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Expression and characterization of lactate dehydrogenase from Schistosoma japonicum

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Abstract

Some effective antiparasitic drugs for chemotherapy schistosomiasis, such as praziquantel and artemether, are proved to act on lactate dehydrogenase (LDH) in vitro, but detailed molecular action mechanisms of these have not yet been elucidated. To this end, the sequence encoding LDH of Schistosoma japonicum (SjLDH) was amplified from the cDNA library of the adult parasite. Sequence analysis revealed an open reading frame (ORF) of 999 bp, encoding 332 amino acids with a molecular weight of 36.120 kDa. It has three transmembrane regions and may be located in the cytoplasm membrane. The encoding ORF was inserted into pET-28a vector and expressed in Escherichia coli (E. coli) induced by IPTG. A 36-kDa protein with 6×His-tag was purified by metal affinity column and identified by SDS-PAGE and Western blot, which had high LDH activity of 379 U/mg. Characterization such as Michaelis constant (K m) and maximum velocity (V max), optimum pH, and temperature of the protein were assayed. It provides a model for drug screening on SjLDH.

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Acknowledgement

This study was funded by the “211 Project” Found of Guangdong Province, China.

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Authors and Affiliations

  1. Department of Parasitology, Medical College, Sun Yat-sen University, Guangzhou, 74 Zhongshan 2nd Road, 510089, Guangdong, China

    Gang Lu, Xuchu Hu, Zhaiyu Peng, Hongyan Xie, Yanwen Li, Zhongdao Wu & Xinbing Yu

  2. Department of Parasitology, Hainan Medical College, Xueyuan Road, 571101, Haikou, China

    Gang Lu

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  1. Gang Lu
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  2. Xuchu Hu
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  3. Zhaiyu Peng
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  4. Hongyan Xie
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  5. Yanwen Li
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  6. Zhongdao Wu
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  7. Xinbing Yu
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Correspondence to Xinbing Yu.

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Lu, G., Hu, X., Peng, Z. et al. Expression and characterization of lactate dehydrogenase from Schistosoma japonicum . Parasitol Res 99, 593–596 (2006). https://doi.org/10.1007/s00436-006-0152-9

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  • DOI: https://doi.org/10.1007/s00436-006-0152-9

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