Enhanced rhamnolipid production in Pseudomonas putida through systematic medium optimization and metabolic...

Area of Science:

• Biotechnology
• Microbial Engineering
• Biosurfactant Production

Background:

• Rhamnolipids are valuable biosurfactants with diverse industrial uses.
• Current production methods face challenges due to low yields and high costs.
• Pseudomonas putida is a promising host for biosurfactant synthesis.

Purpose of the Study:

• To enhance rhamnolipid production in *P. putida* KT2440.
• To optimize culture medium composition for increased rhamnolipid yield.
• To apply metabolic engineering strategies to boost precursor supply for rhamnolipid synthesis.

Main Methods:

• Response surface methodology was used for medium optimization with mixed carbon and nitrogen sources.
• Transcriptional analysis was performed to understand metabolic pathway flux.
• Metabolic engineering involved overexpressing key genes in rhamnose and fatty acid synthesis pathways.

Main Results:

• Medium optimization increased rhamnolipid titer by 306.7% (from 1.5 to 6.1 µg/L).
• Engineered strain (accAD-fabD-R) achieved 40.2 µg/L in a 5L bioreactor.
• Record-high rhamnolipid production levels were achieved in engineered *P. putida*.

Conclusions:

• A combined approach of medium optimization and metabolic engineering is effective for high-efficiency rhamnolipid production.
• Enhanced precursor supply is crucial for overcoming bottlenecks in rhamnolipid synthesis.
• *P. putida* shows significant potential for industrial-scale biomanufacturing of rhamnolipids.