized a novel lysophospholipase (SE-LysoPL) from this pathogen, finding it crucial for its virulence and host cell invasion.
Area of Science:
- Microbiology
- Molecular Biology
- Pathogen Characterization
Background:
- Spiroplasma eriocheiris is the causative agent of tremor disease in Chinese mitten crabs.
- The molecular mechanisms underlying S. eriocheiris pathogenicity, including its ability to invade host cells, are not well understood.
Purpose of the Study:
- To molecularly characterize a novel lysophospholipase (lysoPL) from Spiroplasma eriocheiris.
- To investigate the role of this enzyme in the pathogenicity of S. eriocheiris.
Main Methods:
- Cloning, sequencing, and expression of the Spiroplasma eriocheiris lysophospholipase gene (SE-LysoPL) in E. coli.
- Enzymatic assays to determine the hydrolytic activity of recombinant SE-LysoPL.
- Western blot analysis to detect SE-LysoPL localization within the pathogen.
- Antibody neutralization test to assess the impact on S. eriocheiris virulence.
Main Results:
- The SE-LysoPL gene was successfully cloned, sequenced, and expressed.
- Purified recombinant SE-LysoPL exhibited lysophospholipase and esterase activity.
- SE-LysoPL was localized to both membrane and cytoplasmic fractions of S. eriocheiris.
- Neutralization with an anti-SE-LysoPL antibody significantly reduced S. eriocheiris virulence in vitro.
Conclusions:
- This study is the first to identify and characterize a Spiroplasma eriocheiris protein with lysophospholipase and esterase activities.
- SE-LysoPL plays a significant role in the pathogenicity of Spiroplasma eriocheiris, likely contributing to host cell invasion and damage.
- Targeting SE-LysoPL may offer a strategy to control Spiroplasma eriocheiris infections.
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