Abstract ▼ AI-Helper

The appearance of antibiotic-resistant bacteria presents a major hurdle for the development of effective antibiotics. Antimicrobial peptides (AMPs) have been studied extensively as new alternative materials. AMP is the antimicrobial substance found in animals and plants and has a strong antimicrobia...

The appearance of antibiotic-resistant bacteria presents a major hurdle for the development of effective antibiotics. Antimicrobial peptides (AMPs) have been studied extensively as new alternative materials. AMP is the antimicrobial substance found in animals and plants and has a strong antimicrobial activity even in small amounts. The two major biological functions of AMP are antiviral and antioxidant. Animal cell lines and yeasts have been used to produce recombinant proteins. However, those expression systems present several disadvantages when producing recombinant proteins, such as complex management of cultural facilities and contamination. On the other hand, plants can be grown with water and sunlight and are not polluted. The chloroplast expression system in plants has been used as an attractive platform for the production of high amounts of therapeutic proteins such as antibody and vaccine. However, protein degradation or low solubility should be considered before producing recombinant proteins. In this study, the recombinant protein was produced by fusion with a SUMO to obtain high yield of the proteins. Stomoxyn from Stomoxys calcitrans is an amphiphilic peptide with α-helix structure that possesses antibacterial substance. Stomoxyn has been reported to have antimicrobial activity against both gram-positive and gram-negative bacteria. SUMO-stomoxyn fusion proteins expressed in E. coli were purified by nikel-column and then were treated with SUMO protease to remove the SUMO moeity from the fusion proteins. The antimicrobial activity of the purified stomoxyn was investigated in agar-diffusion assay and microtiter broth dilution assay. Stomoxyn was shown to have antibacterial activity against Bacillus subtilis, Staphylococcus xylosus, E. coli JM83, and Acromobacter sp. Using a gene gun, chloroplast expression vector containing EGFP gene was delivered to Tobacco leaves to test whether the EGFP gene was expressed in chloroplasts. The expression of EGFP in chloroplasts suggested the successful delivery and the expression of stomoxyn gene in chloroplasts.

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