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. 2018 Mar 9;17(1):39.
doi: 10.1186/s12934-018-0888-9.

Oxygen effects on rhamnolipids production by Pseudomonas aeruginosa

Feng Zhao  1 , Rongjiu Shi  2 , Fang Ma  3 , Siqin Han  2 , Ying Zhang  4
Affiliations

Oxygen effects on rhamnolipids production by Pseudomonas aeruginosa

Feng Zhao et al. Microb Cell Fact. .

Abstract

Background: Rhamnolipids are the most extensively studied biosurfactants and has been successfully used in various areas from bioremediation to industrial fields. Rhamnolipids structural composition decide their physicochemical properties. Different physicochemical properties influence their application potential. Rhamnolipids can be produced at both aerobic conditions and anaerobic conditions by Pseudomonas aeruginosa. This study aims to evaluate the oxygen effects on the rhamnolipids yield, structural composition, physicochemical properties and the rhl-genes expression in P. aeruginosa SG. Results will guide researchers to regulate microbial cells to synthesize rhamnolipids with different activity according to diverse application requirements.

Results: Quantitative real-time PCR analysis revealed that rhlAB genes were down-regulated under anaerobic conditions. Therefore, strain P. aeruginosa SG anaerobically produced less rhamnolipids (0.68 g/L) than that (11.65 g/L) under aerobic conditions when grown in media containing glycerol and nitrate. HPLC-MS analysis showed that aerobically produced rhamnolipids mainly contained Rha-C8-C10, Rha-Rha-C10-C12:1 and Rha-Rha-C8-C10; anaerobically produced rhamnolipids mainly contained Rha-C10-C12 and Rha-C10-C10. Anaerobically produced rhamnolipids contained more mono-rhamnolipids (94.7%) than that (54.8%) in aerobically produced rhamnolipids. rhlC gene was also down-regulated under anaerobic conditions, catalyzing less mono-rhamnolipids to form di-rhamnolipids. Aerobically produced rhamnolipids decreased air-water surface tension (ST) from 72.2 to 27.9 mN/m with critical micelle concentration (CMC) of 60 mg/L; anaerobically produced rhamnolipids reduced ST to 33.1 mN/m with CMC of 80 mg/L. Anaerobically produced rhamnolipids emulsified crude oil with EI24 = 80.3%, and aerobically produced rhamnolipids emulsified crude oil with EI24 = 62.3%. Both two rhamnolipids products retained surface activity (ST < 35.0 mN/m) and emulsifying activity (EI24 > 60.0%) under temperatures (4-121 °C), pH values (4-10) and NaCl concentrations less than 90 g/L.

Conclusions: Oxygen affected the rhl-genes expression in P. aeruginosa, thus altering the rhamnolipids yield, structural composition and physicochemical properties. Rhamnolipids produced at aerobic or anaerobic conditions was structurally distinct. Two rhamnolipids products had different application potential in diverse biotechnologies. Although both rhamnolipids products were thermo-stable and halo-tolerant, aerobically produced rhamnolipids possessed better surface activity, implying its well wetting activity and desorption property; anaerobically produced rhamnolipids exhibited better emulsifying activity, indicating its applicability for enhanced oil recovery and bioremediation of petroleum pollution.

Keywords: Di-rhamnolipids; Emulsifying activity; HPLC–MS; Mono-rhamnolipids; Pseudomonas aeruginosa; rhl-genes.

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Figures

Fig. 1
Fig. 1
Expression of rhlAB genes and rhlC gene in P. aeruginosa SG under aerobic conditions or anaerobic conditions. The expression levels were determined by quantitative real-time PCR. 16S rRNA gene was used as endogenous control
Fig. 2
Fig. 2
Fourier Transform infrared spectroscopy analysis of rhamnolipids products of strain SG: a rhamnolipids produced at aerobic conditions, b rhamnolipids produced at anaerobic conditions
Fig. 3
Fig. 3
Liquid chromatogram results of rhamnolipids products of strain SG: a rhamnolipids produced at aerobic conditions, b rhamnolipids produced at anaerobic conditions
Fig. 4
Fig. 4
Relationship graphs between the surface tension and concentration of rhamnolipids products: a rhamnolipids produced at aerobic conditions, b rhamnolipids produced at anaerobic conditions
Fig. 5
Fig. 5
Emulsifying activity of the two types of rhamnolipids produced at aerobic or anaerobic conditions
See this image and copyright information in PMC

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