Comparative study on antimicrobial activity of mono-rhamnolipid and di-rhamnolipid and exploration of cost-effective antimicrobial agents for agricultural applications.

BACKGROUND: Chemical pesticides have defects in crop diseases control, such as narrow antimicrobial spectrum, chemicals residue risk and harm to farmland ecosystem. Antimicrobial agents from microbial sources are highly interested in agriculture. Studies showed that rhamnolipid biosurfactants possessed certain antimicrobial activity. The structural differences in rhamnolipid inevitably affect their activities. But the antimicrobial effect of mono-rhamnolipid and di-rhamnolipid is unknown. Rhamnolipid with unique structure can be produced using specific microbial cell factory. RESULTS: Different types of rhamnolipid were produced from different Pseudomonas aeruginosa strains. Rha-C10-C10 and Rha-Rha-C10-C10 were the main homologues in the separated mono-rhamnolipid and di-rhamnolipid, respectively. Both mono-rhamnolipid and di-rhamnolipid exhibited certain antimicrobial activity against the tested microbial strains, especially the fungi and Gram-positive bacteria. But mono-rhamnolipid was superior to di-rhamnolipid, with inhibition zone diameters larger than 25 mm and inhibition rate higher than 90%. The IC50 values of mono-rhamnolipid were lower than 5 mg/L against the tested bacterium and fungus, whereas the IC50 values of di-rhamnolipid were ranged from 10 mg/L to 20 mg/L. Mono-rhamnolipid stimulated the tested strains to generate higher level of intracellular ROS. Mono-rhamnolipid exhibited better antimicrobial activity to the potential agricultural pathogens, such as Alternaria alternata, Pantoea agglomerans and Cladosporium sp. The mono-rhamnolipid crude extract of strain P. aeruginosa SGΔrhlC can replace the separated mono-rhamnolipid. After 50 times dilution, the fermentation broth of the mono-rhamnolipid producing strain SGΔrhlC exhibited equal antimicrobial effect to mono-rhamnolipid (200 mg/L). Prospects of mono-rhamnolipid were also discussed for antimicrobial applications in agriculture. CONCLUSIONS: This work discovered that mono-rhamnolipid was superior to di-rhamnolipid on antimicrobial activity for agricultural applications. Mono-rhamnolipid is an excellent candidate for agricultural biocontrol. The knockout strain P. aeruginosa SGΔrhlC is an excellent microbial cell factory for high producing mono-rhamnolipid. Its mono-rhamnolipid crude extract and its diluted fermentation broth are cost-effective antimicrobial agents. This work provided new insights to develop green and efficient antimicrobial agents for agricultural applications.

Enhanced production of mono-rhamnolipid in Pseudomonas aeruginosa and application potential in agriculture and petroleum industry.

Differences in the rhamnolipid structures must result in its different activities, thus affecting its application effect. The rhlC gene in Pseudomonas aeruginosa SG was knocked out to construct strain P. aeruginosa SGΔrhlC. Rhamnolipid production was enhanced by 23.3% through knocking out rhlC gene. P. aeruginosa SGΔrhlC produced 14.22 g/L of rhamnolipid using glycerol and nitrate. Five kinds of mono-rhamnolipid but no di-rhamnolipid were produced by strain SGΔrhlC. The main rhamnolipid homologues were Rha-C10-C10, Rha-C10-C12:1 and Rha-C10-C12. Mono-rhamnolipid exhibited better antimicrobial activity to Escherichia coli, Staphylococcus aureus, Aspergillus niger and Penicillium chrysogenum. Rhamnolipid produced from strain SGΔrhlC showed greater emulsifying activity to crude oil with EI24 of 84.73%. Rhamnolipid produced from strain SGΔrhlC efficiently washed oily sludge at 35 °C. High-producing strain P. aeruginosa SGΔrhlC and its produced mono-rhamnolipid are more promising in agriculture and petroleum industry. This study is a step forward to the tailor-made biosynthesis and application of rhamnolipid.