The 46th Academic Congress of Taiwan Association for Food Science and Technology

Expression and purification of recombinant ribosome inactivating protein MAP30 in Lactobacillus plantarum ATIT-018

WC Huang, ZW Chen, JH Lin, SL Hsuan, JP Wang

MAP30 (Momordica anti-viral protein of 30 kDa), a kind of type I ribosome inactivating protein, is isolated from the medicinal plant Momordica charantia (bitter melon). MAP30 was previously found to have anti-HIV (human immunodeficiency virus), anti-tumor, and antimicrobial activities. Thus, it is considered that MAP30 may be a useful therapeutic agent in the treatment of cancer and HIV-infections. The aim of this study was to express recombinant MAP30 in Lactobacillus plantarum ATIT-018 for development of a novel therapeutic delivery vehicle. The MAP30 gene was amplified by polymerase chain reaction from the genomic DNA of M. charantia and cloned into the L. plantarum expression vector. Then, the expression vector was transformed into L. plantarum ATIT-018 by using electroporation. Transformants were inoculated into MRS broth in the presence of chloramphenicol and growth for 8 h at 30°C. The results of SDS-PAGE and western blot showed that the expression of MAP30 in L. plantarum was successful. Recombinant MAP30 could be purified from the soluble intracellular fraction of transformants. In this study, we propose a new method for MAP30 production. The application of L. plantarum ATIT-018 as a novel therapeutic delivery vehicle should be investigated further.